ISSN 0972-1045

Vol. 16, No. 1, January-April, 2014

Appl Hort

Journal of
THE SOCIETY FOR ADVANCEMENT OF HORTICULTURE
JOURNAL OF APPLIED HORTICULTURE
Vol. 16, No. 1, Januar y-April, 2014
CONTENTS
Spatial variability in Ontario Cabernet Franc vineyards I. Interrelationships among
soil composition, soil texture, soil and vine water status
—Andrew G. Reynolds and Javad Hakimi Rezaei 3
Effect of glycinebetaine application on photosynthesis, sugar content, invertase activity
and plant yield of hot pepper (Capsicum annuum L.) under water stress condition
—R.M. Bhatt, N.K. Srinivasa Rao, K.K. Upreti and A.D.D.V.S. Nageswara Rao 24
Linking certain physical characteristics with postharvest needle abscission
resistance in balsam fir
—M.T. MacDonald, R.R. Lada and R.S. Veitch 29
Optimal soil conditions for organic highbush blueberry growth: Assessment
of early results
—B. Hoover, D. Fuglie and R. Miller 32
Comparative efficacy of vermicomposted paper waste and inorganic fertilizer on seed
germination, plant growth and fruition of Cyamopsis tetragonoloba
—M. Karthikeyan, S. Gajalakshmi and S.A. Abbasi 40
AM fungi shields Coleus forskohlii from root rot incidence
—L. Srimathi Priya and K. Kumutha 46
Canopy management in mango (Mangifera indica L.) cv. Alphonso with reference to
flowering, yield and quality characters under ultra high density planting
—B. Gopu, T.N. Balamohan, P. Soman and P. Jeyakumar 50
Comparative evaluation of common bean (Phaseolus vulgaris L.) germplasm for seed
physical and culinary traits
—P. A. Sofi, S. A Wani, M.Y. Zargar, F. A. Sheikh and T. Shafi 54
Physical properties and transmission of papaya ringspot virus
—Isha Bhoyer, Mina D. Koche, Santoshi Pudake and N.B. Ninawe 59
Effect of integrated application of phosphorus and phosphate solubilizing
microorganisms on root colonization, productivity and seed quality of Cucurbita pepo L.
—J. Hamzei and S. Najjari 61
Processing and quality evaluation of blended guava watermelon squash
—J. Shankara Swamy and A.K. Banik 66
Effect of pre-treatment and drying temperature on quality of dehydrated
cauliflower (Brassica oleracea var. botrytis)
—R. Ranjan, M. Longkumer and J. Kabir 71
Planting density and corm size effects on flower yield and quality of
cut-freesia (Freesia hybrid) in Ethiopia
—Tewodros Bezu and Nigusse Kassa 76
Effect of processing and storage on bioactive compounds and antioxidant
activity of carrot juice
—S. Kapoor and P. Aggarwal 80
 Journal

Journal of Applied Horticulture, 16(1): 3-23, 2014 Appl

Spatial variability in Ontario Cabernet Franc vineyards:

I. Interrelationships among soil composition, soil texture, soil
and vine water status

Andrew G. Reynolds* and Javad Hakimi Rezaei

Cool Climate Oenology and Viticulture Institute, Brock University, St. Catharines, ON, Canada L2S 3A1.
*E-mail: areynolds@brocku.ca

Abstract
Spatial variability of vine water status and its relationship to soil moisture (SM) and physical properties was studied in ten vineyard
blocks of Vitis vinifera L. Cabernet Franc in the Niagara Peninsula, Ontario, using geomatic techniques. Soil texture, soil chemical
composition, SM, and leaf water potential (ψ; vine water status), were determined on ≈ 80 sentinel vines per vineyard. Water status
zones were identified in vineyard-specific GIS-generated maps using leaf ψ and SM measurements. SM was temporally consistent for
nine of ten sites (2005-2006), all sites (2006-2007), and eight sites (2005-2007). Vine water status was temporally consistent for two
sites (2005-2006) and three sites (2006-2007), but leaf ψ zones were transient at some sites with temporally variable spatial distribution
(except one site with consistent water status zones 2005-2007). SM and leaf ψ consistently were directly-correlated spatially with %
clay, % organic matter (OM), cation exchange capacity (CEC), soil pH, base saturation (BS), soil K/Ca/Mg. Low SM and water status
zones were related to low % clay, OM, CEC, soil pH, BS, soil K/Ca/Mg zones. This indicate that precision viticulture may be applied
to soil texture, SM, or leaf ψ-based vineyard sub-zones that could relate to differing quality levels.
Key words: Global positioning systems, geographic information systems, soil moisture, leaf water potential

Introduction
Several recent studies have indicated that temporally stable spatial
variation in vineyards exists in terms of vegetative growth, yield,
and fruit composition (Bramley, 2001; Hall et al., 2002). However,
the use of geospatial tools such as geographic information systems
(GIS) coupled with remote sensing techniques for the study of
vineyards is a relatively recent development that has yet seen few
applications. GIS was used to map 2,000 ha of the Loire Valley,
in terms of soil type and rootstock, but the information was not
used to find possible relationships such as those between soil
and wine varietal typicity. In California, GIS was used to map
viticultural regions in terms of phylloxera damage based upon
leaf reflectance (Johnson et al., 1996). These techniques were
also used to relate yields and soluble solids concentrations of
Concord (Davenport et al., 2001), to distinguish between high
and low vigor “management zones” in Cabernet Sauvignon and
Zinfandel vineyards in California (Greenspan and O’Donnell,
2001), and in vineyards throughout Australia (Bramley, 2005).
More recently, geomatic tools were used to assess impact of vigor
zones on anthocyanin and phenolic composition in Oregon Pinot
noir (Cortell et al., 2006).
In Ontario, vineyards are often located on sites that contain
heterogeneous soil types. It was initially hypothesized that soil
texture would play a minor role in the widely accepted terroir
effect, in terms of its determination of yield components, fruit
composition (including aroma compounds), and wine sensory
attributes, and that vine vigor, vine water status, crop size and
fruit environment would play the major roles. This hypothesis
was initially tested using geospatial tools in five commercial
Chardonnay vineyards in the Niagara Peninsula of Ontario in
1998 to 2002, with the conclusion that vine vigor appeared to
impact fruit composition and wine sensory attributes to a greater
degree than soil texture (Reynolds and de Savigny, 2001). A study
in a 4-ha Riesling vineyard was another attempt using geomatic
tools to understand direct and independent soil and vine vigor
effects on yield components, berry, must and wine composition,
and wine sensory attributes (Reynolds et al., 2007; 2010a). Spatial
variability of monoterpenes led to the identification of sub-blocks
that were designated as having potentially higher wine value.
Recent studies in Ontario have definitively linked soil and vine
water status to sensory attributes in Riesling (Willwerth et al.,
2010) and have shown links between Riesling varietal typicity
and both soil texture and vine size (Reynolds et al. 2007; 2010a;
Willwerth et al., 2010). Remote sensing has also been used to
correlate canopy spectral reflectance to vine water status, yield,
vine size, and berry monoterpenes (Reynolds et al., 2010b).
This investigation was initiated to identify the major factors that
contribute to the terroir effect in the vineyards of the Niagara
Peninsula in Ontario. The overall objective of this study was to
determine spatial variability with respect to soil composition,
soil texture, soil and vine water status in ten Cabernet Franc
vineyards in the Niagara Peninsula of Ontario, and to explore
whether relationships existed between these variables. The
specific objectives of this research were: (i) to demonstrate the
influences of soil texture, soil water content, and vine water status
on vine and fruit development within vineyard blocks and to
delineate these terroir effects using GPS/GIS; and (ii) to elucidate
the relationships between soil and vine water status and wine
sensory properties. It was hypothesized that: (i) consistent water
status zones could be identified within vineyard blocks and, (ii)
vine water status would play a major role in fruit composition
4 Spatial variability in Ontario Cabernet Franc vineyards

and sensory characteristics of Cabernet Franc wines, whereas as well as regions of different water status, delineated through
soil type might play a role through its water holding capacity GIS using sensory descriptive analysis (Hakimi Rezaei and
and water supply to the vine. This project therefore had three Reynolds, 2010a,b).
distinct phases; the first phase, described in this paper, examined
spatial relationships among soil characteristics, soil moisture and Materials and methods
vine water status using GPS/GIS technology. The second phase Site selection and site features: Ten commercial vineyard
examined relationships between soil and vine water status and blocks of Cabernet Franc were selected, one each in the ten
vine performance (yield components and vine size) as well as sub-appellations of the Niagara Peninsula including: Niagara
berry composition (including anthocyanins and phenols) using Lakeshore, St. David’s Bench, Creek Shores, Four Mile Creek,
GPS/GIS technology. The third phase consisted of the sensory Niagara River, Lincoln Lakeshore, Beamsville Bench, Short Hills
characterization of wines produced from both sub-appellations Bench, Vinemount Ridge, and Twenty Mile Bench for the project
Table 1. General features of Niagara Peninsula Cabernet Franc vineyards used for elucidation of terroir study, 2005-07. Vineyard site abbreviations
used in the text are indicated
Variable Sites
Château des Charmes Reif Hernder Buis Henry of Pelham
(CDC) (HOP)
VQA sub-appellation St. David’s Bench Niagara River Four Mile Creek Niagara Lakeshore Short Hills Bench
(Lakeshore Plain)
Area of vineyard block (ha) 2.29 0.61 2.63 0.71 2.17
Number of sentinel vines 80 84 70 77 80
Sentinel vines per ha 35 138 27 108 37
Soil series Toledo 7 Chinguacousy 7 Chinguacousy 1 Chinguacousy 19 Beverley 1
(Kingston and Presant, 1989) (Red Phase; CGU.R)
Parent materials Lacustrine Washed reddish Mainly clay Mainly reddish Mainly lacustrine
silty clay hued clay loam loam till hued clay silty clay
till, modified by
lacustrine processes
Soil drainage Imperfect to poor Imperfect Imperfect Imperfect to poor Imperfect
Rootstock 3309 3309 3309 SO4 + 3309 SO4
Vine age at initiation of trial 1992 1999 1998 1988 1999
(year planted)
Vine spacing (m; row x vine) 2.2 x 0.9 3.0 x 1.3 2.8 x 1.25 2.9 x 1.3 2.7 x 1.3
Number of rows; vines per 27 rows 6 rows 58 rows 20 rows 29 rows
row 376 vines/ row 284 vines/ row 137 vines/ row 118 vines/ row 240 vines/ row
Training system Guyot Pendelbogen Guyot Scott Henry Guyot
Floor management Clean cultivated Alternate sod (rows Alternate sod Clean cultivated Alternate sod
alternate between sod
and clean cultivation)
Harbour Estate Morrison Vineyard Cave Spring George Vineyard Vieni Estate
(Harbour)
VQA sub-appellation Creek Shores 20 Mile Bench Beamsville Bench Lincoln Lakeshore Vinemount Moraine
Area of vineyard block (ha) 1.67 0.97 1.54 1.23 1.19
Number of sentinel vines 80 72 75 72 72
Sentinel vines per ha 47 74 49 59 61
Soil series Vittoria 16 (Red Cashel 3 Chinguacousy 14 Chinguacousy 24 Chinguacousy 1
(Kingston and Presant, 1989) Phase; VIT.R) (Loamy Phase; (CGU.L) (Washed Phase; CGU.W)
Parent materials 40-100 cm reddish- 40-100 cm lacustrine 15-40 cm loamy Washed clay loam Mainly
hued sandy textures silty clay over textures over till, modified by clay
over lacustrine silt clay loam clay loam lacustrine loam
loam till till processes till
Soil drainage Imperfect Moderately well Imperfect Imperfect-poor Imperfect
Rootstock Riparia Gloire SO4 101-14 SO4 SO4
Vine age at initiation of trial 1999 1999 1999 1995 1998
(year planted)
Vine spacing 2.7 x 1.5 2.9 x 1.3 2.7 x 1.4 2.7 x 1.4 2.0 x 1.25
(m; row x vine)
Number of rows; vines per 37 rows 18 rows 23 rows 24 rows 30 rows
row 105 vines/ row 155 vines/ row 233 vines/ row 137 vines/ row 135 vines/ row
Training system Scott Henry Scott Henry Guyot Guyot Guyot
Floor management Clean cultivated Clean cultivated Alternate sod Sod every row Alternate sod
 Spatial variability in Ontario Cabernet Franc vineyards 5

in spring of 2005 (Table 1). General features of each vineyard closer grid points have more influence on the calculation of
including VQA sub appellation (Vintners’ Quality Alliance; http:// unknown grid values compared to the points that are further away.
www.vqaontario.com/appellations), area of vineyard, number With regard to power, the exponential option was selected, which
of sentinel vines, soil series, parental material, soil drainage, enables the user to define the exponential rate of decreasing the
rootstock, year of planting, vine spacing, and floor management influence of neighboring points that lie further from the point
were recorded for each vineyard. Soil parent material at the sites being calculated (Proffitt et al., 2006). The lowest value was
ranged from lacustrine silty clay, reddish hued clay, and loamy chosen for the exponential rate. The values in each zone of the
texture to reddish hued sandy texture. Soil drainage was imperfect constructed maps were the minimum values of the range of values
to poor, imperfect or moderately well drained. Area of vineyard for that zone. Where possible, five zones with equal ranges were
blocks varied from 0.6 ha (Reif) to 2.6 ha (Hernder). Vine spacing delineated for each variable in all maps. The range of values for
varied from 2.0 m x 1.25 m (vine x row) at Vieni Estate to 3.0 x each variable was not the same in each vineyard, and therefore
1.3 m at Reif. Training system was Guyot, pendelbogen, or Scott the ranges were of differing magnitudes. Fewer than five zones
Henry. Floor management at some sites was clean cultivation and were delineated in vineyard blocks with low variability. Spatial
at others was sod maintained in alternate rows. Rootstocks were correlation analysis was performed in Vertical Mapper; r values
101-14, 3309 or SO4 and vine age varied from 7 to 18 years. No > 0.8 were assumed to be particularly meaningful in so far as
changes in management were made at these sites during the period these suggested significant spatial correlation between different
of study. Weather data (Table 2) were compiled for each site from variables within seasons, or temporal consistency between like
Weather Innovations Inc. (www.weatherinnovations.com). variables between seasons. For two independent variables
sampled at a density of 72 to 80 observations per site (e.g. soil
GPS and GIS: A Raven Invicta 115 GPS Receiver (Raven moisture), r values, 0.330 and 0.269 were equivalent to p values
Industries, Sioux Falls, SD, USA) with 1.0 to 1.4 m accuracy of < 0.01 and 0.05, respectively; for a density of 20 observations
was used to delineate the shape of each vineyard block as well per site (e.g. soil composition, leaf ψ), r values, 0.606 and 0.509
as to geolocate each sentinel vine. The number of sentinel vines were equal to p values of < 0.01 and 0.05, respectively (Steel
varied from 27 to 138 per ha. Bramley (2001) reported use of and Torrie, 1960).
26 sentinel vines per ha in studies in Sunraysia and Coonawarra
in Australia. Using GIS programs MapInfo Professional and For wine making purposes, each vineyard block was separated
Vertical Mapper (Northwood GeoScience, Ottawa, ON, Canada), into three zones of high, medium, and low water status (HWS,
spatial maps of all variables were created, and water status zones MWS, LWS, respectively) based on the mean leaf ψ maps for
were mapped based on vine leaf ψ values (Table 3). The inverse each season. Grapes from each of these water status zones were
distance weighting (IDW) interpolation algorithm was used to harvested separately based on the leaf ψ map of each vineyard
construct the grid files. IDW interpolation algorithm was chosen block in both 2005 and 2006 and were used to make wine.
vs. kriging due to uneven nature of vineyards. In this method, Therefore, from each vineyard block, three wine categories (high,
Table 2. Growing degree days (GDD) and precipitation for ten sites in the Niagara Peninsula, Ontario, 2005-2007. Data courtesy of Weather
Innovations Inc., Chatham, ON
Name of vineyard block Name of sub-appellation Growing degree days (GDD) Precipitation (mm) May - October
2005 2006 2007 2005 2006 2007
Buis Niagara Lakeshore 1490 1417 1579 483.3 NA 247.6
Château des Charmes St. David’s Bench 1681 1474 1646 476.5 461.9 219.8
Harbour Estate Creek Shores 1672 1457 1606 436.3 534.2 221.4
Hernder Four Mile Creek 1505 1471 1572 457.1 566.6 181.6
Reif Niagara River 1604 1449 1539 498.4 631.3 163.8
George Lincoln Lakeshore 1559 1401 1420 555.9 537.7 241.4
Cave Spring Beamsville Bench 1620 1415 1679 410.2 604.1 197.8
Henry of Pelham Short Hills Bench 1552 1412 1591 466.8 507.7 172.2
Vieni Estate Vinemount Ridge 1565 1354 1594 409.7 526.5 286.6
Morrison Twenty Mile Bench 1667 1457 1606 438.4 534.2 221.4
Table 3. Mean leaf water potential (ψ) and soil moisture ranges in ten Cabernet Franc sites, Niagara Peninsula, ON, 2005-2007. Measurements were
made bi-weekly between July to September
Site Leaf ψ (-MPa) Soil moisture (%)
2005 2006 2007 2005 2006 2007
Buis 1.00-1.35 1.11-1.35 1.14-1.45 14.0-20.4 17.6-32.0 17.2-27.6
Château des Charmes 1.20-1.60 1.25-1.50 1.52-1.64 10.9-16.2 19.4-33.5 9.3-24.8
Hernder Estate 1.26-1.59 1.29-1.60 1.37-1.60 7.3-13.4 15.1-28.0 6.1-27.7
Reif 1.10-1.35 1.07-1.34 1.11-1.34 7.6-13.6 11.3-25.6 8.8-21.3
George 1.10-1.46 1.01-1.26 1.16-1.50 11.1-15.8 18.1-29.0 12.4-21.7
Henry of Pelham 1.10-1.45 1.14-1.37 1.31-1.50 12.0-15.6 18.1-29.7 14.0-25.9
Cave Spring 1.20-1.55 1.09-1.24 1.43-1.58 10.7-15.6 21.8-32.7 10.1-20.9
Harbour Estate 0.80-1.09 0.90-1.15 0.93-1.12 9.9-13.4 11.7-18.7 3.5-8.5
Vieni Estate 1.20-1.45 0.82-1.10 1.38-1.59 9.1-15.7 22.2-35.9 10.7-25.2
Morrison 1.21-1.47 0.97-1.24 1.42-1.64 11.0-19.1 21.3-33.9 11.0-20.6
6 Spatial variability in Ontario Cabernet Franc vineyards
medium, and low water status) were made with three replicates
of each in both years.
Soil sampling and composition: Soil samples were collected
from every fourth sentinel vine (7 to 35 vines per ha) with an
auger from within the row, 40 to 50 cm away from the trunk.
Soil was taken from a 0 to 45 cm depth and in total ≈ 350 g of a
homogenized sample was taken. Depending on the area of each
vineyard block, 15 to 20 soil samples were taken. Soil samples
were analyzed for pH, organic matter (OM), phosphorus (P),
potassium (K), magnesium (Mg), calcium (Ca), texture (% sand,
silt, clay), cation exchange capacity (CEC), and base saturation
(BS; as % Ca, Mg, and K) using standard procedures [Canadian
Society of Soil Science (CSSS), 1993].
Soil water status: Soil moisture data (% water by volume) were
taken bi-weekly on five separate dates between late June and early
September in the 2005 to 2007 growing seasons. Soil moisture
was measured at each sentinel vine by time domain reflectometry
using a Fieldscout TDR-300 soil moisture probe (Spectrum
Technologies Inc., East Plainfield, IL, USA). A total of 72 to 80
vines were measured between 0800h and 1800h. Measurements
were taken in the row ≈ 10 cm from the base of each vine trunk
over a 20 cm depth. Most vineyards in the region contain drain
tiles at ≈60 cm depth, which tends to restrict rooting depth, and
therefore this depth was considered adequate for determining
moisture levels. The mean soil moisture for each sentinel vine
was calculated from the five separate readings.
Vine water status: Midday leaf ψ was determined between
1100h and 1600h for fully exposed, mature leaves of similar
physiological stage which showed no visible sign of damage and
had been in full sunlight (Turner, 1988). Determinations were
made on cloudless days only. Each leaf sample was covered in a
plastic bag and sealed immediately after excision at the petiole to
suppress transpiration. The leaf petiole was cut with a sharp razor
blade and then inserted into a pressure chamber Model 3005 Plant
Water Status Console (Soil Moisture Equipment Corp., Santa
Barbara, CA, USA). A total of ≈20 leaves per vineyard block
were used to estimate leaf ψ for each sampling date. Overall, there
were five sampling dates during the growing season; bi-weekly
between late June and early September 2005 to 2007 for each site.
Data were recorded in negative bar units (10 bars = 1 MPa).
Data analysis: Within each vineyard block, high and low water
status zones were identified accordingly based on GIS- generated
maps, and fruit were harvested separately from each zone for
yield components data. Fruit composition data were based on
individual 100-berry samples collected from each sentinel vine
in each site in all three seasons. In each vineyard block, all data
were analyzed by analysis of variance based on high and low
water status categories using SAS statistical package version
8 (SAS Institute; Cary, NC, USA). Correlation analysis was
performed for each vineyard block as well as across all blocks for
each year. Principal components analysis (PCA) was performed
on all data in each of the three years across all vineyard blocks.
Spatial correlation analysis was done by MapInfo and Vertical
Mapper (Northwood GeoScience, Ottawa, ON, Canada) at each
site and each year by analyzing spatial patterns in the gridfiles
for each variable.
Results
Seasonal weather data for 2005-2007: The three seasons varied
in growing degree days (GDD; base 10 oC) and precipitation
(Table 2). The 2005-07 GDD means ranged from 1495 GDD
(Buis, Niagara Lakeshore sub-appellation) to 1578 GDD
(Harbour Estates, Creek Shores sub-appellation). The 2005 season
was warmer than average with GDD averaging 1582 across the
region. Precipitation in 2005 (426 mm; April to October) was
close to average, but the period between May and late July was
dry. The 2006 season was relatively cool overall (1430 GDD)
with mean precipitation of 472 mm that was evenly distributed
throughout the growing season. Mean daily temperatures were
lower than average throughout much of July and August. The
2007 season was drier than the preceding two years, with
precipitation averaging 227 mm across the region, and GDD of
1583. Mean daily temperatures also remained > 20 oC throughout
much of the September.
Spatial variation at the research sites
Soil texture and composition: Parent material ranged from
lacustrine silty clay, reddish hued clay, and loamy textures to
reddish hued sandy textures (Table 1). Sand content varied from
26 to 52% across all sites with the highest content at the Harbour
Estate site, followed by the Buis and Reif sites (Fig. 1), while clay
ranged from 10 to 23%, with the highest % clay at the Château
des Charmes (CDC) and Buis sites (Fig. 2). OM ranged from 1.0
and 6.0% (Fig. 3); values < 3.0% are considered low for mineral
soils (Brady and Weil, 2002). CEC ranged from 6 to 53 (mEq/100
g soil) (Hakimi Rezaei, 2009; map data not shown). CEC values
> 20 are typically considered optimal (Brady and Weill, 2002).
Soil pH ranged from 5.5 and 8.0 (map data not shown). Soil
BS as % Ca (BS-Ca) ranged between 32 to 94% (map data not
shown). It is recommended that BS-Ca be > 75% (Brady and
Weil, 2002). Soil P varied from 6 and 186 mg/kg and K ranged
from 101 to 653 mg/kg (map data not shown). Soil Ca ranged
from 514 to 9898 mg/kg and Mg ranged from 100 to 716 mg/kg
(map data not shown).
Soil and vine water status 2005 to 2007- temporal stability:
Soil moisture in 2005 ranged from 7 to 20%, from 11 to 36% in
2006, and from 4 to 28% in 2007 across all sites (Figs. 4 to 6).
Lowest and highest soil moisture values, respectively, for each
season, were observed at the Hernder and Buis sites (2005), at the
Reif and Vieni sites (2006) and at the Harbour Estate and Buis
sites (2007) (Table 3). Lowest soil moisture values overall were
observed at the Hernder, Reif and Harbour sites. At the Hernder
site, lowest mean soil moisture values were 7.3, 15.1 and 6.1%
in 2005, 2006 and 2007, respectively; whereas the highest mean
soil moisture values were 13.4, 28.0, and 27.7%. At the Reif site,
lowest soil moisture values were 7.6, 11.3 and 8.8% in 2005,
2006 and 2007, while highest values were 13.6, 25.6 and 21.3%.
Likewise, at the Harbour site low soil moisture values were 9.9,
11.7 and 3.5% in 2005, 2006 and 2007, while highest values
were 13.4, 18.7 and 8.5%. Spatial variation in soil moisture was
temporally consistent at nine of ten sites across the 2005 to 2006
vintages (Table 4), and particularly at the Reif site (Fig. 4J,K),
and was temporally consistent at all sites from the 2006 to 2007
vintages, particularly Buis (Fig. 4B,C), CDC (Fig. 4E,F), Reif
(Fig. 4K,L), Cave Spring (Fig. 5H,I), Henry of Pelham (HOP;
 Spatial variability in Ontario Cabernet Franc vineyards 7

Fig. 1. Spatial distribution of sand at all vineyard blocks, Niagara Peninsula, ON; A: Buis; B: Chateau des Charmes; C: Hernder;
D: Reif; E: Harbour Estate; F: George; G: Cave Spring; H: Henry of Pelham; I: Vieni; J: Morrison.

Fig. 2. Spatial distribution of clay at all vineyard blocks, Niagara Peninsula, ON; A: Buis; B: Chateau des Charmes; C: Hernder;
D: Reif; E: Harbour Estate; F: George; G: Cave Spring; H: Henry of Pelham; I: Vieni; J: Morrison.
8 Spatial variability in Ontario Cabernet Franc vineyards
Fig. 3. Spatial distribution of organic matter in all vineyard blocks, Niagara Peninsula, ON; A: Buis; B: Chateau des Charmes;
C: Hernder; D: Reif; E: Harbour Estate; F: George; G: Cave Spring; H: Henry of Pelham; I: Vieni; J: Morrison.
Fig. 5K,L), and Morrison (Fig. 6E,F). An examination of temporal
consistency between the 2005 and 2007 vintages indicated that
eight of ten sites showed significant spatial correlations for soil
moisture.
Leaf ψ ranged from -0.80 to -1.60 MPa in 2005, -0.82 and -1.60
in 2006, and -0.93 to -1.64 MPa in 2007 (Fig. 7 to 9) across all
sites. The highest and lowest leaf ψ values, respectively, were
observed at Harbour Estate and CDC (2005), at Vieni and Hernder
(2006), and at Harbour Estate and CDC (2007) (Table 3). Leaf
ψ values were the basis for the water status categories that were
tested in terms of yield components and berry composition, and
those from which wines were made in 2005 and 2006. Spatial
variation in leaf ψ (Table 4; Figs. 7 to 9) was temporally consistent
at two sites, 2005 and 2006, at Hernder (Fig. 7G,H) and Harbour
(Fig. 8A,B). Three other sites (CDC, Reif, Vieni) had relatively
high r values, suggesting temporal stability (Table 4). In 2006
and 2007, temporal stability was apparent at three locations,
Reif (Fig. 7K,L), Harbour (Fig. 8B,C), and Cave Spring (Fig.
8H,I). Two other sites (George, HOP) had relatively high r
values, suggesting temporal stability. Comparing the 2005 and
2007 seasons, temporal stability was apparent at two locations,
at Vieni (Fig. 9A,C) and Morrison (Fig. 9D,F). Two other sites
(Reif, Harbour Estate) had relatively high r values, suggesting
temporal stability.
Spatial correlation analysis
Soil and vine water status- Niagara-on-the-Lake sites: Both
soil moisture and leaf ψ displayed spatial correlations with various
soil texture, physical properties, and compositional variables
(Table 5). At the Buis site, an inverse relationship was apparent
between BS and leaf ψ (based upon absolute value) in 2006
(Fig. 4B). At the CDC site, inverse correlative relationships were
observed between OM (Fig. 3B), CEC, BS, P, K, and Ca (Hakimi
Rezaei, 2009; map data not shown) vs. leaf ψ in 2005 (Fig. 7D),
as well as a positive correlation between leaf ψ and Mg (map data
not shown). Soil moisture at the Hernder location in 2006 (Fig.
4H) displayed spatial correlations with soil pH, BS, Ca, and Mg
(map data not shown). Apparent spatial correlations were also
observed between leaf ψ in 2005 and 2006 (Fig. 7G,H); these
included positive correlations with % sand (Fig. 1C), negative
ones with CEC (map data not shown), and positive ones (2006;
leaf ψ only) with P and K (Table 5; map data not shown). At the
Reif site, % sand (Fig. 1D) was inversely correlated with soil
moisture (2005, 2006; Fig. 4J,K), as were P and K (map data not
shown). A multitude of spatial correlations were also apparent
between leaf ψ (2005, 2006; Fig. 7J,K) vs. OM (Fig. 3D), P, K,
and Mg (map data not shown; all positive correlations) and those
involving clay, OM (Fig. 3D), CEC, BS, and Ca (map data not
shown; all inverse correlations).
Jordan, Vineland, and Beamsville sites: At the George site, soil
moisture in 2005 (Fig. 5D) correlated spatially with P (Table 5;
map data not shown), while leaf ψ in 2005 (Fig. 8D) correlated
with % clay (Fig. 2F), OM (Fig. 3F), BS, and Ca (map data not
shown). At the Cave Spring location, soil moisture in 2005 and
2006 (Fig. 5G,H) correlated directly with soil P and K (map data
not shown), and in 2005 only with soil pH, CEC, BS, and Ca
(map data not shown). Inverse relationships were also observed
between soil moisture and % sand (Fig. 1G; both seasons), and
Mg (2005 only; map data not shown). Leaf ψ (2005, 2006; Fig.
8G,H) correlated with % sand (Fig. 1G; 2006) and inversely
 Spatial variability in Ontario Cabernet Franc vineyards 9

Fig. 4. Spatial distribution of soil moisture (%), at four vineyard sites, Niagara Peninsula, ON; A to C: Buis; 2005 (A); 2006 (B); 2007 (C).
D to F: Chateau des Charmes; 2005 (D); 2006 (E); 2007 (F). G to I: Hernder; 2005 (G); 2006 (H); 2007 (I). J to L: Reif; 2005 |(J);
2006 (K); 2007 (L). In each map, the value of each zone represents the corresponding lower limit for that zone.
10 Spatial variability in Ontario Cabernet Franc vineyards

Fig. 5. Spatial distribution of soil moisture (%), at four vineyard sites, Niagara Peninsula, ON; A to C: Harbour Estate; 2005 (A); 2006 (B);
2007 (C). D to F: George; 2005 (D); 2006 (E); 2007 (F); G to I: Cave Spring 2005 (G); 2006 (H); 2007 (I). J to L: Henry of Pelham;
2005 (J); 2006 (K); 2007 (L). In each map, the value of each zone represents the corresponding lower limit for that zone.
 Spatial variability in Ontario Cabernet Franc vineyards 11

Table 4. Temporal stability of soil moisture and leaf water potential in ten
Cabernet Franc vineyards, Niagara Peninsula, Ontario, 2005-2007
Site Soil moisture Leaf ψ
2005 vs. 2006
Buis -0.34 0.14
Chateau des Charmes 0.56** 0.40
Hernder 0.42** 0.83**
Reif 0.84** 0.39
George 0.50** -0.28
Henry of Pelham 0.53** 0.03
Cave Spring 0.45** 0.22
Harbour Estate 0.50** 0.65**
Vieni 0.59** 0.44
Morrison 0.52** 0.24
2006 vs 2007
Buis 0.82** -0.40
Chateau des Charmes 0.78** -0.09
Hernder 0.59** 0.04
Reif 0.87** 0.84**
George 0.45** 0.45
Henry of Pelham 0.71** 0.47
Cave Spring 0.68** 0.50*
Harbour Estate 0.41** 0.66**
Vieni 0.62** 0.19
Morrison 0.71** 0.17
2005 vs. 2007
Buis -0.14 -0.45
Chateau des Charmes 0.69** -0.05
Hernder -0.10 0.11
Reif 0.67** 0.36
George 0.44** 0.08
Henry of Pelham 0.59** -0.40
Cave Spring 0.62** -0.12
Harbour Estate 0.57** 0.47
Vieni 0.66** 0.66**
Morrison 0.55** 0.65**
*, **: Significant at P < 0.05 or 0.01, respectively. Significant inverse
correlations are not indicated.
pH, BS, P, K, Ca, and Mg (map data not shown). Those spatial
correlations at the Morrison location involving soil moisture (Fig.
6D,E) included relationships with CEC (2006), soil pH (2006),
BS (2005, 2006), and Ca (2006)(all map data not shown); inverse
relationships included those with OM (Fig. 3J; 2006), P (map
data not shown; 2005), and K (map data not shown; 2005, 2006).
Leaf ψ in 2005 (Fig. 6D) was directly spatially correlated with
BS and Ca (map data not shown), and inversely with P, K, and
Mg (map data not shown).
Soil texture and composition
Niagara-on-the-Lake sites: In terms of soil texture and
Fig. 6. Spatial distribution of soil moisture (%), at two vineyard sites,
composition, there were many noteworthy spatial correlative
Niagara Peninsula, ON; A to C: Vieni; 2005 (A); 2006 (B); 2007 (C). D relationships in the ten vineyards (Table 5). Soil textural
to F: Morrison; 2005 (D); 2006 (E); 2007 (F). In each map, the value of components were associated with numerous variables, and those
each zone represents the corresponding lower limit for that zone.
relationships with OM may have had implications for soil and
with CEC, pH, and P (2006; map data not shown). Soil moisture vine water status. At the Buis site, % sand (Fig. 1A) showed high
at the HOP location in 2005 (Fig. 5J) displayed direct spatial spatial correlations with OM (Fig. 3A), P, and K (map data not
correlations with % clay (Fig. 2H), pH, BS, and Ca (map data not shown) and was inversely correlated with % clay (Fig. 2A), soil
shown). Leaf ψ (2005; Fig. 8J) was spatially correlated with CEC, pH, BS, Ca, and Mg (map data not shown). Percent clay (Fig. 2A)
pH, BS, and Ca (map data not shown), and inversely correlated showed positive spatial correlations with CEC, soil pH, Ca, and
with OM (Fig. 3H). Soil moisture at the Vieni site in 2005 (Fig. Mg (map data not shown) but was negatively correlated with OM
6A) displayed an inverse spatial correlation with OM (Fig. 3I). (Fig. 3A), P, and K (map data not shown). An example of maps
Leaf ψ in 2005 (Fig. 9A) was spatially correlated with % sand from this location comparing spatial relationships between soil
(Fig. 1I), and inversely correlated with % clay (Fig. 2I), CEC, moisture, % clay, CEC, soil Ca, BS, K and Mg is shown in Fig.
12 Spatial variability in Ontario Cabernet Franc vineyards

Fig. 7. Spatial distribution of leaf water potential (-MPa) at four vineyard sites, Niagara Peninsula, ON; A to C: Buis; 2005 (A); 2006 (B);
2007 (C). D to F: Chateau des Charmes; 2005 (D); 2006 (E); 2007 (F). G to I: Hernder; 2005 (G); 2006 (H); 2007 (I). J to L: Reif;
2005 (J); 2006 (K); 2007 (L) In each map, the value of each zone represents the corresponding lower limit for that zone.
 Spatial variability in Ontario Cabernet Franc vineyards 13

1.23

Fig. 8. Spatial distribution of leaf water potential (-MPa) at four vineyard sites, Niagara Peninsula, ON; A to C: Harbour Estate;
2005 (A); 2006 (B); 2007 (C). D to F: George; 2005 (D); 2006 (E); 2007 (F). G to I: Cave Spring; 2005 (G); 2006 (H); 2007 (I). J
to L: Henry of Pelham; 2005 (J); 2006 (K); 2007 (L).In each map, the value of each zone represents the corresponding lower limit
for that zone.
14 Spatial variability in Ontario Cabernet Franc vineyards
Fig. 9. Spatial distribution of leaf ψ (-MPa) at two vineyard sites, Niagara
Peninsula, ON; A to C: Vieni; 2005 (A); 2006 (B); 2007 (C). D to F:
Morrison; 2005 (D); 2006 (E); 2007 (F). In each map, the value of each
zone represents the corresponding lower limit for that zone.
10A-E. At the CDC site, % sand (Fig. 1B) was correlated spatially
with OM (Fig. 3B) and inversely correlated with % clay (Fig. 2B)
and soil moisture in 2005 (Fig. 4D). An example of maps from this
location illustrating spatial relationships between soil moisture,
% clay, CEC, soil Ca, BS, K and Mg is shown in Fig. 10F-J. At
the Hernder site, sand was inversely correlated spatially with
OM (Fig. 3C). At the Reif site, % clay (Fig. 2D) was positively
correlated spatially with CEC, soil pH, BS, and Ca (map data
not shown), but was negatively correlated with OM (Fig. 3D),
P, K, and Mg (map data not shown). At all sites, several spatial
correlations involved soil OM (Fig. 3A-D), CEC, soil pH, and
BS, as well as P, K, Ca, and Mg (map data not shown).
Jordan, Vineland, and Beamsville sites: Spatial correlation
analysis at the Harbour site showed that % clay was negatively
correlated with % sand (Table 5; Fig. 1E, S3E). At the George
site, % sand (Fig. 1F) was highly negatively correlated spatially
with a multitude of variables including % clay (Fig. 2F), OM
(Fig. 3F), soil pH, BS, K, Ca, and Mg (map data not shown).
As expected, % clay spatial relationships were inverse to those
with % sand. An example of maps from this location comparing
spatial relationships between soil moisture, % clay, CEC, Ca, BS,
K and Mg is shown in Fig. 11A-E. At the Cave Spring site, %
sand (Fig. 1G) was negatively correlated spatially with % clay
(Fig. 2G), CEC, pH, BS, P, K, and Ca (map data not shown), and
positively correlated with Mg (map data not shown). An example
of maps from this site comparing spatial relationships between
soil moisture, % clay, CEC, Ca, BS, K and Mg is shown in Fig.
11F-J. At the HOP site, % sand (Fig. 1H) was positively correlated
spatially with OM (Fig. 3H) and P (map data not shown) and
negatively correlated with % clay (Fig. 2H), pH, CEC, BS, and
Ca (map data not shown). As expected, relationships involving
% clay were inverse to those with sand, with the exception of a
lack of correlation with P. At the Vieni site, % sand (Fig. 1I) was
negatively correlated spatially with % clay (Fig. 2I), OM (Fig. 3I),
pH, CEC, BS, Ca, and Mg (map data not shown). Relationships
involving % clay were the inverse of those with % sand, with the
exception of a lack of correlation with OM. At the Morrison site,
sand (Fig. 1J) was spatially correlated with OM (Fig. 3J) and P
(map data not shown) and inversely with % clay (Fig. 2J) and
pH (map data not shown). At all sites, many spatial relationships
involved OM (Fig. 3F-J), pH, CEC, and BS, as well as P, K, Ca,
and Mg (map data not shown).
Correlation analysis: Correlation analysis of soil factors for all
sites in 2005 indicated that leaf ψ was positively correlated with %
clay, OM, soil pH, BS, Ca and Mg, and was negatively correlated
with % sand (Table 6). Soil moisture had positive correlation with
CEC, BS, and Ca, but was negatively correlated with K. Mg was
positively correlated with % clay, OM, CEC, pH, BS, and Ca, but
was negatively correlated with % sand, P and K. Ca was positively
correlated with % clay, CEC, pH and BS, and was negatively
correlated with % sand; K was positively correlated with %
sand and P, and negatively correlated with BS; P was negatively
correlated with % clay and BS but had a positive correlation
with % sand. BS had a positive correlation with % clay, CEC
and pH, and was negatively correlated with % sand. Soil pH was
positively correlated with CEC and % clay and negatively with
% sand; OM and CEC both negatively correlated with % sand
and positively with % clay; % clay negatively correlated with %
sand. In 2006 and 2007, soil moisture was negatively correlated
with % sand and was positively correlated with % clay, CEC,
pH, BS, Ca and Mg, while leaf ψ correlated negatively with %
sand (2007 only), and positively with % clay, CEC, pH, BS, K,
Ca and Mg (2007 only).
Principal components analysis: PCA conducted on the 2005
leaf ψ, soil moisture, and soil composition data explained 59.6
% of the variability in the data in the first two dimensions (Fig.
12A). PC1 accounted for 45.0 % of the variability and was
most heavily loaded in the positive direction with soil moisture,
leaf ψ, % clay, soil pH, OM, CEC, BS, Ca, and Mg. The small
 Spatial variability in Ontario Cabernet Franc vineyards 15

Fig. 10. Spatial variation in two east Niagara Cabernet Franc vineyards. A to E: Buis Vineyard; F to J: Chateau des Charmes.
A, F: Soil moisture (%); B, G: Clay (%), inset: cation exchange capacity (mEq/100 mL); C, H: Soil Ca (mg/kg), inset: base
saturation (% as Ca); D, I: Soil K (mg/K); E, J: Soil Mg (mg/kg). Values represent the lower limit within each zone.

Fig. 11. Spatial variation in two west Niagara Cabernet Franc vineyards. A to E: George Vineyard; F to J: Cave Spring Vineyard.
A, F: Soil moisture (%); B, G: Clay (%), inset: cation exchange capacity (mEq/100 mL); C, H: Soil Ca (mg/kg), inset: base
saturation (% as Ca); D, I: Soil K (mg/K); E, J: Soil Mg (mg/kg). Values represent the lower limit within each zone.
16 Spatial variability in Ontario Cabernet Franc vineyards

Fig. 12. Principal components analysis of leaf ψ, soil moisture, and soil composition data from ten Cabernet Franc sites, Niagara
Peninsula, ON, A: 2005. Percentages of variation represented by the data set are 45.0% (PC1) and 14.6% (PC2); B: 2006. Percentages
of variation represented by the data set are 44.5% (PC1) and 16.7 (PC2); C: 2007. Percentages of variation represented by the data
set are 43.8% (PC1) and 17.1% (PC2).
angles between the eigenvectors suggested that these variables K. The third PC explained another 13.9 % of the variation (data
were closely correlated. Percent sand was heavily loaded in the not shown). PCA conducted on these data in 2006 explained 61.2
negative direction. PC2 accounted for 14.6 % of the variability % of the variability in the data in the first two dimensions (Fig.
and was most heavily loaded in the positive direction with P and 12B). PC1 accounted for 45.0 % of the variability and was most

Table 5. Spatial correlations 2005-2007—soil composition, soil physical properties, soil moisture, leaf water potential. Abbreviations: OM: organic
matter; CEC: cation exchange capacity; BS: base saturation; SM: soil moisture
Sand Clay Soil pH Soil OM CEC BS-Ca P K Ca Mg
Buis
Clay -0.88**
Soil pH -0.74** 0.76**
Soil OM 0.73** -0.82** -0.66**
CEC -0.48 0.55* 0.39 -0.44
BS-Ca -0.59* 0.43 0.76** -0.29 -0.10
P 0.50* -0.68** -0.65** 0.70** -0.08 -0.27
K 0.64** -0.85** -0.71** 0.70** -0.30 -0.44 0.74**
Ca -0.82** 0.79** 0.91** -0.61** 0.63** 0.70** -0.37 -0.64**
Mg -0.67** 0.73** 0.85** -0.61** 0.63** 0.43 -0.62** -0.64** 0.82**
SM 05 0.17 -0.37 0.13 0.28 -0.10 0.34 0.40 0.45 0.13 -0.07
SM 06 -0.33 0.42 0.21 -0.45 0.75** -0.02 0.02 -0.35 0.45 0.38
SM 07 -0.19 0.25 0.08 -0.40 0.60** -0.15 0.08 -0.22 0.28 0.27
Leaf ψ 05 -0.01 0.06 -0.22 0.07 0.01 -0.01 0.33 -0.23 -0.02 -0.39
Leaf ψ 06 0.28 -0.21 -0.48 0.14 0.01 -0.71** 0.04 0.22 -0.51* -0.34
Leaf ψ 07 0.12 -0.48 -0.14 0.33 -- 0.22 0.53* 0.67** -0.07 -0.17
Chateau des Charmes
Clay -0.79**
Soil pH -0.08 -0.43
Soil OM 0.57* -0.47 0.20
CEC -0.10 -0.26 0.79** 0.03
BS-Ca -0.16 -0.31 0.96** 0.17 0.89**
P 0.20 -0.30 0.51* 0.77** 0.56* 0.62**
K 0.25 0.07 -0.25 0.66** -0.20 -0.18 0.50*
Ca -0.12 -0.25 0.83** 0.05 0.99** 0.92** 0.58* -0.19
Mg 0.18 0.15 -0.78** -0.31 -0.77** -0.86** -0.76** -0.09 -0.80**
SM 05 -0.65** 0.40 0.07 -0.38 -0.07 0.08 -0.20 -0.30 -0.04 -0.04
SM 06 -0.30 0.31 -0.15 -0.14 -0.42 -0.23 -0.36 -0.03 -0.40 0.38
SM 07 -0.48 0.53* -0.25 -0.22 -0.34 -0.23 -0.28 0.03 -0.32 0.22
Leaf ψ 05 0.07 0.07 -0.41 -0.50* -0.55* -0.56* -0.84** -0.55* -0.57* 0.72**
Leaf ψ 06 0.30 -0.15 -0.32 -0.13 -0.19 -0.35 -0.28 -0.43 -0.22 0.31
Leaf ψ 07 -0.42 0.59* -0.18 0.21 -0.24 -0.14 0.15 0.21 -0.23 0.05
 Spatial variability in Ontario Cabernet Franc vineyards 17

Table 5 contd.
Sand Clay Soil pH Soil OM CEC BS-Ca P K Ca Mg
Hernder
Clay 0.45
Soil pH -0.46 -0.23
Soil OM -0.60** -0.34 -0.56*
CEC -0.22 -0.22 0.36 -0.03
BS-Ca 0.02 -0.23 0.98** -0.53* 0.33
P 0.11 0.21 -0.42 0.47 -0.54* -0.38
K 0.17 0.04 -0.26 0.43 -0.52* -0.25 0.92**
Ca -0.03 -0.03 0.93** -0.43 0.65** 0.93** -0.47 -0.35
Mg -0.12 -0.4 0.60** -0.16 0.12 0.69** 0.06 0.15 0.59*
SM 05 0.21 0.18 0.31 -0.13 -0.24 0.29 0.02 0.15 0.14 0.18
SM 06 -0.33 -0.25 0.60** -0.09 0.03 0.66** -0.05 0.01 0.54* 0.57*
SM 07 -0.50* 0.07 -0.03 0.03 0.29 -0.04 -0.14 -0.29 0.14 -0.02
Leaf ψ 05 0.58* 0.25 -0.23 -0.12 -0.64** -0.33 0.27 0.36 -0.48 -0.36
Leaf ψ 06 0.52* 0.38 -0.09 -0.02 -0.75** -0.13 0.60** 0.68** -0.35 -0.07
Leaf ψ 07 -0.31 0.23 -0.03 0.06 -0.14 -0.1 -0.33 -0.43 -0.1 -0.3
Reif
Clay 0.20
Soil pH 0.09 0.98**
Soil OM -0.14 -0.93** -0.94**
CEC -0.03 0.90** 0.94** -0.86**
BS-Ca 0.03 0.96** 0.99** -0.93** 0.98**
P -0.16 -0.75** -0.73** 0.90** -0.58* -0.71**
K -0.49 -0.89** -0.82** 0.81** -0.64** -0.76** 0.76**
Ca -0.03 0.91** 0.95** -0.87** 0.99** 0.98** -0.60** -0.65**
Mg 0.24 -0.62** -0.66** 0.81** -0.65** -0.71** 0.87** 0.45 -0.66**
SM 05 -0.57* -0.35 -0.29 0.43 -0.09 -0.22 0.57* 0.57* -0.1 0.26
SM 06 -0.55* -0.38 -0.27 0.38 -0.06 -0.21 0.61** 0.68** -0.08 0.29
SM 07 -0.50* -0.31 -0.21 0.32 0.02 -0.15 0.59* 0.65** 0.001 0.27
Leaf ψ 05 0.23 -0.22 -0.25 0.52* -0.21 -0.29 0.75** 0.14 -0.22 0.86**
Leaf ψ 06 -0.07 -0.80** -0.75** 0.80** -0.58* -0.71** 0.86** 0.85** -0.60** 0.72**
Leaf ψ 07 -0.11 -0.94** -0.92** 0.95** -0.81** -0.90** 0.88** 0.87** -0.83** 0.80**

heavily loaded in the positive direction with soil moisture, leaf ψ,
% clay, pH, CEC, BS, Ca, and Mg. Again, small angles between
the eigenvectors suggested that these variables were closely
correlated. Percent sand was again heavily loaded in the negative
direction. PC2 accounted for 16.7 % of the variability and was
most heavily loaded in the positive direction with OM, P, and K.
The third PC explained another 11.6 % of the variation (data not
shown). PCA conducted on these data in 2007 explained 60.9
% of the variability in the data in the first two dimensions (Fig.
12C). PC1 accounted for 43.8 % of the variability and as in 2005
and 2006 was most heavily loaded in the positive direction with
soil moisture, leaf ψ, % clay, pH, CEC, BS, Ca, and Mg. Small
angles between the eigenvectors suggested that these variables
were closely correlated. Percent sand was once again heavily
loaded in the negative direction. PC2 accounted for 17.1 % of the
variability and was most heavily loaded in the positive direction
with OM, P, and K. The third PC explained another 10.5 % of
the variation (data not shown).
Discussion
This investigation was initiated with the purpose of identifying the
major factors that contribute to the terroir effect in the vineyards
of the Niagara Peninsula in Ontario. It was hypothesized,
consistently with Seguin (1986), that the main factors might be
soil-texture based. It was also hypothesized, consistent with van
Leeuwen and Seguin (1994) and van Leeuwen et al. (2004) that
the terroir effect would be strongly based upon soil moisture, vine
water status, or both. These hypotheses were for the most part
proven, primarily with respect to soil moisture. In the majority of
situations, distinct spatial patterns in soil texture, soil moisture,
and leaf ψ were demonstrated. Moreover, the spatial patterns were
in most cases (soil moisture) and occasionally (leaf ψ) temporally
stable, and any temporal variations in their spatial patterns were
likely influenced by the volatile precipitation patterns that are
typical of the region. Finally, there were clear spatial correlations
between soil moisture, leaf ψ, and many soil physical and
composition variables, including soil texture (% sand and clay),
soil pH, OM, CEC, BS, P, K, Ca, and Mg.
Spatial variability
Soil moisture: Based on the range of soil moistures obtained
at each site and in each year, it was possible to identify soil
water status zones at each vineyard block; therefore the part
of hypothesis that temporally stable soil water status zones
could be identified was supported by the data in all three years.
18 Spatial variability in Ontario Cabernet Franc vineyards

Table 5 contd.
Sand Clay Soil pH Soil OM CEC BS-Ca P K Ca Mg
Harbour
Clay -0.91**
Soil pH 0.72** -0.56**
Soil OM -0.18 0.26 0.08
CEC -0.60** 0.60** -0.42 0.47*
BS-Ca -0.70** 0.66** -0.16 0.54* 0.46*
P 0.73** -0.68** 0.46* -0.01 -0.10 -0.72**
K 0.22 -0.18 0.23 -0.07 0.39 -0.27 0.64**
Ca -0.76** 0.71** -0.31 0.50* 0.76** 0.92** -0.56** 0.03
Mg 0.34 -0.12 0.68** 0.31 -0.18 0.20 -0.09 -0.07 0.05
SM 05 0.06 0.07 -0.13 -0.62** -0.33 -0.50* 0.02 -0.01 -0.50* ----a
SM 06 -0.40 0.31 -0.33 -0.62** 0.02 0.17 -0.46 -0.06 0.17 -0.23
SM 07 ----a ---- 0.16 -0.53* -0.32 -0.68** 0.37 0.30 -0.62** 0.04
Leaf ψ 05 -0.42 0.51* -0.35 -0.46 0.31 -0.02 -0.14 0.36 0.14 -0.35
Leaf ψ 06 -0.23 0.21 0.01 0.01 0.69** 0.15 0.17 0.61** 0.40 -0.25
Leaf ψ 07 -0.06 0.03 -0.05 -0.18 0.51* -0.02 0.16 0.77** 0.29 0.02
George
Clay -0.88**
Soil pH -0.91** 0.79**
Soil OM -0.51* 0.50* 0.22
CEC -0.33 0.41 0.27 0.64**
BS-Ca -0.89** 0.82** 0.89** 0.35 0.10
P -0.35 0.35 0.64 -0.44 -0.15 0.53*
K -0.68** 0.70** 0.70** 0.22 0.35 0.57* 0.60**
Ca -0.86** 0.84** 0.82** 0.65** 0.67** 0.80** 0.29 0.61**
Mg -0.77** 0.81** 0.83** 0.07 0.22 0.67** 0.67** 0.82** 0.61**
SM 05 -0.23 0.07 0.38 -0.22 -0.14 0.34 0.66** 0.43 0.18 0.32
SM 06 -0.71** 0.72** 0.80** 0.24 0.50* 0.72** 0.68** 0.78** 0.78** 0.75**
SM 07 -0.17 0.31 0.41 -0.51* -0.16 0.28 0.78** 0.48 0.07 0.60**
Leaf ψ 05 -0.48 0.66** 0.27 0.73** 0.26 0.53* -0.05 0.46 0.54* 0.31
Leaf ψ 06 -0.14 0.06 0.29 -0.62** -0.75** 0.32 0.48 -0.05 -0.23 0.27
Leaf ψ 07 0.47 -0.30 -0.54* -0.27 -0.75** -0.30 -0.30 -0.54* -0.66** -0.39

Moreover, the hypothesis that the spatial variation would be stable
temporally was also proven by the data. This hypothesis carried
with it the assumption that soil water status zones as well as vine
water status zones would be stable temporally. Stable water status
zones might give opportunity for selective harvest of different
sections of each block, if water status could be linked to fruit
composition. Since this variation is often reflected in yield and
fruit composition, it might be to the winemaker’s advantage for
these zones to be individually harvested, with the potential of
separating high quality grapes from lesser quality ones (Bramley,
2001; 2002; 2005; Bramley et al., 2003). Therefore, it could be
possible to produce premium quality wine from a portion of a
vineyard block rather than blending all the fruit into a lower
quality product.
Soil texture is an important factor that affects soil water retention.
The available soil moisture ranges from 30 mm/m of soil depth
for sands to 160 mm/m for clays (Goldberg et al., 1971). The
capacity of soil to store water depends on root zone depth and soil
water holding capacity. Infiltration rate also has significant effect
on water supply (Smart and Coombe, 1983). Soil moisture values
varied among vineyards as well as within vineyards in all three
years. The lowest soil moisture values were observed at three
sites (Hernder, Reif and Harbour) (Table 3). The low soil moisture
values at these three sites were attributable to shallow soil
profiles, higher content of gravel, and sandy loam soil textures,
respectively, all of which do not allow for high water retention
in the soil profile. The Hernder site had a clay loam-based soil
texture but with a shallow soil profile with low water-holding
capacity (Table 1). The Reif site also contained a clay loam soil
texture but with considerable gravel that facilitated rapid soil
drainage. The Harbour site, with 48% sand, had a sandy loam
soil texture that provided low soil moisture retention. Highest soil
moisture values in 2005 were at the Buis site in a range of 14.0 to
20.4%; in 2006 the highest soil moisture values were observed at
the Vieni site with a range of 22.2 to 35.9%, and in 2007 the Buis
site again had the highest soil moisture with a range of 17.2 to
27.6%. The Buis site had a deep loam soil with higher ability to
hold water in the soil profile, while the Vieni site also had a clay
loam till-based soil with high water-holding capacity. Overall, soil
moisture values were higher in 2006 at all sites in comparison
with 2005 and 2007 due to higher precipitation (Table 2).
It is worthy of note that Coipel et al. (2006) showed that soil
texture and composition were not crucial to the terroir effect but
soil depth was critical in terms of how it impacted vine water
and nitrogen status. Shallow soils generally led to vines with
low water status and low nitrogen, but also ultimately produced
small berries that were high in Brix and anthocyanins. This was
among the first studies to underscore the interactive effects of
soil texture, vine water status, and nitrogen status. Our results
likewise showed that the deeper, coarse-textured soils typically
had high leaf ψ values whereas the shallow, fine-textured clay
and clay loam soils had low leaf ψ values.
 Spatial variability in Ontario Cabernet Franc vineyards 19

Table 5 contd.
Sand Clay Soil pH Soil OM CEC BS-Ca P K Ca Mg
Cave Spring
Sand
Clay -0.55*
Soil pH -0.78** 0.40
Soil OM 0.27 -0.65** -0.20
CEC -0.80** 0.62** 0.93** -0.46
BS-Ca -0.79** 0.60** 0.91** -0.46 0.94**
P -0.84** 0.29 0.72** -0.28 0.74** 0.71**
K -0.56* -0.08 0.37 0.04 0.32 0.40 0.82**
Ca -0.80** 0.61** 0.93** -0.46 0.99** 0.96** 0.76** 0.36
Mg 0.83** -0.56* -0.82** 0.41 -0.83** -0.95** -0.77** -0.56* -0.87**
SM 05 -0.73** 0.15 0.56* -0.20 0.56* 0.54* 0.82** 0.69** 0.57* -0.59*
SM 06 -0.61** 0.14 0.38 -0.20 0.45 0.31 0.75** 0.56* 0.44 -0.33
SM 07 -0.60** 0.28 0.50* -0.25 0.57* 0.46 0.70** 0.49 0.57* -0.47
Leaf ψ 05 0.09 0.11 -0.55* 0.02 -0.46 -0.37 -0.28 -0.13 -0.46 0.22
Leaf ψ 06 0.57* 0.05 -0.56* -0.15 -0.50* -0.33 -0.52* -0.25 -0.47 0.23
Leaf ψ 07 0.62** -0.36 -0.25 0.48 -0.43 -0.30 -0.71** -0.49 -0.42 0.32
Henry of Pelham
Clay -0.71**
Soil pH -0.65** 0.85**
Soil OM 0.57* -0.72** -0.21
CEC -0.60** 0.92** 0.93** -0.86**
BS-Ca -0.58* 0.82** 0.97** -0.82** 0.96**
P 0.61** -0.03 -0.08 0.25 -0.04 -0.10
K -0.39 0.14 -0.09 0.04 -0.09 -0.23 -0.05
Ca -0.60** 0.90** 0.93** -0.87** 0.99** 0.97** -0.06 -0.12
Mg -0.02 0.01 -0.38 0.15 -0.22 -0.45 0.10 0.76** -0.26
SM 05 -0.11 0.51* 0.50* -0.27 0.49* 0.51* 0.26 -0.39 0.50* -0.42
SM 06 0.05 0.13 0.20 -0.05 0.14 0.20 0.14 -0.33 0.15 -0.30
SM 07 -0.03 0.24 0.32 0.08 0.16 0.23 0.37 -0.16 0.17 -0.32
Leaf ψ 05 -0.47* 0.36 0.60** -0.70** 0.62** 0.68** -0.44 -0.22 0.64** -0.47*
Leaf ψ 06 0.69** -0.50* -0.40 0.29 -0.34 -0.32 0.38 -0.48 -0.32 -0.15
Leaf ψ 07 0.52* -0.29 -0.28 0.31 -0.31 -0.32 0.48 -0.27 -0.29 -0.12
Leaf water potential: The role of vine water relations as an stress appeared earlier and was more severe. The highest and
important driver of the terroir effect was established by both lowest mean leaf ψ values, respectively, were observed at the
Seguin (1986) and van Leeuwen et al. (2004, 2009). It is likely Harbour (range –0.80 to -1.09 MPa) and CDC sites (range -1.20
that the effects of climate and soil on fruit composition are to -1.60 MPa ) in 2005, at the Vieni (range -0.82 to -1.10 MPa)
mediated through their influence on vine water status (Van and Hernder sites (range -1.29 to -1.69 MPa ) in 2006, and at
Leeuwen et al., 2004). Generally, coarse-textured gravelly soils the Harbour (range -0.93 to -1.12 MPa) and CDC sites (range
with exceptional drainage or shallow, fine-textured soils with -1.52 to -1.64 MPa ) in 2007 (Table 3). The potential for water
low growth potential will lead to mild water stress in red wine stress was always more intense at the CDC (three year range -
cultivars, which consequently result in higher soluble solids and 1.20 to -1.64 MPa) and Hernder (three year range -1.26 to -1.60
anthocyanins, and lower berry weights, vine size, and TA (van MPa) sites. The lowest leaf ψ values at CDC (2005, 2007) were
Leeuwen, 2010; van Leeuwen and Seguin, 1994; van Leeuwen possibly due to low precipitation in combination with the heavy
et al., 2004; 2009). Work with Cabernet Franc in St. Emilion lacustrine clay loam soil texture which, even with relatively high
specifically underscored the importance of low leaf ψ during soil moisture in the soil profile (14 and 17.2%), water was below
the veraison to harvest period in terms of ultimate wine quality. wilting point (< -1500 kPa) and therefore unavailable (Kingston
Those sites with low vine water status had fruit with highest Brix, and Presant, 1989). The low leaf ψ values at the Hernder site
anthocyanins, and phenols (Van Leeuwen and Seguin, 1994). were likely due to the shallow soil profile and the clay loam soil
texture that had low available moisture in the profile (Kingston
The leaf ψ values measured at the different sites are in the
and Presant, 1989).
range commonly reported for irrigated grapevines in California
(Williams and Matthews, 1990). High soil water availability at Grapevines growing in deep coarse sands or gravel may have
some sites likely reduced vine water stress by increasing leaf ψ roots penetrating to depths > 6 m (Smart and Coombe, 1983).
values. Leaf ψ values varied within all vineyard blocks, enabling High leaf ψ values at the Harbour site could be due to the sandy
vine separation into high, medium and low water status categories loam soil texture as well as the deep soil profile that permitted
for each vineyard block for all three years, and therefore the vigorous vine growth and a deep rootsystem. The deep roots of
hypothesis that temporally-stable water status zones could be these vines allowed water absorption from deeper soil layers;
delineated was proven by the data in all three years. In 2005 therefore vines at this site did not face water stress in any of the
and 2007, which were dry and hot years, the potential for water three years. Chardonnay vines that received irrigation at 100% of
20 Spatial variability in Ontario Cabernet Franc vineyards

Table 5 contd.
Sand Clay Soil pH Soil OM CEC BS-Ca P K Ca Mg
Vieni
Clay -0.90**
Soil pH -0.61** 0.79**
Soil OM -0.51* 0.31 -0.18
CEC -0.60** 0.69** 0.96** -0.21
BS-Ca -0.65** 0.83** 0.98** -0.11 0.91**
P -0.43 0.44 0.35 0.57* 0.30 0.37
K -0.40 0.17 0.03 0.66** 0.11 0.04 0.75**
Ca -0.63** 0.78** 0.99** -0.18 0.98** 0.98** 0.33 0.06
Mg -0.83** 0.78** 0.51 0.71** 0.46 0.57* 0.78** 0.66** 0.52*
SM 05 0.16 -0.02 0.36 -0.52* 0.42 0.28 -0.04 -0.13 0.36 -0.21
SM 06 0.29 -0.19 0.08 -0.37 0.10 -0.02 -0.15 -0.23 0.05 -0.35
SM 07 -0.25 0.36 0.52* 0.04 0.50* 0.46 0.57* 0.27 0.50* 0.36
Leaf ψ 05 0.64** -0.68** -0.74** -0.30 -0.73** -0.72** -0.82** -0.65** -0.73** -0.78**
Leaf ψ 06 0.16 0.12 -0.08 -0.22 -0.28 0.001 -0.35 -0.71** -0.12 -0.27
Leaf ψ 07 0.85** -0.79** -0.43 -0.75** -0.39 -0.48 -0.63** -0.54* -0.44 -0.94**
Morrison
Clay -0.84**
Soil pH -0.55* 0.58*
Soil OM 0.52* -0.72** -0.47
CEC -0.42 0.36 0.81** -0.42
BS-Ca -0.05 -0.13 0.48 -0.21 0.83**
P 0.50* 0.24 0.45 0.19 0.15 -0.12
K -0.13 -0.08 -0.26 0.59* -0.51* -0.63** 0.67**
Ca 0.28 0.15 0.67** -0.34 0.96** 0.94** 0.02 -0.57*
Mg -0.23 0.62** 0.09 -0.27 -0.29 -0.73** 0.07 0.18 -0.52*
SM 05 0.33 -0.20 0.01 -0.05 0.35 0.52* -0.62** -0.63** 0.45 -0.30
SM 06 -0.11 0.15 0.56* -0.50* 0.70** 0.73** -0.14 -0.63** 0.73** -0.35
SM 07 -0.15 0.27 0.69** -0.31 0.60** 0.42 0.05 -0.36 0.52* 0.06
Leaf ψ 05 0.33 -0.32 0.07 -0.22 0.47 0.78** -0.57* -0.82** 0.63** -0.62**
Leaf ψ 06 0.03 -0.19 0.33 0.13 0.40 0.40 0.23 -0.08 0.40 -0.31
Leaf ψ 07 0.41 -0.22 0.33 -0.17 0.35 0.50* -0.35 -0.68** 0.39 -0.18
*, **: Significant at p < 0.05 or 0.01 (boldfaced values), respectively. a Correlation coefficients were non-determinable.
evapotranspiration had leaf ψ values of -1.0 MPa (Williams and Zones of elevated soil and petiole K have been associated
Araujo, 2002), which suggests that vines at the Harbour site had with zones of high TA, and patterns in K:Mg ratios have been
adequate water availability, similar to that of irrigated vines. The associated with spatial variability in yield and Brix (Bramley,
high leaf ψ values at Vieni in 2006 were due to high soil moisture 2001). Zones within vineyards with high % clay may frequently
in that year; in fact in 2006 the Vieni site had the highest soil be those zones with high soil moisture, as well as higher CEC,
moisture among all ten sites. Although leaf ψ differed spatially BS, K, Mg, Ca, and perhaps other elements, but the implications
within each of the vineyard blocks as well as across vineyards, the of these relationships for yield and wine quality will ultimately
range of leaf ψ values remained very consistent in most vineyard depend upon the potential for those high clay zones to drain
blocks in all years, even with the different weather conditions. sufficiently (Bramley, 2001).
The minimal temporal variation within the season suggests that
Temporal stability
the data density was sufficient.
Soil moisture: The relevance of soil moisture data is strongly
Soil texture and composition: Spatial patterns were observed dependent upon the soil texture and the depth of the rootsystem.
for all sites in terms of soil texture and composition variables, The majority of the rootsystems of grapevines are found in the
and several of these were related to soil and vine water status. top one meter of soil (van Zyl and Weber, 1981) (most of them
Relationships among soil textural components (% sand and in the upper 30 to 50 cm), although a grapevine rootsystem may
clay), soil physical properties (CEC, BS, OM, pH), and major exceed 6 m in depth under some conditions (Smart and Coombe,
elements (P, K, Ca, Mg) are widely known. For instance, as % 1983). The vineyard blocks in this study were all non-irrigated
clay increases in a soil, generally soil moisture, as well as CEC, sites and would be expected to have roots growing deeply into
BS, K, Ca, and Mg likewise will increase (Brady and Weil, 2002). the soil profile. However, the soil water table is relatively high
In limestone-based soils in Ontario, this is often accompanied by in the Niagara region and drainage tiles are typically placed at
increases in pH and P (Kingston and Presant, 1989). OM is also a ≈60 cm depth; hence most rootsystems, particularly those in
a contributing factor to soil moisture. Percent clay also may be lacustrine clays, are in the top 30 to 60 cm. Soil moisture zones
inversely correlated spatially with yield and berry weight, and were temporally stable, particularly at the Reif site for 2005 and
directly with anthocyanins and phenols (Bramley, 2001). Spatial 2006 (Table 4). From 2006 to 2007, soil moisture zones were
relationships between yield and berry composition vs. soil and again temporally stable at all sites, most particularly Buis, Reif
petiole elemental composition have likewise been demonstrated. and CDC. Eight of ten locations were also temporally stable
 Spatial variability in Ontario Cabernet Franc vineyards 21

Table 6. Overall correlations and p values of soil factors for ten Cabernet Franc sites Niagara Peninsula, ON. 2005-07. Abbreviations: OM= organic
matter; CEC= cation exchange capacity; BS= base saturation; SM= soil moisture
Sand Clay OM CEC Soil BS P K Ca Mg SM ψ SM ψ SM ψ
(%) (%) (%) (meq/ pH (%Ca) (ppm) (ppm) (ppm) (ppm) (%) (-MPa) (%) (-MPa) (%) (-MPa)
100g) 2005 2005 2006 2006 2007 2007
% Sand 1.000 -0.895 -0.218 -0.641 -0.526 -0.615 0.205 0.175 -0.613 -0.623 -0.135 -0.597 -0.654 -0.179 -0.262 -0.541
<.0001 0.0055 <.0001 <.0001 <.0001 0.0092 0.0266 <.0001 <.0001 0.0871 <.0001 <.0001 0.0223 0.0004 <.0001
% Clay 1.000 0.156 0.629 0.511 0.610 -0.280 -0.101 0.589 0.647 0.013 0.726 0.693 0.386 0.376 0.662
0.0489 <.0001 <.0001 <.0001 0.0003 0.2013 <.0001 <.0001 0.8709 <.0001 <.0001 <.0001 <.0001 <.0001
% OM 1.000 0.087 0.037 0.049 0.092 0.135 0.039 0.403 -0.013 0.216 0.174 0.110 0.153 0.221
0.2728 0.6447 0.5385 0.2476 0.0889 0.6258 <.0001 0.8689 0.0060 0.0265 0.1639 0.0403 0.0029
CEC 1.000 0.768 0.755 -0.030 -0.146 0.989 0.347 0.268 0.429 0.436 0.237 0.366 0.366
(meq/100 g) <.0001 <.0001 0.7082 0.0648 <.0001 <.0001 0.0006 <.0001 <.0001 0.0024 <.0001 <.0001
Soil pH 1.000 0.891 -0.092 -0.149 0.815 0.341 0.139 0.358 0.315 0.185 0.339 0.281
<.0001 0.2465 0.0595 <.0001 <.0001 0.0786 <.0001 <.0001 0.0184 <.0001 0.0001
BS 1.000 -0.169 -0.160 0.813 0.347 0.190 0.505 0.404 0.302 0.471 0.370
(% Ca) 0.0326 0.0421 <.0001 <.0001 0.0158 <.0001 <.0001 <.0001 <.0001 <.0001
P (ppm) 1.000 0.609 -0.021 -0.450 0.029 -0.130 0.037 -0.050 -0.039 0.146
<.0001 0.7872 <.0001 0.7114 0.0991 0.6406 0.5276 0.6077 0.0505
K (ppm) 1.000 -0.154 -0.255 -0.183 0.049 -0.074 0.219 0.028 0.231
0.0511 0.0011 0.0199 0.5375 0.3481 0.0070 0.7088 0.0019
Ca (ppm) 1.000 0.292 0.269 0.392 0.386 0.218 0.358 0.322
0.0002 0.0006 <.0001 <.0001 0.0051 <.0001 <.0001
Mg (ppm) 1.000 0.038 0.436 0.612 0.015 0.204 0.326
0.6289 <.0001 <.0001 0.8451 0.0062 <.0001
SM (%) 2005 1.000 -0.095 1.000 0.218 1.000 0.418
0.2266 0.0052 <.0001
ψ (-MPa) 2005 1.000 - 1.000 - 1.000

when comparing 2005 and 2007. Therefore, the hypothesis that
soil moisture zones would be consistent and stable temporally
within vineyard blocks was supported by the data from 2005
to 2006, and was also sufficiently proven by the 2006 to 2007
data. The reason for any lack in temporal stability could be in
part due to the overall volatility of weather conditions in the
region, and the particularly low precipitation in 2005 and 2007.
The moisture variation in the upper 30 cm of soil can be high,
and small volumes of precipitation can consequently result in
high soil moisture readings. Therefore, in years of high rainfall,
heavy clay soils with low infiltration rates will have high soil
moisture in the upper portions of the soil profile, whereas the
lower layers might be drier. On the other hand, during seasons
with low precipitation, the upper portions of the soil profile may
be dry and low soil moisture values are typically obtained, even
though lower layers of soil may contain moisture.
Leaf water potential: Perhaps the first published use of
geomatic tools to map vine water status showed some clear
spatial correlations between berry carbon isotope concentration
(б13C) and stem ψ (Van Leeuwen et al., 2006). This supported
data showing relationships between predawn leaf ψ and berry
б13C (Gaudillère et al., 2002). In the present study, leaf ψ zones
were only occasionally temporally stable, e.g. at the Harbour and
Hernder sites from 2005 to 2006, and at the Harbour and Reif sites
from 2006 to 2007 (Table 3). At the Harbour site, leaf ψ zones
were stable over all three years, possibly as a consequence of the
likelihood of deep root systems afforded by the sandy soil at that
site. Therefore, the hypothesis that water-status zones would be
consistent within vineyard blocks was at best partially supported
by the data. Considering that soil texture was stable at each site,
and water holding capacity of each soil was also consistent, the
only difference was the amount of precipitation in each year. It is
safe to assume that since the average volume of water in the soil
profile changes between years so does vine water status likewise
change. There may, however, be relevant variables associated with
soil texture and soil water holding capacity that may vary spatially
and temporally. In a study on spatial variability in a Riesling
vineyard in Ontario, specific areas of the vineyard that produced
high yields or high concentrations of monoterpenes were transient
and that their spatial distribution varied temporally (Reynolds
et al., 2007; 2010a). Willwerth et al. (2010) demonstrated
temporally-stable spatial inverse correlations between Riesling
monoterpenes and leaf ψ. Nonetheless, our data suggest that
caution must be exercised in using leaf ψ measurements as the
basis for precision viticulture, since spatial distribution of leaf
ψ may vary temporally, which makes selected harvest based on
leaf ψ values challenging. It is very likely that a greater density
of sampling may have led to more widespread temporal stability
in leaf ψ patterns throughout the vineyards.
In situations where leaf ψ does not vary temporally, this
knowledge might allow implementation of specific viticultural
practices in high water status situations (canopy management,
crop reduction) or in drought situations (e.g. deficit irrigation)
(Van Leeuwen et al., 2009). There is also potential for establishing
temporally-stable zones of different flavor potential (Willwerth
et al., 2010). In Cabernet Franc, 2-methoxy-3-isobutylpyrazine
(IBMP) is ubiquitous worldwide, and a substantial soil-based
influence has been demonstrated (less IBMP in gravel soils)
22 Spatial variability in Ontario Cabernet Franc vineyards
(Peyrot des Gachons et al., 2005; Roujou de Boubée et al., 2000).
The norisoprenoid β-damascenone has a substantial impact on
wine aroma; by itself (apple notes), by enhancement of odor
activity (increased fruity notes) of ethyl cinnamates and ethyl
caproate, and by suppression of odor activity of IBMP in cultivars
such as Cabernet Franc; its concentration varies according to soil
type (Pineau et al., 2007). Cysteine precursors of odor-active thiol
compounds were closely linked to N status in Sauvignon blanc,
and high N zones within vineyards can potentially increase its
varietal typicity (Choné et al., 2006).
Precision viticulture: For the results of this study to be useful,
the patterns of variation within vineyard blocks must be constant
from year to year. Although the absolute values of yield and berry
composition for a vineyard may vary from vintage to vintage,
the patterns of variation within blocks are normally stable
(Bramley, 2005). In this study, variation in soil composition, soil
moisture, leaf ψ, yield components and fruit composition was
demonstrated in all vineyard blocks either by statistical analysis or
by interpolation of GIS maps. The patterns of variation, however,
were not temporally consistent from year to year for all variables
at all sites. Precision viticulture is dependent on the existence
of stable variability in patterns that can be managed effectively.
If such variability does not exist, then a uniform management
system is less expensive and more effective. To mitigate against
possible variability, vines could be planted and trained within
zones of similar soil texture, therefore reducing the need to
manage them differentially afterwards. By differential planting,
one can impose uniform management, which is more economical
than uniformly planting and differentially managing (Bramley,
2005). Although studies comparing uniform planting/differential
management vs. differential planting/uniform management appear
not to exist in the literature, our data suggests that longer periods
of study would help to elucidate these trends.
There are implications from these geomatic studies for precision
viticulture, if spatial variability in vine vigor and yield can be
found to be highly correlated, and if spatial variation in yield
or other variables are temporally consistent within individual
vineyard blocks. Precision viticulture is an appropriate means to
make use of this vineyard variability for commercial purposes.
Precision viticulture can be used for many purposes, from
increasing precision of vine nutrition to the designation of sub-
blocks for separate wine products. In terms of managing crop
nutrition, petioles may be sampled using geo-referenced sampling
points by GPS, whereby a vineyard manager is enabled to supply
proper nutrients necessary for individual vines rather than uniform
fertilization of entire vineyard (Bramley et al., 2003). As to
designating sub-blocks, yield may vary with the percentage and
position of clay in the soil profile (Bramley, 2001). In a specific
example, spatial variation in yield in a vineyard in Coonawarra,
Australia, was shown to be temporally stable over a 3-year period,
whereby the low yielding areas corresponded with areas where
the clay subsoil occurred close to the surface; and these areas
were more prone to waterlogging in wet years. Different sensory
characteristics were produced in wines made from these different
zones (Bramley, 2001; 2002).
Under the conditions of this study, midday leaf ψ measured
several times throughout the season was a reliable indicator of
vine water status and correlated closely with soil moisture. Soil
moisture zones were temporally consistent at nine of ten sites
from 2005 to 2006 and at all ten sites from 2006 to 2007. Vine
water status zones (based on leaf ψ) were temporally consistent
at two sites from 2005 to 2006 and at two sites from 2006 to
2007. Specific areas of vineyards with high and low water status
appeared to be transient at most sites and their spatial distribution
varied temporally (except Harbour Estate that showed consistent
water status zones from 2005 to 2007). Soil moisture and leaf
ψ were inversely correlated with % sand and directly correlated
with % clay, CEC, soil pH, BS, and K, Ca, and Mg. Spatial
correlation analyses between soil moisture and leaf ψ, and several
soil texture and composition variables, including OM, CEC, pH
and BS, K, P, Ca and Mg were consistent but also demonstrated
a few site-specific relationships. These data suggest that low soil
moisture and low vine water status zones in vineyards are related
to corresponding areas of low % clay, OM, CEC, soil pH, BS,
and soil K, Ca, and Mg. These data further suggest that precision
viticulture techniques may be utilized in this region to soil texture,
soil moisture, or vine water status-based vineyard sub-zones that
could further relate to differing quality levels.
Acknowledgements
Authors wish to thank the Natural Sciences and Engineering
Research Council of Canada and the Wine Council of Ontario
for funding. The participation of all grape growers is also
acknowledged.
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Received: October, 2013; Revised: March, 2014; Accepted: March, 2014
 Journal

Journal of Applied Horticulture, 16(1): 24-28, 2014 Appl

Effect of glycinebetaine application on photosynthesis, sugar

content, invertase activity and plant yield of hot pepper (Capsicum
annuum L.) under water stress condition

R.M. Bhatt*, N.K. Srinivasa Rao, K.K. Upreti and A.D.D.V.S. Nageswara Rao
Indian Institute of Horticultural Research, Hessaraghatta Lake Post, Bangalore 560 089, India.
*E-mail: rmbt@iihr.ernet.in

Abstract
A study was conducted to evaluate the response of hot pepper (Capsicum annuum L.) to foliar applied glycinebetaine (GB) under water
stress condition. Three varieties of hot pepper e.g. Arka Lohit, Pusa Jwala and Arka Haritha were subjected to water stress at flowering
stage. The plants applied with GB had the greater plant height, leaf area, fruit fresh and dry mass under water deficit conditions. GB
application increased the PN under water deficit condition. It was attributed to an improvement in stomatal conductance under water
stress. There was a varietal difference in invertase activity and total sugar contents to GB application under water stress. Higher yield
and better water use efficiency (WUE) were found in GB applied plants. The plants treated with GB 10 days before and at the time
of imposing water stress (T2) responded better. The results suggested that exogenous GB ameliorates the negative effects of water
stress in hot pepper.
Key words: Carbon exchange rate, drought, glycinebetaine, hot pepper, plant yield

Introduction plant-water relation, carbon exchange rate and invertase activity

Water availability is considered the major limiting factor in
vegetable crop production, and high yields are dependent on
adequate water supply. It was observed that under water deficit
conditions, plant responds in different ways and follow different
strategies for its survival. For example, plants show many
morphological and physiological alterations to acclimatize to
unfavourable environment (Sakamoto and Murata, 2002). One
of the most common stress responses in plants is accumulation of
different types of compatible organic solutes (Serraj and Sinclair,
2002). But not all plants can produce osmolytes in sufficient
quantities to combat water stress. In many crop plants, the natural
accumulation of glycinebetaine (GB) is lower than sufficient
to ameliorate the adverse effects of dehydration (Subbarao et
al., 2001). Therefore, protective capacity of GB under various
stress conditions has prompted numerous investigations in order
to increase GB content in plants through genetic engineering
(Sakamoto and Murata, 2000; 2002). The exogenous application
of GB has been suggested as an alternative approach to improve
crop productivity under water stress (Makela et al., 1998;
1999) as it can increases the crop tolerance to water stress (Ma
et al., 2007; Hussain et al., 2008). Yang and Lu (2005) found
that the exogenous application of GB to low-accumulating or
non-accumulating plants may help reduce adverse effects of
environmental stresses. Application of GB has been shown to
protect functional proteins, vital enzymes and photosynthetic
machinery (Xing and Rajashekar, 1999) and has been found
to improve the crop water productivity under limited and well
watered conditions (Hussain et al., 2008). There was not much
information available on the accumulation of glycinebetaine in
relation to water stress in hot pepper (Capsicum annuum L.).
The present study was conducted to evaluate the effect of GB on
under water stress condition in hot pepper.
Materials and methods
Pot experiment: The seedlings of hot pepper (C. annuum L.)
varieties. Arka Lohit and Pusa Jwala were raised in seedling
trays containing coco peat. One month old seedlings were
transplanted in the plastic pots (30 cm dia). The plants were
irrigated regularly and the recommended package of practices
were followed to grow the plants. Uniformly grown plants were
divided into four groups of 25 each. The plants were subjected
to water stress by withholding irrigation at flowering stage (35
days from transplanting) and GB treatments (0.1%) through
foliar application as: T1 = plants treated with GB 10 days before
imposing water stress, T2 = plants treated with GB 10 days before
and at the time of imposing stress, T3 = no treatment during stress,
and T4 = irrigated.
Field experiment: The seedlings of hot pepper varieties i.e. Pusa
Jwala and Arka Haritha were raised in seedling trays as mentioned
in experiment 1. One month old seedlings were transplanted in the
field with a spacing of 60 x 50 cm (row to row and plant to plant).
Water stress was imposed by withholding irrigation at flowering
stage (35 days from transplanting). The GB was applied as defined
in experiment 1. The experiments were conducted in completely
randomized design. All the data were analyzed statistically using
Agris Stat software.
Plant water relation: A portion of the leaf used for the gas
exchange parameters was frozen for a week, thawed and sap
was used for leaf water potential (ψl) determination in leaf water
potential system CR7.
Morphological attributes: Plant height, total leaf area, fruit
 Response of drought stressed hot pepper (Capsicum annuum L.) plant to foliar-applied glycinebetaine
number, fruit fresh weight and total plant dry matter (TDM) were
taken in control and stressed plants before releasing water stress.
The leaf area (LA) of the plant was measured using leaf area meter
(LI-3000). The plant parts were separated and dried in an oven at
80 oC for 72 h to calculate the total dry matter (TDM) of individual
plant. The per plant yield was calculated at the end.
Gas exchange parameters and water use efficiency: The
observations on PN and gs were measured during the water stress
period on fully expanded leaves (5th leaf from top) between 1000
and 1130 hr using ADC open portable photosynthesis analysis
system (model LCA 3, Analytical Development Corporation,
Huddesdon, UK). During the observations, the ambient
temperature varied from 34 to 37 oC, irradiance from 1200 to
1600 μmol m-2 s-1 and the CO2 between 365 and 375 ppm. Leaf
water use efficiency (WUE) was calculated as PN/E (where, PN =
net photosynthetic rate, E = transpiration rate ).
Leaf invertase activity: 500 mg of leaf samples were
homogenized in 2 mL ice cold 100 mM acetate buffer pH 5.0 and
centrifuged at 2500 g for 20 min at 4 oC. Soluble acid invertase
was assayed by incubating 0.2 mL of aliquot (supernatant) with
0.8 mL of 0.1 M sucrose for 30 minutes at 30 oC. Reaction was
stopped by adding 1 mL Somogyi’s copper reagent and boiled
for 10 minutes. Sucrose was added to control sample just before
boiling. Blank sample had no sucrose. Reaction mixture was
cooled and 1 mL arsenomolybdate was added. The absorbance
was read at 630nm. The enzyme activity was estimated as
described by Morris and Arthur (1984). The soluble protein
was determined by Lowry method (Lowry et al., 1951) using
bovine serum albumin as the standard.
Sugar content: Reducing sugars were analysed according to the
procedure of Somogyi (1952). 100 mg dry mass of leaves were
extracted with 5 mL hot 80% ethanol twice and the supernatant
separated out by centrifugation was condensed on a water bath
at 80 oC. The residue was dissolved in 100 mL of water. To 2 mL
of solution, 1 mL of alkaline copper tartrate reagent was added
and the contents were boiled in a water bath for 10 minutes.
After cooling to room temperature, 1 mL of arsenomolybdic
acid reagent was added to it and volume was later adjusted to
10 mL with water. The absorbance of blue colour formed was
recorded at 620 nm. The amount of reducing sugar present in
sample was measured using glucose as standard and expressed
as mg/g d.wt. Total sugars were estimated by hydrolyzing 10 mL
of sugar solution employing 1.0 mL of HCl and keeping solution
overnight. After adjusting pH to 6.5 with NaOH, the total sugar
was estimated with same way as described for reducing sugars.
Non-reducing sugars were calculated by subtracting the reducing
sugars from the total sugar content.
Results and discussion
Pot experiment
Plant water relation: The ψl varied from -1.2 to -1.4 MPa in
Arka Lohit and -1.2 to -1.3 MPa in Pusa Jwala under irrigated
condition, and markedly decreased (-1.7 to -1.9 MPa) under
stress (Fig 1). The decrease in ψl under stress was more in Pusa
Jwala as compared to Arka Lohit. But in GB applied plants, ψl
was relatively higher (less negative) in both the cultivars (-1.4
– 1.7 MPa) under stress.
25
Fig. 1. Leaf water potential (-Mpa) as affected by glycinebetaine under
water stress in hot pepper (T1 = plants treated with GB 10 days before
imposing water stress, T2 = plants treated with GB 10 days before and
at the time of imposing stress, T3 = no treatment during stress and T4
= irrigated)
Photosynthesis and its characteristics: Under water-deficit
condition, PN decreased, with a greater effect seen in Pusa Jwala
than in Arka Lohit when compared with non stress condition
(Fig. 2). However, the PN was higher in the foliar applied GB
plants under water deficit condition, indicating the improvement
in PN under stress. The effect was more pronounced in T2 where
plants were treated with GB 10 days before and at the time of
imposing stress. Among the varieties, the positive effect was
more noteworthy in Arka Lohit. In the GB applied plants, the gs
was also greater than untreated plants under stress. Though the
GB increased the gs in both the varieties, the effect was higher
in Arka Lohit than Pusa Jwala.
Invertase activity and sugars: The invertase activity was greater
in Pusa Jwala than Arka Lohit irrespective of treatments (Fig.
3). In both the varieties, invertase activity was not significantly
affected by water stress. However, the GB application increased
the invertase activity by 2 to 4 times in Pusa Jwala and 3 to 8
times in Arka Lohit as compared to untreated plants under stress.
The effect of GB was noteworthy in T2. A differential response
of sugars to water stress was observed in the hot pepper varieties
(Table 1). In Arka Lohit, the sugar contents decreased under
stress, while in Pusa Jwala there was an increase in total sugar
level under stress (T3).
The GB application increased the sugar level in Arka Lohit under
stress and the effect was more pronounced in T2. However, in
Pusa Jwala, the sugar level further decreased in GB applied
plants under stress. There was no definite trend as far as the
accumulation of reducing and non-reducing sugars in GB applied
plants under water stress is concerned.
Table 1. Effect of glycinebetaine on reducing, non-reducing and total
sugars in hot pepper under water stress (T1 = plants treated with GB
10 days before imposing water stress, T2 = plants treated with GB 10
days before and at the time of imposing stress, T3 = no treatment during
stress, and T4 = irrigated)
Cultivar Sugar (mg g-1) T1 T2 T3 T4 S.Em
Arka Reducing 1216.2 1101.5 905.5 913.6 37.84
Lohit Non-reducing 119.3 232.5 73.6 185.8 17.58
Total 1225.5 1333.9 979.1 1099.4 38.44
Pusa Reducing 880.2 1343.1 1606.0 795.9 96.09
Jwala Non-reducing 112.7 74.6 187.8 168.0 12.92
Total 992.9 1417.7 1793.8 963.9 98.39
26 Response of drought stressed hot pepper (Capsicum annuum L.) plant to foliar-applied glycinebetaine

compared to Pusa Jwala. LA production in Arka

Haritha was almost double in the GB applied
plants than the untreated plants (T3) under stress.
The TDM production was also influenced by GB
under water stress (Table 3).
In both the varieties, the TDM was significantly
greater in the treated plants under water stress.
In Arka Haritha, the TDM was 53.0 to 55.0%
and in Pusa Jwala 29.0 to 41.0% higher than
the plants without GB application under stress.
However, there was no effect of GB on the pattern
of dry matter (DM) partitioning to different plant
parts. But GB application influenced the DM
accumulation in different plant parts under stress
as shown by greater values in fruit, stem and
leaves in GB applied plants compared to untreated
plants under stress in both the varieties.
Water use efficiency: Water use efficiency
(WUE) was determined at 10 days and 25 days
stress (Fig. 4). In general, WUE irrespective
of treatments was higher at 10 days stress and
decreased with the progress of stress in both the
cultivars. However, in the GB applied plants, the
WUE was higher compared to untreated plants
(T3) in both cultivars. The plant response was
better in T2 in both the cultivars.
Plant yield : A considerable reduction in plant
Fig. 2. Photosynthesis and stomatal conductance as affected by glycinebetaine during stress
in hot pepper (T1 = plants treated with GB 10 days before imposing water stress, T2 = plants yield was observed in both the varieties (Arka
treated with GB 10 days before and at the time of imposing stress, T3 = no treatment during Haritha, 39.6% and Pusa Jwala 66.6%) under
stress and T4 = irrigated) water stress (Fig. 5). However, the plant yield
was higher in GB treated plants in both cultivars
under stress and it was more effective in T2. Arka Haritha was more
responsive than Pusa Jwala to GB application under stress.
The present results indicated that the exogenous application of GB
improves the morpho-physiological and biochemical performance
of hot pepper plants under water stress. There was an increase
in TDM production in all the plant parts in GB applied plants
of hot pepper under stress indicating the better plant growth
in GB treated plants. It was evident from our findings that the
Table 2. Effect of glycinebetaine application on morphological
attributes in hot pepper under water stress
Cultivar Treatment Plant Leaf Fruit Fruit fresh
height area number weight
(cm) (cm )2
(g fruit-1)
Arka T1 56.0 2379.60 136.1 237.60
Fig. 3. Invertase activity as influenced by glycinebetaine under water Haritha T2 57.0 2506.41 85.5 147.85
stress in hot pepper (T1 = plants treated with GB 10 days before imposing
water stress, T2 = plants treated with GB 10 days before and at the time T3 45.0 1058.17 80.0 65.70
of imposing stress, T3 = no treatment during stress and T4 = irrigated) T4 61.5 2727.73 124.5 269.71
S Em 1.7 188.43 7.0 23.03
Field experiment Pusa T1 40.5 1259.98 100.0 88.73
Morpho-physiological attributes: The influence of GB Jwala T2 41.0 1853.68 87.0 87.72
application was studied in Pusa Jwala and Arka Haritha under T3 39.0 1242.54 34.0 34.87
the field conditions. The morpho-physiological attributes such T4 42.5 2566.77 102.0 97.68
as plant height, leaf area (LA), fruit numbers and fruit fresh S Em 0.4 156.39 7.9 7.15
weight (FFW) were greater in GB applied plants (T1 and T2) as
T1 = plants treated with GB 10 days before imposing water stress, T2 =
compared to the untreated (T3) under water stress (Table 2). The plants treated with GB 10 days before and at the time of imposing stress,
positive effects of GB were more noteworthy in Arka Haritha as T3 = no treatment during stress, and T4 = irrigated.
 Response of drought stressed hot pepper (Capsicum annuum L.) plant to foliar-applied glycinebetaine 27

Table 3. Total dry mass (g/plant) and its partitioning (g/plant) to different
plant parts in glycinebetaine applied plants under water stress. (T1 =
plants treated with GB 10 days before imposing water stress, T2 = plants
treated with GB 10 days before and at the time of imposing stress, T3
= no treatment during stress, and T4 = irrigated)
Cultivar Treatment Leaf dry Stem dry Root dry Fruit dry Total dry
weight weight weight weight weight
(g) (g) (g) (g) (g)
Arka T1 18.746 24.694 3.190 25.320 71.950
Haritha T2 16.140 22.924 3.460 30.622 73.145
T3 9.983 15.522 2.382 18.827 46.714
T4 21.816 29.190 4.677 42.791 98.474
SEm 1.260 1.420 0.240 2.540 5.280
Pusa T1 12.894 13.879 2.151 15.533 44.455
Jwala T2 10.038 14.141 1.941 21.918 48.038
T3 9.144 11.816 1.449 11.711 34.119
T4 17.548 16.202 2.887 24.323 60.960
SEm 0.950 0.440 0.150 1.450 2.770 Fig. 4. Effect of glycinebetaine on water use efficiency in hot pepper
under water stress (T1 = plants treated with GB 10 days before imposing
water stress, T2 = plants treated with GB 10 days before and at the time
GB application increased PN rate under water stress, and it was of imposing stress, T3 = no treatment during stress and T4 = irrigated)
more effective when applied 10 days before and at the time of
inducing stress (T2). It is well known that generally PN rate under
water stress may be reduced either by stomatal closure and/or
photosynthetic apparatus damage. In the present study, there
was a considerable increase in gs in GB treated plants under
stress and can be attributed for a higher PN rate under stress.
However, the role of photochemical capacity in increasing PN of
GB-treated water-stressed plants cannot be ruled out. Increased
gs by the GB application was observed in tomato and turnip plants
when subjected to drought (Makela et al., 1999). Yang and Lu
(2005) also found that GB application in salt stressed plants of
maize improved the PN and such an improvement was associated
with an improvement in gs and the PSII efficiency. The higher ψl
of hot pepper in GB treated plants under water deficit indicated
the maintenance of leaf water balance and leaf turgidity in GB Fig. 5. Effect of glycinebetaine on plant yield (g plant-1) in hot pepper
treated plants (Fig 1). Our results support the earlier findings that under water stress (T1 = plants treated with GB 10 days before imposing
GB application maintained better leaf water status in the plants water stress, T2 = plants treated with GB 10 days before and at the time
of imposing stress, T3 = no treatment during stress and T4 = irrigated)
(Xing and Rajashekar, 1999; Ma et al., 2007). The GB application
improves the WUE in hot pepper and can be attributed to the higher In conclusion, exogenous GB application resulted in better plant
PN rate under stress. Apart from the PN and gs, the biochemical growth, PN, WUE and plant yield of hot pepper under water deficit
capacity was also affected differentially in hot pepper cultivars by condition. The results suggests that the improved photosynthetic
the GB application under stress. GB treatment increased the sugar capacity in hot pepper may be associated with an improvement in
content under water deficit in cv. Arka Lohit, but it was not the stomatal conductance, maintenance of better plant water relation
same in cv. Pusa Jwala, indicating the genotypic variability in sugar and increase in the biochemical capacity in GB treated plants
response to water stress in GB applied plants. The differential trend under water stress.
in sugar accumulation in relation to GB application under water
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Received: January, 2014; Revised: March, 2014; Accepted: April, 2014
 Journal

Journal of Applied Horticulture, 16(1): 29-31, 2014 Appl

Linking certain physical characteristics with postharvest

needle abscission resistance in balsam fir

M.T. MacDonald, R.R. Lada* and R.S. Veitch

Christmas Tree Research Center, Dalhousie Agricultural Campus, Bible Hill, Nova Scotia, Canada, B2N 5E3.
*E-mail: raj.lada@dal.ca

Abstract
Balsam fir trees are the most popular choice for Christmas trees in Atlantic Canada and a major export commodity, despite postharvest
needle abscission challenging the industry’s viability. The objective of this study was to determine if any needle or branch biophysical
and/or morphological characteristics may be linked with needle abscission resistance (NAR) in balsam fir. A total of 17 different
parameters were measured in branches of clones that belonged to low, medium, or high needle abscission resistant groups. Of the
parameters measured, branch diameter, initial mass, needle density, break strength, and needle retention duration were significantly (P
< 0.05) different between genotype groups. It was found that high NAR genotypes had a 9.1% smaller diameter, 25.0% lower initial
mass, 33.2% lower needle break strength, 32.4% lower needle density, and 91% longer needle retention than low NAR clones. Of
these factors, needle density was the best predictor for needle retention duration (R2 = 47%). Identification of these parameters is an
important first step to understand physiological and genetic linkage for development of Christmas trees with high NAR.
Key words: Abiesbalsamea, break strength, Christmas tree, conifer, needle density, needle retention, senescence, xylem pressure
potential

Introduction Most of the research to date has focused on physiological

Balsam fir is the most popular Christmas tree species used
in Atlantic Canada, with over 3 million trees harvested each
year contributing approximately $72 million to the economy
(MacDonald, 2010). However, postharvest needle abscission
poses a major problem and may result in low consumer
satisfaction and a shift in preference towards artificial trees. In
order to help solve postharvest abscission problems, several
recent studies have focused on identifying and understanding
key factors linked with abscission.
There is considerable variation in needle retention characteristics
between balsam fir genotypes. Four separate screening experiments
of over 100 unique genotypes identified that complete abscission
occurs between 6 and 60 days after harvest (MacDonald and Lada,
2008; MacDonald et al., 2012, unpublished results). Though
variation in abscission is high between balsam fir genotypes,
variation is relatively small within the same genotype tested in
successive years. As a result, genotypes have been classified
according to their needle abscission resistance (NAR), which is
the length of time a branch resists abscission after harvest. Three
broad classifications have been established: low NAR (abscission
< 20 days), moderate NAR (abscission between 21 and 40 days),
and high NAR (abscission > 41 days) MacDonald (2010). A
comparison between NAR classes allows for identification of key
characteristics related to abscission. For example, ethylene was
recently identified as a signal molecule for postharvest abscission
in balsam fir (MacDonald et al., 2010; 2011). Comparison
of ethylene evolution and sensitivity in balsam fir of low and
high NAR determined that low NAR trees release ethylene at a
50% higher rate and are more sensitive to low (i.e. < 10 ppm)
concentrations of ethylene (MacDonald et al., 2012).
mechanisms, such as changes in ethylene or abscisic acid, or
abscission mitigating technologies such as cold acclimation,
ethylene inhibition, and light emitting diodes (MacDonald et al.,
2010b; Thiagarajan et al., 2012; Veitch et al., 2012). There is little
information available regarding biophysical and morphological
characteristics that may be linked to NAR. MacDonald (2010)
tested a theory among producers that trees with short needle
lengths retain needles longer, but found no significant relationship
between needle length and NAR. However, this result does not
preclude the possibility of other physical factors that may be
better indicators of NAR. Identification of these factors is of
practical use as a method to choose trees with superior needle
retention for purchase or breeding, and is of scientific relevance
as it could also raise questions as to why certain factors are linked
to postharvest needle abscission. Thus, the objective of this study
was to identify any differences in biophysical and morphological
characteristics of low, moderate, or high NAR genotypes and
potentially link those characteristics to the length of time until
complete postharvest abscission.
Materials and methods
Experimental design: A total of 45 branches, cut from the most
recent 2 years of growth at 1.5 m above ground, were collected
from a balsam fir clonal orchard at the Tree Breeding Centre,
Department of Natural Resources, Debert, NS (45° 25’ N, 63°
28’ W) on July 13, 2010. The experiment followed a completely
randomized design. NAR classification was used as the explanatory
variable at 3 levels: low, moderate, and high (MacDonald, 2010).
Fifteen branches were randomly selected from each NAR class
and each branch served as a replicate. Handling and storage of
branches was meant to simulate conventional industry procedures
30 Linking certain physical characteristics with postharvest needle abscission resistance in balsam fir

for tree harvest. Thus, branches were not provided any water and Results
were stored at 20°C for 16 hours during the day, then at 15°C for
8 hours at night. A relatively low light intensity of 80 μmol m-2 Of the 17 physical characteristics measured, only 5 were found to
s-1 was provided constantly throughout the experiment simulating be significantly different between different NAR genotypes. The
household or covered storage conditions. two branch characteristics linked to NAR were branch diameter
and initial mass (Table 1). High NAR branches had a 9.1%
Branch related response variables: Response variables smaller diameter and 25.0% lower initial mass than low NAR
directly related to branch included diameter, length, mass,
branches. The three needle characteristics linked to NAR were
internode distance, fragrance, flushing, and xylem pressure
break strength, needle density, and NRD (Table 2). High NAR
potential (XPP). Branch diameter was measured at the cut end
branches had a 33.2% lower break strength, 32.4% lower needle
and length was determined as the distance from the cut end to
density, and 91.3% higher NRD than low NAR branches.
the end of the primary branch. Branch mass was measured once
immediately after sample collection (initial mass) and again Table 1. Branch characteristics of low, moderate, and high needle
abscission resistant balsam fir genotypes. An asterisks indicates that
after complete abscission (final mass). Internode distance was a characteristic was significant at α = 0.05 and letter groupings were
the average distance between each node of first and second determined using least significant difference multiple means comparison.
year growth. Flushing was a count of new growth areas on the Means for each NAR class were calculated from 15 replicates.
branch. Fragrance was determined by smelling a branch and Physical NAR Classification
comparing it to needle extracts on a scale of 0 to 5. A value of Characteristic Low Moderate High
zero was assigned to water control and values 1, 2, 3, 4, and 5 Branch diameter (mm)* 6.53a 6.02b 5.94b
corresponded needle extract concentrations of 2.5, 5, 7.5, 10, Branch length (cm) 32.00 33.10 31.0
Initial mass (g)* 29.38a 27.47a 22.04b
and 12.5 ppm, respectively. Finally, XPP was determined using a Final mass (g) 5.86 5.55 5.14
pressure bomb and recording the pressure required to force water Internode distance (cm) 10.60 10.5 9.9
out of the xylem. XPP was determined immediately after harvest Fragrance 3.00 3.0 2.0
(XPP0), after complete abscission (XPPf), and then calculated Flushing 9.00 10.0 8.0
as the difference between initial and final values (ΔXPP = XPPf XPP0 -0.76 -0.66 -0.59
– XPP0). XPPf -0.77 -0.58 -0.50
ΔXPP -0.01 0.08 0.09
Needle related response variables: Response variables directly
Table 2. Needle characteristics of low, moderate, and high needle
related to the needles included needle retention duration (NRD), abscission resistant balsam fir genotypes. An asterisks indicates that
angle of attachment, needle proportion, needle density, needle a characteristic was significant at α = 0.05 and letter groupings were
length, relative water content, and break strength. NAR was determined using least significant difference multiple means comparison.
Means for each NAR class were calculated from 15 replicates.
determined as the number of days required to complete abscission.
Angle of attachment was the average angle of needles from three Physical NAR Classification
Characteristic Low Moderate High
positions along the primary branch, assuming that the direction
Break strength (N)* 1.87a 1.61ab 1.25b
of the cut end was 0°. Needle proportion was the weight of shed RWC (%) 81.1 85.5 83.9
needles as a percentage of total branch fresh mass. Needle density Density (needles/cm)* 34.0a 28.0b 23.0c
was the average number of needles in a 1 cm length averaged Needle length (mm) 15.42 14.71 15.95
from three positions along the primary branch. Needle length Needle proportion (%)* 80.1 79.8 76.6
was the average length of 5 randomly selected needles. Relative Angle of attachment (°) 102.0 104.0 107.0
NRD (days)* 15.0c 21.2b 28.7a
water content was calculated from measurements of dry needle
mass, fresh needle mass, and turgid needle mass according to Plotting all biophysical and morphological parameters against
the formula (fresh needle mass – dry needle mass) / (turgid NRD revealed several weak relationships (R2< 10%). However,
needle mass – dry needle mass) x100. Finally, break strength there was a significant negative relationship (P = 0.003) between
was determined as the force required to remove a needle from needle density and NRD, which suggests a higher needle density
the branch. A clip, which was attached to electronic spring scale, may result in poorer needle retention (Fig. 1).
was fastened to a needle. The branch was pulled away from the
spring scale and force was recorded using Logger Pro 3.1 (Vernier
Software and Technology, Beaverton, OR), which allows force to
be plotted against time. The amount of force required to remove
a needle is represented as a peak on the force-time graph.
Statistical analysis: Data were subjected to an analysis of
variance using SAS v9.1 to determine significant differences (α
= 0.05) in characteristics of low, moderate, and high NAR balsam
fir. Statistical assumptions of normality, constant variance, and
independence were valid in all parameters except break strength.
Break strength required a square root transformation to induce
constant variance and normality. When statistical significance
Fig.1. Relationship between needle density and time required for
was detected, multiple means comparison was conducted with complete abscission. The relationship is best described with NRD =
least significant difference (LSD). 441(Density)-0.9. N = 45.
 Linking certain physical characteristics with postharvest needle abscission resistance in balsam fir
Discussion
The link between NAR and NRD was expected, since NAR
classifications were based on NRD screenings. Previous average
NRDs for low, moderate, and high NAR classes were 14.9,
32.1, and 47.3 days, respectively (MacDonald and Lada, 2008;
MacDonald, 2010). The NRD values determined from this
experiment were similar for the low NAR genotypes, but much
lower for moderate and high NAR genotypes. The likely cause
for this discrepancy may be the collection date as branches were
collected in July for this experiment as compared to October for
previous work and it has been suggested that a period of cold
acclimation beginning in autumn increases needle retention in
conifers (MacDonald and Lada, 2008; Mitcham-Butler, 1988;
Thiagarajan, 2012; Thiagarajan et al., 2012).
Branch diameter, mass, and needle density were significantly
lower in high NAR branches compared to low NAR branches.
It is possible that each of these characteristics is linked to
decreased water use, which has long been suggested as a key
trigger for postharvest needle loss in conifers (Chastagner and
Riley, 2003). However, final XPP values of approximately -0.5
to -0.8 MPa were much higher (more positive) than threshold
values of -3 to -4 MPa found in related species (Chastagner and
Riley, 2003; Mitcham-Butler et al., 1988). Also, there was no
significant decline in XPP over the duration of the experiment.
The XPP observations in this experiment were consistent with the
hypothesis that postharvest needle abscission in balsam fir must
be due to a factor other than declining water status (MacDonald,
2010). The exact reason that diameter, mass, and needle density
may be linked to NAR is not known. It is possible that lower
needle density may have reduced water loss, thereby delaying
postharvest needle abscission. If this is so, XPP would have
declined significantly in low NAR genotypes, but a lack of a
significant relationship between XPP and NAR suggests a weak
link between postharvest dehydration and NAR.
The other parameter linked with NAR was break strength,
although the relationship was rather counterintuitive. A decrease in
break strength is normally associated with accelerated abscission
(Brummell et al., 1999). In our experiment, however, high break
strength was consistently associated with low NAR genotypes.
It’s obvious from the early abscission of low NAR genotypes that
break strength must rapidly decrease after harvest, but there is
currently no explanation for high initial break strengths compared
to high NAR genotypes.
31
Overall, increased needle retention was observed in branches with
a lower stem diameter, mass, break strength, and needle density.
These results identify several factors not previously known to
be associated with postharvest abscission in balsam fir and also
offer a potential screening tool to producers, who may selectively
breed trees with the aforementioned characteristics. In addition,
if consumers are made aware of these characteristics it will allow
for more informed Christmas tree selection.
References
Brummell, D.A., B.D. Hall and A.B. Bennett, 1999. Antisense
suppression of tomato endo-1,4-β-glucanase Cel2 mRNA increases
the force required to break fruit abscission zones but does not affect
fruit softening. Plant Mol. Biol., 40: 615-622.
Chastagner, G.A. and K.L. Riley, 2003. Postharvest quality of noble and
Nordmann fir Christmas trees. HortScience, 38: 419-421.
MacDonald, M.T. 2010. Physiological Significance of Ethylene in Needle
Abscission of Root-detached Balsam Fir (Abiesbalsamea L.). Ph.D.
Diss., Université Laval, 2010. 161 pp.
MacDonald, M.T. and R.R. Lada, 2008. Cold acclimation can benefit
only the clones with poor needle retention duration (NRD) in balsam
fir. HortScience, 43: 1273.
MacDonald, M.T., R.R. Lada, M. Dorais, S. Pepin, Y. Desjardins and A.I.
Martynenko, 2011. Ethylene exposure duration affects postharvest
needle abscission in balsam fir (Abiesbalsamea L.). HortScience,
46: 260-264.
MacDonald, M.T., R.R. Lada, A.I. Martynenko, M. Dorais, S. Pepin
and Y. Desjardins, 2010. Ethylene triggers needle abscission in
root-detached balsam fir. Trees, 24: 879-886.
MacDonald, M.T., R.R. Lada, A.I. Martynenko, M. Dorais, S. Pepin
and Y. Desjardins, 2012. Is there a relationship between ethylene
evolution, ethylene sensitivity, and needle abscission in root-detached
balsam fir? Acta Hort., 932: 405-412.
Mitcham-Butler, E.J., L.E. Hinesley and D.M. Pharr, 1988. Effect of
harvest date, storage temperature, and moisture status on postharvest
needle retention of Fraser fir. J. Environ. Hort., 6: 1-4.
Thiagarajan, A. 2012. Physiology of Low Temperature-modulated
Postharvest Needle Senescence and Abscission in Balsam Fir
(Abiesbalsamea L.). Ph.D. Diss., Université Laval, 2012. 159 pp.
Thiagarajan, A., R. Lada, S. Pepin, S.F. Forney, Y. Desjardins and M.
Dorais, 2012. Characterization of phytohormonal and postharvest
senescence responses of balsam fir (Abiesbalsamea (L.) Mill.)
exposed to short-term low temperature. Trees, 26: 1545-1553.
Veitch, R.S., R.R. Lada and M.T. MacDonald, 2012. Effect of light
emitting diodes (LEDs) on postharvest needle retention in balsam
fir (Abiesbalsamea L.). J. Appl. Hort., 14: 13-17.
Received: December, 2013; Revised: February, 2014; Accepted: March, 2014
 Journal

Journal of Applied Horticulture, 16(1): 32-39, 2014 Appl

Optimal soil conditions for organic highbush blueberry growth:

Assessment of early results

B. Hoover, D. Fuglie and R. Miller*

Department of Biology, Eastern Mennonite University, 1200 Park Road, Harrisonburg, VA-22802.
*E-mail: millerrj@emu.edu

Abstract
To ascertain optimal soil conditions for creating an organic and sustainable blueberry operation, 160 highbush blueberry plants
representing five different cultivars (Duke, Bluecrop, Jersey, Chandler, and Bluegold) were planted at Knoll Acres Farm, Harrisonburg,
Virginia in 2009 within four soil treatment plots (horse manure, sheep manure, pine straw, and Planters Choice mulches). To define
optimal growth conditions, selected soil characteristics and plant vigor assessments including photosynthesis and respiration activities
as well as plant growth measurements were recorded. Statistical analyses indicated that soil treatments of pine straw and Planters
Choice mulches produced significantly higher plant growth values than horse and sheep manure mulches. Among the five cultivars,
Chandler bushes thrived the best, based on growth parameters except for bush height. Including cost/benefit considerations, pine straw
mulch was the most economical and effective treatment among four mulches tested.
Key words: Soil, organic, sustainable, mulch, Duke, Bluecrop, Jersey, Chandler, Bluegold, Vaccinium corymbosum, Ericaceae

Introduction et al., 2005), protection against age-related neurological defects

Knoll Acres, a small blueberry farm located in the Shenandoah
Valley near Harrisonburg, Virginia, aims to establish a small,
sustainable, commercial organic blueberry operation and thereby
discern and model best organic horticultural practices in this area
of Virginia. Blueberries are members of the Ericaceae family.
Commercial blueberry types in the United States include southern
lowbush, rabbiteye, and northern highbush blueberry. Highbush
blueberries (Vaccinium corymbosum) are the most widely
cultivated species in North America and best fit the growth zone
of this area of Virginia (Retamales and Hancock, 2012).
Besides cranberries, blueberries are the only commercially grown
endemic fruit crop in the United States (Pritts and Hancock,
1992). In 2009, about 40,500 hectares (100,000 acres) of
blueberries were planted in North America which represents a
50% increase over the prior 5 years. World-wide, outside of North
America, about 27,000 hectares is under blueberries production
(Retamales and Hancock, 2012). The market and demand for fresh
blueberries is high; US blueberry consumption in 2004 doubled
the consumption rate twenty years earlier. Americans consume
200 million pounds of blueberries each year and the highbush
industry in North America is currently worth more than 100
million dollars (Pritts and Hancock, 1992). The area of Virginia
is currently not a major source of blueberry production. However,
based on a few small successful commercial growers in this area,
and a specialty crop profile paper from Virginia Cooperative
Extension (Bratsch and Pattison, 2009) blueberry production in
this part of the Shenandoah Valley is a viable option.
Many consumers are aware of the health benefits that the fresh
blueberries provide (Lewis and Ruud, 2005). These include
basic nutrient properties, antioxidant activity (Kratchanova et
al., 2008; Philpott et al., 2009), anti-ageing properties (Wilson
et al., 2006), cancer prevention (Bagchi et al., 2004; Matchett
(Joseph et al., 2007), urinary tract health, protection against
diabetes (McDougall and Stewart, 2005), and cardiovascular
health (Kalt et al., 2008). The polyphenolics and anthocyanins,
found in ripened blueberries, are the primary health promoters and
protective agents (Nichenametla et al., 2006) and in comparison
to many other fruits tend to contain relatively higher levels of
protective anthocyanins (Suojalainen and Keranen, 1961). Berry
content of anthocyanins varies among blueberry genotypes
(Scalzo et al., 2008). Between the picking of a ripe blueberry
and its consumption, the storage and processing of this fruit
may affectively lessen the anthocyanin content and antioxidant
capacity (Brownmiller et al., 2008; Scibisz and Mitek, 2009;
Trost et al., 2008; Nikkah et al., 2007).
Several studies have demonstrated the superiority of organic
versus conventionally raised blueberries, but this research needs
confirmation and development to convince a potential blueberry
grower that organic production techniques are superior to
conventional ones (McCullum-Gomez et al., 2009). Wang et al.
(2008) compared harvested blueberry chemical characteristics
from multiple sites and found that organically grown blueberries
(Bluecrop) have elevated levels of sugars, total phenolics,
total anthocyanins, and antioxidant activity, when compared
to conventionally grown blueberries. Missing data from this
study elicit questions for the potential organic grower: What is
the difference in productivity (yield) between the organic and
the conventional blueberry plants? What mechanism accounts
for the increased antioxidant activity in the organic versus
conventionally grown blueberries? Is it possible that organic
(versus conventional) practices can produce blueberries that are
healthier, higher in quality and quantity, and more cost effective
for the grower? The superior antioxidant quantities (elevated
anthocyanins and phenolics) of organic blueberries have not been
linked to specific growing practices or to the effect of specific
soil profiles or foliar nutrient levels.
 Optimal conditions for highbush blueberry growth
Nationwide attention has been given to growing blueberries
in an organic system through several important publications
(Drummond et al., 2009; Kuepper and Diver, 2004). As interest
and grower involvement in organic production have increased,
the organic arsenal of effective insecticides and fungicides
continue to be researched and expanded (Kamminga et al., 2009).
Most of the published papers, providing economic guidance
on growing blueberries, are regionally focused for the large
conventional acreage growth, rather than the smaller organic
producer. Examples include northern highbush blueberries in
Oregon (Eleveld et al., 2005), California (Takele, 2005; Takele
et al., 2007), and Pennsylvania (Demchak et al., 2001) as well as
southern highbush blueberries in Georgia (Fonsah et al., 2006)
and Kentucky (Woods, 2008).
A commercial operation is sustainable only to the degree that it is
profitable. Costs and steps to initiate a small commercial organic
blueberry production facility and its potential profitability have
not been fully explored or documented. While economic studies
of large commercial (non-organic) blueberry production have
been published detailing costs of machine harvesting, it is difficult
to extrapolate these findings to smaller organic production
efforts. A study on highbush blueberry production in New
Jersey, investigating best organic practices in cultivar selection,
weed and insect management, and usage of selected organic
fungicides and insecticides, has been very helpful to the organic
grower (Sciarappa, 2008). However, this study did not connect
economics and plant productivity to their organic practices. More
recently, however, Julian et al. (2012) connected establishment
costs of organic northern highbush blueberries involving different
mulches and fertilization approaches with the productivity of two
cultivars, Duke and Liberty.
Blueberries tolerate a wide range of soils. A natural blueberry
soil has low fertility, high polyphenol content, and more than
4% organic matter. However, any good loam soil with some
amendments will be suitable for blueberries. Loams with an
organic matter content of 3-15% are excellent (Gough, 1994).
Increased organic matter in the lower levels of the soil enhances
downward growth of blueberry roots (Shoemaker, 1978). Given
high organic matter content, blueberry optimal growth and
production occur in acidic soils with a low pH range of 3.8 to
5.5 (Pritts and Hancock,1992). Blueberry plants thrive on organic
fertilizers (Kuepper and Diver, 2004), which tend to slowly
release nutrients creating a stable ecosystem.
The purpose of this investigation was to identify optimal soil
treatments and cultivar selections for the Shenandoah Valley,
based on soil quality measures and indicators of plant vigor. Since
substantial berry production does not occur in the first two years
after planting, this paper reports on planting methodology, soil
characteristics, and plant vigor measures during the early years.
These data provide the basis to predict optimal horticultural
practices that enhance a sustainable blueberry operation.
Materials and methods
Experimental design and plantings: To assess the effects of
varying soil mulch treatments on the growth of five different
highbush blueberry cultivars, a block design (Fig. 1) was created
with four soil treatment plots: horse manure and sawdust mulch
(HM), sheep manure and hay mulch (SM), pine needles and
33
shredded pine bark mulch (PS), and a commercial Planters Choice
(PC) mulch based on bovine manure, sawdust, and fodder.
At Knoll Acres in November and December of 2009, planting
holes (approximately 50 x 25 cm), at 1.5 meter intervals in the
middle of the row, were hand dug (Fig. 2). One hundred and sixty
bare-rooted 3 year old dormant blueberry plants representing five
cultivars: Duke (40 plants), Bluecrop (39), Jersey (40), Chandler
(25) and Bluegold (16) were planted in the moistened holes, using
a mix of soil, shredded pine bark, and peat moss as covering
material (Fig. 3). The Duke, Bluecrop, and Jersey plants were
obtained from Miller Nurseries, New York. The Chandler and
Bluegold bushes were obtained from Finch’s Blueberry Nursery,
NC. After planting, the blueberry plants were top-mulched with
7-9 cm of shredded pine bark and left to over-winter. Although
most of the plants thrived in the following spring and summer
(Figs. 4 and 5), the initial planting was enhanced by replacing a
few plants that were not thriving.
Soil sampling and assays: Soil quality measures were based on
soil respiration and rate of water infiltration. Soil respiration, based
on a prescribed USDA method (USDA, 2001), is one measure of
biological activity and organic decomposition. Specifically, this
is a measurement of carbon dioxide (CO2) released from the soil
surface due to aerobic microbial respiration, plant root and faunal
respiration, and eventually from the dissolution of carbonates
in soil solution. Using an enclosed ring chamber (15.54 cm in
diameter and 7.6 cm high), positioned over a sample area, trapped
air was drawn through a Draeger tube apparatus to estimate the
amount of CO2 produced and released from the soil surface within
a given time frame. The rate of CO2 release was expressed as
CO2-C kg/ha/day (1 pound / acre = 1.12085116 kg/ha). Soil water
infiltration, based also on a prescribed USDA method (USDA,
2001), involved applying 444 mL of water (2.54 cm layer) into
a 15.24 cm diameter metal ring driven into the soil and allowing
the water to drain freely into the enclosed ring of soil. After an
initial infiltration was done to wet the soil, the infiltration time
(in seconds) for the second water sample was recorded.
To assay the effectiveness of different fertilizing techniques, soil
samples were obtained from the four soil treatment plots: HM,
SM, PS, and PC. A soil sample was obtained by mixing five
cores taken from a single area 30 cm in diameter. Samples were
obtained near plants in each bed with six different samples taken
from each soil treatment plot. Soil samples were dried overnight
and filtered through a screen to remove the large particles. The
screened and dried soil samples were tested using commercial
semiquantitative soil test assays (LaMotte, 2001) to determine
micronutrient and macronutrient concentrations as well as the
pH values.
The four experimental mulches (horse manure, sheep manure,
pine straw, and Planters Choice) were also sampled and analyzed
for macro- and micronutrient content, pH, and percentage of
organic matter at a commercial soil laboratory (Virginia Tech
Soil Laboratory, Blacksburg, Virginia).
Cultivar sampling: Measurement data were collected at Knoll
Acres in the fall of 2010 about ten months after the bushes were
planted (see Fig. 5) in November and December of 2009 as
three year old bare-rooted stock from commercial nurseries.To
determine plant growth data, all 160 plants representing the five
34 Optimal conditions for highbush blueberry growth

highbush blueberry cultivars were assayed. For the photosynthesis (PSH) where PS, PB, and PSH denote the numbers of primary
measures, selected Jersey and Bluecrop plants from the different stalks, average number of primary branches per stalk, and average
treatment plots were sampled. primary stalk length in cm.
Growth parameters: Overall bush height and diameter, length Photosynthesis and transpiration measurements: Rates of
of the primary stalk, primary stalk diameter at 10 and 25 cm, photosynthesis and transpiration were measured for two cultivars,
and the number of primary stalks and primary branches were Jersey and Bluecrop, with a LiCOR 6400, which simultaneously
measured. Four growth parameters, considered as measurements measures photosynthesis (via net CO2 uptake) and transpiration
of plant vigor, included: average primary stalk diameter (in mm), (Long et al., 1996). This instrument allows for non-invasive
bush height (in mm), volume of a plant cylinder (in which the field measurements by isolating individual leaves inside a 2 x 3
volume of a single plant was measured by placing the plant in cm clamped chamber that measures the difference in incoming
a virtual cylinder and calculating the formula V=Πr2h), and a and outgoing CO2 and H2O. Mature highbush blueberry leaves
relative bushiness value (B), based on the formula B = (PS) (PB) average 5 cm long and 3 cm wide and completely fill the LiCOR
Fig. 1. Plot design and layout for organic blueberry cultivars. Four different mulch treatment plots: Horse Manure (horse manure and sawdust mulch);
Sheep manure (sheep manure and hay mulch); Pine Straw (pine needle and shredded pine bark mulch); Planters Choice (commercial cow manure
& fodder mulch). Five highbush blueberry cultivars [number of plants] comprise 160 plants: Duke [40], Bluecrop [39], Jersey [40], Chandler [25],
and Bluegold [16].
Row # Plants Compost Organic Plots
Letter /row Type
Duke Duke Duke Duke Duke Duke Duke Duke Duke Duke Duke Duke Duke Duke
A 14 #01 A #02, A #03, A #04, A #05, A #06, A #07, A #08, A #09, A #10, A #11, A #12, A #13, A #14, A
Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop
B 14
Horse #01, B #02, B #03, B #04, B #05, B #06, B #07, B #08, B #09, B #10, B #11, B #12, B #13, B #14, B
Manure Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey
C 13 #01, C #02, C #03, C #04, C #05, C #06, C #07, C #08, C #09, C #10, C #11, C #12, C #13, C
Chandler Chandler Chandler Chandler Chandler Bluegold Bluegold Bluegold Bluegold Bluegold Bluegold Chandler Chandler
D 13 #01, D #02, D #03, D #04, D #05, D #01, D #02, D #03, D #04, D #05, D #06, D #06, D #07, D
Duke Duke Duke Duke Duke Duke Duke Duke Duke Duke Duke Duke Duke
E 13 #15 E #16 E #17 E #18 E #19 E #20 E #21, E #22, E #23, E #24, E #25, E #26, E #27, E
Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop
F 12
Sheep #15, F #16, F #17, F #18, F #19, F #20, F #21, F #22, F #23, F #24, F #25, F #26, F
Manure Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey
G 11 #14, G #15, G #16, G #17, G #18, G #19, G #20, G #21, G #22, G #23, G #24, G
Chandler Chandler Chandler Bluegold Bluegold Bluegold Bluegold Bluegold Chandler Chandler Chandler
H 11 #08, H #09, H #10, H #07, H #08, H #09, H #10, D #11, D #11, H #12, H #13, H
Duke Duke Duke Duke Duke Duke Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop
I 12
Pine #28, I #29, I #30, I #31, I #32, I #33, I #27, I #28, I #29, I #30, I #31, I #42, I
Straw Jersey Jersey Jersey Jersey Jersey Jersey Chandler Chandler Chandler Chandler Chandler
J 11 #25, J #27, J #285, J #29, J #30, J #31, J #14, J #15, J #16, J #17, J #18, J
Duke Duke Duke Duke Duke Duke Bluecrop Bluecrop Bluecrop Bluecrop Bluecrop
K 11 #34, K #35, K #36, K #37, K #38, K #39, K #32, K #33, K #34, K #35, K #36, K
Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Jersey Duke #43,
L 11
Planters #32, L #33, L #34, L #35, L #36, L #37, L #38, L #39, L #40, L #41, L L*
Choice Chandler Chandler Chandler Chandler Chandler Chandler Chandler Bluecrop Bluecrop *Small plant, few
M 9 #19, M #20, M #21, M #22, M #23, M #24, M #25, M #43, M #44, M roots
Bluegold Bluegold Bluegold Bluegold Bluegold
N 5 #12, N #13, N #14, N #15, N #16, N
160

Fig. 2. Planting hole for planting bare-rooted 3 year old blueberry plants.
Planting holes about 50 cm in diameter and 25 cm deep were initially
dug in the center of the row at 1.5 meter intervals. A mixture of soil, peat
moss, and shredded pine bark were used as planting media. See the pile
of planting media to the left side of the planting hole.
Fig. 3. Newly planted bare-rooted 3 year old blueberry plant. Plants were
planted in November/December 2009 while they were dormant. Planting
media (soil:peatmoss:shredded pine bark at a ratio of 1:1:1) was firmed
around the bare rooted plant and then thoroughly soaked with water.
 Optimal conditions for highbush blueberry growth 35

Fig. 4. Young organic Bluegold blueberry bush at Knoll Acres loaded Fig. 5. Young organic blueberry plants about 10 months (September
with blossoms about 5 months after planting. Most of the blossoms were 2010) after planting.
stripped from the plant during this first summer to enhance vegetative
and root growth by restricting the flow of plant energies into fruit plots were very high with all values less than one minute (Table
production. 1). Between the blueberry rows (the middles, covered with native
chamber. LiCOR 6400 infra-red gas analyzer measurements grasses) storm water runoff was rapid with little resultant erosion
of photosynthesis and transpiration are widely reported in the due to the vegetative covering.
literature for various plant species (Hunt, 2003; Flexas et al.,
Soil respiration measurements (CO2-C kg/ha/day) varied with
2002). Photosynthetic values were expressed as μmol/CO2/m2/s;
time and in comparing the mulch treatments (Table 1). Initial
transpiration values were expressed as mmol H2O/m2/s. Bluecrop
respiration measures (November 2009) in the four organic plots
and Jersey bushes were assayed by measuring a minimum of two
varied from a low of 16.3 in the PS to a high of 24.3 in the SM
leaves per plant and two plants per soil treatment.
plot. The average reading across all of the organic plots was 19.7.
Statistical analysis: Data were assessed using SPSS Version 4 Typically values ranging from 18-36 are considered to reflect
software. Group descriptive statistics means and standard error medium soil activity. However, a year later (October, 2010) the
of the means (SEM) were determined with the SPSS program. values increased three-fold with a low of 34.6 in the PS and a
Potential group statistical differences for growth parameters (P high of 84.8 in the PC plot. The average value (in 2010) across all
< 0.05) were determined with One Way ANOVA and Student- plots was 59.6, which corresponded to what is considered to be
Neuman-Keuls post hoc testing to determine differences between ideal soil activity with adequate organic matter and populations
soil treatments and/or cultivars. of active microorganisms. Differences in soil mulch mixes result
in variations in soil organic matter (SOM) and populations of
Results and discussion organisms, which are keys to soil respiration. Mulches with a
Soil treatments and assays: The blueberry planting plots at low carbon to nitrogen (C:N) ratio produce higher CO2 rates than
Knoll Acres consist of a cherty silt loam on what was originally residues with a high C:N ratio. Increases in SOM improve soil
an eroded slope of about 35%. The surface layer is brown cherty aggregation and porosity and therefore aeration and soil moisture
silt loam about 15 cm thick. The subsoil extends about 150 content, factors that enhance CO2 production rates.
cm or more. Between the depths of 15 to 32 cm it is a brown The macro- and micronutrient values determined directly from
clay loam; at depths greater than 32 cm, it is yellowish red and
red clay. Naturally, this soil is slightly acidic with low organic Table 1. Soil quality parameters: respiration and water infiltration. Values
represent averages of three-four samples per plot
matter content and low natural fertility. The subsoil has moderate
Organic Plot Test Date Respiration Water
permeability and moderate available water. Surface runoff of
(CO2-C kg/ha/ Infiltration
water is rapid. About a decade ago, these plot areas were wooded. day) (seconds)
They were then cleared and used as native pasture prior to their Horse Manure November, 2009 18.5 3.4
usage as blueberry planting plots. October, 2010 46.7 nd
The organic plots at Knoll Acres are located on a south-east Sheep Manure November, 2009 24.3 7.5
hillside with a 35% drop in elevation. To maximize consistent October, 2010 72.4 nd
sun exposure to the blueberry plants, rows were designed to run Pine Straw November, 2009 16.3 6.0
from north to south, which formed an oblique angle with the October, 2010 34.6 nd
primary fall of the hill. Consequently, the potentiality of soil Planters Choice November, 2009 19.8 13.6
erosion and water runoff following rainstorms were concerns. October, 2010 84.8 nd
However, due to the incorporation of large amounts of organic Respiration values in 2010 reflect higher metabolic activity than in
materials from the added mulches, the resultant high porosity of 2009, although soil temperatures were similar. Rapid water infiltration
reflects high porosity of the soil plots. Water infiltration records time
the soil effectively absorbs storm water with little or no runoff in for 2.54 cm (1 inch) layer of water to be absorbed by prior wetted soil
the blueberry rows. Water infiltration rates in all of the organic (nd = not determined)
36 Optimal conditions for highbush blueberry growth

mulch samples (Table 2) reflected very high nutrient values. Table 2. Nutrient composition of mulches
Using these organic mulches as soil amendments thus positively Parameter Horse Sheep Pine Planters
influenced the available nutrients in the soil plots for the blueberry manure manure straw Choice
plants. A negative factor, especially in the sheep manure and P (ppm) 528 1101 338 544
Planters choice mulches, was their high pH value (8.6 and 8.7, K (ppm) 2066 1642 428 1999
respectively) which consequently elevated the pH of the mulch Ca (ppm) 1363 2940 1744 1916
amended soil plot far above the desired pH of 5. In contrast, the Mg (ppm) 418 1077 379 616
acidity of the added pine straw mulch (5.5) helped to stabilize the Zn (ppm) 5.9 8.3 10.9 11.2
soil pH near the desired level in the PS plot. The high percentage
Mn (ppm) 28.6 33.2 67 33.2
of organic matter in the mulches (Table 2) also positively
Cu (ppm) 0.2 0.1 0.2 0.2
influenced the organic percentage in the various soil plot raising
their organic percentages from an average of 2-3% organic Fe (ppm) 7.7 4.5 8.3 5.5
matter to an average greater than 12% organic matter across all B (ppm) 1 1.7 0.7 1.6
the four plots with PS plot showing the lowest percentage and pH 7.2 8.6 5.5 8.7
SM plot the highest (data not shown). The percentage of organic Organic Matter (%) 45.2 33.1 34.3 55.1
material in the soil plots is reflected in parallel relative humus Est/ CEC (meq/100g) 15.5 27.7 15.3 19.7
values (Table 3). Base Sat. (%) 100 100 84.5 100
Soil assays from the four soil treatment plots contained adequate Ca Sat. (%) 43.8 52.9 56.9 48.4
levels of macronutrients (nitrogen, phosphorus, and potassium) Mg Sat. (%) 22.1 32 20.4 25.7
with no significant differences between them (Table 3). The K Sat. (%) 34.1 15.1 7.2 25.9
levels of micro and macronutrients in PS plot displayed the
Table 3. Soil nutrients and parameters present in multiple soil
most variance in comparison to the other treatments. Except samples from each mulch treatment plot
for sulfate levels, PS plot contained consistently lower levels of Soil nutrient Mulch Treatment Plots
soil nutrients than other plots. PS plot had a significantly lower Horse Sheep Pine Planters
amount of calcium (542 ± 127 ppm) while SM plot contained a Manure Manure Straw Choice
significantly higher level of calcium (3500 ppm) in comparison to Nitrate N (ppm) 65±7a 75±0a 48±13a 71±4a
the other plots. PS had a significantly lower amount of magnesium Phosphorus (ppm) 96±4a
100±0 a
92±8 a
100±0a
(8 ± 1 ppm) than the other soil plots. SM and PC plots contained Potassium (ppm) 78±10 a
108±16 a
- 78±9a
statistically higher levels of humus than HM and PS plots. Each Calcium (ppm) 1700±300b 3500±0c 542±127a 1400±0b
treatment contained statistically different pH levels. SM plot had Magnesium (ppm) 64±10 b
80±0 b
8±1 a
62±12b
the highest pH level of 5.92 ± 0.23 and PS plot, the lowest (3.87 Sulfate (ppm) 458±173 a
750±250 1167±167 620±233a
a a

±.04). The plot pH trends reflected the contrasting pH values of 2.1±0.3 a

4.6±0.2c 1.7±0.2a 3.3±0.8b
Humus
the incorporated mulches. Sheep manure mulch had a pH of 8.6
pH 4.86±0.14b 5.92±0.23d 3.87±0.04a 5.43±0.23c
and pine straw mulch with a pH of 5.5 (Table 2).
Values are shown as averages ± SEM. N values range from 3 to 6 and
Growth parameters- Comparing soil mulch treatments: PC represent different composite soil samples taken during the fall of
and PS plot bushes had statistically greater heights (73.4±2.6 2010; each individual soil sample consists of a mix of several soil cores
taken from the top 6-7 inches. Differing superscript letters, along the
cm and 73.4±2.6 cm, respectively) than HM and SM (Table rows, indicate statistically significant mulch group differences when P
5). Blueberry bushes in PS plot had the largest average stalk < 0.05 based on One-Way ANOVA with Student-Neuman-Keuls post
diameters (8.20±1.60 mm) compared to all the other plots. hoc testing
Table 4. Rates of photosynthesis and transpiration compared between Jersey and Bluecrop cultivars across four treatment plots, horse manure, sheep
manure, pine straw, and Planters Choice mulches
Parameter Cultivar Horse Manure Sheep Manure Pine Straw Planters Choice Average
Photosynthesis Jersey 9.8 ± 1.4 5.6 ± 0.8 13.0 ± 1.9 13.0 ± 5.3 10.3 ± 1.6a
μmol CO2/m2/s Bluecrop 4.1 ± 0.6 2.5 ± 1.2 5.1 ± 0 4.9 ± 0.4 4.3 ± 0.4b
Transpiration Jersey 2.5 ± 0.1 1.4 ± 0.1 2.4 ± 0.5 3.3 ± 1.3 2.4 ± 0.4 a
mmol H2O/m2/s Bluecrop 1.7 ± 0.6 1.0 ± 0.5 1.7 ± 0.2 1.6 ± 0.1 1.5 ± 0.1b
Values shown are group averages ± SEM. Jersey photosynthesizes at higher rates than Bluecrop in all treatments. Jersey bushes, planted in Planters
Choice plot, have an overall higher rate of photosynthesis than others.
Table 5. Plant growth parameters for all cultivars compared across soil mulch treatments, 2010
Plant parameter Soil mulch treatments
Horse Manure Sheep Manure Pine Straw Planters Choice
N Mean ± SEM N Mean ± SEM N Mean ± SEM N Mean ± SEM
Plant height (cm) 54 62.5 ± 2.2 (a) 47 56.2 ± 2.8 (a) 23 73.4 ± 2.6 (b) 36 62.5 ± 2.2 (a)
Volume cylinder (cm3) x1000 54 182 ± 15 (a) 47 125 ± 16 (a) 23 381 ± 37 (c) 36 283 ± 20 (b)
Bushiness 54 385 ± 35 (a) 47 322 ± 33 (a) 23 627 ± 131 (b) 36 1171 ± 140 (c)
Primary stalk diameter (mm) 54 6.73 ± 0.19 (a) 47 6.56 ± 0.23 (a) 23 8.20 ± 1.60 (c) 36 7.50 ± 1.51 (b)
N = number of bushes measured in each soil treatment plot. Expressed values represent averages and standard error of the mean for that group.
Differing lower case letters across the columns for a given plan parameter indicate statistically different subsets (P <0.05) based on one-way ANOVA
with Student Neuman-Keuls post hoc testing.
 Optimal conditions for highbush blueberry growth
Similarly, PS plants had the largest plant cylinder volume (381±37
mm3x 1000) in comparison to all other cultivars. Regarding plant
bushiness, PC bushes were significantly higher (1171± 140) than
all other cultivars. PS and PC plants were each significantly higher
in all parameters in comparison to both HM and SM plants. PS
bushes had significantly larger average stalk diameters and plant
cylinder volumes, and PC plants were greatest in height and
bushiness (Table 5).
Growth parameters- Comparing cultivars: Bush height
comparison resulted in Bluecrop having a significantly greater
height (77.85±2.6 cm) than other cultivars (Fig. 6). Analyses
of primary stalk diameter comparisons indicated that Chandler
bushes were significantly thicker (8±0.36 mm) than the other
cultivars (Fig. 7). Additionally Bluecrop had significantly larger
primary stem diameters than either Duke or Bluegold. Plant
Fig. 6. Bush height comparison across five cultivars. Measurements were
taken in the fall of 2010. Asterisk indicates statistically significantly
different group (P < 0.05). Height is a significant indicator of plant
health, suggesting Bluecrop bushes fair the best.
Fig. 7. Mean primary stalk comparison across five cultivars, fall of
2010. Differing lower case letters indicate statistically different subsets
(P < 0.05). Stalk diameter is a significant indicator of plant hardiness,
suggesting Chandler bushes are the hardiest.
37
vigor in terms of an overall plant bushiness assessment resulted
in Chandler (976.38 ± 172.8) being significantly greater than
Duke (241.44 ± 26.16) and Bluecrop (459.64 ± 40.16), but
comparatively similar to Jersey (690.9±119.4) and Bluegold
(799.34 ± 116.64) (Fig. 8). Comparative assay of the mean volume
of plant cylinders resulted in Duke (114683 ± 14639 cm3) being
smaller than any of the other cultivars (Fig. 9). These statistical
analyses of plant growth parameters resulted in the following
ranking from superior to inferior cultivar per organic plot:
Chandler, Bluecrop, Jersey, Bluegold, and Duke, respectively. The
optimal cultivar was Chandler, which was superior in all, but one
parameter. Bluecrop had a significantly larger height.
Photosynthesis and transpiration measurements: Results
from a Student Independent T-test indicated that Jersey bushes
have overall higher rates of photosynthesis than Bluecrop bushes
Fig. 8. “Bushiness” comparison across five cultivars, fall of 2010.
Measurements were taken in the fall of 2010. Differing lower case
letters indicate statistically different subsets (P < 0.05). “Bushiness”
is a significant indicator of plant health, suggesting Chandler bushes
are thriving the best in terms of plant stalk height, number of primary
branches, and number of primary stalks.
Fig. 9. Average volume of plant cylinder comparison across five cultivars,
fall of 2010. Asterisk indicates statistically significantly different group
(P < 0.05). Since bush volume is a significant indicator of plant vigor,
data suggest Chandler bushes thrive better overall.
38 Optimal conditions for highbush blueberry growth
(See last column in Table 4). There was no significant difference
between the soil treatments of each cultivar. Though PS and PC
trended toward higher photosynthesis and respiration values
than the average (Table 4), these trends were not found to be
statistically significant. Thus we cannot definitely say that any
one mulch enhances or decreases the photosynthetic or respiration
characteristics of a given cultivar. In retrospect, due to reading
variations for a given plant, the number of sample readings should
have been increased at least 10 fold to be able realistically to
determine potential differences due to different soil treatments.
Pine straw and Planters Choice are optimal mulches for highbush
blueberries grown in the Shenandoah Valley. Reason, why other
mulch treatments were not as beneficial, could be the conditions
of the raw components of the horse and sheep manure, causing
the bushes planted in the HM and SM plots to respond poorly
in all growth parameters as indicated by Table 5. The manure
based mulches may have been too raw at time of deposition
and therefore promoted the higher levels of nitrate and calcium
present in the soil. An excess of nitrate or calcium could have
detrimental effects, including growth retardation of blueberry
plants. While, the high pH levels in Planters Choice and sheep
manure mulches potentially mitigated against optimal growing
conditions in the soils of their respective treatment plots, the PC
plot contained other elements that apparently enhanced plant
growth. Thus while an acidic pH 4.5-5.0 may be more optimal
in promoting blueberry growth, other nutritional factors such as
increased percentage of organic content or elevated humus levels
may alleviate the negative effects of a higher pH (>5.5) and still
provide and promote quality growth. Although the soil nutrients
for the PS plot were lower than the others, the more acidic pH of
that plot (3.87) may have compensated in promoting plant growth
with minimal soil nutrient content.
Though PC bushes were determined to be optimal in height
and bushiness, cost/benefit consideration should be taken
before planting. Planters Choice mulch was locally purchased.
Applications of Planters Choice mulch were applied to the
PC rows in June 2009 and again in September 2009 at overall
purchase cost of $126. In contrast, pine straw was acquired from
neighborhood pine trees without charge; the only expense was
the hauling and time spent loading the pine needles. Analysis of
bushes grown in the PS plot showed that this mulch promotes
optimal growing conditions based on both stalk diameter and
volume of plant cylinder parameters. Thus pine straw mulch
provided the most economic and effective soil treatment to
enhance highbush blueberry growth at Knoll Acres.
Results indicate Chandler to be the superior cultivar in all growth
parameters except for height. This could be due to the intrinsic
characteristics of Chandler highbush blueberries. Chandler bushes
have the longest ripening period, largest berries, and harden
quickly. Resource allocations to these could explain the growth
parameter patterns present, including the shorter overall height
in Chandler bushes when compared with Bluecrop.
The measurement of in-field photosynthesis rates indicate that
Jersey bushes are photosynthesizing at higher capacities than
Bluecrop bushes. This suggests that there could be an intrinsic
difference in photosynthetic function across each cultivar.
However, Table 4 values indicate that soil treatments play no
role in enhancing or decreasing the rate of photosynthesis in each
cultivar. A more inclusive study across each cultivar is needed to
determine this more fully.
At the time of the initial writing of this report, the planted
blueberry bushes had experienced two growing seasons during
which flower buds were removed to enable optimal plant growth.
No berries were harvested. Our plant growth measures and foliar
values (photosynthesis and respiration readings) demonstrated
that treatment with horse manure or sheep manure was suboptimal
in enhancing plant vigor. In contrast plots treated with pine straw
or Planters Choice optimized plant vigor. Based on the first berry
harvest which occurred the following summer (data not shown
in this paper), harvested berry yields paralleled these reported
growth and foliar measures. Per bushberry productivity in the
pine straw and Planters Choice plots averaged more than two-fold
greater than bushes in the horse manure and sheep manure plots.
Thus early growth measures and foliar readings in blueberry
bushes can accurately predict future productivity.
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Received: January, 2014; Revised: March, 2014; Accepted: April, 2014
 Journal

Journal of Applied Horticulture, 16(1): 40-45, 2014 Appl

Comparative efficacy of vermicomposted paper waste and

inorganic fertilizer on seed germination, plant growth and
fruition of Cyamopsis tetragonoloba

M. Karthikeyan, S. Gajalakshmi* and S.A. Abbasi

Centre for Pollution Control and Environmental Engineering, Pondicherry University, Chinnakalapet,
Puducherry-605014, India. *E-mail: dr.s.gajalakshmi@gmail.com

Abstract
The aim of the present study was to assess the influence of vermicompost generated from the paper waste spiked with cow dung slurry
on the germination, plant growth and fruition of cluster bean. Two kinds of treatments were studied: (i) vermicast was applied to the
soil at the rates of 5, 7.5,10 t ha-1 and (ii) amounts of essential nutrients equivalent to those present in the vermicast treatments in
inorganic form was amended to the soil. There was a control with only soil without any nutrient supplement. The finding is in contrast
to the reports on the beneficial impacts of vermicast on plant growth. In the present study, the inorganic fertilizer treatment exhibited
better seed germination and plant growth than the equivalent vermicast treatments. The results indicate that the dose of vermicompost
used in the present study was not sufficient to satisfy the nutrient demand of plant species studied. Additional fertilization would have
improved the crop productivity.
Key words: Vermicompost, paper waste, plant growth, Cyamopsis tetragonoloba.

Introduction
Paper waste generation has been continually increasing over
the past years due to increasing population, industrialization,
urbanization and literacy. In India, the paper consumption is about
8.5 kg per capita per year and it is 0.81-5.8 % of municipal solid
waste (MSW) (Gupta and Garg, 2009; www.indiastat.com). Due
to the absence of waste segregating practices, the paper waste is
dumped along with all other kinds of waste in open and poorly
managed landfills, which is very common practice in most of the
cities in India. The improper disposal of this degradable waste
may lead to long term threat to the environment and public health,
such as the risk of ground water pollution due to leachate seepage,
fugitive greenhouse gas emission contributing to climate change
and odour pollution caused by non-methane organic compounds,
which is direct harassment to adjacent communities (Zhang et al.,
2012). Also, open dumping of wastes facilitates the breeding for
disease vectors such as flies, mosquitoes, cockroaches, rats, and
other pests (CPCB, 2000).
Paper and cardboard have a relatively high heating value,
similar to wood, and this energy utilized via incineration can
be transformed into electricity (Villanueva and Wenzel, 2007).
However, incineration is not very much practiced in India due
to lack of awareness and absence of waste segregating practices.
The paper waste when mixed with other moist organic waste
and inert material reduces its calorific value (Negi and Suthar,
2013). In recent years, due to shortage of raw material, waste
paper is preferred for paper production. Also, recycling of paper
consumes only 40% of the energy in comparison to the process
based on other raw materials (Gupta et al., 1998). However, the
paper recycling industries prefer to use imported waste paper
because of its better quality in terms of fibre strength and also
due to inadequate domestic supply owing to the unorganized
collection of waste paper within the country. In addition, the yield
from imported waste paper can be as high as 90%, whereas the
widely available agro-pulp and wood pulp-based waste paper in
India gives yield less than 50% (IPMA, 1996). Nevertheless, if
waste papers are segregated at the source itself, it could be the
input material for paper recycling units. However, due to lack of
efficient waste management service, the paper waste invariably
finds its way to the MSW at the end.
The huge generation of these paper wastes can be treated by
vermicomposting which convert the waste into useful end product
that can be used as a soil amendment (Sinha et al., 2010). Unlike
recycling and incineration, the biological composting process
is not affected either by quality of waste paper or when mixed
with other organic wastes. Processing of paper waste through
vermicomposting may provide an answer to the minimization
of waste accumulation and also to widespread deteriorated
agricultural land due to rampant use of inorganic fertilizers. It is
well established that vermicompost application have beneficial
impact on soil physical, chemical and biological properties and
can increase the germination, plant growth and yield in both
natural and agricultural ecosystem (Edwards and Bohlen, 1996).
These beneficial effects have been attributed to improvement
in soil properties and structure, to greater availability of
mineral nutrients to plants (Edwards, 1998). In addition to this,
vermicompost contain plant growth regulating components,
including plant growth hormones and humic acids that are
reported to be responsible for increased germination, growth
and yields of plants, in response to vermicompost applications or
substitutions, independent of the nutrients they contain (Tomati
et al., 1988; Muscolo et al., 1999; Atiyeh et al., 2002; Arancon
et al., 2003; 2006).
However, the vermicompost generated from waste paper causes
 Comparative efficacy of vermicomposted paper waste and inorganic fertilizer on Cyamopsis tetragonoloba 41

apprehension towards the beneficial impact on plant growth due exhibited radical extension of >3 mm. Counts of germinated seeds
to the low nutrient content of this substrate. Therefore, an attempt were made daily up to eight days to determine the germination
was made to investigate the beneficial impact of vermicompost rate in terms of germination percentage (GP) and germination
generated from the paper waste (VC) on the germination, plant value (GV) by method described by Djavanshir and Pourbeik
growth and yield of cluster bean, a vegetable crop. This plant (1976). The GV is used as a comparative index to statistically
was chosen due to their drought tolerance which reduces the assess the effects of the treatments. After germination, one plant
error due to the other environmental factors. Moreover as it is a per compartment was maintained. A separate nursery was also
leguminous plant, influence of vermicast and inorganic fertilizers maintained with all VC and IF treatments. Healthy plants from
on nodules formation and its growth can be revealed. In addition, the nursery were transplanted to containers where seeds failed
to evaluate the possible non-nutrient (i.e., hormones and other to germinate. Adequate amount of water was provided during
growth regulating components) dependent effect of vermicast the experiment. Weeding was done manually. In few instances
over inorganic fertilizers, all essential nutrients present in the when pests were seen, organic pesticides such as neem extract
vermicast were supplied in inorganic form (IF) to the plant, and and cow urine were used.
the response of plant to the different fertilizers is briefed in this The plant height, length of shoots and roots, number of leaves,
paper. stem diameter, number of nodules present in the root and their
size, biomass of shoot and root and its dry weight were recorded
Material and methods with randomly collected samples at each week. Leaf chlorophyll
Study area: The experiment was conducted at Pondicherry a, b and carotenoids content were estimated photometrically by
University, Puducherry, India, located on the east coast of Indian using N,N-di-Methyl formamide (DMF) as extractant (Moran and
peninsula (latitude 11°56’ N and longitude 79°53’ E). The climate Porath, 1980). Throughout the study, the disease incidence, day
of the experimental site is typical maritime of tropical climate of flowering and number of flowers produced were recorded.
with a disymmetric rainfall. The average annual rain fall is about Analytical methods: Soil pH was measured in suspension of
1300 mm with 57.25 mean rainy days ((http://port.puducherry. 1:2 (v/w) by using Digison™ digital pH meter 7007. The total
gov.in/Port_data/MetData.htm) ), and around 60% of the total organic carbon content was measured by modified dichromate
rainfall is received during period of October to December through redox method according to Heans (1984). The total nitrogen
the north-east monsoon. content of the samples was determined by modified Kjeldahl
Treatments: The experiment was set up in 0.4 m3 size wooden method (Kandeler, 1993) using Kel Plus™ semi-automated
containers lined up with HDPE sheets. These containers were digester and distillation units. Inorganic N (N-NH4+ and N-NO3- )
filled with low fertile barren land soil collected inside the was extracted in 2M KCl solution (1:10 weight: volume) and
Pondicherry University campus to reduce the errors due to ammonia content in the suspensions were determined by modified
previous soil practices. The experimental soil was characterized as Table 1. Characterization of vermicast and soil used in this study
sandy loam soil and its physico-chemical properties are shown in Parameters Vermicompost Soil
Table 1. The experiment was conducted during the Kharif season Chemical properties
which is best time for sowing the cluster beans in south India. pH 7.83 6.30
Pusa Navabahar variety was used which is locally available in Total organic carbon (g kg-1) 259.60 8.87
the experimental area. The vermicompost was generated from the Total nitrogen (g kg-1) 11.70 2.66
paper waste spiked with cow dung slurry by employing an epigeic Plant available form of
species, Eudrilus eugeniae Kinberg. The VC was applied to the Nitrogen (g kg-1) 6.21 0.39
plant growth containers at the rate of 5, 7.5 and 10 t ha-1. In another Phosphorus (mg kg-1) 71.40 4.05
set, an equivalent amount of all major and minor nutrients present Potassium (g kg-1) 1.98 0.40
in vermicompost was supplied as inorganic chemical form to Sulphur(mg 100g-1) 0.40 0.54
check the efficiency of vermicast over the inorganic fertilizer. Calcium (g kg-1) 15.90 8.27
In the IF treatment, the primary nutrients N, P and K, secondary Magnesium (g kg-1) 7.13 0.09
nutrients Ca, Mg and S, and micronutrients of Fe, Mn, Cu, Boron (mg kg-1) 7.28 26.90
Zn, B, Mo and Cl were applied to an equivalent amount of 5, Copper (mg kg-1) 16.9 5.08
7.5 and 10 t ha-1 VC treatment. The chemical fertilizers were Iron (mg kg-1) 208.30 59.90
applied in the form of urea, di-ammonium phosphate, potash, Manganese (mg kg-1) 63.80 45.10
CaCO3, MgO, Na2B4O7, CuSO4, FeSO4, MnSO4 and ZnCl2. Zinc (mg kg-1) 94.10 55.00
Besides these treatments, one more set was maintained without Molybdenum (mg kg-1) BDL BDL
any supplementation, as control i.e. only soil. The nutrients Physical properties
were supplied in two phases. The first phase was at the time of Dry weight (%) 49.60 94.70
sowing which comprised half of the total nutrients. The second Bulk density (g cm-3) 0.24 1.28
supplementation of nutrients was done at the time of flowering Practical density (g cm-3) 1.21 2.72
of plants. Water-holding capacity (%) 118.00 36.90
Germination, plant growth and yield characteristics: Two Electrical conductivity (mhos cm-1) 2.96 0.12
seeds per container, 72 seeds per treatment were sown in all Total porosity (%) 80.08 52.9
the containers. Seeds were considered germinated when they Air filled porosity (%) 68.14 46.3
42 Comparative efficacy of vermicomposted paper waste and inorganic fertilizer on Cyamopsis tetragonoloba
indophenol blue technique with Labindia™ UV- 1201 model UV-
Vis spectrophotometer (Bashour and Sayegh, 2007). The nitrate
concentration in the extract was measured by Devada’s alloy
method using Kel Plus™ distillation unit. Inorganic N contents in
samples are a measure of N available to plants (Jones, 2001).
Extractable potassium, calcium and sodium were determined using
a Flame photometer (Elico™ CL378) after extraction with neutral
1N ammonium acetate solution. Extractable magnesium, boron,
copper, iron, manganese, zinc, molybdenum were determined
using a ICP-AES (JobinYvon – Ultima 2) by extracting sample/
solution ratio of 1:25 with Mehlich 3 extraction solution (Mehlich,
1984). The same extract was used to determine the extractable
phosphorus according to the ammonium molybdate-ascorbic
acid method (Knudsen and Beegle, 1988). Mineral S in soil was
extracted with 0.0125M CaCl2 solution (ratio of soil: solution,
1:4), and analyzed with a turbidimeter after the addition of BaCl2
which generates insoluble BaSO4 (Bashour and Sayegh, 2007).
Bulk density was measured on undisturbed cores of soil and
graduated cylinder method was used for vermicast. The particle
density was determined by volumetric flask method (Bashour
and Sayegh, 2007). The total and water filled porosity were
calculated from the particle and bulk density values of respective
samples using standard formula (Carter and Gregorich, 2008).
Water holding capacity (WHC) was obtained by water retention
of samples filled in perforated base cylinders immersed in water
after draining (Margesin and Schinner, 2005). The electrical
conductivity (EC) was measured in suspension of 1:2 (v/w) by
using EI™ 611E EC meter.
Statistical analysis: The data were analyzed statistically with
software SPSS 16 package and subjected to either two-way
ANOVA or MANOVA. Comparisons between means were tested
with an LSD test. Selection of methods for different parameters
were chosen based on the number of observations and number
of independent variables for particular parameter.
Results and discussion
Seed germination: In both VC and IF treatments, maximum
GV was observed on fourth day since the experiment began and
then a decreasing trend was observed as the days progressed.
The lowest GV was reported on the eighth day of the experiment
(Table 2). The results of germination in terms of GV did not
show significant difference with different forms or amount of
nutrients applied. However, both forms of fertilizer and amount
of nutrient applied were significantly higher than the control
treatment (P<0.001) (Table 4). In the VC treatment maximum
GP of 88.6% was observed with both 7.5 and 10 t ha-1 treatment
and maximum GV of 40.0 was also recorded in these treatments.
Table 2. Effect of vermicast and inorganic fertilizer on germination of
cluster bean’s seed in terms of germination value (GV)
Treatment Amount Number of days from start of experiment
4 5 6 7 8 treatments induce suppression in nodules formation in soybean,
Vermicompost 5 t ha-1 24.70 17.27 15.33 11.26 8.62
7.5 t ha-1 40.00 27.46 20.41 16.01 12.26
10 t ha-1 40.00 29.38 21.79 16.01 12.26
Inorganic 5 t ha-1 40.00 27.46 20.41 16.01 12.26
fertilizer 7.5 t ha-1 42.91 29.38 20.41 14.99 13.06
10 t ha-1 40.00 33.44 24.70 18.14 13.89
Control Nil 2.50 2.64 2.74 4.26 3.47
The VC treatment at dose of 5 t ha-1 showed lowest GP and GV
values among both VC and IF treatments. In the IF treatment, an
increasing trend in GP was observed with increasing dosage of
fertilizer application. In this treatment, GP of 88.6, 91.4 and 94.3%
was recorded with 5, 7.5 and 10 t ha-1 treatment on the eighth
day and, a maximum GV of 42.91 was in 7.5 t ha-1 treatment.
In both VC and IF treatments, maximum seed germination was
recorded in 10 t ha-1 treatment followed by lower dose of nutrient
application. However containers amended with VC had shown
lesser germination rate than their respective equivalent inorganic
treatments (Table 3).
Other studies on influence of VC on seed germination either
showed inhibitory or stimulatory effect with low concentration of
VC substitution in growth media. The discrepancy in the response
to vermicompost depends on plant species which reacts differently
to the concentration of vermicompost application (Atiyeh et al.,
2000; Edwards et al., 2004). The different organic substrate
used for vermicomposting also changes the quality of vermicast
(Zaller, 2007). Ievinsh (2011) reported inhibitory effect similar
to the present study on beetroot, beans and pea at low dose of
vermicompost generated from cow dung (5-10%), but germination
dramatically increased with the increase of vermicompost
concentration. The higher germination in IF treatments may be
due to nitrate which would have been converted from the urea
applied as nitrogen source. This constituents are breakers of
dormancy and also stimulator of germination (Egley and Duke,
1985; Hilhorst and Karssen, 2000). Since, the nitrate readily
leaches from surface soil by irrigation water, the concentration
of nitrate would have declined as the days progressed, and this
may be the reason for higher germination at initial days followed
by steep decline till the end.
Plant growth: There was no significant difference found in
results pertaining to stem diameter, length, fresh weight and dry
weight of shoot and root and number of leaves with different
treatments. There was significant difference only with number of
nodules between different treatments (Table 4). However, there
was a differential impact of the vermicast and inorganic fertilizer
treatment on all the morphological parameters (Table 3). Since
the data gathered were from three leaves stage to maturity, there
was a larger difference within the samples. This might be the
reason for the insignificant difference in data. In this experiment,
the plants which were treated with IF grew faster than the VC
treated plants. The highest stem diameter (14.4 mm), shoot length
(134 cm), number of leaves (72), fresh weight (182.1 g ) and dry
shoot weight (52.4 g ) was recorded in the plants treated with IF
treatment equivalent to 10 t ha-1 paper waste VC. Plants treated
with VC at the rate of 10 t ha-1 showed a maximum of 14.1 mm
of stem diameter; 121 cm shoot length, 65 numbers of leaves, 180
g of shoot fresh weight and 57.3 g of shoot dry weight.
Many authors have reported that high dose of chemical fertilizer
chick pea and lupins (van Schreven, 1959; Carroll and Gresshoff,
1983; Harper and Gibson, 1984; Nie, 1989). The finding of the
present study contradicts with the previous reports on suppression
of nodules formation with IF treatment. There was no significant
difference in the number and size of nodules between VC and IF
treatment. The results indicates that there was no suppression or
stimulation of nodules formation with both VC and IF treatment.
 Comparative efficacy of vermicomposted paper waste and inorganic fertilizer on Cyamopsis tetragonoloba 43

Table 3. Plant growth, leaf pigments, flowering, disease incidence, stunted growth and plant death of cluster bean treated with vermicast and inorganic
fertilizer at the end of the experiment
Parameters Vermicompost Inorganic fertilizer Control
5 t ha-1 7.5 t ha-1 10 t ha-1 5 t ha-1 7.5 t ha-1 10 t ha-1
Plant growth
Stem diameter (mm) 10.9 12.8 14.1 11.9 13.1 14.4 11.2
Shoot length (cm) 96.8 118 121 108 119 134 76.8
Root length (cm) 59.2 72.1 83.4 55.7 89.1 50.1 50.2
Number of leaves 59 62 65 51 68 72 42
Number of nodules 28 41 45 38 38 37 16
Shoot dry weight (g) 37.1 47.6 57.3 39.8 43.2 52.4 45.3
Root dry weight(g) 4.78 6.82 7.22 7.53 9.39 12.7 3.21
Leaf pigments
Chlorophyll a (mg g-1) 1.83 1.92 1.87 1.84 2.24 2.29 1.32
Chlorophyll b (mg g-1) 1.61 1.63 2.05 1.51 2.12 2.28 1.45
Total chlorophyll (mg g-1) 3.44 3.55 3.92 3.35 4.35 4.57 2.77
Carotenoids (mg g-1) 0.29 0.46 0.46 0.36 0.36 0.37 0.26
Flowering
Number of flowers per plant 1.4 3.1 3.6 3.2 4.1 4.4 0.7
Disease incident, plant death and stunted plants
Number of infected plant 19 28 21 8 14 12 16
Number of plant died 0 1 0 1 0 0 0
Number of stunted plant 0 2 1 1 6 11 11

Table 4. Calculated F-values using two-way ANOVA and MANOVA to study the effect of different fertilizer and amount of application on germination
and plant growth parameters of cluster bean
Treatment Germination Stem Shoot Root
GV diameter length length
Type of fertilizer 1.032n.s 0.034n.s 0.009n.s
Amount 0.915n.s 0.579n.s 0.971n.s
Type of fertilizer x Amount 0.352n.s 0.096n.s 0.075n.s
*P<0.05, **P<0.01, ***P<0.001, n.s - not significant.
The reason may be that very low amount of nutrient was supplied
to the plants in both VC and IF treatment. In addition to this,
slow releasing property of vermicast might be lowering nutrient
availability to the plants. In the VC treated containers, it was
observed that applied VC did not disintegrate till the end of the
experiment period. This stable nature of this VC may have slowed
down the nutrient release to plant. It might be the reason for lower
plant growth in the VC treatment than equivalent IF treatment.
Photosynthetic pigments: Although, photosynthetic pigments
such as chlorophyll and carotenoid in the leaves of plants
amended with vermicast and inorganic fertilizer highly correlated
with amount of nutrient application (Table 3), they were not
significantly different (Table 5), probabaly due to the same
reasons discussed above. In general, IF application was more
effective than respective VC treatment. Lower rate of nutrient
application and their poor availability due to the slower releasing
property of vermicast may be the reason for lower photosynthetic
pigments in this treatment compared to IF treated plants. In the
VC treatment, the maximum total chlorophyll content (3.92 mg
g-1) was recorded with the dose of 10 t ha-1 followed by 3.46 and
3.44 mg g-1 with 7.5 and 5 t ha-1 treatments, respectively. In case
of IF treatment, the maximum total chlorophyll content (4.57
mg g-1) was recorded in 10 t ha-1 treatment followed by 4.35
and 3.35 mg g-1 with 7.5 and 5 t ha-1 IF treatments. The control
showed lowest chlorophyll content (2.76 mg g-1) at the end of the
Number of Number of Shoot dry Root dry
leaves nodules weight weight
0598n.s 0.149n.s 0.360n.s 0.034n.s 1.999n.s
2.477n.s 1.086n.s 0.788n.s 0.507n.s 0.962n.s
0.856n.s 0.147n.s 0.499n.s 0.022n.s 0.223n.s
experiment period. Carotenoids pigment exhibited similar trend of
results. The increasing nutrient availability with increasing dose
of fertilizer application can be attributed to the formation of leaf
pigments (Mengel and Kirkby, 1987; Shadchina and Dmitrieva,
1995; Ruza, 1996; Tejada et al., 2007).
Disease incidence, plant death and stunted growth: In the first
two months of the experimental period many plants were infected
with bacterial blight, Alternaria fungal infection and whiteflies.
As a consequence some plants died and some did not grow
normally (Table 3). The observed disease incidence and plant
mortality was not significantly different between different forms
of nutrient application (vermicast and inorganic fertilizer) (Table
5). The number of stunted plants was maximum in the beginning
of the experiment. After a month, all the plants grew well.
Inorganic treatment showed more stunted plants in comparison
to the VC treatment. In IF treatment a total of 18 stunted plants
were recorded, whereas in the case of VC treatment at different
doses, two or less stunted plants were recorded.
Effect on flowering: The IF treated plants produced more number
of flowers in comparison to the equivalent VC treated plants
(P <0.001) (Table 5). A maximum was recorded with 10 t ha-1
treatment with both VC and IF treatments. Increasing dose of both
fertillizer application showed the trend with maximum number of
flowers 10 t ha-1> 7.5 t ha-1> 5 t ha-1> control. The plants grown
44 Comparative efficacy of vermicomposted paper waste and inorganic fertilizer on Cyamopsis tetragonoloba
Table 5. Calculated F-values using two-way ANOVA and MANOVA to study the effect of different fertilizer and amount of application on flowering,
photosynthetic pigments and diseases incident of cluster bean
Treatment Flowering Photosynthetic pigments
Number of Chlorophyll a Chlorophyll b
Flowers
Type of fertilizer 21.04*** 2.061n.s 1.959n.s
Amount 17.00*** 0.531n.s 2.830n.s
Type of fertilizer x Amount 1.316n.s 0.384n.s 0.895n.s
*P<0.05, **P<0.01, ***P<0.001, n.s - not significant.
in the soil treated with 10 t ha-1 of both VC and IF showed early
flowering than the other treatments. Increase in availability of
nutrient in this treatment could be the reason for the higher and
early flowering.
Effect on yield: In all the treatments no vegetables were
produced. The reason may be due to inadequate availability of
minor and trace nutrients to the plants in all these treatments. In
this study, low fertile soil collected from barren land was used
to minimize the errors due to the previous soil practices in the
experimental results. The soil was characterized as very low
nutrient soil. Therefore, the growth and yield of plants relied
on the applied nutrient either in the form of VC or IF. The trace
nutrient applied would have been exhausted thereby impeding
the fruiting in all the treatments.
The results obtained from the experiment shows that application
of VC generated from the paper waste had no beneficial impact on
growth of cluster bean plant. Moreover, the IF treatment exhibited
better seed germination and plant growth than the equivalent
VC treatment. This indicates that the slow releasing property
of VC might have led to depletion of nutrient availability to the
plants. In this experiment, increasing dose of both vermicast
and inorganic fertilizer improved the germination rate and
plant growth parameters. However, the application of inorganic
fertilizer did not suppress the formation and growth of root
nodules, which is contradictory to the previous reports. There
was no significant difference in number and size of nodules with
different treatments. In addition to this, the plants did not fructify
in all the treatments.
Acknowledgments
Senior author thanks to University Grants Commission,
Government of India for the research fellowship under the RGNF
scheme.
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 Journal

Journal of Applied Horticulture, 16(1): 46-49, 2014 Appl

AM fungi shields Coleus forskohlii from root rot incidence

L. Srimathi Priya and K. Kumutha

Department of Agricultural Microbiology, Tamil Nadu Agricultural University, Coimbatore– 641 003, Tamil Nadu, India.
*E-mail: agrisriya@gmail.com

Abstract
This study was taken up to determine the combined effect of arbuscular mycorrhizal (AM) fungi and plant growth promoting rhizobacteria
(PGPR) in controlling root rot caused by Macrophomina phaseolina in Coleus forskohlii. AM root colonization was up to 70-73 per cent
under combined inoculation of Scutellospora sp + Pseudomonas fluorescens + Trichoderma viride and 44-45 per cent under individual
inoculation. A correlation analysis indicated that more the AM root colonization (73 per cent) less the root rot (28 per cent) incidence.
The activity of the defense enzymes viz., peroxidase, polyphenol oxidase and superoxide dismutase was found to be high at 30 days
after inoculation of the pathogen in the co-inoculated treatments. Another correlation study between AM colonization and enzyme
activity, showed low root rot index. There was a loss in the alkaloid content due to pathogen infection, yet, the combined treatments
recorded a threefold increase in disease suppression.
Key words: AM fungi, plant growth promoting rhizobacteria, Coleus forskohlii, colonization, root rot index, peroxidase, polyphenol
oxidase, superoxide dismutase, alkaloid.

Introduction phaseolina; Trichoderma viride (TV 1) + M. phaseolina; SSP 3

Plant response to colonization by mycorrhizal fungi can range
from dramatic growth promotion to growth supression and the
factors affecting this response include mycorrhizal dependency of
the host crop, nutrient status of the soil, and the inoculum potential
of the mycorrhizal fungi (Dalpe and Monreal, 2004). Besides,
their ability to increase absorption surface of the roots and making
the immobile ions available for growth, their antagonistic activity
against the root rot pathogens is remarkable since it decreases
disease susceptibility and increases tolerance against biotic and
abiotic stress.
Coleus forskohlii is a medicinal herb, susceptible to root rot
disease and as a result, the tubers get infected by pathogen
like Macrophomina phaseolina that leads to reduction in its
forskohlin content. Though many chemical formulations and
many management practices are used to overcome this problem, a
biocontrol practice is a must to support sustainable farming. With
this necessity, studies were taken up using various inoculants like
Pseudomonas fluorescens, Trichoderma viride and AM fungi to
exploit the potential of AM fungus and Pseudomonas towards
controlling the root pathogen M. phaseolina.
Materials and methods
An experiment was carried out at Department of Agricultural
Microbiology, TNAU to study the effect of AM inoculation
(Scutellospora sp.) with two PGPR isolates (Pseudomonas sp.)
on Coleus forskohlii against M. phaseolina. Pots of 30 x 28 cm
size with 10 kg soil having pH-8.18; EC-0.89 dSm-1; available N -
219 kg/ha; P2O5 - 14.3 kg/ha and K2O - 293.4 kg/ha was subjected
to the following treatments. Treatments were replicated thrice.
Design used was completely randomized block.
Treatments: Scutellospora sp. SSP 3 + M. phaseolina; P.
fluorescens (CPF 1) + M. phaseolina; P. fluorescens (CPF 2) + M.
+ P. fluorescens (CPF 1) + T. viride (TV 1) + M. phaseolina; SSP
3 + P. fluorescens (CPF 2) + T. viride (TV 1) + M. phaseolina;
M. phaseolina alone.
Inoculants: The pathogen used for the study was M. phaseolina
obtained from the Department of Plant pathology, Tamil
Nadu Agricultural University, Coimbatore. The pathogen was
multiplied in sand maize medium for 15 days (Riker and Riker,
1993) at the rate of 10 g per kg soil (i.e. 100 g/pot) before
planting. The Pseudomonas isolates (CPF 1 and CPF 2) and the
isolate SSP 3 (Scutellospora sp.) isolated from the rhizosphere
of C. forskohlii were used as inocula for treatments. The AM
fungus at the rate of 50 g pot-1 (containing 300-400 spores 100
g-1 inoculum) and talc based bio-control agent T. viride (TV 1)
obtained from the Department of Plant pathology, Tamil Nadu
Agricultural University (at the rate of 100 g pot-1) were applied
in soil before planting. Pseudomonas suspension (108 cells mL-1
broth) was inoculated (50 mL broth pot-1) at the time of planting.
Two to three leaved cuttings of C. forskohlii were used for
planting. The control treatment was maintained with pathogen
inoculation alone.
Observations: Root samples were taken after 45 days of planting
and AM colonization was assessed in roots (Phillips and Hayman,
1970). Based on the disease incidence, root rot index was
calculated (Kesavan and Choudhary, 1977) at 45th DAP.
Root rot index: The root rot index was calculated by employing
the formula given below:
Summation of individual scores
Root rot index = × 100
Maximum grade x Total number of plants
Assay of defense related proteins and chemicals induced by
bio-inoculants
Peroxidase and polyphenol oxidase: The peroxidase enzyme
activity was assayed using pyrogallol and the intensity of
 AM fungi shields Coleus forskohlii from root rot incidence 47

formation of yellow colour due to the enzyme activity was Table 1. Effect of combined inoculation of AM fungus and PGPR
measured at 430 nm spectrophotometrically (Hammerschmidt et organisms on root rot index and AM root colonization in C. forskohlii
inoculated with M. phaseolina at 45 days after inoculation
al., 1982). For estimation of polyphenol oxidase enzyme activity,
Treatments Root rot AM root
catechol was used as the substrate and the activity was measured index colonization
as change in absorbance at 495 nm spectrophotometrically. (%) (%)
T1: Scutellospora sp. (SSP 3) 38.2 66.8
Superoxide dismutase (SOD): SOD activity was determined
T2: Pseudomonas fluorescens (CPF 1) 40.6 48.2
by measuring inhibition of the photochemical reduction of NBT T3: Pseudomonas fluorescens (CPF 2) 53.3 44.6
(Nitro Blue Tetrazolium) and the absorbance was read at 560 nm T4: Trichoderma viride TV 1 34.2 45.5
(Beauchamp and Fridovich, 1971). T5: SSP 3 + CPF 1+ TV 1 28.5 73.5
T6: SSP 3 + CPF 2 + TV 1 29.5 70.0
Forskohlin estimation: Forskohlin was estimated at harvest Pathogen inoculated Control 79.3 40.0
using HPLC. Fresh root tissue (200 mg) was powdered, soaked in LSD (P=0.05) 2.0 2.3
2 mL of absolute methanol (HPLC grade) for 24 h and centrifuged
at 10,000g for 10 min at 4 oC. The supernatant obtained was Table 2. Correlation analysis between AM colonization in roots of C.
forskohlii, root rot index and defense enzymes activity under pot culture
concentrated by vacuum centrifugation for 5 h. The concentrate condition inoculated with M. phaseolina
was dissolved in HPLC grade methanol and then filter sterilized Enzyme activities AM root colonization
using membrane filter (0.2 μm). Forskohlin estimation was done Root rot index -0.700
using High Performance Liquid Chromatographic system (HPLC) Peroxidase activity 0.803
employing a mixture of acetonitrile and water (50 : 50 v/v) as the Poly phenol oxidase activity 0.819
mobile phase and 250 x 4.6 mm C 18 column (Octadecyl silane - Super oxide dismutase activity 0.944
5μ size) as the stationary phase. The flow rate was 1.5 mL/min and Catalase activity 0.884
the wavelength was 220 nm. Forskohlin - 100 ppm concentration colonization and root rot index was found to be negative (r =
was used as the standard. From the area of the peak obtained in -0.719) (Table 2).
the graph, the content of forskohlin present in the samples was
calculated. The biometric observations were recorded along with Enzyme activities: Significant peroxidase activity was found at
enzyme related studies in the roots of C. forskohlii as well as in 20 days after inoculation (DAI) in all the treatments, where the
the rhizosphere soil. The data were subjected to statistical analysis combined inoculation of SSP 3 + CPF 1 + TV 1 registered 2-3 fold
by variance (P= 0.05) with mean separation by Least Significant increase in peroxidase activity, which was 48-87 per cent higher
Difference (LSD), as per the methods detailed by the Panse and than the individual inoculations. The induction of polyphenol
Sukhatme (1978). The analysis for microbial population was oxidase was higher (85.5-112.2 per cent increase over control) in
based on the log and arc sine transformed values. combined inoculations, while among the individual inoculations,
SSP 3 recorded a significant increase over pathogen inoculated
Results control. The defense related enzyme activity in the infected roots
was maximum at 20 DAI. The ‘Native page’ analysis revealed
AM colonization: Treatments significantly influenced the root that the induction of peroxidase as well as polyphenol oxidase
colonization percentage than when compared to the control. Inspite iosenzyme was prominent under combined inoculations (Fig.
of the presence of M. phaseolina the inoculants were not only 2). The SOD activity showed 2-3 fold increase over pathogen
able to reduce the root rot but also influence the colonization rate inoculated control (T8) because of the combined inoculation
positively. AM fungi SSP3 under combination with P. fluorescens of SSP 3 +CPF 1 +TV 1. Correlation analysis between AM
and T. viride (73.5 and 70 % in T5 and T6 respectively) still showed colonization and the enzyme activities showed positive ‘r value’
remarkable impact than when inoculated in single (66.8%) (Fig. (Table 2).
1).
Forskohlin content: Root rot infection led to reduction of the
Root rot index: A significant reduction in root rot index was forskohlin, but the loss of the alkaloid due to the pathogen severity
observed in all the treatments. T. viride performed better by was mitigated by the combined inoculation of AM fungus with
reducing the root rot index up to 34.2 %, followed by AM T. viride and Pseudomonas (Table 3, Fig. 5).
inoculation (38.2%) and Pseudomonas CPF 1 (40.6%). T. viride Table 3. Effect of combined inoculation of AM fungus and PGPR
and AM inoculation expressed significant reduction of root organisms on forskohlin content in roots of C. forskohlii inoculated
rot index over the control. In combination, these inoculants’ with M. phaseolina
performance was superior, which recorded the lowest root rot S No. Treatments Forskohlin content (%)
index of 28-29% (Table 1). A simple correlation between AM 1. Pathogen inoculated control 0.001
2. SSP 3 + CPF 1 + TV 1 + Pathogen 0.004
3. SSP 3 + CPF 2 + TV 1 + Pathogen 0.002

Fig. 1. Colonization of Scutellospora spp. SSP 3 in infected roots of C.
forskohlii; a & b: Hyphae with vesicles; a spore of Scutellospora
Discussion
Generally AM plants have high concentrations of ‘P’ than non
AM plants. This leads to the reduction in root exudates by altering
the membrane permeability and hence the pathogen population
may get reduced. This favours the negative correlation of AM
colonization and root rot index (Table 2). The reduction in disease
48 AM fungi shields Coleus forskohlii from root rot incidence
Fig. 2. Native PAGE analysis profile of peroxidase (a) and polyphenol
oxidase (b) isoforms induced due to combined inoculation of AM fungus
and PGPR organisms on 20 DAI in C. forkohlii; Lane (In order from
the right end): T1 - Scutellospora spp. SSP 3; T2 - P. fluorescens CPF
1; T3- P. fluorescens CPF 2; T4 - T. viride TV 1; T5 - SSP 3 + CPF 1 +
TV 1; T6 - SSP 3 + CPF 2 + TV 1
Fig. 3. Effect of combined inoculation of AM fungus and PGPR
organisms on AM root colonization and SOD acivity in C. forskohlii
inoculated with M. phaseolina
severity could be explained by morphological alterations in
host plants or by physiological changes due to concentration of
phenols, aminoacids etc. (Bagyaraj, 1989) or by the elicitation
of certain defense related molecules during AM colonization
(Gianinazzi et al., 1996). Increased peroxidase activity was
noticed in a number of interactions involving plant pathogenic
fungi, bacteria and viruses (Chen et al., 2000). Accumulation
of peroxidase has been correlated with induced systemic
Fig. 4. Fig. 4. Siderophore production by Pseudomonas isolates; a.
From the left, PFC6, CPF1 and control; b. From the left, PFC6, CPF1
and control
Fig 5. Effect of combined inoculation of AM fungus and PGPR organisms
on forskohlin content C. forskohlii inoculated with M. phaseolina
resistance in several plants (Hammerschmidt et al., 1982) and
with deposition of phenolic materials into plant cell wall during
resistance interactions (Graham and Graham, 1991), which
serve as a barrier that prevents the pathogenic fungal spread.
Loganathan (2002) reported that application of bioformulation
mixture induced several isoforms of polyphenol oxidase (PPO)
in cabbage and cauliflower against Plasmodiophora brassicae,
Sclerotinia sclerotiorum and Rhizoctonia solani. Enhanced PPO
activity in a mixture of biocontrol agents have been reported
by several workers. Accumulation of PPO was higher in the
combination of Pseudomonas strains (EPB22 + Pf-1) treated
banana plants and PPO activity was significantly enhanced in rice
plants pretreated with PGPR isolates (Radjacommare, 2005). This
oxidative enzymes alter substances related to pathogen infection
and thereby reduce the infection. Palma et al. (1993) studied the
presence of two isoenzymes (Cu-Zn SOD and Mn-SOD) in the
red clover roots due to the inoculation of G. mosseae in soil,
as a result of which, there was an increase in O2(-) radicals in
root. Arines et al. (1993) showed the presence of an additional
protein in the mycorrhizal extracts of red clover which was a plant
induced SOD. These oxidative enzymes are grouped as active
oxygen species (AOS) detoxifying enzymes; their production is
associated with stress for water as well as for pathogen infections.
Pathogenesis affects the biological oxidations in plant tissues
 AM fungi shields Coleus forskohlii from root rot incidence
which alter the concentrations of various substrates and products
of these reactions which in turn, are closely associated with plants’
defense mechanisms.
In this study, there was a positive correlation between AM root
colonization and isoenzyme activity in the infected roots, which
proved the enzyme induction induced systemic resistance in
plants (Fig. 3). The correlation values between the defense
enzymes (peroxidase, polypheol oxidase and SOD) and the AM
colonization percentage were positive (r = 0.8 to 0.9), which was
due to the influence of proteins produced during biotic stress in
the roots of the plants (Table 2). Several other reports showed the
role of AM fungus in accumulating the secondary metabolites in
crops like cucumber (Akiyama and Hayashi, 2002). C. forskohlii
being the nursery raised crop, AM inoculation offers better scope
to produce the disease free healthy seedlings and to reduce the use
of chemicals. C. forskohlii has gained popularity by virtue of its
exclusive constituent ‘forskohlin’ which has shown positive effect
in treating glaucoma, congestive heart failure, intestinal spasms,
insomnia, convulsions, hypertension, asthma and certain type
of cancers, by its ability to increase the synthesis of 3’5’ cyclic
ester of AMP (cAMP), by the action of adenylate cyclase enzyme
(Vishwakarma et al., 1988; De Souza, 1991).
Volatile and non volatile antibiotics and chitinase produced by
T. viride may inhibit the root pathogens (Ezziyanni et al., 2007).
It is now, the fluorescent Pseudomonas sp. that are considered
as an alternative to agrochemicals for controlling plant diseases
and increasing plant development (Gloria and Leda, 2006). Their
antagonistic property is attributed to the production of compounds
like, siderophore and hydrogen cyanide. Further these results were
supported by the field study, where in combined inoculation of
T. viride with G. mosseae gave the best results with less root rot
index of 32.28% in C. forskohlii (Boby and Bagyaraj, 2003).
The study revealed that there was no reduction in AM colonization
in plants receiving Pseudomonas/Trichoderma inoculants
along with the root-rot pathogen. The study also showed
the compatability of AM fungi with Trichoderma as well as
Pseudomonas. Forskohlin content was higher in the combined
inoculation of AM with PGPR organisims.
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Received: September, 2013; Revised: February, 2014; Accepted: March, 2014
 Journal

Journal of Applied Horticulture, 16(1): 50-53, 2014 Appl

Canopy management in mango (Mangifera indica L.) cv. Alphonso

with reference to flowering, yield and quality characters under
ultra high density planting

B. Gopu*, T.N. Balamohan1, P. Soman2 and P. Jeyakumar3

Department of Fruit Crops, Horticultural College and Research Institute, Tamil Nadu Agricultural University,
Coimbatore (TN), India. 1Horticultural College and Research Institute for Women, Trichy, Tamil Nadu Agricultural
University, Coimbatore (TN), India, 2Jain Irrigation Systems Ltd, Jalgaon (MH), India, 3Department of Crop
Physiology, Tamil Nadu Agricultural University, Coimbatore (TN), India. *E-mail: gopu16388@gmail.com

Abstract
An experiment was conducted to study the effect of different pruning levels on flowering, yield and quality characters in Alphonso
mango under Ultra High Density Planting from 2010-2011 at Jain Irrigation Systems Pvt. Limited (JISL) Farms, Udumalpet, Tripur
District, Tamil Nadu. The treatments included control, light pruning, moderate pruning, heavy pruning, 50 per cent removal of past
season growth and total removal of past season growth and imposed on five-year-old uniform sized Alphonso trees grown under a close
spacing of 3 x 2 m. The minimum number of days taken for first flowering and 50 per cent flowering were recorded by the control.
The highest number of panicles per tree and the maximum number of panicles produced per sq.m canopy area were recorded in the
control. However, highest percentage of hermaphrodite flower per panicle and per cent fruit set were found in the treatment T5 (50 per
cent removal of past season’s growth and tipping). Fruit and yield characters were influenced by different pruning levels. Treatment
T2 (light pruning) recorded the highest mean fruit weight, fruit length, fruit volume, fruit pulp weight and stone weight. However,
treatment T3 (moderate pruning) registered highest fruit circumference. Highest pulp to stone ratio was observed in T4 (Heavy pruning)
followed by T2 (light pruning). Highest number of fruits per tree and yield per tree were observed in control. Highest total soluble
solids, total sugars and non reducing sugars of the fruit were observed in T6 (total removal of past season’s growth). The maximum
acidity and ascorbic acid content were observed in control. Maximum total carotenoid content was recorded in T3 (moderate pruning)
and reducing sugars in T4 (heavy pruning).
Key words: Mango pruning, flowering, fruit set, fruit yield and quality.

Introduction flowering and good fruit quality as well as to rehabilitate mature

Mango (Mangifera indica L.) is one of the most important tropical
fruits of the world and is commonly known as the ‘King of fruits’.
Besides delicious taste and excellent flavour, mango is rich in
vitamins and minerals. Though many reasons are attributed for
low productivity, poor canopy management is considered as one
of the major limiting factors in mango production. Being an
evergreen tree, mango is seldom pruned in India, which leads
to over-crowding of branches resulting in poor penetration of
sunlight causing low productivity coupled with inferior quality
fruits (Rathore, 2009). There are several reasons for pruning
perennial fruit trees and if done drastically may influence several
physiological processes directly or indirectly. These effects result
from alteration in biochemical system within the tree and also
helps to restore the balance between root system and the above
ground parts. These operations are followed for maintaining tree
height, canopy spread and density which is required for effective
spraying which results in better fruit quantity and quality (Singh
et al., 2010c).
In general, management of canopy architecture deals with
positioning and maintenance of trees frame work in relation to
optimum productivity of quality fruits (Pathak, 2009). Charnvichit
et al. (1994) reported that, pruning operations to control tree
size are scarce and studies are mainly targeted to obtain early
trees (Medina-Urrutia and Nunez-Elisea, 1997). Yeshitela et al.
(2003) studied that pruning at the point of apical bud attachment
induced re-flowering, more rapid fruit development and more
fruits per panicle. For inducing flowering on old Alphonso mango
trees, severe pruning was more effective than mild pruning (Srihari
and Rao, 1998). On the other hand, Chen et al. (1996) observed that
heavily pruned trees of mango resulted in delayed flowering than
those of severely pruned ones. Davenport (2006) reported that
the main advantage of annual tip pruning was to provide reliable
synchronized flowering year after year in trees thus making them
to remain in the same size for many years. Waghmare and Joshig
(2008) made a study to regulate the vegetative flush for induction
of uniform flowering in ‘Alphonso’ mango and reported that the
sex ratio was maximum in 2.5 cm pruning immediately after
harvesting. Singh et al. (2010b) found that the pruning intensity
at moderate level took the least days to 50 per cent flowering.
In mango cv. Amrapali, the size of the fruits was improved with
the severity of pruning treatments (Pratap et al., 2003). Rao and
Shanmugavelu (1976) reported that the Mulgoa trees not yielding
for many years, yielded exceeding well after sever pruning. Singh
et al. (2010c) reported that TSS was the highest in the severely
pruned trees, while TSS: acid ratio were higher in the lightly
pruned trees. Keeping in view of above mentioned facts, the
present investigation was carried out to study how flowering,
 Canopy management in mango under ultra high density planting
yield and quality characters are changed after pruning in mango
cv. Alphonso under Ultra High Density Planting.
Materials and methods
The study was undertaken at Jain Irrigation Systems Pvt.
Limited Farms, Elayamuthur, Udumalpet during 2010-2011. The
experiment was laid out in a randomized block design having
six treatments and four replications. The trial was laid out in a
five-year-old orchard having one hundred and forty four uniform
sized trees spaced at 3 x 2 m. Each treatmental unit consisted
of six trees. The trees were maintained under uniform cultural
practices during the investigation period. The pruning was done
in last week of June 2010 and the pruning intensities were: T1
(control: tipping of previous season’s growth), T2 (light pruning:
retention of 70 cm from the base of the past season’s growth), T3
(moderate pruning: retention of 60 cm from the base of the past
season’s growth), T4 (heavy pruning: retention of 50 cm from
the base of the past season’s growth), T5 (severe pruning: 50 per
cent removal of past season’s growth and tipping) and T6 (very
severe pruning: total removal of past season’s growth). Pruning
was done by using shears after the harvesting of fruits. Data
were recorded on days taken for first flowering and 50 per cent
flowering, number of panicles produced per sq.m canopy area,
number of panicles per tree and percentage of hermaphrodite
flowers was calculated by using the given formula:
Number of hermaphrodite flowers
Hermaphrodite flowers = per panicle
(%) x 100
Total number of flowers per panicle
The percentage of fruit set was calculated at pea size stage as
follows:
Number of fruits at pea size
Fruit set (%) = x 100
Number of flowers per panicle
The fruit physical parameters such as mean fruit weight, fruit
length, fruit circumference, fruit volume, pulp weight, stone
weight, pulp to stone ratio were recorded. Yield data was recorded
at the time of harvesting. Fruit quality parameters such as total
soluble solids was determined by using hand refractometer,
titrable acidity and ascorbic acid as per the method of AOAC
(1975), total carotenoids by the method suggested by Roy (1973),
total sugars by the method suggested by Hedge and Horreiter
(1962), reducing sugars was estimated as per Somogyi (1952)
and non-reducing sugars was calculated as the difference between
the estimated total and reducing sugars. Data collected on flower,
yield and quality attributes were subjected to statistical analysis
as per the methods suggested by Panse and Sukhatme (1985).
Results and discussion
Evergreens, unlike deciduous trees, do not normally store large
reserves of manufactured foods and the growth is more closely
related to currently available leaf surface obtained after pruning.
In the present investigation, severity of pruning delayed the
flowering. The control, T5 (50 per cent removal of past season’s
growth and tipping) and T2 (light pruning) recorded early flowering
and 50 per cent flowering while it was delayed in severely pruned
treatments (Table.1). Thus, the shoots with desired maturity gave
rise to early flowering. The late commencement of flowering in
severely pruned trees than the unpruned ones may be explained on the
51
basis that pruned trees put forth new vegetative growth immediately
after pruning and almost the entire amount of carbohydrates which
otherwise favour the flower bud formation/initiation, might have
been utilized in the vegetative growth of the tree, thereby delaying
the flowering. Similar results were obtained by Jannoyer (2009) in
mango.
In an evergreen tree like mango, proper canopy management is
essential to encourage sufficient number of panicles per sq.m of
canopy area and number of panicles per tree, so that the higher
productivity could be achieved. In the present study, the number
of panicles per sq.m canopy area and number of panicles per tree
were higher in control (Table.1). Gopikrishna (1979) reported that
the reduction in number of flowers in severely pruned branches
might be due to loss of potential bearing wood of the tree. The
severely pruned trees showed lesser number of panicles per sq.m
canopy area as well as per tree due to heavy vegetative growth.
This was expected because of lesser number of shoots observed
with higher pruning level when compared to control (T1) and
light pruning treatments. Similar results were recorded by Singh
et al. (2009) in mango.
Percentage of hermaphrodite flowers per panicle had direct
relationship with fruit set and fruit yield. The pruning intensities
significantly improved the percentage of hermaphrodite flowers
per panicle and the lowest percentage of hermaphrodite flowers
per panicle was found in light pruned trees including control
(Table.1). Waghmare and Joshi (2008) attributed that low
percentage of hermaphrodite flowers is due to the development
of lower temperature regime in denser canopies. The highest
percentage of hermaphrodite flowers per panicle was found in T5
(50 per cent removal of past season’s growth and tipping) followed
by heavy pruning (T4). Highest percentage of hermaphrodite
flower per panicle in the pruned trees might be due to removal
of excess shoots, which leads to more light interception and
movement of assimilates to fewer growing points. Besides in
mango, the flowers arise mostly at terminals i.e., very near to sink.
There was every possibility of increase in drawal of more nutrients
from the source towards the sink.
Mango generally produces more number of flowers in the panicles
but the per cent fruit set is relatively low. Hence, knowledge on
the fruit setting ability is very essential for crop management
practices. The maximum per cent fruit set was noticed in T5 (50
per cent removal of past season’s growth and tipping) followed
by T1 (control). However, the per cent fruit set was the least in
severely pruned treatment T6 (total removal of past season’s
growth) (Table.1). Poor fruit set in severe pruned trees might be
due to removal of the potential food synthesizing young shoots.
Improvement in fruit size due to pruning was observed in
mango (Fivaz and Stassen, 1997). In the present study, severe
pruning resulted in decrease in mean fruit weight, length, fruit
circumference and fruit volume. The highest mean fruit weight,
length and fruit volume were observed in the treatments with
light pruning (T2) followed by moderate pruning (T4). It was least
in severely pruned treatment T6 (total removal of past season’s
growth) (Table 2). The reduction in weight, length and volume of
fruit were due to the removal of biomass through severe pruning.
Similar results were obtained by Pratap et al. (2009) in mango.
The highest fruit pulp weight, peel weight and stone weight were
52 Canopy management in mango under ultra high density planting

Table 1. Effect of pruning on flowering characters in mango cv. Alphonso

Treatments Days taken Days taken for Number of panicles Number of Percentage of Fruit set
for first 50 per cent produced per sq.m panicles hermaphrodite (%)
flowering flowering canopy area per tree flower per panicle
T1 168.66 190.89 26.26 160.80 6.91 0.261
T2 172.27 192.43 24.49 124.55 5.78 0.239
T3 188.75 204.33 19.81 85.22 12.34 0.204
T4 192.64 211.84 18.01 73.08 14.02 0.209
T5 171.59 192.76 25.98 140.80 16.53 0.276
T6 197.83 208.58 15.79 54.50 12.28 0.167
LSD (P=0.05) 4.72 7.52 0.55 2.33 0.37 0.005
Table 2. Effect of pruning on fruit characters in mango cv. Alphonso
Treatments Mean fruit Fruit Fruit Fruit Fruit pulp Fruit Stone Pulp to
weight length circumference volume weight peel weight weight stone
(g) (cm) (cm) (cc) (g) (g) (g) ratio
T1 217.10 8.78 21.93 207.98 139.22 34.96 35.93 0.258
T2 252.66 9.18 22.45 243.91 160.02 42.00 46.25 0.289
T3 241.19 8.99 22.74 232.20 157.68 38.40 41.43 0.262
T4 226.68 8.85 21.70 218.81 140.25 40.16 45.50 0.324
T5 231.24 8.86 21.74 222.67 146.16 36.14 33.83 0.231
T6 201.87 8.61 20.81 195.70 126.96 37.25 36.59 0.288
LSD (P=0.05) 5.80 0.25 0.46 6.66 3.33 1.32 0.95 0.008

observed in light pruned trees. However, the pulp weight was Table 3. Effect of pruning on number of fruits per tree and yield per tree
the least in T6 (total removal of past season’s growth) and the (kg) in mango cv. Alphonso
stone weight was least in T5 (50 per cent removal of past season’s Treatments Number of fruits per tree Yield per tree (kg)
T1 81.62 19.96
growth and tipping) (Table 2). Generally, for better sink, better T2 56.55 15.09
source is essential which is very much ensured in light pruning T3 42.37 10.30
than the severe pruning. T4 44.61 10.35
T5 54.33 12.36
Fruit yield in mango is mainly influenced by fruit set per cent. T6 38.50 7.50
In the present study, the number of fruits per tree and yield per LSD (P=0.05) 1.57 0.28
tree during the period of experiment was generally higher in the
control than the trees subjected to pruning (Table 3). This clearly fruits attain better colour and quality. In present study, the
points that the pruning has the supressive effect on the yield. The highest total soluble solids, total sugars and non reducing sugars
reason for more fruit yield in control is due to the retention of of the fruit were observed in T6 (total removal of past season’s
more number of past season shoots as against removal of many growth) where the light penetration was at its maximum (Table
such shoots in the pruning treatments. Singh et al. (2010b) in 4). Besides, lesser number of fruits in the severe pruning, (T6)
mango and Sheikh and Hulmani (1993) in guava also had similar led to less competition among the fruits, finally resulted in better
results. Moreover, as this study was conducted on young mango fruit quality. The results confirmed the earlier reports in mango by
trees of five years, the effect of new shoots produced consequent Venkatesan (2006), Pratap et al. (2009) and Singh et al. (2010a).
to pruning treatments on its flowering potential, fruit setting could Similarly, lowest acidity was observed in T5 (50 per cent removal
not be assessed. Hence, the real effect of pruning on the yield of past season’s growth and tipping), while, the highest acidity
of mango needs to be assessed by continuing the experiment for was recorded in control (Table 4). Similar result were observed
another 2 to 3 years. in mango by Singh (2010a).
Any management practice system, besides increasing the Canopy management in mango cv. Alphonso under UHDP
productivity, should also aim at the production of better quality maximized the yield and maintained the optimum canopy size
fruits. This is more true in the case of canopy management without overlapping. The results indicated that control (tipping
practices, wherein the main objective is to permit better aeration off) encouraged emergence of more flower producing shoots
and light for the inner parts of the trees, so that the developing resulting in better yield (19.96 kg/tree). However, canopy with
Table 4. Effect of pruning on fruit quality characters in mango cv. Alphonso
Treatments TSS Titrable acidity Ascorbic acid Total carotenoid Total sugars Reducing sugars
(oBrix) (%) (mg 100g-1) (mg 100 g-1) (%) (%)
T1 16.99 0.371 41.05 12.07 12.28 4.71
T2 16.68 0.307 39.47 13.79 12.14 4.54
T3 17.57 0.268 37.89 16.11 12.97 4.60
T4 17.30 0.333 36.31 12.90 12.55 4.75
T5 16.38 0.256 38.68 15.50 12.04 4.34
T6 18.40 0.320 40.26 15.42 13.72 4.68
LSD (P=0.05) 0.40 0.009 0.83 0.34 0.39 0.16
 Canopy management in mango under ultra high density planting
overlapping will be of a great concern in control (T1) to keep
the tree well within the manageable limit. To achieve a targeted
yield of 23-25 tonnes/ha/year, treatment T1 (control) and T6 (total
removal of past season’s growth) may be followed in alternate
rows so that the yield as well as canopy spread are taken into
consideration. Pandey and Singh (2008) also reported alternate
pruning method for sustainable production in mango cv. Amrapali.
However, one or more confirmation trials are to be taken up to
arrive firm conclusion.
Acknowledgement
We thank M/s. Jain Irrigation Systems Limited, Udumalpet unit
for provding facilities to conduct the experiment using Ultra High
Density Planting in mango.
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S. Subhadrabandhu, 1994. Effect of paclobutrazol on canopy size
control and flowering of mango cv. Nam Dok Mai Twai No 4, after
hard pruning. Acta Hort., 291: 60-66.
Chen, H., H. Zheng, H.B. Chen and H.L. Zheng, 1996. A preliminary
study of pruning to delay flowering in mango trees. J. S. China
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Davenport, T.L. 2006. Pruning strategies to maximize tropical mango
production from the time of planting to restoration of old orchards.
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and fruit manipulation have on the yield and quality of ‘Sensation’
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Gopikrishna, N.S.I. 1979. Studies on effect of pruning on vegetative
growth, flowering and fruiting in ‘Sardar’ guava (Psidium guajava
L.). M.Sc (Agri.) Thesis. University of Agricultural Science,
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Hedge, J.E. and B.T. Horreiter, 1962. Carbohydrate Chemistry. 17 (Eds
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on mango trees cv. Tommy Atkins at high densities. Proc. Fla. State
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tree size, flowering and yield of mature ‘Tommy Atkins’ mango trees.
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mango (Mangifera indica L.) cv. Amrapali in high density plantation.
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Subtropical Fruits, CISH, Lucknow, India. p. 42-48.
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and yield in mango cv. Amrapali under high density planting. Indian
J. Hort., 60(4): 339-342.
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Received: November, 2013; Revised: February, 2014; Accepted: March, 2014
 Journal

Journal of Applied Horticulture, 16(1): 54-58, 2014 Appl

Comparative evaluation of common bean (Phaseolus vulgaris L.)

germplasm for seed physical and culinary traits

P.A. Sofi1*, S.A. Wani2, M.Y. Zargar1, F.A. Sheikh1 and T. Shafi1
1
Regional Research Station (SKUAST-K), Wadura, Sopore-193201, J&K. 2Directorate of Research, SKUAST-K,
Shalimar-191121, J&K, India, *E-mail: parvazesofi@gmail.com

Abstract
The amount of water absorbed during soaking by dry beans before cooking may be a reliable indicator of the amount of time required
to render them soft and palatable to eat. The present study was undertaken in kharif 2012 at Regional Research Station Wadura. Fifty
diverse germplasm accessions (local and exotic) representing different growth habits and market classes were compared with Shalimar
Rajmash-1, a high yielding bush variety released by SKUAST-K, for 12 seed morphological and physical characters namely seed colour,
seed brilliance, seed shape, seed coat pattern, dry seed weight, soaked seed weight, seed length, seed breadth, seed coat proportion,
water absorption ratio, hydration capacity and swelling capacity. There was a broad range of variation in the traits studied as revealed
by the range and coefficient of variation (%). The CV was highest for swelling capacity (18.62) followed by water absorption (16.281),
hydration capacity (13.61), soaked seed weight (10.712), dry seed weight (3.056) and coat proportion (1.221). However, CV was very
low for seed length and seed breadth owing to low variation in these traits. The correlation between different traits was also worked
out and revealed that highest correlation was recorded between dry weight and soaked weight (0.874) followed by hydration capacity
and swelling capacity (0.720), seed dry weight and hydration capacity (0.710), dry weight and water absorption (0.308), indicating
that the seeds with greater cotyledon mass absorbed more water and that greater water absorption leads to greater swelling. However,
negative correlations were recorded between coat proportion and water absorption (-0.550) and between dry weight and coat proportion
(-0.325). Seed physicochemical traits including the traits used in present study could be effectively used for comparing large set of
germplasm lines for cooking qualities as the varieties that have high hydration and swelling capacities are usually fast to cook.
Key words: Common bean, hydration capacity, swelling capacity, water absorption

Introduction
Common bean (Phaseolus vulgaris L.) is one of the most
important pulse crop in India. It is regarded as “Grain of hope”
as it is an important component of subsistence agriculture and
feeds about 300 million people in tropics and 100 million people
in Africa alone. Besides it is emerging as an important income
generation especially in Central America where beans are No. 1
income generators among field crops. Globally, with 21 million
tonnes produced from about 26 million hectares, it accounts
for about half of the total pulse production. In India, common
bean is grown over an area of about 6 million hectares with a
production of about 2.5 million tones (FOA, 2010). In Kashmir
valley common bean is a niche crop relished for its taste and is an
integral part of culture and agriculture. Dry mature seeds provide
a relatively greater amount of higher quality protein ranging from
17 to 32% which makes them an excellent complement for diets
rich in cereals (Moraghan and Grafton, 2001). Therefore, beans
have a major role in human diet especially in developing countries
where they are considered a low cost protein source. The quality
of beans especially the cooking time, taste and freedom from
flatulence are key parameters for consumer acceptability as well
as marketability of common bean varieties. The proceedure of
evaluation of common bean genotypes for cooking time by CIAT
in Cali, Colombia, is based on a cooking time index derived from
a bardrop cooker (Jackson and Varriano-Marston, 1981). Although
a useful and reliable technique, it is laborious and time consuming
for large number of samples. It has been suggested that the amount
of water dry beans absorb during soaking before cooking may
be a reliable indicator of the amount of time required to render
them soft and palatable to eat. Hence, the water absorption of a
genotype may be a useful and rapid indirect selection method to
screen germplasm for cooking time. A large number of studies
have been undertaken to asses the variation among the genotypes
for various seed traits including water absorption for screening
material for seed culinary properties (Krista and Hosefield, 1991;
Santalla et al., 1999 and Vakali et al., 2009). The present study
was undertaken to asses the variation in seed morphological and
physical characteristics in selected common bean genotypes in
Kashmir valley in view of the fact that the niche status of this
valuable crop is more due to its cooking quality and taste than
its production potential. A large number of genotypes have been
identified for their yielding ability but their acceptance by farmers
will depend on their quality parameters especially the physical
appearance and culinary properties.
Materials and methods
The present study was undertaken in Kharif 2012 at Regional
Research Station Wadura. About 750 germplasm accessions of
common bean were evaluated in an augmented block design in
view of large number of accessions (data not presented). Fifty
diverse germplasm accessions (23 local landraces, one released
variety and 26 exotic accessions procured from CIAT (Columbia),
NORDIC (Sweden) and IPK (Germany), representing different
 Evaluation of common bean (Phaseolus vulgaris L.) for seed physical and culinary traits
growth habits and market classes were selected for the present
study. The performance of the genotypes was compared with
Shalimar Rajmash-1 (SR-1), a high yielding bush variety
released by SKUAST-K, for 12 seed morphological and physical
characters namely seed colour, seed brilliance, seed shape, seed
coat pattern, dry seed weight, soaked seed weight, seed length,
seed breadth, seed coat proportion, water absorption ratio,
hydration capacity and swelling capacity. Seed water absorption
parameters were calculated as per the procedure of Bishnoi and
Khetarpaul (1993) as follows.
Dry and soaked seed weight: The dry seed weight as well as
soaked seed weight were computed on 100 seed basis.
Coat proportion: Seed coat proportion was determined on 20 seeds
per plot, as the weight ratio between coat and cotyledon expressed
in percentage, after removing the seed coat from the cotyledons,
both after soaking and keeping them for 24 h at 105 oC.
Water absorption ratio: Measured as the amount of water
which the dried seeds absorbed during soaking for 24 hours in
double distilled water. The moisture contents of the dry bean
samples were equilibrated before analysis of water absorption by
storing them for 2 weeks in sealed plastic containers at ambient
temperatures and relative humidity. The percent water absorption
was determined by first soaking 30 seeds for 24 h in distilled water
at room temperature and dividing the difference in weight before
and after soaking by the dry weight of the 30-seed sample.
Swelling coefficient: All the seed samples were used two months
after harvesting. Bean samples (50 g per variety) were soaked
in double distilled water for 24 h at room temperature. After
soaking, the increase in water volume was recorded and the
swelling coefficient was determined as the percentage ratio of
increase in the volume of water in bean seeds both before and
after hydration.
Hydration capacity: The hydration capacity was expressed as
hydration absorption per seed and was determined by dividing
the mass gained from seeds by the number of seeds present in
sample (g of water per g of seeds).
Swelling capacity: The swelling capacity per seed was calculated
as the volume gained from the seeds (mL of water per g of seeds)
divided by the number of seeds.
The results were analysed through OPSTAT software developed
Fig. 1. Frequency distribution of water absorption, hydration capacity and swelling capacity among the bean accessions
55
by CCS HAU, Hisar for assessment of variation and trait
correlations in the material studied.
Results and discussion
The accessions selected for the present study were purposefully
included to represent the diverse market classes and the most of
the accessions represented common landraces being cultivated
across diverse niches of the crop. The accessions represented
diversity of different morphological seed characteristics such
as seed colour, seed brilliance, seed shape and seed coat pattern
(Table 1). Most of the accessions were brown and red with
brilliant seeds, mostly kidney shaped and plain seed coat.
Perusal of Table 2 reveals the variation in mean performance for six
seed traits related to consumer acceptability and culinary properties.
Highest dry seed weight (100 seed basis) was observed in WB-457
(74.45 g) and lowest was recorded in WB-245 (22.60 g). Highest
soaked weight was recorded for WB-440 (167.80 g) whereas the
lowest was recorded for WB-245 (29.80 g). Coat proportion was
lowest in WB-195 (8.17 %) and highest in WB-46-2 (25.14 %).
Water absorption was highest in WB-195 (183.09 %) and lowest
in WB-75 (17.53 %). Similarly, hydration capacity was highest
in WB-457 (1.02 g water/g seed) and lowest in WB-245 (0.07 g
water/g seed). Swelling capacity was highest in g water/g seed195
(1.56 mL/g seed) and lowest in WB-75 (1.03 mL/g seed).
Frequency distribution graphs (Fig. 1) in respect of water
absorption %, hydration capacity and swelling capacity revealed
that most of the genotypes (41) had water absorption capacity
ranging from 40-120 %. Only seven genotypes had water
absorption capacity above 120%. Similarly in case of hydration
capacity, 31 genotypes had hydration capacity ranging from 0.3-
0.7 and only two genotypes had hydration capacity above 0.7.
In case of swelling, 45 genotypes had swelling capacity in the
range of 0.97-1.37 and only one genotype had swelling capacity
above 1.5. The graphs in case of water absorption % and hydration
capacity were fairly normal while as in case of swelling capacity,
it was skewed (Fig. 1).
There was a broad range of variation in the traits studied as revealed
by the range and coefficient of variation (Table 3). The CV was
highest for swelling capacity (18.62) followed by water absorption
(16.281), hydration capacity (13.61), soaked seed weight (10.712),
dry seed weight (3.056) and coat proportion (1.221). However,
CV was very low for seed length and seed breadth owing to low
56 Evaluation of common bean (Phaseolus vulgaris L.) for seed physical and culinary traits

Table 1. Origin, pedigree and variability in seed morphological traits in common bean
Accession Pedigree Origin Colour Brilliance Shape Coat pattern
WB-6 G-51420 Columbia Red Brilliant Oval Plain
WB-22 G-51416 Columbia Red Brilliant Kidney Plain
WB-30 PHA-12327 Columbia Cream Medium Oval Plain
WB-46-2 PAS-65-1 Tral Black Brilliant Kidney Mottled
WB-46-3 PAS-65-2 Tral Purple Brilliant Kidney Plain
WB-67 G-3601 USA Red Brilliant Kidney Plain
WB-75 PAS-174 Khag Brown Dull Cuboidal Plain
WB-83 PAS-193 Baramulla Purple Brilliant Cuboidal Plain
WB-93-1 BG-22512 Spain Black Brilliant Kidney Plain
WB-112 PAS-110 Pulwama Purple Brilliant Kidney Plain
WB-131 PAS-54 Uri Brown Medium Kidney Plain
WB-195 PAS-11 Baramulla Choclate Brilliant Kidney Plain
WB-216 RB-39 Nigeria Pink Medium Kidney Plain
WB-245 PAS-248 Poonch Brown Brilliant Cuboidal Mottled
WB-250 PAS-226 Baramulla Red Brilliant Oval Mottled
WB-257 PAS-256 Baramulla Red Brilliant Kidney Plain
WB-261 PAS-261 Kishtwar Red Medium Cuboidal Plain
WB-360 PHA-12663 Turkey White Brilliant Kidney Plain
WB-363 PHA-13576 Russia Chocolate Brilliant Cuboidal Plain
WB-368 PHA-12645 Turkey White Medium Cuboidal Plain
WB-380 PHA-12707 Holland Brown Medium Kidney Plain
WB-413 PHA-5942 Ukraine Brown Brilliant Kidney Mottled
WB-439 PHA-12202 Russia Brown Medium Oval Mottled
WB-440 PHA-7140 Spain Brown Medium Kidney Plain
WB-441 PHA-12266 Russia Brown Medium Kidney Mottled
WB-444 PHA-7549 Spain White Brilliant Kidney Plain
WB-455 PHA-7141 Spain White Brilliant Kidney Plain
WB-457 PHA-13575 Russia Brown Medium Kidney Mottled
WB-467 NG-13964 Sweden Brown Brilliant Kidney Plain
WB-485 NG-21237 Sweden Chocolate Dull Kidney Mottled
WB-497 NG-13858 Sweden Yellow Brilliant Kidney Plain
WB-874 PAS-450 Baramulla Red Medium Oval Mottled
WB-893 PAS-469 Budgam Black Brilliant Kidney Plain
WB-923 PAS-499 Baramulla Brown Medium Kidney Mottled
WB-931 G-37 Canada white Brilliant Oval Mottled
WB-943 G-370 Turkey Purple Brilliant Kidney Mottled
WB-951 G-558 Turkey Purple Brilliant Kidney Plain
WB-954 G-678 Syria Purple Brilliant Kidney Plain
WB-956 G-680 Syria White Brilliant Kidney Plain
WB-966 G-1295 Columbia Red Brilliant Kidney Plain
WB-970 G-1426 Ukraine Cream Brilliant Kidney Plain
WB-1006 PAS-521 Baramulla Chocolate Brilliant Kidney Mottled
WB-1035 EB-7 Nepal Brown Brilliant Kidney Plain
WB-1146 PAS-657 Baramulla Brown Medium Kidney Plain
WB-1181 PAS-707 Kupwara Yellow Brilliant Cuboidal Plain
WB-1182 PAS-708 Kupwara Brown Dull Kidney Plain
WB-1184 PAS-710 Kupwara Yellow Brilliant Kidney Plain
WB-1185 PAS-711 Kupwara Brown Medium Kidney Plain
WB-1186 PAS-712 Kupwara Cream Brilliant Kidney Plain
SR-1 CR x Local red Released variety Red Brilliant Kidney Plain

variation in these traits. Compared to the relased variety SR-1,
most of the accessions had desirable attributes of the eight seed
traits studied. Variability in seed physical traits and culinary
traits have also been reported by various workers (Krista and
Hosefield, 1991; Santalla et al., 1999 and Vakali et al., 2009)
in common bean using coat proportion and water absorption as
indicative traits.
The correlation between different traits (Table 4) revealed that
highest correlation was recorded between dry weight and soaked
weight (0.874) followed by hydration capacity and swelling
capacity (0.720), seed dry weight and hydration capacity (0.710),
wet weight and swelling capacity (0.588), dry weight and water
absorption (0.308) indicating that the seeds with greater cotyledon
mass absorbed more water and that greater water absorption
 Evaluation of common bean (Phaseolus vulgaris L.) for seed physical and culinary traits 57

Table 2. Mean performance for eight seed characteristics in common bean

Entry Dry Wet Seed Seed Coat Water Hydration Swell
weight weight length breadth percentage absorption capacity capacity
(g) (g) (cm) (cm) (%) (%) (g water/g seed) (mL/g)
WB-6 45.77 80.88 1.40 0.95 22.25 54.86 0.35 1.05
WB-22 52.65 75.20 1.50 0.70 15.95 42.83 0.22 1.09
WB-30 45.35 84.40 1.50 1.00 12.79 86.11 0.39 1.22
WB-46-2 40.85 67.60 1.55 0.80 25.14 73.16 0.27 1.19
WB-46-3 36.70 58.80 1.35 0.70 13.60 57.49 0.22 1.03
WB-67 46.25 106.60 1.60 0.75 13.51 130.7 0.60 1.43
WB-75 30.80 36.20 1.30 0.80 15.33 17.53 0.05 1.03
WB-83 31.85 40.10 1.10 0.80 18.45 25.90 0.08 1.15
WB-93-1 40.10 56.60 1.30 0.70 10.60 41.14 0.16 1.19
WB-112 32.30 54.80 1.55 0.70 19.71 51.08 0.22 1.17
WB-131 46.20 87.40 1.60 0.85 13.73 89.17 0.43 1.27
WB-195 35.50 90.50 1.60 0.70 8.17 183.09 0.55 1.56
WB-216 37.85 93.20 1.50 0.90 14.16 119.81 0.55 1.37
WB-245 22.60 29.80 1.30 0.70 20.71 31.85 0.07 1.01
WB-250 45.30 85.60 1.20 0.70 16.12 88.96 0.40 1.14
WB-257 56.35 94.20 1.65 0.80 11.67 60.07 0.38 1.10
WB-261 35.15 47.60 1.40 0.80 16.78 35.41 0.12 1.03
WB-360 64.25 103.42 2.00 1.00 22.41 60.93 0.39 1.08
WB-363 37.85 55.20 1.25 0.80 19.92 45.84 0.17 1.04
WB-368 44.57 87.40 1.15 1.00 9.84 96.09 0.43 1.33
WB-380 36.50 48.60 1.60 0.70 22.35 33.15 0.12 1.06
WB-413 61.80 99.40 1.65 0.90 11.67 60.84 0.38 1.11
WB-439 62.40 124.40 1.30 1.15 14.14 99.35 0.62 1.36
WB-440 65.65 167.80 1.75 1.00 10.13 155.59 1.02 1.44
WB-441 66.75 139.40 1.90 0.90 10.76 108.83 0.75 1.23
WB-444 57.60 115.80 1.90 1.00 15.19 101.04 0.58 1.26
WB-455 56.70 96.50 1.80 0.75 16.16 70.19 0.40 1.08
WB-457 75.40 162.70 1.90 1.00 12.70 115.78 0.87 1.47
WB-467 28.20 47.40 1.20 0.65 14.34 68.08 0.21 1.22
WB-485 43.45 61.40 1.50 0.80 15.63 41.31 0.18 1.24
WB-497 53.20 130.20 1.55 0.80 11.52 144.73 0.77 1.55
WB-874 36.45 92.06 1.30 0.75 12.29 152.56 0.56 1.46
WB-893 35.95 56.80 1.25 0.80 13.38 57.99 0.21 1.17
WB-923 53.50 103.20 1.50 0.70 14.73 92.89 0.50 1.28
WB-931 51.35 94.70 1.30 1.00 19.01 84.42 0.43 1.09
WB-943 54.95 99.20 1.75 0.90 14.31 80.52 0.48 1.41
WB-951 59.10 107.60 1.75 0.85 9.10 80.37 0.48 1.36
WB-954 42.55 75.40 1.60 0.85 12.99 77.20 0.33 1.18
WB-956 42.45 88.20 1.60 0.75 14.74 107.77 0.46 1.25
WB-966 53.15 99.40 1.55 0.90 10.86 87.01 0.46 1.05
WB-970 41.25 60.15 1.50 0.70 14.29 45.81 0.19 1.02
WB-1006 35.35 75.40 1.40 0.80 14.06 113.29 0.40 1.38
WB-1035 52.85 106.80 1.70 0.90 13.11 102.08 0.54 1.33
WB-1146 38.80 69.20 1.40 0.70 10.69 78.35 0.31 1.26
WB-1181 47.10 86.80 1.20 0.85 9.21 83.43 0.39 1.10
WB-1182 32.80 42.85 1.50 0.65 19.51 30.64 0.10 1.08
WB-1184 34.55 68.00 1.15 0.65 13.23 96.81 0.33 1.12
WB-1185 43.90 80.04 1.40 0.80 13.99 82.32 0.36 1.22
WB-1186 27.30 53.20 1.55 0.60 10.15 94.87 0.26 1.34
SR-1 49.75 67.90 1.65 0.70 18.85 36.43 0.18 1.08
Table 3. Mean, range and CV (%) for eight seed culinary traits Table 4. Correlation between eight seed traits in common bean
Trait Mean Range CV (%) Correlation between traits Correlation coefficient
Dry seed weight (g) 43.939 22.60 - 66.75 3.06 Dry weight and wet weight 0.874**
Soaked seed weight (g) 77.793 29.8 - 167.80 10.71 Dry weight and coat proportion -0.325**
Dry weight and water absorption 0.308**
Seed length (cm) 1.483 1.10 - 2.00 0.03
Coat proportion and water absorption -0.550**
Seed breadth (cm) 0.804 0.60 - 1.15 0.02
Coat proportion and hydration capacity -0.561**
Coat proportion (%) 13.764 2.70 - 25.14 1.22 Hydration capacity and swelling capacity 0.720**
Water absorption (%) 70.984 25.90 - 154.92 16.28 Seed dry weight and hydration capacity 0.710**
Hydration capacity (g water/g seed) 0.316 0.07 - 1.02 13.81 Seed dry weight and swelling capacity 0.245
Swelling capacity (mL/seed) 1.204 1.01 - 1.56 18.63 Wet weight and swelling capacity 0.588**
58 Evaluation of common bean (Phaseolus vulgaris L.) for seed physical and culinary traits
leads to greater swelling. However, negative correlations were
recorded between coat proportion and water absorption (-0.550)
and between dry weight and coat proportion (-0.325). The negative
correlation between the traits as reported above is due to the fact
that seeds with thicker seed coats are invariably impermeable to
water and impede water imbibition by dry seeds during soaking
process. Mavromatis et al. (2012) also made a comparative study
in common bean for seed physicochemical traits and concluded
that these traits could be effectively used for comparing large
set of germplasm lines for cooking qualities as the varieties that
have high hydration and swelling capacities are usually fast
to cook (Jackson and Varriano-Marston, 1981; Castillo et al.,
2008). However, the lines that have lower hydration and swelling
capacities usually have longer storage life.
The present study revealed significant variation among the bean
accessions for seed quality traits. The variation can be utilized
through selection to identify high yielding genotypes with better
seed culinary traits. The correlations identified in the study can be
used to develop effective selection index in view of the diversity
and complexity of seed quality traits.
Acknowledgement
The first author gratefully acknowledges the financial support
of J&K State Council for Science and Technology (Grant # 25-
ST/2010).
References
Bishnoi, S. and N. Khetarpaul, 1993. Variability in physicochemical
properties and nutrient components of different pea cultivars. Food
Chemistry, 47(4): 371-373.
Castillo, R., A. Almiral, J. Valero and F. Casanas, 2008. Protected
designation of origin in beans (Phaseolus vulgaris L.): towards
an objective approach based on sensory and agromorphological
properties. Journal Science Food Agriculture, 88: 1954-1962.
FAO, 2010. FAOSTAT. www.fao.org
Jackson, G. and E. Varriano-Marston, 1981. Hard-to-cook phenomenon
in beans: Effects of accelerated storage on water absorption and
cooking time. Journal Food Science, 46: 799-803.
Krista, C. and G.L. Hosfield, 1991. Genotype × Environmental effects on
food quality of common bean: resource-efficient testing procedures.
Journal American Society Horticulture, 116: 732-736.
Mavromatis, A., I. Arvanitoyannis, V. Chatzitheodorou, A. Kaltsa,
I. Patsiaoura and C.T. Nakas, 2012. A comparative study among
landraces of Phaseolus vulgaris L. and P. coccineus L. based on
molecular, physicochemical and sensory analysis for authenticity
purposes. Scientia Horticultureae, 144: 10-18.
Moraghan, J. and K. Grafton, 2001. Genetic diversity and mineral
composition of common bean seed. Journal Science Food
Agriculture, 81(4): 404-408.
Santalla, M., M. Fueyo, A. Rodino, I. Montero and A. de Ron, 1999.
Breeding for culinary and nutritional quality of common bean
(Phaseolus vulgaris L.) in intercropping systems with maize (Zea
mays L.). Biotechnology Agronomy Society Environment, 3(4):
225-229.
Vakali, C., F. Papathanasiou, I. Papadopoulos and E. Tamoutsidis, 2009.
Prelimenary results on a comparative study evaluating landraces of
common bean (Phaseolus vulgaris L.) under organic agriculture in
a protected area in Greece. Proceedings of 2nd Scientific Conference
within the framework of the 9th European Summer Academy on
Organic Farming, Lednice na Moravě, Czech Republic, June 24
- 26, 2009.
Received: March, 2013; Revised: January, 2014; Accepted: March, 2014

Journal of Applied Horticulture, 16(1): 59-60, 2014
Physical properties and transmission of papaya ringspot virus
Isha Bhoyer, Mina D. Koche*, Santoshi Pudake and N.B. Ninawe
Department of Plant Pathology, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola-444104 (Maharashtra). India.
*E-mail: mdkoche@gmail.com
Abstract
Experiment was conducted in vitro to see the different physical properties and transmission of papaya ring spot virus with different aphid
species. The virus was found to be inactivated between temperature 50 to 55°C and between the dilutions of 10-3 to 10-4. It remained
viable upto 24 hours at temperature 28 to 30°C and 5 days at 6 to 8°C temperature. The virus was transmissible by five aphid species
Aphis gossypii (Glover), Aphis craccivora (Koch), Acyrthosiphon pisum (Buczacki S. and Harris K.), Dactynotus carthami (Hille Ris
Lambers), Aphis nerii (Boyer de Fonscolombe) in non persistent manner.
Key words: Papaya, ringspot virus, physical properties, aphid
Introduction
Papaya ringspot is one of the most economically important
diseases of papaya and widely prevalent in all papaya growing
states in India including Maharashtra and causes severe losses
in papaya cultivation. During the survey of papaya in various
locations of Vidarbha region reported that papaya ringspot virus
(PRSV) caused 80 to 100 per cent damage to papaya cultivar
Honeydew (Lokhande et al., 1992). In Marathwada region, 79 per
cent of disease incidence reported due to papaya ringspot virus
(Yemewar and Mali, 1980). Papaya ringspot virus P-infection is
typically characterized by the production of ringspot symptoms
on fruits of infected papaya plant (Jensen, 1949). In addition to
ringspot symptoms, PRSV produces a range of other symptoms
such as leaf mosaic and chlorosis, water soaked oily streaks on
the petiole and upper part of trunk, distortion of young leaves that
sometimes results in shoestring like symptoms, stunting of infected
plants and flower abortion. Consequently, fruit production can be
severely decreased and fruit sugar level reduced by 50 % or more.
The fruit size and quality is much affected resulting decrease in
market value.
The losses caused due to papaya ringspot virus infection, can be
minimized by aquiring knowledge about the vector responsible
for viral disease transmission, host range of papaya ringspot virus,
physical properties of virus, susceptible/ resistant papaya cultivars
to PRSV. Keeping in view the significance of transmission
mode of PRSV, its host range, physical properties, response of
different available varieties of papaya to PRSV infection, aphid
transmissibility of PRSV, the present investigation on PRSV strain
prevalent in this area was carried out.
Materials and methods
Source of inoculum: The young leaves of papaya plants showing
mosaic, leaf distortion, shoestring symptoms were collected from
Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola.
Thermal inactivation point (TIP): Standard leaf extract was
prepared in 1:1 ratio of infected leaf tissues of diseased plant to
0.1 M phosphate buffer, pH 7.5 containing sodium sulphite 0.5%.
Aliquots of 2 mL standard leaf extract were pipetted into a set of
Journal
Appl
12 thin walled test tubes of about 1 cm diameter. Each test tube
was then individually exposed to temperature starting from 40°C
to 95 °C for 10 minutes in a metallic constant temperature water
bath. All the treated extracts were then inoculated on the leaves of
healthy young papaya plants at six leaves stage by conventional
leaf rub method. Similarly, a set of 10 plants of assay host
were inoculated with untreated extract which served as control.
Numbers of local lesions per leaf were worked out.
Dilution end point (DEP): The standard leaf extract prepared from
infected leaves of papaya was diluted in 10 fold series of dilutions
viz., 1:10 (10-1), 1:100 (10-2), 1:1000 (10-3), 1:10000 (10-4), 1:100000
(10-5), 1:1000000 (10-6) by adding required quantity of sterilized
distilled water. A set of 5 plants were inoculated separately for each
dilution treatments. Standard leaf extract without dilution served
as control. Observations were recorded periodically for expression
of symptoms for each dilution treatment.
Longevity in vitro (LIV): In order to know the in vitro longevity
of the virus, standard leaf extract was kept at room temperature
28 to 30°C and second lot in refrigerator at 6° to 8°C temperature.
The plants inoculated with extract immediately after extraction
period served as control. Whereas, the sap from test tubes stored
at room temperature and refrigerated condition was inoculated at
a fixed period intervals i.e. 0, 4, 8, 12, 24 h and average number
of local lesions were per leaf were recorded on 2, 3, 4, 5, 6 and
7 days.
Aphid transmission: Five aphid species viz., Aphis gossypii
(Glover), A. craccivora (Koch), Acyrthosiphon pisum (Buczacki
S. and Harris K.), Dactynotus carthami (Hille Ris Lambers) and
A. nerii (Boyer de Fonscolombe) were collected directly from
cotton, cowpea, safflower and calatropis respectively. In all the
transmission tests, only adult apterous aphids were used. Adult
apterous aphids from each species were given a pre-acquisition
fasting period of 1 hour under darkness by keeping them in
sterilized petridishes. Pre-acquisition fasting period was followed
by an acquisition feeding period of 5-10 minutes on young PRSV
infected leaves of papaya plant (cv. Honeydew). Then the aphids
were gently picked with moistened camel hair brush No. 0 and
collected in another sterilized petridish and then transferred to
60 Physical properties and transmission of papaya ringspot virus
(oC)
Fig. 1. Thermal inactivation point of papaya ringspot Fig. 2. Dilution end point of papaya ringspot Fig. 3. Longevity in vitro of papaya ringspot
virus (5 plants inoculated in each treatment). virus (5 plants inoculated in each treatment). virus (5 plants inoculated for each treatment).
the young healthy six leaves stage test plants of papaya with the
help of another moistened camel hair brush.
For aphid transmission study 10 healthy seedlings of papaya were
used. 10 viruliferous aphids were released on each test plant and
were allowed to feed overnight and then killed by spraying 0.02
per cent dimethoate insecticide on test plants. All the test plants
were maintained in an insect proof cage house and observations
were recorded on their transmission ability and types of symptoms
developed on the test plants.
Results and discussion
Physical Properties: The result of physical properties revealed
that the virus isolate could not withstand to the temperature of 55
°C and the plants inoculated with this treatment failed to produced
any symptoms of the disease (Fig. 1). Above this temperature i.e.
from 60 to 95 °C, no symptoms were observed on test plants.
This indicated that thermal inactivation point of the virus under
investigation lies between 50 to 55°C temperature. The present
virus could retain the infectivity up to 50 °C temperature only.
Wu et al. (1983) also got similar observations on PRSV and he
reported the TIP of PRSV in between 50 to 55°C. However, Rosa
and Lastra (1983) reported the TIP in case of PRSV 57°C and 52
to 54°C, respectively. This may be due to strainal variations.
The crude sap diluted to 10-1 to 10-3 (1:10 to 1:1000) dilution
produced mosaic, leaf distortion and shoestring symptoms, the
typical symptoms produced by papaya ringspot virus (PRSV), on
all the test plants inoculated (Fig. 2). Whereas sap diluted to 10-4
(1:10000) dilution or more failed to produce any symptoms of the
disease on inoculated test plants. The results thus indicated that the
dilution end point of papaya ringspot virus under study was found
between 10-3 to 10-4 dilution. Dethe (2000) reported that dilution
end point (DEP) of all the tested strains of PRSV viz., PRSV-S,
PRSV-M1, PRSV-P, PRSV-Y was found between 10-3 to 10-4.
Papaya ringspot virus (PRSV) under investigation retained
infectivity for 24 hours at room temperature (28 to 30°C) and
upto 5 days at 6 to 8°C temperature. Virus lost its infectivity
within 2 days at room temperature and after 5 days at 6 to 8°C
temperature (Fig. 3). Dethe (2000) reported that the longevity in
vitro of all tested strains viz., PRSV-S, PRSV-M1, PRSV-P, PRSV-
Y was found upto 28 hours. The results of aphid transmission of
the virus causing Papaya ringspot virus (PRSV) are depicted in
Table 1. It was found that five aphid species viz., Aphis gossypii
(Glover), A. craccivora (Koch), Acyrthosiphon pisum (Buczacki
Table 4. Aphid transmission study of papaya ringspot virus. Test plants
showed mosaic, leaf distortion, shoestring symptosm with all aphid
speceis
Aphid species Transmission (%) Incubation period (days)
Aphis gossypii 90 14
Aphis craccivora 80 13
Acyrthosiphon pisum 90 13
Dactynotus carthami 100 14
Aphis nerii 70 14
and Harris), Dactynotus carthami (Hille Ris Lambers), A. nerii
(Boyer de Fonscolombe) were able to transmit the virus in non-
persistant manner from papaya to papaya. Dactynotus carthami
was found to be most efficient and showed (100%) transmission
of PRSV followed by A. gossypii (90%), Acyrthosiphon pisum
(90%), A. craccivora (80%) and A. nerii (70%). All test plants
used in aphid transmission study showed a typical mosaic, leaf
distortion, shoestring symptoms during the incubation period of 13
to 14 days. Mali (1987) reported that PRSV was transmitted by A.
gossypii, A. craccivora, Acyrthosiphon pisum, A. nerii, D. sonchi,
MelanoA. sacchari and Rhophalosiphum maidis whereas, Myzus
persicae was most efficient vector. Dethe (2000) reported that the
four strains viz., PRSV-S, PRSV-M1, PRSV-P and PRSV-Y were
found to be transmissible by Acyrthosiphon pisum, Dactynotus
carthami, Myzus persicae, A. gossypi and Aulacarthum solani in
non persistent manner.
References
Dethe, D.W. 2000. Further Studies on Papaya Ringspot Virus (PRSV)
Isolates Occurring on Papaya. M.Sc. Thesis (Unpub.) MAU,
Parbhani, India.
Jensen, D.D. 1949. Papaya virus diseases with special reference to papaya
ringspot. Phytopathol., 39: 191-211.
Lokhande, N.M., P.G. Moghe, A.D. Matte and B.J. Hiwase, 1992.
Occurrence of papaya ringspot virus (PRSV) in Vidarbha region of
Maharashtra. J. Soils and Crops, 2(2): 36-39.
Mali, V.R. 1987. Research Review Report on Plant Pathology (Virology).
Maharashtra Agric. Univ. Joint Agresco Rept. 1987: 14-16.
Rosa, M.De and L.R. Lastra, 1983. Purification and partial characterization
of papaya ringspot virus. Phytopath. zeitschrift, 106(4): 329-336.
Wu, F.C., X.X. Peng and S.H. Xu, 1983. Preliminary studies on
identification, purification and properties of papaw ringspot virus in
South China. Acta Phytopathologia Sinica, 13(3): 21-28.
Yemewar, S.I. and V.R. Mali, 1980. On the identity of sap transmissible
virus of papaya in Marathwada. Indian J. Mycol. Plant Pathol.,
10(2): 155-160.
Received: October, 2013; Revised: March, 2014; Accepted: April, 2014
 Journal

Journal of Applied Horticulture, 16(1): 61-65, 2014 Appl

Effect of integrated application of phosphorus and phosphate

solubilizing microorganisms on root colonization, productivity
and seed quality of Cucurbita pepo L.

J. Hamzei* and S. Najjari

Department of Agronomy and Plant Breeding, Faculty of Agriculture, Bu-Ali Sina University, Postal Code: 6517833131,
Hamedan, Iran. *E-mail: j.hamzei@basu.ac.ir

Abstract
Phosphorus is a major nutrient and its deficiency limits plant growth of pumpkin (Cucurbita pepo L.). The investigation was aimed
at studying integrated application of phosphorus on growth and production of pumpkin. Co-inoculation of phosphate solubilizing
microorganisms (PSM) (mycorrhiza and bacteria) with and without seed inoculations, and P chemical fertilizer at 0, 25, 50, 75 and
100% of recommended fertilizer were applied in a factorial experiment in randomized complete block design with three replications.
Data indicate that PSM and P fertilizer show significant effects on all traits. Maximum oil yield (41.80 g m-2) and linoleic acid (68.30%)
were obtained with PSM and 50% of the recommended P fertilizer. Seed yield was significantly increased in response to inoculation
of PSM in the presence of low levels of P fertilizer. However, maximum mycorrhizal colonization obtained in 25% recommended P
fertilizer. A high level of P fertilizer had a negative effect on the activity of PSM. On the other hand, a low level of phosphorus with
PSM has a simulative impact on root colonization and productivity of pumpkin and favoured the activities of PSM.
Key words: Cucurbita pepo L., fertilizer, linoleic acid, mycorrhiza fungi, oil percentage, symbiosis.

Introduction
Chemical fertilizer application and inappropriate energy
production methods have harmful effects on biological cycle of
nutrients and have destroyed stability of farming systems and thus
encourage the application of biological fertilizers for restoration
(Kannayan, 2002). Nowadays attention to biofertilizers has
been increased due to developments of countries, high costs of
chemical fertilizers and attention towards sustainable agricultural
systems (Ehteshami et al., 2007). Medicinal pumpkin (Cucurbita
pepo L.) is widely used in traditional and industrial pharmacology
(Ghaderi et al., 2008). Pumpkin seed oil contains several essential
fatty acids that help to maintain healthy blood vessels, and nerves
(Horveth and Bedo, 1998; Applequist et al., 2006). The oil content
of pumpkin seed varies from 30-50% (Stevenson et al., 2007) and
the composition of fatty acids is dependent on several factors.
Pumpkin seed oil, especially when produced organically, is used
in pharmacology and alternative medicine (Wagner, 2000).
Phosphorus (P) is one of the major nutrients which limits plant
growth when deficient. Most of the soils throughout the world
are P deficient (Batjes, 1997) and therefore require P to replenish
the P demand by crop plants. P is an essential element for cell
division, root development and seed formation (El-gizawy and
Mehasen, 2009). It causes early ripening in plants, decreases seed
moisture, improves crop quality and is most sensitive nutrient
to soil pH. To circumvent the phosphorus deficiency in soils, P
fertilizers are applied (Zaidi and Khan, 2006).
Vesicular arbuscular mycorrhiza (VAM) fungi encourage the plant
roots to rapidly absorb solubilized phosphorus. VAM fungi play
an important role in changing fixed or insoluble P into soluble
P, which can be used by plant freely. Thus, inoculation with
VAM fungi has been found to increase the availability of P and
other nutrients in crop plants. They exert these effects through
symbiotic associations with plant roots, colonizing cortical
tissues and extending hyphae into the rhizosphere (Hetrick et
al., 1996). It is documented that inoculation of sorghum plants
with VAM fungi helps to absorb enough micronutrients through
chelate formation with sidrophores (Aliasgharzad et al., 2009).
Inoculum production from mycorrhizal fungi and using it in
proper environmental conditions is a significant environmental
friendly way to help plant growth and development through
the enhancement of P absorbance (Mehrvarz et al., 2008).
The significance of this practice, especially under low fertility
conditions, has been evident.
Phosphate solubilizing bacteria are also considered to be among
the most effective plant assistants to supply P at a favorable level.
Among the soil bacterial communities, Pseudomonas sp. and
Bacillus species could be referred to as the most important strains
(Akmakc et al., 2006; Mehrvarz et al., 2008). The population of
phosphate solubilizing bacteria depends on cultural activities
and different soil properties involving physical and chemical
properties, organic matter, and soil phosphorus content (Kim et
al., 1989).
This study was designed to evaluate the effects of co-inoculation
of VAM fungi with P solubilizing bacteria and effectiveness of
different P levels on growth and production of pumpkin.
Materials and methods
Experimental site and design: The study was conducted at the
Research Farm of Bu-Ali Sina University, Hamedan, Iran, during
growing season of 2010-11. Physical and chemical properties of
the site soil are summarized in Table 1. Factorial experiment was
62 Effect of integrated application of phosphorus and phosphate solubilizing microorganisms on Cucurbita pepo
carried out based on randomized complete block design (RCBD)
with 3 replications. Treatments consisted of co-inoculation
of phosphate solubilizing microorganisms (mycorrhiza fungi
and phosphate solubilizing bacteria) at two level with (MP1)
and without seed inoculums (MP0), and phosphorus chemical
fertilizer at five levels, 25% (C1), 50% (C2), 75% (C3), 100%
of recommended fertilizer (C4) and control (C0: no chemical
fertilizer). All treatments were applied before planting. The
phosphate solubilizing bacteria used for this experiment
were supplied by the Green Bio-tech Co., which consisted of
Pseudomonas putida (strain p13) and Bacillus lentus (strain p15).
The bacterial strains are endemic to the farm soils of Iran. VAM
fungus Glomus intraradices (University of Technology, Munich,
Germany) was used in the experiments. All seeds were sown soon
after the microbial inoculations. P chemical fertilizer as triple
super phosphate was utilized as strip under seeds according to
experimental treatments. Each plot consisted of 5 rows with 140
cm space between rows and 30 cm distances between plants in
the rows. During the different stages of plant growth, practices
such as weeding, irrigation and pest control were performed for
all plots.
Measurements: During the flowering stage, the percentage of
mycorrhizal colonization of roots was measured using a gridline
intersection technique, according to Phillips and Hayman (1970).
The oil content of seeds was measured according to Soxhlet
method (AOAC, 1980). Measurements of oil yield were calculated
by multiplying two factors, i.e. grain yield and oil percentage
(Siam et al., 2008). Also, at the flowering stage, agronomic
traits viz., number of leaves per plant, number of branches,
number of nodes per plant, and leaf dry weight were measured.
Leaf chlorophyll was also measured during phenological stages.
At the harvesting time, 2 m2 from each experimental unit was
harvested and based on the fruit number per plant, fruit weight
average, seed number per fruit, 100 seed weight, fruit yield and
seed yield were determined.
Statistical analysis: The statistical analyses of data were carried
out by ANOVA. Comparison of the means was made using LSD
(Least Significant Difference) method at the probability level
of 5%.
Results
Analyses of variance showed that number of leaves per plant,
number of nodes per plant, number of branches, leaf dry weight
and leaf chlorophyll were affected by P chemical fertilizer
treatment (Table 2). Also, the interaction effect of P solubilizing
Table 1. Soil physical and chemical characteristics of the experimental site
Sample depth (cm) Soil texture pH OC (%) EC (mMohs/cm) P (ppm) K (ppm) Zn (ppm) Fe (ppm) Cu (ppm) Mn (ppm)
0-30 Silty clay loam 7.7 0.72 1.45
Table 2. Analysis of variance for the effect of phosphorus treatments on growth traits of pumpkin
Variables df Number of leaves plant-1 Number of nodes plant-1 Number of branches
Replication 2 0.09ns 0.24ns
MP 4 5.39 ns
1.45 ns
C 1 34.49** 11.38**
MP×C 4 7.04 *
6.04 *
Error 18 1.58 1.49
CV ( %) 6.85 8.98
ns: non significant;**,* significant at P=0.01 and P=0.05, respectively; MP: mycorrhiza fungi + P solubilizing bacteria; C: P chemical fertilizer.
microorganisms (MP) and P fertilizer (C) for number of leaves
per plant, number of nods per plant, number of branches and leaf
chlorophyll was significant at 5% probability level (Table 2).
Mean comparisons indicated that highest values of number of
leaves per plant (22.33), number of nodes per plant (15) and
number of branches (2.63) were obtained with application
of mycorrhiza fungi + P solubilizing bacteria and 50% of
recommended P fertilizer (MP 1C 2). The lowest value was
observed in control (MP0C0) (Table 3). Results showed that, 75%
P fertilizer produced maximum leaf dry weight, in the flowering
stage. With decreasing P chemical fertilizer from 100 to 25%
recommended fertilizer, phosphate solubilizing microorganisms
significantly increased the leaf chlorophyll content. The maximum
leaf chlorophyll (51.65 SPAD) was obtained in phosphate
solubilizing microorganisms and 25% recommended P fertilizer
plots (MP1C1), while the minimum amount of leaf chlorophyll
(47.00 SPAD) was obtained in control treatments (MP0C0). It
seems that P solubilizing microorganisms, i.e. mycorrhizal fungi
and P solubilizing bacteria are very important components of the
treatments that enhance P solubility and other elements.
There was a significant difference in mycorrhiza + P solubilizing
bacteria (MP) and P chemical fertilizer treatments in yield and
yield components traits (Table 4). Indeed, the interaction effect of
inoculation and phosphorus treatments on yield components and
fruit yield wasn’t significant. The higher value of seed number
per fruit (235.72 seeds), 100 seed weight (15.08 g) and fruit
weight average (1.77 kg) was obtained in 50% of P recommended
fertilizer and the lowest values of seed number per fruit (182.22
seeds) and 100 seed weight (12.66 g) was observed in 100% P
recommended fertilizer treatment (C4). Also, inoculation of P
solubilizing microorganisms increased yield components and fruit
yield. Moreover, inoculated plants (by Mycorrhiza and bacteria)
were more successful than non- inoculated plants that can suggest
a synergic effect between bacteria and the fungus (Table 5). The
results showed that with increasing use of P chemical fertilizers
from 0 to 100%, fruits number per plant increased. According to
our findings (Table 5), fruit yield had a range from 5.69 kg m-2
in 50% P recommended fertilizer (C2) to 3.03 kg m-2 in control
treatment (C0). Also, seed inoculums had about 35% more fruit
yield compared to non-seed inoculums. Moreover, according to
the analyses of variance (Table 4), the effects of treatments on
pumpkin seed yield were significant. The maximum seed yield
of 135.57 g m-2 obtained in MP1C1 (seed inoculums + 25% P
recommended) which was not significantly different from MP1C2
and MP1C3 treatment (data not shown), and the minimum seed
yield of 60.71 g m-2 was obtained in MP0C0 (control treatment).
8.2 220 1.06 4 0.94 5
Leaf dry weight Leaf chlorophyll
0.12ns 0.31ns 0.68ns
0.05 ns
12.59 ns
15.17**
1.43** 42.78** 4.68*
0.39 **
6.26ns
5.70*
0.05 5.83 1.37
12.29 10.24 2.40
 Effect of integrated application of phosphorus and phosphate solubilizing microorganisms on Cucurbita pepo 63

Table 3. Mean comparisons of the interaction effects of co-inoculation P solubilizing microorganisms and P chemical fertilizer on growth traits of
pumpkin
Treatments Number of leaves plant-1 Number of nodes plant-1 Number of branches Leaf chlorophyll
Non P Solubilizing Non fertilizer 15.33 9.67 1.27 47.00
Microorganisms 25% recommended 16.63 13.50 1.58 47.08
application 50% recommended 17.83 14.50 1.83 49.83
75% recommended 21.86 14.55 2.23 48.45
100% recommended 18.17 14.83 2.40 48.25
P Solubilizing Non fertilizer 16.17 13.00 1.30 49.03
Microorganisms 25% recommended 17.07 14.00 1.42 51.65
application 50% recommended 22.33 15.00 2.63 50.40
75% recommended 21.33 14.75 2.58 48.50
100% recommended 17.17 12.50 1.83 48.15
LSD 5% 2.16 2.10 0.40 2.01
Table 4. Analysis of variance for the effect of phosphorus treatments on yield and yield components of pumpkin
Variables df Seed number per 100 seed weight Fruit number per Fruit weight Seed yield Fruit yield
fruit plant average
Replication 2 414.93ns 2.55ns 0.05ns 0.05ns 122.38ns 2.10*
MP 4 11231.32 **
18.67 **
0.31 **
0.31 **
13594.51 **
29.62**
C 1 2639.55* 7.18* 0.33** 1.34** 1024.17** 6.09**
MP×C 4 594.41ns 1.41ns 0.02ns 0.08ns 520.96* 1.53ns
Error 18 902.81 2.11 0.01 0.05 175.32 0.55
CV, % 13.98 10.59 9.79 16.13 14.40 15.95
ns: non significant;**,* significant at P=0.01 and P=0.05, respectively; MP: mycorrhiza fungi + P solubilizing bacteria; C: P chemical fertilizer.
Table 5. Mean comparisons of the main effects of co-inoculation P solubilizing microorganisms and P chemical fertilizer on yield and yield components
of pumpkin
Treatments Seed number per fruit 100 seed weight Fruit number per plant Fruit weight average Fruit yield
PSM application 234.23a 14.52a 1.51a 1.69a 5.67a
No PSM application 195.53b 12.94b 1.31b 1.27b 3.68b
No fertilizer 218.00ab 12.92c 1.14b 1.19c 3.03c
25% of recommended dose 229.46a 14.72ab 1.29b 1.57ab 4.55b
50% of recommended dose 235.72a 15.08a 1.30a 1.77a 5.18ab
75% of recommended dose 208.96ab 13.29bc 1.63a 1.57ab 5.69a
100% of recommended dose 182.22b 12.66c 1.69a 1.31bc 4.91ab

Table 6. Analysis of variance for the effect of phosphorus treatments on mycorrhizal colonization and seed quality of pumpkin
Variables df Oil percentage Oil yield Mycorrhizal colonization Oleic acid Linoleic acid
Replication 2 0.15ns 79.18ns 8.15ns 0.41ns 1.44ns
MP 4 77.95 **
2552.83 **
11080.33 **
11.38 **
9.28*
C 1 39.60** 252.73** 138.58** 1.10** 4.19*
MP*C 4 28.40 **
99.99 ns
79.69 ns
0.71 **
1.47ns
Error 18 5.74 42.45 28.95 0.15 1.49
CV, % 6.14 18.11 21.11 1.75 1.80
ns: non significant;**,* significant at P=0.01 and P=0.05, respectively; MP: mycorrhiza fungi + P solubilizing bacteria; C: P chemical fertilizer.

Table 7. Mean comparisons of the main effects of co-inoculation P solubilizing microorganisms and P chemical fertilizer on mycorrhizal colonization
and seed quality of pumpkin
Treatments Oil yield Mycorrhizal colonization Linoleic acid
PSM application 45.18a 44.70a 68.13a
No PSM application 26.73b 6.26b 67.02b
No fertilizer 25.33b 17.32b 66.31b
25% of recommended dose 38.60a 29.50a 67.16ab
50% of recommended dose 41.80a 28.33a 68.30a
75% of recommended dose 39.40a 27.33a 68.09a
100% of recommended dose 34.58a 25.16a 68.05a
As seen in Table 6, application of P solubilizing microorganisms in 25% recommended P fertilizer. P fertilizer consumption
and P fertilizer levels had significant effect on quality traits significantly increased linoleic acid, oil yield and mycorrhizal
of pumpkin plants. Also, interaction between P solubilizing colonization. The Pumpkin plants showed positive response
microorganisms and P fertilizer on oil percentage and oleic acid
to P solubilizing microorganism’s application for oil yield,
was significant. Table 7 shows that the highest values of oil
yield (41.80 g m-2) and linoleic acid (68.30 %) were obtained mycorrhizal colonization and linoleic acid traits. The highest oil
with application of 50% P recommended fertilizer. However, percentage (44.44 %) and oleic acid (23.68 %) were obtained
the maximum mycorrhizal colonization of 29.50% was obtained from 50% P fertilizer and inoculated plants (Fig. 1).
64 Effect of integrated application of phosphorus and phosphate solubilizing microorganisms on Cucurbita pepo
Fig. 1. The interaction effect of P chemical fertilizer levels and co-
inoculation of P solubilizing microorganisms on oil percentage and
oleic acid of pumpkin.
Discussion
In the present study, it is shown that significant positive effects
were obtained with P solubilizing microorganisms and P fertilizer
on growth and production traits of pumpkin.
Improved performance of plant with application of P solubilizing
microorganisms for growth and yield was probably due to the
absorption of more nutrients by roots. It is known that mycorrhiza
fungi and P solubilizing bacteria can increase P uptake by plants
through their phosphatase activity, a better exploration of the
soil by hyphae and the uptake of fixed soil P, which is otherwise
unavailable to plant roots. Phosphate solubilizing microorganisms
secrete organic acids, and enzymes that act on insoluble phosphates,
and convert it into soluble form, thus, providing it to plants
(Ponmurugan and Gopi, 2006). In addition, the microorganisms
involved in P solubilization can enhance plant growth by enhancing
the availability of other trace element such as iron, zinc, etc. (Ngoc
et al., 2006; Akhtar and Siddiqui, 2009).
Based on our data, treatments of P solubilizing microorganisms
with 50% of P fertilizer increased the number of leaves per plant,
number of nodes per plant and number of branches. Similarly it
has been shown, that biofertilizer with 50% of chemical fertilizers
led to an increase in plant growth, plant height, branch number,
fresh and dry weight of safflower in comparison to application
of chemical fertilizers alone. It is reported that, soil chemical
and biological characteristics were improved by biofertilizers.
In addition, due to the use of low doses of chemical fertilizers,
agricultural production would be free from contaminants (EL-
Habbasha et al., 2007; Salimpour et al., 2010).
Seed yield was significantly increased in response to inoculation
of P solubilizing microorganisms combined with P fertilizer
compared to control. It seems that application of biofertilizer
combined with chemical fertilizer is an important approach in
maintaining and improving the soil fertility, increasing fertilizer
use efficiency and improving crop yield. Interaction effect was
higher with co-inoculation phosphate solubilizing microorganisms
and low levels of chemical fertilizer. Thus, it can be concluded
that the high rates of P chemical fertilizers application, lead to
antagonistic interaction with mycorrhiza and bacteria.
In inoculated plants (by mycorrhiza and bacteria), the leaf
chlorophyll content was significantly increased compared to non-
inoculated plants. In this respect, it is indicated that mycorrhizal
fungi symbiosis increased the rate of photosynthesis (Augé,
2001). Concentration of chlorophyll in mycorrhizal plants was
higher than control plants resulting production of larger grains
and enhanced economical yield. It is reported that grain yield
and dry matter production in barley increased by utilization
of P solubilizing bacteria in the presence P chemical fertilizer.
Advantages of P solubilizing bacteria on enhanced plant growth
have been previously observed on different crops (Kim et al.,
1998; Fernandez et al., 2007).
In this study, co-application of P solubilizing microorganisms
and P fertilizer (up to 50% recommended dose) reduced the seed
oil percentage and oleic acid. Also, the oil percentage and oleic
acid were increased with increase in P fertilizer application from
0 to 100% of recommended dose. Biologically active isopernoid
requires acetyl-COA, ATP and Nicotinamide adenine dinucleotide
phosphate (NADPH) for synthesis. Hence, the biosynthesis of
essential oil is dependent on inorganic P content in the plant
(Kapoor et al., 2004). Our results suggest that application of P
solubilizing microorganisms in combination with 50% P chemical
fertilizers has a great potential to increase oil content of pumpkin.
Application of P solubilizing microorganisms and P fertilizer
produced high oil yield. Oil yield is the main purpose of oil seeds
cultivation of medicinal pumpkin.
Based on our findings, with increase in seed oil content, linoleic
acid increased, indicating a positive correlation between oil
content and linoleic acid (Gholipouri and Nazarnejad, 2007). The
relative amount of oleic acid is always negatively correlated with
the relative amount of linoleic acid (Murkovic et al., 1996). This is
due to the precursor-product relationship of these two fatty acids.
Results showed that amount of pumpkin linoleic acid were higher
than oleic acid. The high content of linoleic acid is an important
nutritional aspect of pumpkin seed oil. Linoleic acid is an essential
fatty acid for humans as it is required for the formation of cellular
membranes, vitamin D and various hormones.
Based on our findings it seems that seed inoculation with P
solubilizing microorganisms, in general, has a simulative impact
on growth, yield and its components, productivity and seed
quality of pumpkin. Moreover, there is a synergic relationship
between bacteria and fungi. Further, data suggest that low
levels of phosphorus fertilizer favour the activities of fungi and
bacteria.
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Received: January, 2014; Revised: March, 2014; Accepted: April, 2014
 Journal

Journal of Applied Horticulture, 16(1): 66-70, 2014 Appl

Processing and quality evaluation of blended guava watermelon

squash

J. Shankara Swamy* and A.K. Banik

Department of Postharvest Technology, Bidhan Chandra Krishi Viswa Vidhyalaya, Mohanpur, West Bengal-741 252,
India. *E-mail: shankara.swamy@gmail.com

Abstract
Guava fruit juices are pleasant when diluted with other tropical fruit juices due to its too acidic or strongly flavoured and less coloured
nature, thus blending offers the opportunity to adjust sugar and acid ratios and eliminates some defects in juice quality or nutritional
attributes by proper combination of juices and further adjustments in ingredients. Guava-watermelon squash at different ratio (50:50,
75:25, 25:75) of pulp blending level containing 40 oBrix TSS and 1% of acidity were prepared with incorporation of different
concentrations of xanthan gum, an exocellular polysaccharide produced by obligately aerobic bacteria Xanthomonas campestris, to
investigate the effect of different ingredients on the product quality and stability during 180 days of storage. There were little changes
in quality parameters, TSS, pH, titratable acidity, ascorbic acid during the storage and 0.5% w/w of xanthan gum gave stability to the
product during storage. Blended guava-watermelon squash (75:25) having 0.3% of xanthan gum, 40 oBrix TSS, 1% acidity showed
highest overall acceptability during the storage period.
Key words: Blended guava-watermelon squash, blended fruit beverages, xanthan gum, non-enzymatic browning and stability.

Introduction obtained by mashing in a grinder with filtered water in proportions

Guava (Psidium guajava L.), called as apple of tropics, is
one of the most common fruits in India. In West Bengal, it is
commercially cultivated near gangetic alluvial zone North &
South 24 Parganas, Nadia, Murshidabad district and lateritic zone
of Paschim Medinipur and Birbhum district covering an area of
nearly 8.27 thousand ha (Anonymous, 2011). Guava is available
in plenty during the rainy season and its disposal becomes
a serious problem. Its utilization is very little in processing
industry. Only jam, jelly is made from its fruits, but jam and
jelly manufactured from guava pulp are not acceptable like other
fruit products because of gritty texture. Hence, big industries
do not manufacture it. Its excellent flavour and nutritive value
have a great potential in beverage industry. Guava fruit juice
is too acidic, strongly flavoured, less coloured thus its dilution
with other tropical fruits such as watermelon to impart colour
and flavour increase consumer acceptability. Blending juices by
choosing the individual components at different levels have been
suggested for acceptable product development (Huor et al., 1980).
Keeping the above facts in view, the present study was undertaken
to evaluate physico-chemical, sensory and microbiological
attributes of blended guava-watermelon squash supplemented
with soluble dietary fibre in the form of xanthan gum at different
concentrations during storage.
Materials and methods
Guava (cv. Allahabad Safeda) was procured from local Barajaguli
market of Kolkata. Fruits with the same level of maturity, ripening
and firmness, free from blemishes and bruises were carefully
selected for the study. The ripe cremish white guava, cut into
slices (2-2.5 cm), were allowed for heat treatment at 74-75 oC for
(2-5 min) to inactivate enzymes which cause browning. The pulp
of 1:3 (guava slices/water, w/v) (water used for the heat treatment
also used for the juice extraction) was passed through a muslin
cloth. At this stage the total soluble solids were measured by
using hand refractometer. Seedless pulp of watermelon (var. Arka
Manik) fruits was used for juice extraction by mashing in grinder.
The blended guava-watermelon squash was prepared by mixing
calculated amounts of blended juice of guava and watermelon,
sugar, citric acid, xanthan gum, preservative (KMS) and water.
Squash from blended guava-watermelon fruits juice was adjusted
with 25% pulp/juice, 40 % TSS and 1% acidity (as citric acid)
with varied level of xanthan gum (0.1, 0.2, 0.3, 0.4, and 0.5%) as
outlined in recipe (Table 1). Sugar syrup was prepared by heating
the mixture of cane sugar, water and at boiling stage citric acid
was added and boiled for 2-3 minutes to get consistent product.
The strength of sugar was determined with the help of hand
refractometer. The final total soluble solids were adjusted by
adding extra syrup. The prepared syrup was filtered through
muslin cloth to remove impurities. Prior to use the pure xanthan
gum (Loha Chemie Pvt. Ltd. Mumbai, India) was dispersed
uniformly in water and kept aside for 60 minutes to accomplish
hydration. The xanthan gum in the form of dispersion was added
to the blended guava and watermelon juice. The hot syrup and
fruit pulps/juice having calculated amount of xanthan gum
were mixed on weight basis. The blended guava-watermelon
squash treatments were heated to 85 oC and preservative KMS
(potassium metabisulphite) 350 ppm was added to final product
to prevent spoilage during the storage. The prepared blended
guava-watermelon squash was poured into pre-sterilized bottles
of 200 mL capacity and sealed airtight. Bottles were sterilized in
boiling water for 20 min, cooled immediately and stored at room
temperature (18-25 oC) for further observations.
Blended guava-watermelon squash were analyzed for pH with
 Processing and quality evaluation of blended guava-watermelon squash 67

Table 1. Recipes for blended guava-watermelon squash scoring method (Amerine et al., 1965). The prepared product
Recipe Blending levels TSS Acidity Xanthungum was observed for mold growth by visual methods at monthly
(%) guava : (0Brix) (%) levels intervals throughout the storage period. In this experiment,
watermelon (%)
factorial completely randomized design (factorial CRD) was
B1X1 50 : 50 40 1 0
B 1X 2 50 : 50 40 1 0.1
adopted. The data was analyzed and main interaction effects were
B 1X 3 50 : 50 40 1 0.2 presented (Sundararaj et al., 1972). Six different combinations
B 1X 4 50 : 50 40 1 0.3 of xanthan gum under the CRD for guava squash examined for
B 1X 5 50 : 50 40 1 0.4 recipe standardization.
B 1X 6 50 : 50 40 1 0.5
B 2X 1 75 : 25 40 1 0 Results and discussion
B 2X 2 75 : 25 40 1 0.1
B 2X 3 75 : 25 40 1 0.2 Significant chemical changes were noticed in different guava-
B 2X 4 75 : 25 40 1 0.3 watermelon squash blending levels and interaction effect with
B 2X 5 75 : 25 40 1 0.4 the xanthan gum levels throughout storage period (Table 2).
B 2X 6 75 : 25 40 1 0.5
B 3X 1 25 : 75 40 1 0 Maximum increase in total soluble solids (40 to 41.95 oBrix)
B 3X 2 25 : 75 40 1 0.1 was noticed in the blending level B2 (75:25 pulp). This might be
B 3X 3 25 : 75 40 1 0.2 due to increase in total soluble sugars caused by hydrolysis of
B 3X 4 25 : 75 40 1 0.3 polysaccharides like starch, cellulose and pectin substances into
B 3X 5 25 : 75 40 1 0.4 simpler substances. Minimum increase (from 40-41.38 oBrix) in
B 3X 6 25 : 75 40 1 0.5 total soluble solids was noticed in the blending level B3 (25:75
Toshniwal digital pH meter (Model DI 707), total soluble pulp). Changes in watermelon squash and interaction effect due
solids with hand refractometer (Erma hand refractometer) and to various xanthan gum and pulp levels have been recorded by
acidity by titration method. Ascorbic acid was determined by Shankaraswamy and Banik (2012). Maximum changes in TSS
2, 6-dichlorophenol indophenols titration method at every 30 (40-43.03 oBrix) was noticed in treatment X1B1. A minimum
days interval during 180 days of storage (Ranganna, 1986). The change in TSS (40-40.66 oBrix) and acidity (1-0.91%) was
viscosity in blended Guava-watermelon squash was determined noticed in treatment X6B3 (0.5% of xanthan gum 25:75 levels of
over a wide range of temperature (30-50 oC) as well as at constant pulp (guava: watermelon). Interaction effect in different xanthan
concentration (40 oBrix) by using the viscometer bath (Model No. gum and blending levels during 180 days of storage period were
- SVB, S.L. No. - S/01 Simco Brand, Kolkata, West Bengal) and significant in chemical changes (Table 3). Maximum changes
capillary viscometer tube (Cannon fenske viscometer) during in TSS (from 40-43.92 oBrix), acidity (1-0.63%) was noticed in
the 180 days storage period (Ranganna, 1986). Blended guava- treatment D6B2 (at 180 days with 75:25 levels of pulp (guava:
watermelon squash product was evaluated at 180 days of storage watermelon) and minimum was observed in D1B2 (40-40.17
for sensory attributes such as appearance, aroma and flavour, taste o
Brix). Difference in the pulp level, xanthan gum levels, different
and overall acceptability by a panel of 8 judges by numerical storage period and their interaction were significant in changing
Table 2. Changes in TSS, pH, acidity, ascorbic acid at different levels of blended guava-watermelon squash
Blending TSS pH Acidity Ascorbic acid
levels (%) (oBrix) (%) (mg/100mL)
Initial Final Initial Final Initial Final Initial Final
B1(50 : 50) 40 41.91 3.38 3.76 1.0 0.884 19.4 15.07
B2 (75 : 25) 40 41.95 3.14 3.37 1.0 0.783 30.9 20.56
B3 (25 : 75) 40 41.38 3.59 3.75 1.0 0.882 12.8 7.84
LSD (P=0.05) 0.01 0.03 0.001 0.01
Interaction [X (Xanthangum levels) x B (Blending levels)] during 180 days of storage period
X1 B1 43.03 3.684 0.85 11.53
X1 B2 42.85 3.518 0.72 17.48
X1 B3 42.30 3.798 0.85 6.51
X2 B1 42.10 3.565 0.84 13.28
X2 B2 41.99 3.465 0.74 19.19
X2 B3 41.62 3.794 0.86 7.25
X3 B1 41.94 3.753 0.88 14.59
X3 B2 41.94 3.347 0.75 19.66
X3 B3 41.27 3.765 0.86 7.30
X4 B1 41.73 3.711 0.89 16.80
X4 B2 41.78 3.336 0.80 20.40
X4 B3 41.27 3.724 0.88 7.87
X5 B1 41.59 4.137 0.89 17.04
X5 B2 42.12 3.323 0.93 21.77
X5 B3 41.20 3.665 0.90 7.97
X6 B1 41.08 3.756 0.93 17.16
X6 B2 41.03 3.248 0.83 24.88
X6 B3 40.66 3.676 0.91 10.11
LSD (P=0.05) 0.06 0.019 0.01 0.05
68 Processing and quality evaluation of blended guava-watermelon squash

TSS. Maximum changes in TSS (from 40-45.30) was noticed Table 3 continued
in treatment X1D6B1 (0% of xanthan gum, 50:50 levels of pulp
X2 D3 B2 41.73 3.41 0.80 20.42
(guava: watermelon), at 180 days of storage period). Minimum
X2 D3 B3 40.96 3.81 0.88 7.55
increase in TSS (from 40-40.03) was noticed in treatment X6D1B3
X2 D4 B1 42.56 3.60 0.82 12.20
(0.5% of xanthan gum, 25:75 levels of pulp (guava: watermelon).
Similar results were observed in squash prepared from mango- X2 D4 B2 41.96 3.43 0.68 17.46
papaya blended juice (Kalra et al., 1991), kiwi fruit (Thakur and X2 D4 B3 41.96 3.83 0.85 5.43
Barwal, 1998) and aonla (Reddy and Chikkasubbanna, 2008). X2 D5 B1 43.06 3.61 0.79 9.20
X2 D5 B2 42.80 3.59 0.62 13.42
Table 3. Changes in TSS, PH, Acidity, Ascorbic acid content of X2 D5 B3 42.86 3.84 0.81 4.48
Watermelon blended guava squash with 30, 60, 90, 120, 150, 180 days
with 3 levels of blending X2 D6 B1 43.86 3.64 0.77 8.50
Interaction TSS pH Acidity Ascorbic acid X2 D6 B2 44.49 3.64 0.60 11.60
(Days x Blending) (oBrix) (%) (mg/100 mL) X2 D6 B3 43.16 3.88 0.80 3.88
D1 B1 40.88 3.45 0.96 18.05 X3 D1 B1 40.49 3.42 0.96 17.96
D1 B2 40.17 3.21 0.94 28.70 X3 D1 B2 39.96 3.16 0.94 29.20
D1 B3 40.21 3.60 0.97 11.48 X3 D1 B3 40.20 3.60 0.96 11.26
D2 B1 41.00 3.54 0.92 17.26
X3 D2 B1 41.26 3.47 0.93 17.23
D2 B2 40.77 3.29 0.87 25.61
D2 B3 40.54 3.67 0.93 10.44 X3 D2 B2 40.50 3.27 0.86 24.83
D3 B1 41.51 3.62 0.89 15.72 X3 D2 B3 40.40 3.71 0.91 10.58
D3 B2 41.57 3.36 0.80 22.51 X3 D3 B1 41.56 3.53 0.88 15.35
D3 B3 40.91 3.72 0.88 8.51 X3 D3 B2 41.76 3.36 0.73 21.13
D4 B1 42.28 3.78 0.86 14.32
X3 D3 B3 40.70 3.75 0.85 7.58
D4 B2 42.26 3.39 0.74 18.76
D4 B3 41.60 3.76 0.85 6.43 X3 D4 B1 42.00 3.80 0.85 14.66
D5 B1 42.85 3.95 0.84 13.10 X3 D4 B2 42.43 3.37 0.74 17.60
D5 B2 43.03 3.46 0.69 14.96 X3 D4 B3 41.70 3.81 0.83 5.83
D5 B3 42.26 3.80 0.83 5.40 X3 D5 B1 42.86 3.98 0.83 12.56
D6 B1 43.46 4.24 0.81 11.95
X3 D5 B2 42.70 3.42 0.68 13.60
D6 B2 43.92 3.51 0.63 12.84
D6 B3 42.79 3.84 0.81 4.75 X3 D5 B3 42.00 3.84 0.83 4.51
LSD (P=0.05) 0.06 0.02 0.02 0.05 X3 D6 B1 43.49 4.30 0.80 9.77
Interaction TSS pH Acidity Ascorbic X3 D6 B2 44.30 3.47 0.58 11.62
(X x D x B) during (oBrix) (%) acid X3 D6 B3 42.63 3.85 0.82 4.06
180 d of storage (mg/100 mL)
X4 D1 B1 40.38 3.43 0.96 18.61
X1 D1 B1 40.80 3.47 0.95 17.18 X4 D1 B2 40.00 3.14 0.96 30.13
X1 D1 B2 40.51 3.33 0.88 25.67 X4 D1 B3 40.06 3.58 0.99 11.73
X1 D1 B3 40.51 3.65 0.97 10.61
X4 D2 B1 41.00 3.46 0.94 18.14
X1 D2 B1 41.39 3.55 0.87 14.74
X4 D2 B2 40.56 3.26 0.94 24.86
X1 D2 B2 41.29 3.41 0.80 23.10
X4 D2 B3 40.33 3.61 0.93 10.70
X1 D2 B3 41.30 3.73 0.90 8.28
X4 D3 B1 41.40 3.51 0.90 16.54
X1 D3 B1 42.12 3.62 0.85 12.33
X4 D3 B2 41.43 3.36 0.80 22.66
X1 D3 B2 42.00 3.49 0.77 18.42
X4 D3 B3 40.86 3.70 0.89 9.15
X1 D3 B3 42.00 3.78 0.84 6.88
X4 D4 B1 42.00 3.69 0.87 16.44
X1 D4 B1 43.92 3.75 0.82 9.68
X4 D4 B2 42.00 3.37 0.77 17.86
X1 D4 B2 43.51 3.56 0.67 15.01
X4 D4 B3 41.33 3.76 0.86 5.96
X1 D4 B3 42.60 3.83 0.82 4.89
X4 D5 B1 42.53 3.92 0.84 16.06
X1 D5 B1 44.67 3.82 0.80 8.16
X4 D5 B2 42.83 3.40 0.70 14.06
X1 D5 B2 44.67 3.61 0.61 12.41
X4 D5 B3 42.26 3.81 0.83 4.88
X1 D5 B3 43.36 3.88 0.79 4.42
X4 D6 B1 43.06 4.24 0.81 15.43
X1 D6 B1 45.30 3.87 0.79 7.10
X1 D6 B2 45.10 3.68 0.58 10.30 X4 D6 B2 43.86 3.46 0.63 12.80
X1 D6 B3 44.01 3.91 0.78 3.98 X4 D6 B3 42.76 3.86 0.81 4.80
X2 D1 B1 40.28 3.45 0.95 18.03 X5 D1 B1 40.29 3.48 0.97 18.33
X2 D1 B2 40.22 3.31 0.91 26.53 X5 D1 B2 40.23 3.15 0.97 30.33
X2 D1 B3 40.23 3.65 0.95 11.56 X5 D1 B3 40.23 3.55 0.97 11.83
X2 D2 B1 41.00 3.53 0.87 17.37 X5 D2 B1 40.89 3.64 0.95 18.03
X2 D2 B2 40.76 3.38 0.84 25.72 X5 D2 B2 41.16 3.26 0.89 26.56
X2 D2 B3 40.56 3.75 0.89 10.60 X5 D2 B3 40.63 3.66 0.95 10.70
X2 D3 B1 41.86 3.55 0.86 14.36 X5 D3 B1 41.10 3.76 0.90 18.56
 Processing and quality evaluation of blended guava-watermelon squash 69

Table 3 continued Differences in pH were significant (P=0.05) with different pulp

X5 D3 B2 41.76 3.31 0.84 25.36 levels. Increasing trend of pH was noticed throughout storage
X5 D3 B3 40.73 3.68 0.91 9.24
period. Maximum increase (3.38- 3.76) in pH was noticed in
the blending level B1 (50: 50 pulp). Minimum changes (3.14 to
X5 D4 B1 41.86 4.02 0.89 16.15
3.37) in pH was noticed in the blending level B2 (75: 25 pulp) and
X5 D4 B2 42.70 3.36 0.79 18.86
in treatment X5D5B1 (3.38-4.49). The increase in pH of guava-
X5 D4 B3 41.06 3.66 0.88 6.37
watermelon blended squash during storage could be attributed
X5 D5 B1 42.36 4.49 0.85 16.10
to acid hydrolysis of polysaccharides and non-reducing sugars
X5 D5 B2 43.33 3.41 0.75 14.63 to hexose sugars (reducing sugars) or complex formation in the
X5 D5 B3 42.00 3.66 0.85 4.87 presence of metal ions as reported in aonla juice (Gajanana,
X5 D6 B1 43.06 5.40 0.81 15.10 2002). There was a declining trend in acidity of guava blended
X5 D6 B2 43.56 3.42 0.74 14.86 watermelon squash throughout storage period. Pulp levels and
X5 D6 B3 42.43 3.76 0.83 4.82 xanthan gum and storage period and there interaction effects were
X6 D1 B1 40.10 3.43 0.97 18.70 significant throughout storage period. Minimum changes in the
X6 D1 B2 40.10 3.14 0.97 30.36 acidity in the treatment X6D1B3 (1 to 0.98) and D1B3 (1 to 0.97)
X6 D1 B3 40.03 3.57 0.98 11.86 was noticed, respectively. Difference in the pulp level, xanthan
X6 D2 B1 40.46 3.60 0.96 18.03 gum levels and at different storage period and their interaction
X6 D2 B2 40.36 3.17 0.91 28.56 were significant in changing ascorbic acid. Maximum loss in the
X6 D2 B3 40.03 3.60 0.98 11.80 ascorbic acid (30.9-20.56mg/100mL) was noticed in the treatment
X6 D3 B1 41.00 3.75 0.95 17.16 B2 (75:25). Minimum changes in the ascorbic acid were noticed in
X6 D3 B2 40.73 3.22 0.87 27.10 the treatment B1 (19.42-15.0mg/100mL) and X6B1 (19.42- 17.16
X6 D3 B3 40.23 3.64 0.91 10.66 mg/100g) (Table 2). The decline in ascorbic acid concentration
X6 D4 B1 41.33 3.85 0.94 16.76 could be due to its thermal degradation during processing and
X6 D4 B2 40.96 3.26 0.8 25.80
subsequent oxidation in storage (Brock et al., 1998). Similar
observations were made in guava squash (Shankaraswamy and
X6 D4 B3 40.86 3.68 0.89 10.12
Banik, 2011), aonla squash (Reddy and Chikkasubbanna, 2008)
X6 D5 B1 41.63 3.88 0.91 16.50
and amla jam (Reddy and Chikkasubbanna, 2009). Increasing
X6 D5 B2 41.86 3.82 0.78 21.63
trend of viscosity observed in the blended guava-watermelon
X6 D5 B3 41.06 3.78 0.88 9.26
squash. Less incorporation of xanthan gum in 75:25 blending
X6 D6 B1 42.00 4.01 0.87 15.83
level gave maximum viscosity compare to other treatments.
X6 D6 B2 42.20 3.36 0.66 15.86 The maximum score for the aroma and flavour (3.07) and taste
X6 D6 B3 41.73 3.76 0.84 6.98 (2.50) with highest overall acceptability (2.08) was observed in
LSD (P=0.05) 0.15 0.05 0.02 0.11 the treatment B2 (75:25) (Table 4). Results indicate that addition
Table 4. Organoleptic score and relative variation in viscosity of blended guava-watermelon squash
Blending level Aroma and flavor Colour and Teste Over all Viscosity (Pa-sec)
(%) appearance acceptability 30 oC 40 oC 50 oC
B1(50 : 50) 2.69 1.28 1.89 1.72 2.760 3.570 2.283
B2 (75 : 25) 3.07 1.09 2.50 2.08 2.130 4.010 1.985
B3 (25 : 75) 2.15 1.79 1.46 1.54 2.313 2.300 1.677
CD at 5% 0.15 0.11 0.12 0.11 0.015
X1B1(50 : 50) 1.43 0.72 1.61 1.00 0.531 0.981 0.715
X1B2 (75 : 25) 1.71 0.76 2.43 1.17 0.503 6.904 0.071
X1B3 (25 : 75) 1.18 0.83 1.31 0.90 0.353 0.384 0.003
X2B1(50 : 50) 2.10 1.18 2.13 2.17 1.073 1.928 0.961
X2B2 (75 : 25) 3.16 1.06 3.05 3.11 0.947 1.242 0.832
X2B3 (25 : 75) 1.35 2.06 1.76 1.93 0.698 0.340 0.762
X3B1(50 : 50) 2.73 1.75 2.22 2.52 2.716 3.651 1.668
X3B2 (75 : 25) 3.20 1.25 2.97 3.07 2.547 3.574 1.553
X3B3 (25 : 75) 2.25 2.16 1.71 2.31 2.361 2.673 1.423
X4B1(50 : 50) 3.05 1.53 2.13 1.97 3.012 3.977 2.674
X4B2 (75 : 25) 3.32 1.18 2.70 2.22 2.877 2.520 2.423
X4B3 (25 : 75) 2.75 2.11 1.33 1.80 2.741 1.464 2.027
X5B1(50 : 50) 2.96 1.25 1.65 1.45 4.071 4.725 3.005
X5B2 (75 : 25) 3.26 1.15 1.95 1.81 1.123 4.511 2.816
X5B3 (25 : 75) 3.28 1.81 1.36 1.26 3.367 4.262 2.321
X6B1(50 : 50) 3.58 1.27 1.57 1.20 5.158 6.161 4.677
X6B2 (75 : 25) 3.76 1.16 1.91 1.12 4.780 5.307 4.214
X6B3 (25 : 75) 3.95 1.75 1.30 1.02 4.359 4.678 3.528
LSD (P=0.05) 0.36 0.28 0.29 0.27 0.037
70 Processing and quality evaluation of blended guava-watermelon squash
of xanthan gum 0.3% positively imparts stability and acts as
emulsifier to the blended guava-watermelon squash. Squash
prepared from 75:25 blending level was highly preferred thus
it has added advantage in utilizing more guava fruits during the
rainy season.
Acknowledgment
The author is thankful to Dr. P.P. Thirumalaisamy, Scientist,
DGRC, Junagadh, Gujarat for suggestions and correction of
manuscript.
References
Amerine, M.A., R.M. Pangbron and E.B. Roesler, 1965. Principles of
Sensory Evaluations of Food. Academic Press, New York, 123-
125.
Anonymous, 2011. Economic Review (2010-2011). Government of West
Bengal, Statistical Appendix.
Brock, V.D., L. Ludikhuyze, C. Weemaes, L.A. Van and M. Hendrickx,
1998. Kinetics for isobaric isothermal degradation of L-Ascorbic
acid. J. Agric. Food Chem., 46(5): 22-25.
Gajanana, K. 2002. Processing of Aonla (Emblica officinalis Gaertn.)
fruits. Thesis submitted to UAS, GKVK, Bangalore, India.
Huor, S.S., E.M. Ahmed, R.D. Carter and R.L. Huggart, 1980. Colour
and flavour qualities of white grapefruit: watermelon juice mixtures.
Journal Food Sci., 45: 1419-1421.
Kalra, S.K., D.K. Tandon and B.P. Singh, 1991. Evaluation of mango-
papaya blended beverages. Indian Food Packer, 45(1): 33-36.
Ranganna, S. 1986. Handbook of Analysis of Fruit and Vegetable
Products. Tata McGraw-Hill Publishing Company Limited, New
Delhi, India.
Reddy, H.A. and V. Chikkasubbanna, 2008. Studies on the storage
behaviour and organoleptic evaluation of amla squash. J. Asian
Hort., 4(3): 206-212.
Reddy, H.A. and V. Chikkasubbanna, 2009. Storage behaviour of amla
syrup. Asian J. Hort., 4(1): 5-9.
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evaluation guava squash. J. Appl. Hort., 13: 82-84.
Shankaraswamy, J. and A.K. Banik, 2012. Effect of xanthan gum on
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Sundararaj, N., S. Nagaraju, M.N. Venkataramana and M.K. Jagannath,
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Received: November, 2013; Revised: March, 2014; Accepted: March, 2014
 Journal

Journal of Applied Horticulture, 16(1): 71-75, 2014 Appl

Effect of pre-treatment and drying temperature on quality of

dehydrated cauliflower (Brassica oleracea var. botrytis)

R. Ranjan, M. Longkumer and J. Kabir*

Department of Post Harvest Technology of Horticultural Crops, Faculty of Horticulture, Bidhan Chandra Krishi
Viswavidyalaya, Mohanpur, Nadia, West Bengal, 741252, India. *E-mail: j_ kabir@rediffmail.com

Abstract
Cauliflower curd were pre-treated with hot water blanching + 0.125% KMS, with/without microwave blanching for 5 minutes and
were dehydrated at three levels of temperature viz., 65, 60 and 55 oC at different treatment combinations. Considering the dehydration
characters and quality after dehydration and storage it was found that T2 (hot water blanching + 0.125% KMS + microwave blanching
+ drying at 65 oC) was the best treatment followed by T4 (hot water blanching + 0.125% KMS + microwave blanching + drying at
60 oC) and T5 (hot water blanching + 0.125% KMS + drying at 55 oC). In T2, time taken for complete dehydration (445 minutes) and
moisture content (3.62%) was least. Further, the moisture content after 6 month of storage was also less (9.63%), drying rate (135.74%)
and dehydration ratio (10.70) was medium after dehydration. Ascorbic acid retention was maximum during storage in the treatment.
Sensory evaluation also supported the superiority of this treatment.
Key words: Cauliflower, pretreatment, drying temperature, quality, dehydration.

Introduction
India is the second largest producer of cauliflower (Brassica
oleracea var. botrytis) in the world with a total annual production
of 4.8 million metric tonnes amounting to 30% of the total world
production (Anon., 2003). Cauliflower contains considerable
amount of protein, carbohydrate, fat, vitamin B, vitamin C and is
fairly rich in minerals like phosphorus, potassium, sodium, iron,
magnesium etc. (Chatterjee, 1993).
The edible portion of cauliflower constitutes approximately
67.3% of the vegetable (Abhay et al., 2006). Fresh cauliflower
is delicate in handling and highly perishable in nature. In peak
season, due to market glut cauliflower are sold at a very low price
resulting in heavy losses to the growers. In India, it has been
estimated that postharvest losses of cauliflower is to the extent
of about 28.6 to 35.1 per cent (Pal et al., 2002). Fresh cauliflower
cannot be stored for longer period due to its poor shelf life, but
good compact heads can be cold stored for 3-4 weeks at 0 °C
with 85-90% relative humidity (Madhavi and Ghosh, 1998). Only
limited quantity is processed and preserved in different form for
future consumption. By drying the fresh cauliflower, it is quickly
transformed into a dried stable material, volume and weight about
10 times less than the original fresh material. It is simple, low cost
and economical method of preservation. Dehydrated cauliflower
is used as an important ingredient in several food preparations
including instant soups, kheer, instant mix and to make stuff
in prontha (semi friedcake) (Abhay et al., 2006). The demand
of dried cauliflower is ever increasing because its export is in
progress. Household drying of cauliflower is an age-old common
practice in villages. However it develops rancid smell because of
prolonged drying under shade.
Osmotic dehydration of cauliflower as influenced by temperature,
salt concentration, ratio of brine to material and time was studied
by Vijayanand et al. (1995). Dehydration of cauliflower by hot
air has not been tested extensively as in other vegetables (Von
Loesecke, 1998). The meager information available regarding
dehydration of cauliflower indicated the problem of retention
of texture, colour, flavour and rehydration characteristics of
rehydrated product (Srivastava and Sulebele, 1975; Raina et al.,
1982 and Abhay et al., 2006).
It has been reported that blanching along with pretreatment
with sodium hydroxide, calcium hydroxide, sodium sulphate,
potassium metabisulphite has been effective in maintaining
the physical and chemical characters of dehydrated product of
different vegetables (Srivastava and Sulebele, 1975; Bawa and
Saini, 1986; Mulay et al., 1994; Rouf et al., 2003). Further,
temperature of dehydration is a critical factor for good quality
dehydrated product (Abhay et al., 2006). The high temperature
and considerable oxidation of ascorbic acid associated with hot
air drying is reported to cause a brown colour (Ranganna and
Setty, 1968).
Thus, considering the above facts, the present investigation
was undertaken to standardize the pretreatment with different
temperature of dehydration and to study the quality of dehydrated
product.
Materials and methods
The cauliflower curd cut pieces (2 × 2 cm) were divided into six
lots and subjected to different combinations of pre-treatments
viz., hot water blanching (97°C for 1 min) + soaking in 0.125%
potassium metabisulphite (KMS) solution for 10 minutes and hot
water blanching (97°C for 1 min) + soaking in 0.125% potassium
metabisulphite (KMS) solution for 10 minutes and were then
dehydrated in hot air cabinet drier at three different temperatures
i.e., 65, 60 and 55oC. The drying was carried out at an air velocity
of 4.7 mm/sec. The trays were weighed on digital balance at
regular interval (5-10 minutes interval during first 4 hours then
at an interval of 25 to 30 minutes) until the product attained more
72 Effect of pre-treatment and drying temperature on quality of dehydrated cauliflower

or less constant weight. Dried sample was collected from the tray,
cooled to room temperature and each treatment was sealed in zip
lock (self sealing, 200 gauge) polyethylene bags, which facilitated
storage study at different month interval upto 6 months. All the
polyethylene bags were stored in air tight plastic container having
sachet of silica gel. The moisture content and ascorbic acid of
the sample was estimated in three replications before and after
dehydration and also during storage using standard procedure
(Ranganna, 1991).
Sensory evaluation was conducted by a panel of judges for colour,
flavour, texture, odour and overall acceptability using 9 point
hedonic scale with rating of 1 for extremely disliked and 9 for
extremely excellent (Dasgupta et al., 1999). Statistical analysis
was done according to factorial completely randomized design
using standard statistical procedure.

Results and discussion

Fig. 1. Moisture content at different time intervals in treatments
Relationship between the moisture content (% db) vs time of
different treatments has been shown in Fig. 1. The initial moisture T4) and 55°C (655 minutes in T5 and 625 minutes in T6). Hence, at
a temperature level of 65 °C, the drying time ranged from 445-495
content was high in all the treatments and it varied from 1158.44%
min, 505-510 minutes for temperature level of 60 °C and 655-625
in T1 (hot water blanching + 0.125 KMS + drying at 65°C) to
minutes for temperature level of 55 °C. The driving force for the
913.71% in T6 (hot water blanching + 0.125% KMS + microwave
mass transfer at the surface increases markedly with temperature,
blanching + drying at 55°C) (Table 2). Moisture content decreased
since mass loss due to evaporation at the surface is the function
rapidly during the first hour of drying in all the treatments
of partial vapour pressure difference between the surface and the
indicating easy escape of moisture during the early period of
convective air which increases the moisture diffusivity at higher
drying due to high moisture content at the initial stage of drying.
temperature, contributing to faster drying at higher temperature
It is believed that in the early stages of drying the material behaves
(Tulsidas et al., 1995).
as though the surface was saturated with water. Dehydration curve
indicated that the time required to achieve desired final moisture Drying rate versus average moisture content indicated that in
(5 to 8%) is influenced predominantly by the temperature of general, with decrease in the average moisture content drying rate
dehydration. Higher temperature of dehydration at 65°C yielded decreased in all the treatments (Fig. 2). In T2 (hot water + 0.125%
a faster drying (495 minutes in T1 and 445 minutes in T2) than the KMS + microwave blanching + drying at 65 °C), the decrease
lower temperature of 60 °C (510 minutes in T3 and 505 minutes in in drying rate with the average moisture content indicated that
Table 1. Initial moisture content in different treatments of dehydrated
cauliflower, before dehydration and their final moisture content after
dehydration and storage
Treatment Initial moisture Final moisture Final moisture
content before content after content after 6
dehydration dehydration months of storage
(% wb) (% wb) (% wb)
T1 92.23 4.08 9.59
T2 91.17 3.62 9.63
T3 90.92 5.26 9.80
T4 90.13 5.02 10.55
T5 91.81 7.48 12.49
T6 91.26 6.24 13.19
LSD (P=0.05) 1.15 0.13 4.40
LSD (P=0.01) 1.61 0.18 N.S.
Fig. 2. Drying rate versus average moisture under different treatments
Table 2. Initial moisture (% db) final moisture content (% db), moisture
reduction (%) drying time and drying rate in different treatments of dehydrated whole drying took place in falling rate period i.e., a declined
cauliflower
trend and no constant rate period of drying was observed. In
Treatment Moisture Moisture Drying Total Overall T1 (hot water + 0.125% KMS + drying at 65°C), T3 (hot water
Initial Final reduction time drying drying rate
(% db) (% db) (%) (min.) time in h (%/h) blanching + 0.125% KMS + drying at 60°C) and T4 (hot water
T1 1158.44 5.56 1152.88 495 8.15 139.74 blanching + 0.125% KMS + microwave blanching + drying
T2 1012.23 5.47 1006.76 445 7.25 135.74 at 60°C) drying rate increased slightly and thereafter showed
T3 1002.15 5.62 996.53 505 8.25 118.39 the declining trend. But in T5 (hot water blanching + 0.125%
T4 886.85 4.96 881.89 500 8.20 105.82 KMS + drying at 55°C) and T6 (hot water blanching + 0.125%
T5 1151.82 8.02 1143.80 655 10.55 104.77 KMS + microwave blanching + drying at 55°C) drying rate
T6 913.71 6.64 907.07 625 10.25 87.07 increased slightly and thereafter almost a constant rate of
 Effect of pre-treatment and drying temperature on quality of dehydrated cauliflower
drying was observed which was followed by decreasing trend.
The characteristics features of cauliflower curd and pretreatment
effect (like KMS and microwave) might be attributed to the
differences in the drying rate with average moisture content in
treatments (T1 to T6). It has been reported earlier (Madamba et
al., 1996) that almost all the drying of biological products takes
place in the decline rate period. i.e., with decrease in average
moisture content drying rate decreased continuously.
Moisture content before dehydration varied from 90.13% (T4 i.e.
hot water blanching + 0.125% KMS + microwave blanching +
drying at 60°C) to 92.23% (T1 i.e. hot water blanching + 0.125%
KMS + drying at 65°C) and after dehydration moisture content
ranged from 3.62% in T2 (hot water blanching + 0.125% KMS
+ microwave blanching + drying at 65°C) to 7.48% in T5 (hot
water blanching + 0.125% KMS + drying at 55°C). The moisture
content (before and after dehydration on wet basis) in almost
all the treatment was within the range of earlier findings (Kaur
and Singh, 1981; Raina et al., 1982; Bawa and Saini, 1986 and
Maldonado et al., 2003).
The high final moisture content of T5 (hot water blanching +
0.125% KMS + drying at 55°C) and T6 (hot water blanching +
0.125% KMS + microwave blanching + drying at 55°C) (7.48%
and 6.24, respectively) was due to comparatively low drying
temperature which was also demonstrated by Abhay et al. (2006).
After six months of storage, moisture content was lower in T1
and T2 compared to other treatments, however, there was no
significant difference between the treatments for the dehydrated
cauliflower (Table 1).
The total moisture reduction (% db) in different treatments
and their respective drying time is shown in Table 3. The total
moisture reduction was observed to be highest (1152.88%) in T1
(hot water blanching + 0.125% KMS + drying at 65°C) followed
by 1143.80% in T5 (hot water blanching + 0.125% KMS + drying
at 55°C), 1006.76% in T2 (hot water blanching + 0.125% KMS +
microwave blanching + drying at 65°C), 996.53% in T3 (hot water
blanching + 0.125% KMS + drying at 60°C), 907.07% in T6 (hot
water blanching + 0.125% KMS + microwave blanching + drying
at 55°C) and 881.89% in T4 (hot water blanching + 0.125% KMS
+ microwave blanching +drying at 60°C).
The drying time taken for the dehydration was maximum in T5
(655 min) to remove 1143.80% moisture from the initial moisture
content of 1151.82% whereas drying time taken was least in T2
(445 min) to remove 1006.76% moisture from the initial moisture
content of 1012.23% (Table 3). Thus, drying time for T2 was
minimum i.e. 445 minutes followed by 495 minutes in T1, 500
minutes in T4, 505 minutes in T3, 625 minutes in T6 and 655
minutes in T5. The difference in drying time can be attributed to
differences in dehydration temperature of 65°C (for T1 and T2),
60°C (for T3 and T4) and 55°C (T4 and T5). Drying time decreased
with increased temperature of drying. Higher temperature causes
the increase in the product temperature and higher vapour
pressure gradient resulting in the increased moisture diffusivity
and accelerated drying (Sarvacos and Rouzeos, 1986). The
treatments T1, T3 and T5 received common pretreatment (hot
water blanching + 0.125% KMS) and only the temperature of
dehydration varied (65, 60 and 55°C temperature, respectively)
as indicated earlier. Thus, the drying time increased in the order
73
of T1 (495 min), T3 (505 min) and T5 (655 min) bearing the
inverse relationship with temperature in these treatments. Similar
behaviour was exhibited by the treatments T2, T4 and T6 which
received same pretreatments (hot water blanching + 0.125%
KMS + microwave blanching) but different drying temperatures
as T2 (445 min, at 65°C), T4 (500 minutes at 60°C) and T6 (625
minutes at 55°C). It is interesting to note that drying time varied
in different pretreated cauliflower samples dehydrated at common
temperatures i.e. T1, T2 at 65°C, T3, T4 at 60°C and T5, T6 at 55°C,
indicating that although the moisture content slightly varied
initially, pretreatment with microwave blanching influenced the
drying time. T2, T4 and T6 took lesser drying time compared to
T1, T3 and T5 (without microwave blanching). Overall drying
rate (%/hr) was least in T6 (87.07%) followed by T5 (104.77%),
T4 (105.82%), T3 (118.35%), T2 (135.74%) and T1 (139.74%) in
that increasing order. It showed that drying rate increased with
increased temperature of drying from 55°C (T6 and T5) to 65°C
(T1 and T2).
The pretreatments followed by dehydration at 3 levels of
temperature affected the dehydration ratio and it ranged from
9.37 in T4 (hot water blanching + 0.125 KMS + microwave
blanching + drying at 60°C) to 12.09% in T1 (hot water blanching
+ 0.125% KMS + drying at 65°) (Table 2). Rehydration ratio after
dehydration on the other hand ranged from 4.60 (T4) to 6.72 (T5).
Similar trend of dehydration ratio to the present findings has also
been reported by Srivastava and Sulebele (1975) and Raina et al.
(1982). Dehydration ratio as reported by Bawa and Saini (1986),
Mishra and Agarwal (2005) was however higher than our findings,
which might be due to varietal differences. Additional microwave
blanching treatments (T2, T4 and T6) lowered the dehydration
ratio in contrast to treatments without microwave blanching
(T1, T3 and T5). The high dehydration ratio of T1 (12.09) at 65°C
compared to lower dehydration temperature treatments at 60
and 55°C is contrary to the reports of Maskan (2000) and Rouf
et al. (2003) in banana and cabbage. Further among treatments
without microwave blanching (T1, T3 and T5) drying ratio of T1
at 65°C was highest i.e. 12.09 followed by T5 (11.58) at 55°C
and T3 (10.43) at 60°C. This result is not consistent with Abhay
et al. (2006) who reported that dehydration ratio increased with
decreased temperature of drying.
Rehydration ratio was high in treatments not receiving microwave
blanching i.e. T1, T3 and T5 (Table 2). Among these treatments
rehydration ratio was maximum in T5 (6.72) followed by T3 (5.21)
and T1 (5.03) indicating that rehydration ratio decreased with
increase in temperature of drying. However, among the treatments
receiving microwave blanching rehydration ratio was highest in
T2 (5.02) with drying temperature of 65°C. In others microwave
blanching treatments i.e. T4 (drying temperature of 60°C) and T6
(drying temperature of 55°C) the rehydration ratio was 4.6 and
4.7 which were more or less similar. Rehydration ratio gradually
decreased throughout the period of storage upto six months.
Highest reduction in the capacity to rehydrate was observed in
T5 (30.13) where the rehydration ratio dropped sharply from
5.21 (initially) to 3.64 after six months which was followed by
T2 (5.02 to 3.63), T4 (4.60 to 3.58), T5 (6.72 to 5.34), T6 (4.7 to
4.04), and T1 (5.03 to 4.37).
Rehydration coefficient increased with decrease in dehydration
temperature (Table 4). After dehydration, treatments without
74 Effect of pre-treatment and drying temperature on quality of dehydrated cauliflower

Table 3. Dehydration ratio in different treatments of dehydrated cauliflower In storage, coefficient of rehydration decreased gradually
and their rehydrated ratio after dehydration and during storage and on 6th month rehydration coefficient remained high in
Treatment Drying Rehydration ratio Reduction (%) T5 (0.55) followed by T6 (0.454), T1 (0.423), T4 (0.402), T3
ratio (storage period in months) (storage period in months)
(0.359) and T2 (0.352).
0 2 4 6 2 4 6
T1 12.09 5.03 4.93 4.60 4.37 1.98 8.54 13.12 Pretreatment with chemicals like KMS has been reported
T2 10.70 5.02 4.96 4.26 3.63 1.19 15.13 27.68 to increase rehydration ratio and coefficient of rehydration
T3 10.43 5.21 4.60 3.82 3.64 11.70 26.67 30.13 (Srivastava and Sulebele, 1975; Mishra and Agrawal, 2005).
T4 9.37 4.60 4.41 3.90 3.58 4.13 15.21 22.17
T5 11.58 6.72 6.07 5.38 5.34 9.67 19.94 20.53
However rehydration ratio and coefficient of rehydration
T6 9.50 4.70 4.53 4.20 4.04 3.61 10.63 14.04 decreases with increase in temperature of dehydration
(Abhay et al., 2006). Dehydrated product sometimes did
Table 4. Rehydration characteristics of different treatment of cauliflower not recover their structural properties after rehydration as a
during storage
result of structural damage during drying and the hysteresis
Treatments 0 months 2 months 4 months 6 months
phenomenon that takes place during rehydration (Magdalini
R.R. CO R. R.R. CO R. R.R. CO R. R.R. CO R.
T1 1 : 503 0.486 1 : 4.93 0.476 1 : 4.60 0.445 1 : 4.37 0.423 and Zacharias, 2001).
T2 1 : 502 0.486 1 : 4.96 0.475 1 : 4.26 0.408 1 : 3.63 0.352 The ascorbic acid content for each treatment varied
T3 1 : 521 0.528 1 : 4.60 0.525 1 : 3.82 0.440 1 : 3.64 0.359
significantly at both 5 and 1% level (Table 5). In treatments
T4 1 : 4.6 0.516 1 : 4.41 0.495 1 : 3.90 0.438 1 : 3.58 0.402
T5 1 : 6.72 0.699 1 : 6.07 0.684 1 : 5.38 0.612 1 : 5.34 0.55 without microwave blanching i.e. in T1 (drying temperature
T6 1 : 4.70 0.529 1 : 4.53 0.507 1 : 4.20 0.470 1 : 4.04 0.484 65°C), T 3 (drying temperature 60°C) and T 5 (drying
R.R. = Rehydration ratio, COR = Coefficient of rehydration temperature 55°C) ascorbic acid content was recorded
Table 5. Ascorbic acid content (mg/100 gm) in different treatments of raw and
36.09 mg/100 g, 55.36 mg/100 g and 96.04 mg/100 g,
dehydrated cauliflower after dehydration and during storage period (MFB) respectively indicating that ascorbic acid content increased
Treatment Initial Ascorbic acid content (mg/100 g) (MFB) with decrease in drying temperature. Ascorbic acid content
(months of storage) deteriorated rapidly and after 6th month, it remained highest
0 2 4 6 in T5 (12.78 mg/100 g) followed by T1 (10.79 mg/100 g),
T1 502 36.09 15.07 12.11 10.79 T6 (8.32 mg/100 g), T3 (7.91 mg/100 g), T4 (6.55 mg/100 g)
T2 404 24.82 17.62 14.31 6.53 and T2 (6.53 mg/100 g). Although ascorbic acid content after
T3 440 55.36 14.28 11.51 7.91
T4 358 59.27 11.32 9.72 6.55
dehydration was low in T1 and T2 after 6 months storage, loss
T5 441 96.04 19.29 15.61 12.78 of ascorbic acid was low (70.10 and 73.69% respectively)
T6 357 67.70 10.65 8.54 8.32 and retention of ascorbic acid was maximum (29.90 and
LSD (P=0.05) 114 14.71 4.97 4.21 3.66 26.31%, respectively). Rate of loss of ascorbic acid decreased
LSD (P=0.01) N.S. 20.62 6.97 5.90 5.13 and retention of ascorbic acid increased with increase of
Table 6. Losses and retention of ascorbic acid content of dehydrated cauliflower dehydration temperature during storage (Table 6).
after dehydration and during storage (MFB)
Treatment Loss (%) Retention (%) Bawa and Saini (1986) and Kadam et al. (2005) also recorded
(months of storage) (months of storage) similar range of ascorbic acid in dehydrated cauliflower.
2 4 6 2 4 6 Losses of vitamins during processing occur either due to
T1 58.24 66.4 70.10 41.76 33.56 29.90 oxidation or by dissolving into water (Vail et al., 1978). Loss
T2 29.00 42.34 73.69 71.00 57.66 26.31 of ascorbic acid to the extent of 63.2% has been reported
T3 74.20 79.20 85.71 25.80 20.80 14.29 by Raina et al. (1982) which is slightly lower than present
T4 80.90 83.60 88.94 19.10 16.40 11.06
T5 79.91 83.74 86.69 20.09 16.26 13.31
finding probably due to different blanching time (Kadam et
T6 84.26 87.38 87.71 15.74 12.62 12.29 al., 2005) and pretreatemnt (Mulay et al., 1994).
Table 7. Sensory quality evaluation after dehydration and after storage period Sensory evaluation of rehydrated product indicated that T2
of six months of dehydrated cauliflower (hot water blanching + 0.125% KMS + microwave blanching
Treatments Storage period (in months) + drying at 65 °C), T3 (hot water blanching + 0.125% KMS +
Colour Flavour Texture Over all drying at 60 °C), T5 (hot water blanching + 0.125% KMS +
acceptability drying at 55 °C) and T4 (hot water blanching + 0.125% KMS
0 6 0 6 0 6 0 6
+ microwave blanching + drying at 60 °C) recorded high
T1 8 6 8 5 8 6 8 6
T2 9 7 9 7 9 6 9 7 score for colour (8-9), flavour (8-9), texture (8-9) and overall
T3 9 6 9 6 9 7 9 6 acceptability (8-9). In storage, the sensory qualities decreased
T4 9 7 9 7 9 7 9 7 rapidly, the overall acceptability of T3, T4 and T5 decreased to
T5 8 6 9 6 9 6 8 6 5, 5 and 6, respectively. However, the colour and flavour and
T6 7 5 8 6 8 7 8 6 overall acceptability of T2 could be retained even after six
microwave blanching (T1, T3 and T5) exhibited lowest coefficient months to acceptable score of 7. Previous reports (Srivastava
of rehydration values in T1 (0.486) at 65°C of drying followed by and Sulebele, 1975; Kaur and Singh, 1981; Raina et al., 1982;
T3 (0.528) at 60 °C and T5 (0.699) at 55°C in that increasing order Bewa and Saini, 1986; and Abhay et al., 2006) also indicated
(Table 4). The trend was similar in microwave blanched treatments that chemical like potassium metabisulphite was effective in
(T2, T4, T6). At same temperature of drying the rehydration coefficient improving quality in respect of colour, flavour, texture and
of microwave blanched treatments was equal or lesser than untreated. overall acceptability.
 Effect of pre-treatment and drying temperature on quality of dehydrated cauliflower
Thus considering the dehydration characters and quality after
dehydration and storage it can be concluded that T2 (hot water
blanching + 0.125% KMS + microwave blanching + drying at
65°C) was the best treatment followed by T4 (hot water blanching
+ 0.125% KMS + microwave blanching + drying at 60°C) and
T5 (hot water blanching + 0.125% KMS + drying at 55°C). In T2,
time taken for complete dehydration (445 minutes) and moisture
content (3.62%) after dehydration was least. Further the moisture
content after six month of storage was also less (9.63%), drying
rate (135.74%/h) and dehydration ratio (10.70) was medium
after dehydration. Ascorbic acid retention was maximum during
storage in the treatment (T2). Sensory evaluation also supported
the superiority of this treatment.
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Received: March, 2013; Revised: December, 2013; Accepted: February, 2014
 Journal

Journal of Applied Horticulture, 16(1): 76-79, 2014 Appl

Planting density and corm size effects on flower yield and

quality of cut-freesia (Freesia hybrid) in Ethiopia

Tewodros Bezu1* and Nigusse Kassa2

1
School of Plant Sciences, College of Agriculture and Environmental Sciences, Haramaya University, Ethiopia.
2
College of Agriculture & Veterinary Medicine, Jimma University, Ethiopia.*E-mail: tedrosneguse@yahoo.co.uk

Abstract
Greenhouse experiment was conducted at Freesia Ethiopia Plc., located at Sululta, Ethiopia, to determine the effects of planting density
and corm size on flower yield and quality of cut-freesia. Planting densities 90, 100 and 110 corms per m2 and corm sizes of 3, 3.5 and
4 cm in circumference were evaluated on two varieties ‘Volante’ and ‘Casino’ using Randomized Complete Block Design in factorial
arrangement (3 x 3 x 2) with three replications. Emergence date, flowering date, cut flower yield and quality parameters were recorded
and analyzed. Consequently, increment of planting density resulted highest number of cut-flowers. Corm size difference positively
influenced the stem length, spike length and cut-flowers yield. Significant interaction effects were also found between corm sizes and
varieties on yield and quality traits. In general, using the biggest corm and highest planting density exhibited superior result for the
greenhouse production of the stated varieties. However, to come up with complete recommendations, further investigations should be
conducted in line with other agronomic packages and varieties of economic viability.
Key words: Cut-flower, freesia corm, freesia hybrid, Ethiopia

Introduction planting spacing in relation to corm size, varietal screening etc.

Floriculture is relatively new sub-sector in Ethiopia as for long
the production of flowers had been limited to few genera/species
of field flowers (like Allium spp.). Even if the country is known
for its potential for fertile soil, abundant water, proximity to
main markets, growing cut-flowers for export purpose is new,
dating back only to 1992 (EHPEA, 2006). The country earned
114 million dollars from the floriculture industry in 2008 (Biruk
et al., 2013). The rapidly growing flower sector in Ethiopia has
now become the fourth foreign-currency generator of the country
next to the top three: coffee, oilseeds and cereals (Gebre, 2011).
In this regard, many local and international investors are getting
involved to take advantage of the manifold potentialities.
Freesia, a genus of about 14 species, belongs to the Iridaceae
family and is native to mountainous areas of South Africa. It
is usually grown as cut-flower. Their appealing shapes make
them suitable for flower arrangement, and their wide range of
colour increases their versatility. The flowers are popularly used
for wedding in which it is the seventh wedding anniversary
flower and in the language of flowers, freesias are the symbol
of innocence and friendship (Teleflora, 2009). It is also used in
making fragrant additions to bouquets and body flavours since
most of the cultivars are highly fragrant. Moreover, Freesias are
also used as a forced pot crop, becoming popular when cultured in
cool houses or in hobby greenhouses (Wang, 2006). Owing to the
existence of the favourable investment environment and abundant
resources, freesia is known to be grown near the capital city, Addis
Ababa, Sululta (25 km from Addis Ababa) since 2007.
Due to the fact that the plant is new to Ethiopia’s agro-ecological
condition, a number of production constraints such as lack of
recommended agronomic practices in plant protection, fertigation,
are being observed on the already established farm. Among these,
problems associated with the plant spacing and planting materials
selection in terms of corm size are of major considerations.
Proper plant spacing is important for providing good open
position for sunlight, availability of moisture and nutrients vital
for successful crop production and quality (Sanjib et al., 2002).
Besides, it affects yield of quality spikes and corms (Singh et al.,
2000). Plant spacing of freesia corms or cormels is dependent on
time of the year, cultivars and size of corms (Cuppen, 2006; Singh
et al., 2011). For cut flower production, growers usually use corms
sizes from 5 to 7 cm in circumference, planting depth and density
of 5 cm and, 96 to 120 corms per m2, respectively (De Hertogh,
2008). Therefore, the study was conducted with objectives of
evaluating the effects of different planting densities, corm sizes
and their interaction on flower yield and quality of freesia.
Materials and methods
Description of the study site: The study was conducted in a
plastic greenhouse at Freesia Ethiopia PLC located at Sululta
District of North Shewa Administrative Zones of Oromia Region,
Ethiopia which is 20 km away from Addis Ababa, geographically,
situated at 9° 11’ latitude and 38° 39’ longitude at an altitude of
2785 m. The inside temperature and relative humidity (RH) were
controlled within the range of 15-20°C and 65-75%, respectively
using a computerized system (Hogendoren systems). The soil
type of the area is Luvisoils.
Experimental materials, design and treatments : Two freesia
hybrid varieties, Volante and Casino, having three different corm
sizes 3, 3.5 and 4 cm in circumference were imported from The
Netherlands and tested with planting density of 90, 100 and 110
corm per m2 at 5 cm planting depth. The experiment was laid
 Planting density and corm size effects on flower yield and quality of cut-freesia
in 3x3x2 (planting densities, corm sizes and varieties) factorial
arrangements using a Randomized Complete Block Design
(RCBD) and replicated three times. Each experimental plot was
0.6 m x 1.2 m=0.72 m2 with 8 rows and spacing of 0.125 m,
0.45 m and 0.5 m between rows, plots and blocks, respectively.
Fertilizer was applied as per the recommendation of soil analysis
result throughout the growing season and other management
practices like supporting, weeding and pest management were
performed whenever necessary. Fertigation was conducted using
a computerized system.
Emergence date, days to flowering, stem length, number of florets,
total cut-flower yield and vase life were recorded and analyzed
from 10 to 16 randomly selected plants depending on the plant
population size per plot, except for cut-flower yield in which the
data were recorded on whole plot basis.
Statistical analysis: The replication-wise mean values of
different entries were subjected to three way factorial experiment
in Randomized Complete Block Design (RCBD) analysis of
variance according to Montgomery (2005) using SAS version
9.1 (SAS Institute, 2001) statistical package after the data were
checked for meeting the various ANOVA assumptions.
Results and discussion
Emergence date: The interaction effects of varieties with corm
sizes were found highly significant (Table 2) while other two
way and three way interaction were non-significant with respect
to emergence date. Volante with bigger corm size emerged
earlier (12 days) than others. While Casino of corm size 3 cm in
circumference emerged late. On the other hand, all corm sizes
of variety Casino took the longest period for emergence. This
might be due to the inherent genetic differences between the
two varieties as far as dormancy period is concerned. On top of
this, within both vaieties, as the corm size increased emergence
date was early. The considerable effect of corm size on days of
emergence may be attributed to the length of dormancy period,
which normally is short in larger corms. These findings are in
agreement with the previously reported investigation in gladiolus
(Uddin et al., 2002).
Days to flowering: The number of days required for flowering
is the major concern in cut-flower production due to the fact that,
it enables the grower to develop production scheme as well as
marketing plan. Highly significant difference between varieties,
corm sizes and planting densities were obtained in relation to
flowering date. In addition, the combined effect between varieties
and corm sizes was also statistically significant. However, three-
way interaction effects among planting densities, corm sizes
and varieties; and two-way interaction effects between planting
densities and varieties; and planting densities with corm sizes
were non-significant. With the increment of corm size within
varieties, the days to flowering showed reduction (Table 2). This
is in line with the finding of Uddin et al. (2002) in gladiolus and
Kapczynska (2008) in Sparaxis tricolor. This might be due to
the fact that flower initiation is dependent on the availability of
food reserves in the corm (Rees, 1992). Moreover, comparing the
two varieties, Volante with corm size 4 cm in circumference was
the earliest in flowering (115 days). On the other hand, Casino
with corm size 3 cm in circumference took the longest time for
77
flowering (136 days). In any of the corm sizes and varieties
combinations, Casino flowered later than Volante. Variation in
days required for spike emergence was due to difference in genetic
composition of the cultivars that responded differently to the
environment (Ahmed et al., 2010). Variation in planting density
was also found to be one of the factors that affected flowering
time (Table 1). Subsequently, as the planting density increased
from 90 to 110 corms per m2, the days to flowering was reduced
from 127 to 123 days. The reason could be the competition for
available nutrients and light, which forced the plants to shorten
their life cycle.
Total cut-flower yield: The combination effects of variety with
corm size revealed significant difference with regard to cut
flowers harvested per unit area. In addition, significant differences
resulted with varying plant densities. However the three way
interactions among variety, planting density and corm size; and
two way interactions between variety and planting density as
well as planting density with corm size were non-significant.
The highest cut-flowers (124 cut-flowers/m2) were harvested
from the biggest corm size (4 cm) of Volante while the least (29
cut-flowers/m2) was from smallest corm size of Casino (Table
2). This result is in agreement with studies conducted on Saffron
(Omidbeigi et al., 2003). As the planting density increased from
90 to 110 corms per m2, the cut-flowers yield also increased
from 61.4 to 76.8 per m2 (Table 1). This result illustrated the
existence of a positive relationship between planting densities
and marketable yield. Obviously, as the number of plants per
unit area increased, more marketable stems per unit area could
be obtained provided that each plant has the capability to produce
more under high competition. These results are in agreement with
those of Mili and Sable (2003).
Stem length: Highly significant variation among corm size was
noticed for stem length. However, both the two way and three-way
interactions among planting densities, corm sizes and varieties
exhibited no significant difference. In addition, main effects of
planting density and variety showed non-significant effect on stem
length. The biggest corm size (4 cm in circumference) produced
higher average stem length (30.5 cm) while least average stem
length (25 cm) was obtained from smallest corm size (3 cm
in circumference) (Table 1). This could be due to more stored
food materials in large sized corms, which helped in early and
rapid vegetative growth (Uddin et al., 2002). On the other hand,
though non-significant differences were found regarding planting
density differences, there was slight increment in the average
stem length with the increasing planting density (Table 1). It is
in agreement with the study on gladiolus grown under polythene
tunnel (Roychowdhury, 1989).
Spike length: Highly significant variability was exhibited in the
average spike length with the main effects of varieties and corm
sizes differences. However, there was non-significant variation
for this trait with respect to two ways as well as three way
interactions. In addition, planting densities influence on spike
length was also statistically non-significant. Casino produced
longer average spike length (12.7 cm) than Volante (8.9 cm).
The biggest corm size (4 cm in circumference) produced longer
spike length (11.8 cm) than medium (3.5 cm in circumference)
and smallest (3 cm in circumference) corms (Table 1). The
observed result could be due to the presence of more reserve
78 Planting density and corm size effects on flower yield and quality of cut-freesia

Table 1. Effect of planting density, corm size and variety on flowering and quality of cut-freezia at Sululta, Ethiopia
Effects and levels Days to flowering Total cut-flowers per m2 Stem length (cm) Spike length (cm)
Planting density
90 corms m-2 127.1a 61.4b 27.2 10.7
100 corms m-2 126.6ab 69.1ab 27.9 10.8
110 corms m-2 123.6c 76.8a 28.9 10.9
LSD (P=0.05) 1.33 11.31 ns ns
Corm size (circumference)
3.0 cm 128.7a 43.8c 25.0c 10.1b
3.5 cm 126.6b 72.7b 28.5b 10.5 b
4.0 cm 121.9 c 90.9a 30.5a 11.8 a
LSD (P=0.05) 1.32 11.31 2.0 1.4
Variety
Volante 118.5b 94.1a 27.5 8.9 b
Casino 133.0a 44.2b 28.5 12.7 a
LSD (P=0.05) 1.1 9.2 ns 1.2
Means followed by different letters within a column are significantly different at 0.01 level of probability; ns= non-significant difference
Table 2. Interaction effect of variety with corm size on emergence date, days to flowering, number of florets per spike and vase life of cut-freesia
Variety Corm size (cm) Emergence date Days to flowering Number of florets Vase life (days) Total flower per m2
Volante 3.0 14.4c 121.3d 6.5e 12.9e 59.0c
3.5 13.9d 118.7e 6.9ae 13.1e 99.0b
4.0 12.2 e
115.4 f
7.3 ab
13.8 bcd
124.0a
Casino 3.0 17.9 ab
136.1 a
7.1 ad
13.9 ac
29.0d
3.5 18.0a 134.6b 7.3ac 14.2a 46.0c
4.0 18.0 a
128.3 c
7.4 a
14.0 ab
58.0c
LSD (P=0.05) 0.2 1.0 0.1 0.1 9.2
Means followed by different letters within a column are significantly different at P=0.01; ns= non-significant difference
food in large sized corms, which may result in early growth and result suggested the presence of the genetic variations between
development, thereby the plant could produce longer stem as well the two varieties. Besides, as the corm size increased, increasing
as spike. Singh (2000) and Uddin et al. (2002) also reported the trend was observed in vase life. This could be due to the presence
same result in gladiolus. Conversely, non-significant difference of more reserved food in the large sized corms that ensured
was observed on the mean value of spike length measurement
vigorous growth and development of the plant, and also helps in
from medium (3.5 cm in circumference) and smallest (3 cm in
prolonging the vase life of the cut-flowers.
circumference) corm.
Number of florets: Significant combined effects between Planting density of 110 corms per m2 exhibited significant
varieties with corm sizes were obtained for the average number improvement in total cut-flower yield. As the corm size
of florets count. However, statistical differences were not detected increased, the early flowering date was recorded. Flower stalk
between planting densities; two way interactions between planting length, number of florets, spike length, vase life, and cut-flowers
densities with varieties; planting densities with corm sizes; and production increased with bigger corm. Therefore, the results
three-way interactions. The interaction effects of varieties with suggest that the use of biggest corm (4 cm in circumference) has
corm sizes on number of florets (Table 2) showed increasing imperative advantages. There was also varietal difference on
trend in the mean number of florets of both varieties with the rise yield and quality parameters evaluated. Volante was superior in
of corm size. It may be due to high food reserve in large sized
terms of total flower yield, earlier in emergence and flowering.
corms (Uddin et al., 2002). This is in accord with Uddin et al.
Conversely, Casino was superior with regard to quality attributes
(2002) and Singh (2000) results who reported higher number of
spikelet (florets) from large sized gladiolus corm. However, there such as spike length, number of florets and vase life. Therefore,
were non-significant differences among all combinations, except the pronounced variability of the two varieties may suggest that
between Volante of corm size 3 cm in circumference (Table 2). they are distinct in their genetic makeup. The combined effect of
Furthermore, the biggest corm size of both varieties produced cultivars and corm sizes were significant on emergence date, days
more florets as compared to the medium and smallest corm sizes. to flowering, average number of floret and total cut-flowers.
In contrast, Volante of corm size 3 cm in circumference produced
the least number of florets. In general, it can be concluded that planting density, variety and
corm size difference have valid effects on flower yield and quality
Vase life: Varieties and corm sizes showed highly significant of cut-freesia and using the biggest corm (4 cm in circumference)
interaction effects on average vase life of cut freesias. Volante of
and highest planting density (110 corms per m2) is advisable for
corm size 3 cm in circumference exhibited least vase life (12.9
cut flower production of the tested varieties. However, to come
days) (Table 2). However, there was non-significant difference
among corm sizes of Casino. Nevertheless, the mean value of up with complete package of practices, informaiton on planting
Casino with corm size 3.5 cm in circumference was found to be depth, environmental influences (temperature, relative humidity,
superior (14.2 days). Additionally, Casino with all corm sizes light etc.), fertigation and varieties of economic viability will be
showed longer vase life as compared to Volante. The observed useful.
 Planting density and corm size effects on flower yield and quality of cut-freesia
Acknowledgement
We thank Freezia Ethiopia Plc and its management team for
sponsoring the overall research project and Jimma University
College of Agriculture and Veterinary Medicine for kind support
during write-up of the report.
References
Ahmed, M.J., T. Bashir, A. Yaqoob, M.S. Jillani and M. Saeed, 2010.
Effect of plant spacing on vegetative and reproductive growth of
gladiolus cultivars. Sarhad J. Agric., 26(4): 539-543
Biruk Mellesse, Nigussie Kassa and Ali Mohammed, 2013. Yield and
quality of statice [Limonium sinuatum (L.) Mill.] as affected by
cultivars and planting densities. African Journal Plant Sciences,
7(11): 528-537.
Cuppen, A. 2006. Feasibility of freesia corm (let)s production in china,
in ganzu province. M.Sc. Thesis, University of HAS Den Bosch,
the Netherlands.
DeHertogh, A. 2008. Growing freesia, <http://www.growertalks.com/
archive/articles/33.asp>
EHPEA (Ethiopia Horticultural Product Exporter Association), 2006.
Addis Ababa, Ethiopia.
Gebre, S. 2011. Underlying causes of business failure of floriculture
investment in Ethiopia. MSc. Thesis, Addis Ababa University,
Ethiopia.
Kapczynska, A. 2008. Effect of corm size on the yield of Sparaxis tricolor
Ker-Gawl grown in the field, <http://psjc.icm.edu.pl/psjc/cgi-bin/
getdoc.cgi?AAAA022762>
Mili, R. and A.S. Sable, 2003. Effect of planting density and nitrogen
levels on growth and flower production of calendula (Calendula
officinalis L.). Indian J. Hort., 60(4): 399-403.
79
Montgomery, D.C, 2005. Design and Analysis of Experiments. 6th ed.
John Wiley and Sons, Inc., USA. pp. 97- 203.
Omidbeigi, R., A. Rezaii, B. Sadegi and M. Zeiaratnia, 2003. The effect
of corm weight in the yield of saffron in nishaboor climate. Paper
collection of third national Iranian congress of saffron. pp. 34-37.
Rees, A.R. 1992. Ornamental Bulbs, Corms and Tubers. C.A.B
International, UK.
Roychowdhury, N. 1989. Effect of plant spacing and growth regulators
on growth and flower yield of gladiolus grown under polythene
tunnel. Acta Hort., 246: 259-264.
Sanjib, S., M.C. Talukdar, S. Sharma, R.L. Misra and M. Sanyat,
2002. Effect of time, spacing, and depth of planting on gladiolus.
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NC. USA.
Singh, A.K., S. Chetan and C. Singh, 2000. Effect of spacing and zinc
on production of corms and cormels in gladiolus. Hort. J., 13(2):
61-64.
Singh, K.P. 2000. Growth, flowering and multiplication in Gladiolus
cultivar ‘Aarti’ as affected by grades of mother corm and cormel. J.
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Singh, G., H.S. Sekhon, G. Singh, J.S. Brar, T.S. Bains and S.
Shanmugasundaram, 2011. Effect of plant density on the growth
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Received: October, 2013; Revised: January, 2014; Accepted: February, 2014
 Journal

Journal of Applied Horticulture, 16(1): 80-84, 2014 Appl

Effect of processing and storage on bioactive compounds and

antioxidant activity of carrot juice

S. Kapoor* and P. Aggarwal

Department of Food Science and Technology, Punjab Agricultural University, Ludhiana-141 004,
*E-mail: swatikapoor74@yahoo.com

Abstract
Fresh carrot juice is one of the widely consumed vegetable juice during winter season. Recipe for ready-to-serve carrot ginger juice
was standardized with the addition of sugar, salt and ginger on the basis of sensory evaluation. The effect of processing and storage
on bioactive compounds and antioxidant activity of control and ready-to-serve carrot ginger juice was studied. Among the various
combinations prepared, 4% sugar, 0.6% salt, 0.8% ginger and 0.05% citric acid showed highest overall acceptability on the basis
of sensory scores and was chosen for further analysis. Total phenolic content was determined by using Folin–Ciocalteu reagent and
antioxidant activity was determined by using DPPH assay. During processing, significant losses were found in bioactive compounds and
antioxidant activity of control and carrot ginger juice. The study revealed that carrot ginger juice was found to retain more antioxidant
activity compared to control juice due to addition of ginger. Storage of six months had no significant effect on TSS and acidity of
processed carrot juices. However, storage led to significant decrease in bioactive compounds and thus decreased antioxidant activity
of carrot products.
Key words: Carrot juice, bioactive compounds, antioxidant activity, processing, storage

Introduction Some studies have shown that absorption of carotenoids from

Processed and refrigerated vegetable juices are a fast-growing
segment within the beverage industry, due to the fact that a
high consumption of vegetables has been associated with the
prevention of cancer, cardiovascular diseases and other ailments.
The beneficial properties of vegetables have been attributed
to the presence of antioxidant compounds such as vitamin C,
carotenoids and phenolic compounds. Carrot based products
are of great health impact because of their high content of
carotenoids, some of which also contribute to vitamin A activity
such as β-carotene. They also act as free-radical scavengers (Bast
et al., 1998; Bramley, 2000). It has been shown that the presence
of α- and β-carotene in blood has a protective effect against
atherosclerosis (D’Odorico et al., 2000). Carrots also contain
substantial amount of vitamin C and phenols. Phenols are known
to interrupt lipid peroxidation induced by reactive oxygen species.
Vitamin C has been reported to prevent free-radical-induced
damage to DNA quenching oxidants, thus, acting as antioxidants
(Oviasogie et al., 2009).
In recent years, carrot juice has become an important food
commodity for these health reasons. Keeping in view the
nutritional and therapeutic value of carrots and to make them
available throughout the year, carrot juice in combination
with other vegetable and fruit juices can be prepared, stored
and evaluated for various characteristics. The importance of
naturally occurring bioactive compounds in the maintenance
of health is raising considerable interest among scientists, food
manufacturers and consumers as the trend of the future is moving
toward functional food with specific health effects. Processed
carrot juice blended with ginger juice serve as a good source of
carotenoids and polyphenols, providing consumers with nutritious
and antioxidant rich ready to serve beverage round the year.
unprocessed foods is low but it increases with mild processing
(Failla and Chitchumroonchokchai, 2005). Thus, processing
can have beneficial effects on human health. On the other hand,
a significant loss of nutrients may occur as a result of thermal
degradation or leaching during blanching, pasteurization and
dehydration as well as during storage, home handling and cooking
(Kalt, 2005). Hence, the present investigation was carried out to
study the effect of processing and storage on bioactive compounds
and antioxidant activity of carrot juice.
Materials and methods
Carrots were procured during the last week of January from the
village Ranuuan, Samrala, Distt. Ludhiana, Punjab.
Carrot juice processing: Fresh carrots were washed thoroughly,
peeled and cut from the top and bottom. Carrot juice was extracted
in a screw type juicer extractor. Carrot juice was optimized with
different sugar concentrations such as 2, 4, 6% and different
ginger concentration (0.8 and 1%); constant salt (0.6%) and citric
acid content (0.05%). Finally the particular level was selected
on the basis of sensory evaluation (Table 1). All the ingredients
were mixed and the juice was heated in a stainless steel vessel
at 85oC for 3 min. Glass bottles of 200 mL capacity were pre-
washed and pre-sterilized in boiling water. Bottles were filled
with hot juice and corked. They were finally heated at 100 oC for
20 min in water and gradually cooled under running tap water.
Control juice was prepared in the same manner without any added
ingredients. Finally the bottles were kept at room temperature
for six months.
Analytical methods: Fresh carrot juice, control and ready-to-
serve processed carrot juice were evaluated for TSS, acidity,
 Effect of processing and storage on bioactive compounds and antioxidant activity of carrot juice
ascorbic acid, total carotenoids, β-carotenoids, lycopene, total
phenolic content and antioxidant activity. TSS, acidity, ascorbic
acid, total carotenoids, β-carotenoids and lycopene content was
determined as per Ranganna (1986). Total phenolic content
was determined by using Folin–Ciocalteu reagent (Swain and
Hillis, 1959). A standard curve was plotted by taking known
amount of gallic acid as reference standard and concentration
was calculated from the standard curve. Free radical scavenging
activity was determined by DPPH (di phenyl picryl hydrazyl)
method according to Brand-Williams et al. (1995) with some
modifications. Methanolic extract of 5 g sample was taken
for antioxidant activity analysis and calculated according to
the following formula. BHT was taken as a standard at a fixed
concentration of 5mg/mL.
Radical Control OD (0 min) – Sample OD (30 min)
scavenging = x 100
activity(%) Control OD (0 min)
The sensory quality of processed carrot juice was assessed for
appearence, flavor, mouthfeel and overall acceptability. Different
attributes of the product were evaluated by a panel of seven judges
using 9-point Hedonic rating scale (Amerine et al., 1965). Results
were analyzed statistically using completely randomized design
as discussed by Singh et al. (1991).
Results and discussion
Optimization of carrot juice recipe on the basis of sensory
evaluation: No significant effect of different levels of sugar,
salt and ginger was found on appearance of the juice. However
there was significant (P≤0.05) difference in the flavor, mouth
feel and overall acceptability due to different levels of sugar
and ginger. On the basis of sensory evaluation carrot juice with
4% sugar, 0.8% ginger and 0.6% salt received highest score in
terms of overall acceptability and was chosen for further studies
(Table 1).
Effect of processing on bioactive compounds and antioxidant
activity of carrot juice
TSS and acidity: The TSS of fresh carrot was found to be 8.20
o
Brix (Table 2). Total soluble solids for carrots ranged from 8.46
to 9.98 oBrix (Sandhu et al., 1988). TSS of fresh juice was same
as that of fresh carrot i.e. 8.20 oBrix. TSS of control juice was
recorded to be 10.00 oBrix whereas TSS of carrot ginger juice was
found to be13.00 oBrix due to addition of sugar. Acidity of 0.05
per cent was found in fresh carrots. Acidity in carrot cultivars was
found to be 0.06 per cent in ‘Sel 21’, 0.04 per cent in ‘PC-34’,
0.06 per cent in ‘Ambala local’ and 0.05 per cent in ‘Nantes’ (Sra
et al., 2011). Heat processed control juice and carrot ginger juice
had 0.06 and 0.08 per cent acidity, respectively. Increase in the
Table 1. Optimization of carrot ginger juice recipe on the basis of sensory evaluation*
Sample Appearance
2% sugar + salt + 0.8% ginger 8.42±0.60
2% sugar + salt + 1% ginger 8.42±0.44
4% sugar + salt + 0.8% ginger 8.57±0.44
4% sugar + salt + 1% ginger 8.57±0.34
6% sugar + salt + 0.8% ginger 8.57±0.44
6% sugar + salt + 1% ginger 8.57±0.53
CD (P≤0.05) NS
*Mean ± SD (Standard deviation) of score by seven penalist
81
acidity of altered juice compared to control sample was due to
the addition of citric acid.
Total carotenoids, β carotene and lycopene: Table 2 depicts the
effect of processing on total carotenoids and β-carotene of carrot
products. Fresh carrots were found to have 13.35 mg/100g total
carotenoid and 10.13 mg/100g β-carotene. The present study
corroborates the findings of Sethi and Anand (1983) who reported
the β-carotene content of raw carrot to be 11.88 mg/100g but
Gopalan et al. (1997) reported lower (6.46 mg/100g) β-carotene
content in fresh carrots. The variation in carotenoids may be due
to variety, maturity, growing conditions, growing season and
the part of the root sampled (Hart and Scott, 1995).There was
a reduction in total carotenoids and β-carotene in carrot juices
after processing. The total carotenoid content of raw juice, heat
processed control and carrot ginger juice was 12.42 mg/100g,
10.10 mg/100g and 10.62 mg/100g, respectively. β-carotene
content of raw juice, processed control and carrot ginger juice was
found to be 9.86, 8.50 and 8.82 mg/100g, respectively. Lycopene
content in raw juice, processed control and carrot ginger juice
was 3.16 mg/100g, 2.93 mg/100g and 2.96 mg/100g, respectively.
The decrease in lycopene content may be due to its co-planarity
structure being very reactive to oxidation (Lin and Chen, 2005).
Similar results were found by Dhaliwal and Hira (2001) who
reported that fresh carrot: beetroot (95:5) had 3.56 mg/100g of β-
carotene which reduced to 2.88 mg/100g after pasteurization, thus
accounting for 19.10 % reduction. The loss of total carotenoids
in juice during preparation may be due to oxidation or thermal
degradation of unsaturated total carotenoids.
Ascorbic acid: Fresh carrots contained 4.31 mg/100g ascorbic
acid (Table 2). According to Alasalvar et al. (2001) vitamin C
content varied between 1.25 and 5.33 mg/100g, being lowest
in white and highest in orange varieties of carrot. Ascorbic
acid content in raw juice, processed control and carrot ginger
juices were 2.24, 1.79 and 1.94 mg/100g, respectively. Losses
of ascorbic acid in heat processed juice may be due to thermal
degradation. Grewal and Jain (1982) also reported 27.62 % loss
of ascorbic acid during processing of carrot juice beverage.
Total phenols: Raw juice contained 34.98 mg/100g galic acid
equivalents (GAE) of total phenols while in control and altered
juice it was found to be 29.95 mg/100g GAE and 34.84 mg/100g
GAE, respectively (Table 2). Teixiera et al. (2009) found that
fresh carrot juice had 334 mg L-1 of total phenol which reduced to
306 mg L-1 after heat treatment at 90o C for 30 sec. Carrot ginger
juice had higher phenolic content than control juice due to the
addition of ginger to carrot juice. Tarko et al. (2010) found that
addition of any of the seasonings to apple chips such as cinnamon,
garlic, ginger, pepper, mint, onion increased the values of total
Flavor Mouth feel Overall acceptability
7.85±0.37 7.85±0.37 8.05±0.23
8.00±0.40 7.85±0.55 8.09±0.23
8.42±0.53 8.28±0.39 8.42±0.18
8.28±0.48 8.00±0.57 8.28±0.26
8.00±0.57 8.00±0.40 8.14±0.20
8.14±0.55 8.14±0.55 8.28±0.31
0.14 0.13 0.14
82 Effect of processing and storage on bioactive compounds and antioxidant activity of carrot juice

phenolic content which explains the higher content of phenolics juice and altered carrot juice decreased non significantly during
in carrot ginger juice. storage period of six months. It decreased from 10.00 to 9.73 oBrix
in control carrot juice and from 13.00 to 12.60 oBrix in carrot
Antioxidant activity: Fresh carrots were found to have 23.83 %
ginger juice. Similar results have also been reported by Aggarwal
scavenging activity (Table 2). Radical scavenging capacity in raw
carrots decreased continuously with increase in reaction time and et al. (1995) who found negligible change in the total soluble
maximum activity was shown at 30 minutes and became stable solids of processed tomato juice from six varieties during six
thereafter (Fig.1). Antioxidant activity of raw carrot juice was months storage. There was a slight increase in the acidity during
found to be 23.39 % (Table 2) and maximum antioxidant activity storage. However, the overall increase was non significant. Carrot
as determined by DPPH assay was shown at 30 minutes (Fig. 1). ginger juice also showed non significant increase in acidity during
After processing the antioxidant activity of control and carrot storage. Dhaliwal and Hira (2001) also observed non-significant
ginger was found to be 21.73 and 23.37 %. Similar results were change in the acidity of pasteurized carrot-beetroot juice during
found by Teixiera et al. (2009) who observed that antioxidant 6 month storage.
capacity of fresh carrot juice was 211 milimol trolox L-1 which Table 3. Effect of storage on total soluble solids and acidity of carrot
decreased to 199 milimol trolox L-1 after heat processing at 90o C juice*
for 30 seconds. Decrease in the antioxidant activity may be due Storage TSS (oBrix) Acidity (%)
to decrease in the levels of bioactive compounds such as ascorbic months Control Carrot ginger Control Carrot ginger
juice juice
acid, total carotenoids and phenolic compounds. Duddone et al.
0 10.00±0.03 13.00±0.02 0.06±0.01 0.08±0.02
(2009) found that antioxidant activity were correlated to phenolic
1 9.94±0.10 13.06±0.05 0.06±0.03 0.09±0.04
compound concentration and Teixeira et al. (2009) found positive
2 9.87±0.11 12.87±0.11 0.07±0.04 0.10±0.02
correlation between ascorbic acid and DPPH values (R2 = 0.807)
3 9.87±0.11 12.87±0.10 0.08±0.02 0.11±0.03
in processed carrot juice. Thus, bioactive compounds in carrot
juice have positive relation with antioxidant activity, as a result 4 9.80±0.03 12.80±0.02 0.08±0.01 0.12±0.03
antioxidant activity increases with increase in concentration of 5 9.73±0.09 12.67±0.11 0.06±0.03 0.10±0.02
bioactive compounds and vice-versa. Carrot ginger juice had 1.64 6 9.73±0.11 12.60±0.00 0.05±0.01 0.10±0.01
% more antioxidant activity than control juice. This increase is F test NS NS NS NS
due to the addition of ginger in carrot juice, as ginger is found to *Values are Mean ± SD (Standard deviation) of three replicates
have good antioxidant properties (Shirin and Jamuna, 2010). Both Total carotenoids, β-carotene and lycopene: Effect of storage
control carrot juice and carrot ginger juice showed maximum on total carotenoids, β-carotene and lycopene of control and carrot
antioxidant activity at 40 minutes (Fig. 1). ginger juice is shown in Table 4. Total carotenoids decreased
Effect of storage on bioactive compounds and antioxidant significantly (P≤0.05) from 10.10 mg/100g to 7.75 mg/100g in
activity of carrot juice control juice and from 10.62 mg/100g to 7.97 mg/100g in carrot
TSS and acidity: As can be seen from Table 3, TSS of control ginger juice, after six months storage at room temperature. β-
carotene content showed significant (P≤0.05) reduction during
storage in both control and carrot ginger juice. In control, juice
β-carotene decreased from 8.50 mg/100g to 5.65 mg/100g and
in carrot ginger juice from 8.82 mg/100g to 5.87 mg/100g.
Dhaliwal and Hira (2001) also observed the decreasing trend in
β-carotene content during six month storage of carrot: beetroot
(95:5) beverage. The decrease in total carotenoids and β-carotene
content may be due to oxidation of highly unsaturated carotenoid
structure (Kidmose et al., 2002). Chen et al. (1996) observed
the stability of carotenoids during storage of carrot juice by
subjecting the carrot juice to storage at different temperatures
and in light and dark for three months. They found that α-
carotene, β-carotene and vitamin A in carrot juice decreased
with increasing storage temperature and light is more destructive
to carotenoids than darkness. Teixeira et al. (2009) found that
Fig. 1. Radical scavenging capacity of fresh and processed carrot juice β-carotene concentration of carrot juice decreased throughout
Table 2. Effect of processing on physico-chemical and bioactive compounds of carrot and carrot juice*
Products TSS Acidity Ascorbic acid Total β-carotene Lycopene Total phenols Scavenging
(oBrix) (%) (mg/100g) carotenoid (mg/100g) (mg/100g) (mg/100g) activity (%)
(mg/100g)
Raw carrot 8.20±0.02 0.05±0.01 4.31±0.05 13.35±0.07 10.13±0.08 3.36±0.04 39.93±0.19 23.83±0.19
Raw juice 8.20±0.02 0.06±0.02 2.24±0.03 12.42±0.07 9.86±0.08 3.16±0.06 34.98±0.22 23.39±0.07
Control juice 10.00±0.03 0.06±0.01 1.79±0.03 10.10±0.06 8.50±0.05 2.93±0.03 29.95±0.25 21.73±0.28
Carrot ginger juice 13.00±0.02 0.08±0.02 1.94±0.05 10.62±0.06 8.82±0.04 2.96±0.03 34.84±0.26 23.37±0.32
CD (P≤0.05) 0.21 NS 0.06 0.11 0.09 0.05 0.15 0.19
*Values are Mean ± SD (Standard deviation) of three replicates
 Effect of processing and storage on bioactive compounds and antioxidant activity of carrot juice 83

the storage following first order kinetics and found that higher phenolics from 29.95 mg/100g to 15.97 mg/100g and from 34.84
degradation of β-carotene was found in heat-treated juices than mg/100g to 19.93 gm/100g in case of carrot ginger juice. These
high intensity pulse electric field treated juices. According to results are in agreement with Klimczak et al. (2007) who reported
Aczel, (1972) and Drdak and Sorman (1979) time of storage and that hydrocinnamic acid in orange juice stored at 18, 28 and 38oC
temperature considerably lowers the retention of β- carotene. decreased by about 13, 22 and 32 per cent, respectively. Similarly,
Lycopene content decreased significantly (P≤0.05) from 2.93 Teixeira et al. (2009) also found that total phenolic content in
mg/100g to 2.33 mg/100g in control juice and from 2.96 mg/100g carrot juice decreased during storage of 56 days.
to 2.42 mg/100g in carrot ginger juice, after six months storage.
Antioxidant activity: During storage period, antioxidant activity
These results are in accordance with the findings of Lin and Chen
of control and carrot ginger juice decreased significantly (P≤0.05)
(2005) who found decrease in lycopene content and its isomers
(Table 5). Scavenging activity in control carrot juice reduced
in tomato juice during storage. According to them, in addition
from 21.73 per cent during zero month to 10.57 per cent after six
to isomerization, oxidative degradation is a major factor causing
months. In carrot ginger juice the scavenging activity decreased
lycopene loss during storage of tomato juice. Also, Sharma and Le
from 23.37 to 13.51 per cent after storage. The decrease in the
Magure (1996) stated that illumination could enhance the reaction
antioxidant activity may be linked to a decrease in total phenolic
rate of lycopene in the presence of air, and the degradation of all-
content and vitamin C (Klimczak et al., 2007). According to
trans-lycopene could be accompanied by the isomerization.
Teixeira et al. (2009) the antioxidant activity of carrot juice
Ascorbic acid: The ascorbic acid content of control and altered depleted with storage time regardless of the processing treatment.
juice decreased significantly (P≤0.05) during storage (Table 5). They found that DPPH values co-related well with vitamin C
With the advancement of storage, ascorbic acid decreased from (R2 = 0.807) which indicates that vitamin C is among one of the
1.79 to 0.57 mg/100g in case of control juice and from 1.94 to antioxidant compounds in carrot juices. Same authors reported
0.72 mg/100g in carrot ginger juice. This decrease in ascorbic that antioxidant capacity and β-carotene are also co-related well
acid may be due to oxidation of ascorbic acid in presence of light. (R2 = 0.788), suggesting that the variation in antioxidant capacity
Similarly, Dhaliwal and Hira (2001) observed significant losses over time can be modulated by carotenoids. Klimczak et al.
of ascorbic acid during storage of carrot juice. Higher losses have (2007) found that antioxidant activity of orange juices decreased
been found at elevated temperature storage. According to Kaur by 18, 45 and 84 per cent after six months of storage at 18, 28
et al. (2004) the rate of ascorbic acid destruction increases with and 38oC, respectively.
increased temperature in the presence of air.
Processing induces significant changes in chemical composition,
Total phenols: The effect of storage on total phenolic content influencing the concentration and bioavailability of bioactive
as investigated during six month storage of control and altered compounds in carrots. It can have both positive and negative
carrot juice has been presented in Table 5. There was significant effects depending on process conditions (Miglio et al., 2008). It is
(P≤0.05) reduction of total phenols during storage of both the therefore desirable to assess the effect of processing on bioactive
juices. In case of control juice there was a decrease in total compounds and antioxidant activity of carrot products. Thus,
Table 4. Effect of storage on total carotenoids, β-carotene and lycopene content of carrot juice*
Storage months Total carotenoids (mg/100g) β-carotene (mg/100g) Lycopene (mg/100g)
Control Carrot ginger juice Control Carrot ginger juice Control Carrot ginger juice
0 10.10±0.06 10.62±0.06 8.50±0.05 8.82±0.04 2.93±0.03 2.96±0.03
1 9.65±0.07 9.83±0.05 7.95±0.05 8.03±0.06 2.82±0.02 2.89±0.03
2 9.26±0.05 9.30±0.03 7.19±0.03 7.37±0.05 2.68±0.04 2.74±0.05
3 8.53±0.03 9.09±0.03 6.70±0.06 6.72±0.07 2.60±0.02 2.56±0.02
4 7.99±0.04 8.33±0.03 6.39±0.05 6.57±0.06 2.54±0.01 2.51±0.03
5 7.82±0.04 8.11±0.06 5.86±0.05 6.14±0.03 2.42±0.04 2.47±0.03
6 7.75±0.06 7.97±0.04 5.65±0.06 5.87±0.06 2.33±0.02 2.42±0.02
CD (P≤0.05) 0.05 0.05 0.05 0.08 0.05 0.06
*Values are Mean ± SD (Standard deviation) of three replicates
Table 5. Effect of storage on ascorbic acid, total phenols and antioxidant activity of carrot juice*
Storage months Ascorbic acid (mg/100g) Total phenols (mg/100g) Antioxidant activity (%)
Control Carrot ginger juice Control Carrot ginger juice Control Carrot ginger juice
0 1.79±0.03 1.94±0.05 29.95±0.25 34.84±0.26 21.73±0.28 23.37±0.32
1 1.31±0.04 1.65±0.03 29.97±0.40 34.93±0.34 18.85±0.35 20.26±0.31
2 1.08±0.04 1.32±0.03 24.82±0.48 29.94±0.27 15.46±0.40 18.15±0.28
3 0.92±0.03 1.05±0.04 20.00±0.48 24.90±0.30 13.22±0.51 16.98±0.23
4 0.79±0.03 0.91±0.02 19.95±0.31 19.89±0.38 13.02±0.46 14.72±0.30
5 0.70±0.02 0.84±0.03 19.94±0.44 19.95±0.39 12.64±0.32 14.30±0.31
6 0.57±0.02 0.72±0.04 15.97±0.23 19.93±0.33 10.57±0.25 13.51±0.29
CD (P≤0.05) 0.06 0.08 0.44 0.40 0.88 0.99
*Values are Mean ± SD (Standard deviation) of three replicates
84 Effect of processing and storage on bioactive compounds and antioxidant activity of carrot juice
from the present investigation we conclude that during processing
79.94-86.76% ascorbic acid, 81.33-85.51% total carotenoids,
86.21-89.46% β-carotene, 92.73-93.68% lycopene, 85.63-98.6%
total phenols and 92.91-99.81% antioxidant activity was retained
in control and ready-to-serve carrot ginger juice, respectively.
More antioxidant activity was found in ready-to-serve carrot
ginger juice as compared to control juice due to addition of ginger
which is known as a good source of antioxidant. During storage,
mean sensory scores decreased non-significantly. No significant
changes were observed in TSS and acidity of control and ready-
to-serve carrot juice.
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Received: May, 2013; Revised: January, 2014; Accepted: February, 2014