Accepted Manuscript

Title: Effect of Green Manures and Zinc Fertilizer Sources on DTPA-

extractable Zinc in soil and Zinc Concentration in Basmati Rice Plants
at Different Growth Stages

Author: AMARPREET SINGH and YASHBIR SINGH SHIVAY

PII: S1002-0160(17)60442-9
DOI: 10.1016/S1002-0160(17)60442-9
Reference: NA

To appear in:

Received date: NA
Revised date: NA
Accepted date: NA

Please cite this article as: AMARPREET SINGH and YASHBIR SINGH SHIVAY, Effect of Green
Manures and Zinc Fertilizer Sources on DTPA-extractable Zinc in soil and Zinc Concentration in Basmati
Rice Plants at Different Growth Stages, Pedosphere (2017), 10.1016/S1002-0160(17)60442-9.

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our
customers we are providing this early version of the manuscript. The manuscript will undergo
copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please
note that during the production process errors may be discovered which could affect the content, and all
legal disclaimers that apply to the journal pertain.
 ACCEPTED MANUSCRIPT

PEDOSPHERE
Pedosphere ISSN 1002-0160/CN 32-1315/P doi:10.1016/S1002-0160(17)60442-9

Effect of Green Manures and Zinc Fertilizer Sources on DTPA-extractable Zinc in soil and
Zinc Concentration in Basmati Rice Plants at Different Growth Stages

AMARPREET SINGH1 & 2 and YASHBIR SINGH SHIVAY1*

1
Division of Agronomy, ICAR-Indian Agricultural Research Institute (IARI), New Delhi 110 012
(India)

t
2

ip
Present address: Division of Crop Production, ICAR-Central Research Institute for Jute and
Allied Fibres, Barrackpore, Kolkata, West Bengal 700 120 (India)
*Corresponding author. E-mail: ysshivay@hotmail.com

cr
us
ABSTRACT

Field experiments were conducted during summer and rainy (kharif) seasons (April-
an
November) of 2009 and 2010 for two consecutive years at the research farm of the Indian
Agricultural Research Institute, New Delhi. The objectives of this study was to determine the
effects of summer green manuring crops [Sesbania aculeata (Dhaincha), Crotalaria juncea
M
(Sunhemp) and Vigna unguiculata (Cowpea)] and zinc (Zn) fertilizers [EDTA-chelated Zn (12%
Zn), ZnSO4.7H2O (21% Zn), ZnSO4.H2O (33% Zn), ZnO (82% Zn) and ZnSO4.7H2O + ZnO
(50% + 50%)] and on the diethylene triamine penta-acetic acid (DTPA) extractable (available)
d

Zn concentration in soil and total Zn content in of Basmati rice cv. ‘Pusa Basmati 1’plants at
periodic intervals. Beneficial effects of summer green manuring crops and Zn fertilizers on
e

DTPA-extractable (available) Zn concentration in soil and total Zn content in dry matter of

pt

Basmati rice at periodic intervals were observed, with significant increases in all the determined
parameters, in comparison with the control (no Zn application or summer fallow). The rates of
increase varied among summer green manuring crops and Zn fertilizers during both the years.
ce

Among the summer green manures, incorporation of Sesbania aculeata led to a significant
increase in mean Zn content in Basmati rice grain (31.5 and 32.4 mg kg-1 grain) and straw (167.5
and 172.1 mg kg-1 straw) compared with Crotalaria juncea, Vigna unguiculata and summer
Ac

fallow treatments. Among the Zn fertilizers, significant improvement in Zn concentration in

Basmati rice dry matter and DTPA-extractable Zn content in soil at various growth stages were
recorded with EDTA-chelated Zn (12% Zn) application, followed by ZnSO4.7H2O (21% Zn),
ZnSO4.H2O (33% Zn), ZnSO4.7H2O + ZnO (50% + 50%), ZnO (82% Zn) and control (no Zn
application). The highest mean Zn concentration in Basmati rice grain (33.9 and 35.7 mg kg-1
grain) and straw (173.5 and 177.4 mg kg-1 straw) was recorded with EDTA-chelated Zn (12%
Zn) application in 2009 and 2010, respectively. The ZnSO4.7H2O (21% Zn) application was the
second best treatment after EDTA-chelated Zn (12% Zn), but was statistically inferior to it. The
lowest values were recorded with control (no Zn application) during both the years of
experimentation. The amount of Zn content in soil has been found to be significant positively
correlated with the Zn concentration of Basmati rice dry matter during both the years of
experimentation. Significantly higher levels of residual fertility in soil after harvest of Basmati

2
 ACCEPTED MANUSCRIPT

rice was observed with application of EDTA-chelated Zn (12% Zn) and Sesbania aculeata
incorporation compared with rest of Zn sources and summer green manures during both the
years.

Key Words: Basmati rice, DTPA-extractable (available) Zn, summer green manures, zinc
concentration in dry matter, zinc sources

INTRODUCTION

The application of green manures to rice soils is considered a good management practice
in any agricultural production system because it can increase cropping system sustainability by

t
ip
reducing soil erosion and ameliorating soil physical properties, by increasing soil organic matter
and fertility levels (Mandal et al., 2003), by increasing nutrient retention and by reducing global

cr
warming potential (Robertson et al., 2000). Both leguminous and non-leguminous plants are

us
used in the production of green manures. Leguminous plants have advantage of adding nitrogen
to soil by symbiotic fixation of atmospheric N through Rhizobia.
an
The importance of leguminous green manure crops in improving soil fertility and soil
physical properties has received increasing attention in recent times (Whitbread et al., 2000; Ray
and Gupta, 2001). The improvement in soil physical conditions as a result of buildup of organic
M
matter by incorporation of green manure or crop residue is associated with a decrease in bulk
density, increase in total pore space, water stable aggregates and hydraulic conductivity of the
d

soil (Tejada et al., 2008a, b). Fast growing leguminous green manures with their adaptability to
e

different rice based cropping pattern and their ability to fix atmospheric nitrogen may offer
pt

opportunities to increase and sustain productivity and income in the rice based cropping system
ce

(Yadvinder-Singh et al., 1991). Rice soils are usually deficient in organic matter because of high
temperature and moisture, which causes rapid decomposition of organic matter (Mohammed et
Ac

al., 2005). Green manures enhance organic matter content in soil and may recycle plant nutrients
including zinc; however, data on this aspect are not available. The positive effect of green
manures on paddy yield has been reported by Sharma and Prasad (1999). Hemalatha et al. (1999)
observed that in-situ incorporation of cowpea before transplanting of rice increased the grain
yield by 18% and straw yield by 16% as well as quality of rice. Similarly, Sesbania aculeata,
Crotalaria juncea and Vigna unguiculata are some of the important leguminous green manuring
crops in rice regions of Asia, but the information on their relative capability in improving soil
fertility is not available and therefore the present study was undertaken.

3
 ACCEPTED MANUSCRIPT

Zinc (Zn) is the most deficient micronutrient in rice soils and its deficiency is prevalent
world-wide, especially in high pH calcareous soils (Adriano, 2001; Cakmak, 2002; Fageria et al.,
2003; Norman et al., 2003; Prasad, 2006; Alloway, 2008; Cakmak, 2008; Prasad et al., 2014). A
recent analysis of two hundred forty one thousand soil samples in India showed that 49% soils
are deficient in Zn (Behera et al., 2009) and Zn deficiency is widespread in north Indian rice-
wheat cropping system belt (Prasad, 2005). Zn deficiency in rice is characterized by brown spots,
which appear first in the younger leaves and later in the lower leaves. In severe Zn deficiency

t
burnt dark brown patches of plants appear in rice fields (Dobermann and Fairhurst, 2000).

ip
Response of rice to Zn has been reported by several workers in India (Srivastava et al., 2006;

cr
Shivay et al., 2007b; Shivay et al., 2010; Pooniya et al., 2012; Shivay and Prasad, 2012;
Amarpreet-Singh and Shivay, 2015), China (Shihua and Wenqiang, 2000) and the USA (Slaton

us
et al., 2005a). Zn is also now recognized as the fifth leading risk factor for human health in
developing Asian countries (IFA, 2007) and efforts are underway for encouraging Zn
an
fertilization not only from the view point of getting higher rice yields but also for increasing Zn
concentration in grain and straw to improve human and animal Zn nutrition (WHO, 2002). Also,
M
the increased Zn concentration in rice straw is of importance from the viewpoint of cattle
nutrition since in developing countries of Asia, rice straw is the major feed for farm cattle
d

(Shivay et al., 2008a). Application of Zn as zinc sulphate (ZnSO4·7H2O) is therefore generally

e

recommended for rice and most other crops in India. Other Zn fertilizers include ZnO and Zn-
pt

EDTA, the latter supplies substantial amount of Zn to the plants without interacting with soil
components (Karak et al., 2005), because the central metal ion Zn2+ is surrounded by chelate
ce

ligands (Mortvedt, 1979). Recent studies have shown that the effectiveness of the Zn fertilizers
in correcting Zn deficiencies depends on the water solubility of Zn fertilizers (Gangloff et al.,
Ac

2000; Gangloff et al., 2002; Gangloff et al., 2006; Slaton et al., 2005a; Shaver et al., 2007;
Shivay et al.(2014); Shivay et al.(2015a); Shivay et al.(2015b). The primary soil factors
controlling the potential bioavailability of metals are soil pH, the accessibility and character of
sorption sites on soil surfaces, the contents of iron and aluminum oxyhydroxides, soil organic
matter, and clay minerals (Gangloff et al., 2002). Some studies have indicated that organic
sources are more effective fertilizers than inorganic ones. Their effectiveness depends on the rate
of their disappearance from the soil solution, which is related to their stability (Alloway, 2008).

4
 ACCEPTED MANUSCRIPT

No information on the relative efficiency of different Zn sources in the presence of large

amounts of fresh organic residues as obtained from green manures is however available.
Basmati (aromatic) rice is premier rice grown in north-western India and Pakistan. This
rice is preferred for their long and slender kernels which expand 3-4 times in length and remain
fluffy on cooking. The varieties of Basmati rice differ greatly in kind and intensity of aroma.
Aroma in rice is due to a chemical diacetyle 1-pyrolline. These type of rices are well known all
over the world, especially in the middle East and South Asia for their long fluffy grains on
cooking, a desirable characteristic for the dish biryani or pulao made by cooking rice with

t
ip
vegetables, mutton or chicken and flavored with special oriental spices (Shivay et al., 2010).
Traditional Basmati rice in the Doon valley of India is tall indicas yielding 1.5 to 2.0 tonnes ha-1

cr
and lodge on heavy fertilization. Attempts were therefore made at the Indian Agricultural

us
Research Institute, New Delhi to develop Basmati rice varieties having high yielding qualities of
semi-dwarf indica rice and also having better quality aromas compared to traditional one.
an
Considering the large demand of Basmati rice in the world, concerted research efforts was made
by the researchers at the Indian Agricultural Research Institute (IARI), New Delhi, India to
develop rice varieties having both Basmati traits and high yield potential. This involved
M
convergent breeding and simultaneous selection at field and laboratory for the complex inherited
key characteristics of Basmati quality (extra-long slender grain, excessive elongation on cooking,
d

aroma and ideal physicochemical properties of starch) and high yield potential of new semi-
e

dwarf rice varieties (Shivay et al., 2010). ‘Pusa Basmati 1’ was the result of this research, which
pt

took about 25 years and now occupies the largest area in north-western India and fetches about
ce

500 million US $ in the international market (Siddiq, 2006). ‘Pusa Basmati1’ is high yielding
Basmati rice which has long and slender kernels as per classification by Jennings et al. (1979).
Ac

Basmati rice varieties developed from IARI now covered about 60% of the total area under rice
in north-western India, which is estimated at about 5 million hectare. .
The present study was undertaken to study the effect of green manuring with legumes and
zinc fertilization on the DTPA-extractable Zn in soil and Zn concentration in rice plant at
different intervals during growth and Zn uptake by Basmati rice.

5
 ACCEPTED MANUSCRIPT

MATERIALS AND METHODS

Site and soil descriptions

The field experiments were conducted for two consecutive years at the research farm of
the Indian Agricultural Research Institute, New Delhi, India during summer-kharif/rainy-seasons
of 2009 and 2010 on a sandy clay-loam soil (typic Ustochrept). The experiments in both the
years were conducted with a fixed lay-out plan on the same site. The institute farm is located at
latitude of 28o40’ N and longitude of 77o12’ E, altitude of 228.6 meters above the mean sea

t
ip
level. The mean annual rainfall of New Delhi is 650 mm and more than 80% generally occurs
during the south-west monsoon season (July-September) with mean annual evaporation 850 mm.

cr
The soils of experimental field had 135.75 kg ha-1 alkaline permanganate oxidizable
nitrogen (N) (Subbiah and Asija, 1956), 16.04 kg ha-1 available phosphorus (P) (Olsen et al.,

us
1954), 292.10 kg ha-1 1 N ammonium acetate exchangeable potassium (K) (Hanway and Heidel,
an
1952) and 0.53% organic carbon (OC) (Walkley and Black, 1934). The pH of soil was 7.5 (1: 2.5
soil and water ratio) (Prasad et al., 2006) and diethylene triamine penta acetic acid (DTPA)
extractable Zn (Lindsay and Norvell, 1978) in soil was 0.67 mg kg-1 of soil. The critical level of
M
DTPA-extractable Zn for rice grown on alluvial soils in the rice-wheat belt of North India varies
from 0.38-0.90 mg kg−1 soil (Takkar et al., 1997) and thus the response of Basmati rice to Zn
d

application was expected on the experimental field. The DTPA-extractable Fe (Lindsay and
e

Norvell, 1978) in soil was 4.53 mg kg-1 of soil.

pt
ce

Treatments details and their allocation

The experiment was conducted in a split plot design, keeping three green manuring crops
Ac

viz. Sesbania aculeata (Dhaincha), Crotalaria juncea (Sunhemp), and Vigna unguiculata
(Cowpea) and one summer fallow treatment as main-plot treatments and five Zn fertilizers viz.
ZnSO4.7H2O (21% Zn), ZnSO4.H2O (33% Zn), ZnO (82% Zn), ZnSO4.7H2O + ZnO (50% +
50%), EDTA-chelated Zn (12% Zn) and a control (no Zn application) in sub-plots and was
replicated thrice.
During summer seasons three green manuring crops viz. Sesbania aculeata, Crotalaria
juncea, and Vigna unguiculata were planted as main plot treatments. A summer fallow treatment
was also considered as the control. After 40 days the summer green manuring crops were

6
 ACCEPTED MANUSCRIPT

incorporated into the soil before transplanting of rice. After incorporation of green manuring
crops, each main plot (with green manure incorporation or otherwise) was divided into six sub-
plots, which received the Zn-fertilization treatments. The experimental field was disk-ploughed
twice, puddled three times with a puddler in standing water and levelled. At the time of final
puddling a single application of 26 kg P ha−1 as single superphosphate and 33 kg K ha−1 as
muriate of potash was broadcasted. Addition of 26 kg P as single super phosphate also supplied
45 kg S ha−1 and would correct S deficiency (if any). It also provides S thus permitting a more
fare comparation to applications involving Zn sulfates. Nitrogen @ 120 kg ha-1 as prilled urea

t
ip
was applied into two equal splits, half at the time of transplanting and remaining half at panicle
initiation stage (40 DAT). In all the Zn treatments uniformly 5 kg Zn ha-1 was applied.

cr
us
Basmati rice transplanting

After execution of all treatment materials in the respective plots, two 25-day-old
an
seedlings of Basmati rice (Oryza sativa L.) variety ‘Pusa Basmati 1’ was transplanted at 20 cm x
10 cm in the first fortnight of July in both the years of study. It is an aromatic (Basmati) variety
M
released from Indian Agricultural Research Institute, New Delhi, India during 1989 for its
commercial cultivation. It is a cross between ‘Pusa 150’ and ‘Karnal Lokal’. It produces long
d

slender grains with good aroma and excellent cooking qualities (Rani et al., 2009; Singh and
e

Singh, 2009; Siddiq et al., 2012). Irrigation channels measuring 1 m wide were placed between
pt

the replications to ensure easy and uninterrupted flow of irrigation water where an individual plot
was independently irrigated from the irrigation channels. Rice crop was grown as per
ce

recommended package of practices and was harvested in the second fortnight of October in both
the years of experimentation. The rice seedlings were allowed to grow until harvest.
Ac

Plant sampling for dry matter accumulation

Plant samples of Basmati rice were collected periodically at the interval of 15 days from
five randomly selected locations in each sub-plot up to 90 days of transplanting and again at
harvest. Plant samples were cut at ground level and were washed initially with tap water
followed by dilute hydrochloric acid (0.05 N), de-ionised water and finally Zn-free double

7
 ACCEPTED MANUSCRIPT

distilled water. The plant samples were dried in a hot air oven at 65±5ºC. At harvest stage, the
plant was separated into grain and straw and analysed for Zn content separately.

Determination of zinc in plant samples

The dried samples of rice were ground for Zn analyses with a milling machine and
sieved. Thereafter, the samples were digested using a di-acid [perchloric acid (HClO4) + nitric
acid (HNO3) in 3:10 ratio] method. After digestion in the aliquot of samples, total Zn was

t
estimated with the help of atomic absorption spectrophotometer (Perkin Elmer; Model-A.

ip
Analyst 100) as described by Prasad et al. (2006).

cr
Soil sampling and determination DTPA-extractable zinc

us
Soil samples were collected at a periodic interval of 15 days from five randomly selected
locations in each plot in submerged condition from the rhizosphere region up to 90 days after
an
transplanting. After the harvest of crop, soil samples were collected at a depth of 0-15 cm.
Composite soil samples were made for each plot and dried in the shade, ground and passed
M
through 80-mesh sieve. Ten grams of soil was taken for estimation of 0.005 M DTPA-extractable
Zn by following the method described by Lindsay and Norvell (1978).
e d

Statistical analyses
pt

All the replicated data obtained from the experiments for consecutive two years of study
ce

were statistically analysed using the F-test as per the procedure given by Gomez and Gomez
(1984). Least significant difference (LSD) values at P=0.05 were used to determine the
Ac

significance of differences between treatment means.

RESULTS
DTPA-extractable Zn in soil
The average amount of DTPA-extractable Zn in soil as affected by summer green
manuring crops’ residue incorporation and various Zn sources is presented in Fig 1 & 2. A
general increase in DTPA-extractable Zn was observed up to 30 days after transplanting (DAT),
after that the decrease in available Zn was noticed in all the treatments during both the years of

8
 ACCEPTED MANUSCRIPT

study. The results show that the periodic changes of DTPA-extractable Zn content during both
the years showed a similar pattern of changes. However, the amount of DTPA-extractable Zn in
soil was significantly affected by summer green manure residue incorporation and by Zn sources
during both the years of experimentation. Significantly higher values of DTPA-extractable Zn
were recorded with green manure incorporation over summer fallow during all the crop growth
stages as well as at harvest (Fig. 1 & 2), irrespective of the type of green manure. Among the
summer green manuring crops, incorporation of Sesbania aculeata recorded significantly higher
levels of available Zn in soil over Crotalaria juncea, Vigna unguiculata and summer fallow

t
ip
during 2009 and 2010, respectively.
Zn fertilization appreciably improved the DTPA-extractable Zn content in soil,

cr
irrespective of the sources applied during both the years of experimentation. A general decrease

us
in available Zn in soil with progress of crop growth was observed in all the treatments during
2009 and 2010. Significantly higher level of available Zn in soil was observed during all the crop
an
growth stages as well as at harvest (Fig. 1 & 2; Table I) with Zn application over control (no Zn
application) during both the years of experimentation. Application of EDTA-chelated Zn (12%
Zn) was found superior over all other Zn sources with respect to maintenance of soil available
M
Zn. ZnSO4.7H2O (21% Zn) was the second best treatment with respect to DTPA-extractable Zn
in soil but was significantly inferior to EDTA-chelated Zn (12% Zn). ZnO was found to be least
d

effective Zn source during both the years of experimentation. Significantly lower DTPA-
e

extractable Zn values in soil were recorded with control (no Zn application). In general, an
pt

increase in the amount of DTPA-extractable Zn concentrations in soil was observed from 15

ce

DAT to 30 DAT, irrespective of the treatments applied. After that, a general decline in the
amount of DTPA-extractable Zn concentrations in soil was observed with the progress of crop
Ac

growth after 30 DAT. All the interactions were non-significant at P=0.05.

Zn concentration in Basmati rice plants

Data pertaining to Zn concentration in Basmati rice plants at different periods of growth
and grain and straw at harvest are given in Fig. 3 & 4 and Table I. Different green manures and
Zn fertilizer sources significantly influenced the Zn concentrations in Basmati rice plants at all
the crop growth stages as well as in grain and straw at harvest during both the years of
experimentation. The results showed that the periodic changes of Zn concentrations in Basmati

9
 ACCEPTED MANUSCRIPT

rice dry matter at all the crop growth stages and grain and straw at harvest during both the years
showed a similar pattern of changes. The Zn concentrations in rice dry matter gradually
increased during the initial growth period (up to 30 DAT) and thereafter decreased, irrespective
of treatments applied during 2009 and 2010, respectively. Significantly higher values of Zn
concentrations in Basmati rice were recorded with green manure incorporation over summer
fallow during all the crop growth stages as well as in grain and straw at harvest (Fig. 3 & 4;
Table I), irrespective of the type of green manure. The Zn concentration in Basmati rice dry
matter and in grain and straw was significantly higher with Sesbania aculeata residue

t
ip
incorporation compared with Crotalaria juncea, Vigna unguiculata and summer fallow
treatments during 2009 and 2010. The performance of summer green manures for improvement

cr
in Zn concentration of Basmati rice was in the order; Sesbania aculeata > Crotalaria juncea >

us
Vigna unguiculata. Lowest values were recorded with summer fallow treatments during both the
years of experimentation. The percentage increase in Zn concentration in grain of Basmati rice
an
with Sesbania aculeata residue incorporation was in the order of 5.70 and 9.45; 15.80 and 21.34;
30.16 and 36.70% over Crotalaria juncea, Vigna unguiculata and summer fallow during 2009
and 2010, respectively. Almost similar trend was also observed in case of Zn concentration in
M
Basmati rice dry matter at all the growth stages as well as in rice straw of during both the years.
In general, application of Zn fertilizers significantly increased the Zn concentration in
d

Basmati rice dry matter at all the growth stages as well as in grain and straw compared with
e

control (no Zn application) during both the years of experimentation, irrespective of its source
pt

applied. Among the Zn fertilization treatments, application of EDTA-chelated Zn (12% Zn)

ce

resulted into statistically higher values of Zn content of Basmati rice dry matter at all the growth
stages as well as in grain and straw compared with all other Zn fertilizer sources and control (no
Ac

Zn application), respectively during 2009 and 2010. The ZnSO4.7H2O (21% Zn) application was
second best treatment with respect to Zn concentration in Basmati rice after EDTA-chelated Zn
(12% Zn), but was statistically inferior to it. The ZnO was found to be least effective Zn source
with respect to Zn concentration in Basmati rice during both the years of experimentation.
Significantly lower values of Zn concentration in Basmati rice at all growth stages as well as in
grain and straw were recorded with control (no Zn application).
A general decrease in Zn concentration in Basmati rice dry matter with progress of crop
growth after 30 DAT was observed in all the treatments during 2009 and 2010. In general, an

10
 ACCEPTED MANUSCRIPT

increase in the Zn concentration in Basmati rice dry matter was observed from 15 DAT to 30
DAT, irrespective of the treatments applied. After that, a general decline in the Zn concentration
in Basmati rice dry matter was observed with the progress of crop growth after 30 DAT during
both the years of experimentation. The performance of Zn sources for improvement in Zn
concentration in Basmati rice dry matter was in the order; EDTA-chelated Zn (12% Zn) >
ZnSO4.7H2O (21% Zn) > ZnSO4.H2O (33% Zn) > ZnSO4.7H2O + ZnO (50% + 50%) >ZnO
(82% Zn). However, all the interactions were non-significant at P=0.05 during both the years of
experimentation.

t
ip
The percentage increase of Zn content in Basmati rice dry matter was recorded relatively
higher in the treatment where EDTA-chelated Zn (12% Zn) was applied during both the years.

cr
The percentage increase in Zn concentration in grain of Basmati rice with EDTA-chelated Zn

us
(12% Zn) application was in the order of 11.88 and 17.82; 15.69 and 22.68; 27.92 and 39.45;
20.64 and 27.04; 60.66 and 82.14% over ZnSO4.7H2O (21% Zn), ZnSO4.H2O (33% Zn), ZnO
an
(82% Zn), ZnSO4.7H2O + ZnO (50% + 50%) and control (no Zn application) during 2009 and
2010, respectively. Almost similar trend was also observed in case of Zn concentration in
Basmati rice dry matter at all the growth stages as well as in straw during both the years.
M

Correlation and regression analysis

d

The results of correlation and regression coefficients {Table II and Figs. 5 (A-C)}
e

showing the relationship between the Zn content in dry matter and Zn content in soil show that
pt

the amount of Zn content in soil has been found to be significantly positively correlated with the
ce

Zn content of dry matter at different periods of crop growth during both the years. This further
justified the beneficial effect of Zn fertilization and use of summer green manuring crops in
Ac

improvement of the DTPA-Zn content in soil and Zn concentration in Basmati rice dry matter.

Available (DTPA-extractable ) Zn in soil after harvest of Basmati rice

Data pertaining to available (DTPA-extractable) Zn in soil after harvest of Basmati rice

are presented in Table I. Incorporation of summer green manure crops and Zn fertilizer sources
had significant influence on soil available Zn after the harvest of Basmati rice. The significantly
higher available Zn in soil was recorded after harvest of Basmati rice with Sesbania aculeata
residue incorporation compared with Crotalaria juncea, Vigna unguiculata and summer fallow

11
 ACCEPTED MANUSCRIPT

treatments during 2009 and 2010. The percentage increases in available Zn in soil after harvest
of Basmati rice with incorporation of preceding Sesbania aculeata residue was in the order of
5.49 and 12.90; 15.66 and 23.52; 26.31 and 40.0% over Crotalaria juncea, Vigna unguiculata
and summer fallow during 2009 and 2010, respectively.
The available Zn in the soil after the harvest of Basmati rice was significantly influenced
due to Zn fertilization treatments. Application of EDTA-chelated Zn (12% Zn) resulted into
higher residual available Zn content over all other Zn sources and control (no Zn application).
The ZnSO4.7H2O (21% Zn) application was second best treatment with respect to available Zn

t
ip
content after EDTA-chelated Zn (12% Zn). The lowest values were recorded with control (no Zn
application) during 2009 and 2010. The percentage increase in available Zn in soil after harvest

cr
of Basmati rice with EDTA-chelated Zn (12% Zn) application was in the order of 22.58 and

us
30.92; 32.55 and 47.67; 52.00 and 71.62; 40.74 and 54.87; 65.21 and 81.42% over ZnSO4.7H2O
(21% Zn), ZnSO4.H2O (33% Zn), ZnO (82% Zn), ZnSO4.7H2O + ZnO (50% + 50%) and control
an
(no Zn application) during 2009 and 2010, respectively. However, all the interactions were non-
significant at P=0.05 during both the years of experimentation.
M
DISCUSSION

DTPA-extractable Zn in soil
e d

The amount of DTPA-extractable Zn in soil at periodical intervals was significantly

pt

influenced by the summer green manure incorporation and Zn sources. A general increase in
DTPA-extractable Zn was observed up to 30 days after transplanting (DAT), after that a decrease
ce

in available Zn was noticed in all the treatments during both the years. The increase in DTPA-
extractable Zn up to 30 days may be due to the acidification of rhizosphere resulting in release of
Ac

H+ from the roots to balance excess intake of cations over anions, and H+ generated in the
oxidation of Fe2+ by root-released O2. Kirk and Bajita (1995) concluded that Fe oxidation
released Zn from highly insoluble fractions and that this Zn was re-adsorbed on Fe(OH)3 and on
organic matter in forms that were acid soluble and therefore accessible to the plants. The
decrease of Zn content in soil (after 30 DAT) might be due to the effect of submergence as well
as depletion by crop. The decline in the availability of Zn in rice soil with time has also been
reported by several investigators (Dutta et al., 1989; Das & Mandal, 1988; Prasad et al., 1991).
In submerged condition there is a formation of large amounts of Fe2+ and Mn2+ due to reduction

12
 ACCEPTED MANUSCRIPT

of its higher valent oxides. As some of the reduced Fe diffuses to more oxic regions of the soils,
it gets oxidized, leading to its precipitation as Fe3(OH)8 (hydrated magnetite) and Zn2+ gets
adsorbed on the hydrous oxides of iron and hence Zn availability to plant decreases (Mandal et
al., 1993; Singh et al., 1999). Brar and Sekhon (1976) summarized the decrease in availability of
Zn in submerged soil as: formation of insoluble franklinite (ZnFe2O4) compound in submerged
soils, formation of very insoluble compounds of Zn as ZnS under intense reducing condition,
formation of insoluble compounds of Zn as ZnCO3 at the later period of soil submergence,
formation of Zn(OH)2 at a relatively higher pH which decreases the availability of Zn and

t
adsorption of soluble Zn2+ by oxide minerals, e.g. sesquioxides, carbonates and clay minerals.

ip
The results show that the periodic changes of DTPA-extractable Zn content during both the years

cr
showed a similar pattern of changes. Significantly higher values of DTPA-extractable Zn were

us
recorded with green manure incorporation over summer fallow during all the crop growth stages
as well as at harvest (Fig. 1 & 2; Table I), irrespective of the type of green manure. Incorporation
an
of green manures supplied significantly higher amount of readily decomposable organic
materials, which improved soil organic matter status. It leads to recycling of Zn (and other
nutrients) into the soil and this might have increased the DTPA-extractable Zn content in soil.
M
Lowest values of DTPA-extractable Zn content in soil at all periodic intervals were recorded
with summer fallow (no green manure residue incorporation) before rice transplanting. The
d

performance of summer green manures with respect to improvement in DTPA-extractable Zn

e

content in soil was in order; Sesbania aculeata > Crotalaria juncea > Vigna unguiculata >
pt

summer fallow. This might be because, highest amount of readily decomposable organic biomass
ce

was added by Sesbania aculeata due to its fast and determinate growth habit compared with
other green manures.
Ac

Zn fertilization appreciably improved the DTPA-extractable Zn content in soil,

irrespective of the sources applied during both the years of experimentation. A general decrease
in soil available Zn with progress of crop growth was observed in all the treatments during 2009
and 2010. Significantly higher level of available Zn in soil was observed during all the crop
growth stages as well as at harvest (Fig. 1 & 2; Table I) with Zn application over control (no Zn
application) during both the years of experimentation. Relatively higher amount of DTPA-
extractable Zn content in soil with Zn application is due to the presence of increased amounts of
Zn in soil solution with applied Zn compared with control (no Zn application). Application of

13
 ACCEPTED MANUSCRIPT

EDTA-chelated Zn (12% Zn) was found superior over all other Zn sources with respect to
maintenance of soil available Zn. The ZnSO4.7H2O (21% Zn) was second best treatment with
respect to DTPA-extractable Zn in soil but was significantly inferior to EDTA-chelated Zn (12%
Zn). The ZnO was found to be least effective Zn source during both the years of
experimentation. Significantly lower DTPA-extractable Zn values in soil were recorded with
control (no Zn application). The performance of Zn sources in terms of DTPA-extractable Zn in
soil was in the order; EDTA-chelated Zn (12% Zn) > ZnSO4.7H2O (21% Zn) > ZnSO4.H2O
(33% Zn) > ZnSO4.7H2O + ZnO (50% + 50%) >ZnO (82% Zn).

t
ip
The relatively higher maintenance of Zn in soil due to applied EDTA-chelated Zn (12%
Zn) may be attributed from the very little or no interaction between soil components preventing

cr
various harmful reactions occurring in soil as compared to soil treated with ZnSO4.7H2O (21%

us
Zn) and other Zn sources which enhances greater fixation, adsorption etc, resulting from the
greater interaction between soil components. Ortiz and Garcia (1998) also reported that the
an
chelated-Zn is fixed less Zn than the sulphate source. These results are in agreement with the
findings of Karak et al. (2005). Further, the incorporation of green manuring crops before
transplanting of Basmati rice improved the organic matter content in soil. The applied Zn might
M
have been complexed with the humic substances present in soil due to organic matter addition
and there might have been lesser Zn fixation by the formation of insoluble Zn complexes. This
d

resulted into increase in the DTPA-extractable Zn content in soil. Improvement in the nutrient
e

use efficiency of the applied fertilizers by transplanted rice after green manure incorporation was
pt

also reported by Yadvinder-Singh et al. (1991). These results also confirmed those reported by
ce

Maftoun and Karimian (1989). Mehdi et al. (1990) also reported that the relative effectiveness of
Zn sources was in order of Zn-EDTA > Zn(NO3)2 > (NH4)2ZnO2 > ZnSO4 > ZnCl2. The results
Ac

might also be due to the fact that EDTA-chelated Zn (12% Zn), ZnSO4.7H2O (21% Zn) and
ZnSO4.H2O (33% Zn) are more water-soluble and therefore readily available, making its effects
visible in the DTPA-extractable Zn content in soil, while ZnO (82% Zn) is sparingly soluble and
is not readily available. Water solubility of Zn sources is considered an important criterion for Zn
availability (Slaton et al., 2005a; Slaton et al., 2005b; Shaver et al., 2007; Shivay et al.(2014);
Shivay et al.(2015a); Shivay et al.(2015b). Mikkleson and Brandon (1975), Nayyar et al.(1990),
Naik and Das (2008), Shivay et al.(2007b), Shivay et al. (2008a) and Jat et al. (2011) also
reported similar findings.

14
 ACCEPTED MANUSCRIPT

Zn concentration in Basmati rice dry matter

Incorporation of summer green manures and Zn fertilizer sources significantly influenced
the Zn concentrations in Basmati rice dry matter at all the crop growth stages as well as in grain
and straw at harvest. The results show that the periodic changes of Zn concentrations in Basmati
rice dry matter at all the crop growth stages as well as in grain and straw at harvest during both
the years showed a similar pattern of changes. The Zn concentrations in rice dry matter gradually
increased during the initial growth period (up to 30 DAT) and thereafter decreased, irrespective

t
ip
of treatments applied during 2009 and 2010, respectively. This is due to the decrease in available
Zn in soil with time because of higher plant uptake up to initial 30 DAT. In waterlogged soil, the

cr
availability of native as well as applied Zn usually decreases due to precipitation of Zn as ZnCO3

us
or ZnS (Reddy and Patrick, 1977). This might also be the reason for decrease in Zn uptake by
rice plants with the passage of time. Significantly higher values of Zn concentrations in Basmati
an
rice were recorded with green manure incorporation over summer fallow during all the crop
growth stages as well as in grain and straw at harvest (Fig. 3 & 4), irrespective of the type of
green manure. Zn concentration in Basmati rice dry matter and in grain and straw was
M
significantly higher with Sesbania aculeata residue incorporation compared with Crotalaria
juncea, Vigna unguiculata and summer fallow. The performance of summer green manures for
d

improvement in Zn concentration in Basmati rice was in the order; Sesbania aculeata >
e

Crotalaria juncea > Vigna unguiculata. Lowest values were recorded with summer fallow
pt

treatments during both the years of experimentation. This might be because, highest amount of
ce

readily decomposable organic biomass was added by Sesbania aculeata due to its fast and
determinate growth habit compared with other green manures.
Ac

Application of Zn fertilizers significantly increased the Zn concentration in Basmati rice

dry matter at all the growth stages as well as in grain and straw compared with control (no Zn
application) during both the years of experimentation, irrespective of its source applied. The
increase in Zn concentration in Basmati rice dry matter at all the growth stages as well as in grain
and straw compared with control (no Zn application) might be due to the presence of increased
amounts of Zn in soil solution by the application of fertilizer Zn that facilitates greater absorption
of Zn compared with that of control (no Zn application). Application of EDTA-chelated Zn (12%
Zn) resulted into statistically higher values of Zn content in Basmati rice dry matter at all the

15
 ACCEPTED MANUSCRIPT

growth stages as well as in grain and straw compared with all other Zn fertilizer sources and
control (no Zn application). The performance of Zn sources for improvement in Zn concentration
in Basmati rice dry matter was in the order; EDTA-chelated Zn (12% Zn) > ZnSO4.7H2O (21%
Zn) > ZnSO4.H2O (33% Zn) > ZnSO4.7H2O + ZnO (50% + 50%) >ZnO (82% Zn). The results
pointed out that the significantly higher Zn concentration in Basmati rice at all growth stages as
well as in grain and straw recorded with EDTA-chelated Zn (12% Zn) application suggests its
greater efficiency for the absorption of the Zn by rice plants as compared to inorganic Zn
sources. This is due to less reaction of chelated Zn with various soil components, resulting in

t
ip
higher concentrations of Zn in soil solution (Das et al., 2002). The comparatively lower recovery
of added Zn when applied as inorganic Zn sources than when applied as EDTA-chelated Zn

cr
(12% Zn) may be attributed to rapid dissociation of Zn2+ from inorganic Zn sources and its

us
subsequent precipitation as ZnCO3 and Zn5(CO3)2(OH)2 (Brar and Sekhon, 1976). These results
are in agreement with the findings of Karak et al. (2005) who reported that chelated Zn was the
an
most efficient source of Zn for lowland rice production. Further, the incorporation of green
manuring crops before transplanting of Basmati rice improved the organic matter content in soil.
The applied Zn might have been complexed with the humic substances present in soil due to
M
organic matter addition and there might have been lesser Zn fixation by the formation of
insoluble Zn complexes. Thus, resulting into increase in the availability of soil applied Zn to rice
d

plants. Improvement in the nutrient use efficiency of the applied fertilizers by transplanted rice
e

after green manure incorporation was also reported by Yadvinder-Singh et al. (1991). These
pt

results also confirmed those reported by Maftoun and Karimian (1989). They concluded that the
ce

greater influence of Zn-EDTA over other sources of Zn in terms of growth and its utilization by
plants is due to less retention and greater transport and movement of chelated Zn to plant roots.
Ac

This could be attributed to lesser fixation in soil of Zn applied as Zn-EDTA than ZnSO4 (Ortiz
and Garcia, 1998). Similar findings were also reported by Chatterjee and Mandal (1985), Mehdi
et al.(1990) and Srivastava et al.(1999). The ZnSO4.7H2O (21% Zn) application was second best
treatment with respect to Zn concentration in grain and straw and its uptake after EDTA-chelated
Zn (12% Zn), but was statistically inferior to it. The ZnO (82% Zn) was found to be least
effective among all the Zn sources. This is due to the fact that the EDTA-chelated Zn (12% Zn),
ZnSO4.7H2O (21% Zn) and ZnSO4.H2O (33% Zn) are more water-soluble and therefore readily
available, making its effects visible in the plants, while ZnO (82% Zn) is sparingly soluble and is

16
 ACCEPTED MANUSCRIPT

not readily available. Water solubility of Zn sources is considered an important criterion for Zn
availability (Slaton et al., 2005a; Slaton et al., 2005b; Shaver et al., 2007). Mikkleson and
Brandon (1975) and Nayyar et al. (1990) also showed that ZnO was inferior to ZnSO4, both in
grain yield and Zn uptake. Similar findings were also reported by Naik and Das (2008), Shivay et
al. (2007b), Shivay et al.(2008b), Jat et al. (2011), Shivay et al.(2014), Shivay et al.(2015a) and
Shivay et al.(2015b). The results of correlation coefficients {Table II and Figs. 5 (A-C)}
showing the relationship between the Zn content in dry matter and Zn content in soil show that
the amount of Zn content in soil has been found to be significant positively correlated with the

t
ip
Zn content of dry matter at different periods of crop growth during both the years. This further
justified the beneficial effect of Zn fertilization and use of summer green manuring crops in

cr
improvement of the DTPA-Zn content in soil and Zn concentration in Basmati rice dry matter.

us
Significantly lower values of Zn concentration in Basmati rice at all growth stages as well as in
grain and straw were recorded with control (no Zn application). This is because of lower amount
an
of soil available Zn present in control plots compared with Zn applied plots.

CONCLUSIONS
M
Following conclusions can be drawn from above investigation:
i. Among the green manuring crops, Sesbania aculeata was found to be best in respect to
d

DTPA-Zn content in soil as well as in Zn concentration in Basmati rice dry matter at all
e

growth stages compared with incorporation of Crotalaria juncea, Vigna unguiculata and
pt

summer fallow treatments. Among the Zn fertilizers, application of EDTA-chelated Zn

(12% Zn) was significantly superior in respect to DTPA-Zn content in soil as well as in
ce

Zn concentration in Basmati rice dry matter at all growth stages compared with all other
Zn sources including control (no Zn application);
Ac

ii. The amount of DTPA extractable-Zn content in soil has been found to be significantly
positively correlated with the Zn content of Basmati rice plants.
iii. In respect to residual soil fertility after Basmati rice harvest, application of EDTA-
chelated Zn (12% Zn) and Sesbania aculeata residue incorporation was found to be
significantly superior over rest of Zn fertilizers and summer green manures.

17
 ACCEPTED MANUSCRIPT

REFERENCES
Adriano, D.C. 2001. Trace Elements in Terrestrial Environments-Biogeochemistry,
Bioavailability and Risks of Metals. 2nd Ed. Springer Verlag, New York, USA.
Alloway, B.J. 2008. Zinc in Soils and Crop Nutrition. International Zinc Association Publication.
International Zinc Association, Brussels, Belgium.
Alvarez, J.M., Almendros, P. and Gonzalez, D. 2009. Residual effects of natural Zn chelates on
navy bean response, Zn leaching and soil Zn status. Plant Soil.317: 277-291.
Amarpreet-Singh and Shivay, Y.S. 2015. Effect of summer green manuring crops and zinc
fertilizer sources on productivity, Zn-uptake and economics of Basmati rice. J Plant
Nutr. DOI: 10.1080/01904167.2015.1009108.
Behera, S.K., Singh, D., Dwivedi, B.S., Singh, S., Kumar, K. and Rana, D.S. 2008. Distribution

t
of fractions of zinc and their contribution towards availability and plant uptake of zinc

ip
under long-term maize (Zea mays L.)-wheat (Triticum aestivum L.) cropping on an
Inceptisol. Australian J Soil Res.46: 83-89.

cr
Brar, M.S. and Sekhon, G.S. 1976. Effect of Fe and Zn on the availability of micronutrients
under flooded and unflooded condition. J Indian Soc Soil Sci.24: 446-451.

us
Brennan, R.F. and Bolland, D.A. 2006. Residual values of soil-applied zinc fertilizer for early
vegetative growth of six crop species. Australian J Experimental Agric.46: 1341-1347.
Cakmak, I. 2002.Plant nutrition research: Priorities to meet human needs for food in sustainable
an
ways. Plant and Soil.247: 3-24.
Cakmak, I. 2008. Enrichment of cereal grains with zinc: Agronomic or genetic biofortification?
Plant Soil.302: 1-17.
Chatterjee, A.K. and Mandal, L.N. 1985. Zinc sources for rice in soil at different moisture
M
regimes and organic matter levels. Plant Soil.87: 393-404.
Das, D.K. and Mandal, L.N. 1988. Effect of puddling and different times of organic matter
application before puddling on the availability of applied iron, manganese, copper and
d

zinc in rice soils. Agrochimica.32: 327-336.

e

Das, D.K., Karak, T. and Karmakar, S.K. 2002. Efficiency of chelated Zn on the maintenance of
Zn in soils in relation to yield and nutrition of rice. In: The 17th world congress of soil
pt

science, 14-21, August 2002, Thailand.

Dobermann, A. and Fairhurst, T. 2000. Rice: Nutrient disorders and nutrient management.
ce

Handbook series. Potash & Phosphate Institute (PPI), Potash & Phosphate Institute of
Canada (PPIC) and International Rice Research Institute. 192 p.
Dutta, D., Mandal, B. and Mandal, L.N. 1989. Decrease in availability of zinc and copper in
Ac

acidic to near neutral soils on submergence. Soil Sci.147: 187-195.

Fageria, N.K. and Baligar, V.C. 2003. Methodology for evaluation of lowland rice genotypes for
nitrogen use efficiency. J Plant Nutr.26: 1315-1333.
Gangloff, W.J., Westfall, D.G., Peterson, G.A. and Mortvedt, J.J. 2002. Relative availability
coefficients of organic and inorganic Zn fertilizers. J Plant Nutr.25: 259-273.
Gangloff, W.J., Westfall, D.G., Peterson, G.A. and Mortvedt, J.J. 2006. Mobility of organic and
inorganic Zn fertilizers in soils. Comm Soil Sci Plant Anal.37: 199-209.
Gangloff, W.J., Westfall, D.G., Peterson, G.A. and Mortvedt, J.J. 2000. Availability of organic
and inorganic Zn fertilizers. Colorado State University Agricultural Experiment Station
Technical Bulletin TB: 1.
Gomez, K.A. and Gomez, A.A. 1984. Statistical procedures for agricultural research. Second
Edition. John Wiley & Sons, New York, USA.

18
 ACCEPTED MANUSCRIPT

Gupta, S.P., Singh, M.V. and Dixit, M.L.2007. Deficiency and management of micronutrients.
Indian J Fertil. 3(6): 57-60.
Hanway, J.J. and Heidel, H. 1952. Soil analysis methods as used in Iowa State College Soil
Testing Laboratory. Bulletin 57, Iowa State College of Agriculture, Iowa, USA. pp 131.
Hemalatha, M., Thirumurugan, V. and Balasubramanian, R. 1999. Influence of organic,
biofertilizer and inorganic forms of nitrogen on rice quality. Inter Rice Res Notes. 24(1):
33.
IFA. 2007. Help eliminate the fifth leading disease risk factor in developing countries. In:
Fertilizer and agriculture, January 2007, Sukalac, K.E. (Ed.), International Fertilizer
Agency, Paris, France. pp. 6.
Jat, S.L., Shivay, Y.S. and Parihar, C.M. 2011. Dual purpose summer legumes and zinc
fertilization for improving productivity and zinc utilization in aromatic hybrid rice (Oryza

t
ip
sativa). Indian J Agron. 56(4): 328-333.
Jennings, P.R., Coffman, W.R. and Kauffman, H.E. 1979. Grain quality. In: Rice Improvement.
International Rice Research Institute (IRRI), Los Banos, Laguna, Philippines. pp. 101-

cr
120.
Karak, T., Singh, U.K., Das, S., Das, D.K. and Kuzyakov, Y. 2005. Comparative efficacy of

us
ZnSO4 and Zn-EDTA application for fertilization of rice (Oryza sativa L.).
Archives Agron Soil Sci 51(3): 253-264.
Karak, T., Singh, U.K., Das, S., Das, D.K. and Kuzyakov, Y. 2005. Comparative efficacy of
an
ZnSO4 and Zn-EDTA application for fertilization of rice (Oryza sativa L.). Archives
Agron Soil Sci. 51(3): 253-264.
Kirk, G.J.D. and Bajita, J.B. 1995. Root induced iron oxidation, pH changes and zinc
M
solubilization in the rhizosphere of lowland rice. New Phytologist.131: 129-137.
Lindsay, W.L., and Norvell, W.A. 1978. Development of DTPA soil test for zinc, iron,
manganese and copper. Soil Sci Soc America J.42: 421-428.
d

Maftoun, M. and Karimian, N. 1989. Relative efficiency of two zinc sources for maize in two
calcareous soils from an arid area of Iran. Agronomie.9: 771-775.
e

Mandal, B., Mandal, L.N. and Ali, M.H. 1993. Chemistry of zinc availability in submerged soils
pt

in relation to zinc nutrition of rice crop. In: Proceedings of the workshop on

micronutrients, Bhubaneswar, Odhisha, India, 22-23 January. pp. 240-253.
Mandal, U.K., Singh, G., Victor, U.S. and Sharma, K.L. 2003. Green manuring: Its effect on soil
ce

properties and crop growth under rice-wheat cropping system. European J Agron.19(2):
225-237.
Mehdi, S.M., Ranjha, A.M. and Hussian, T. 1990. Relative efficacy of various sources of zinc.
Ac

Sarhad J Agric. 6(1): 103-106.

Mikkelson, D.S. and Brandon, D.M. 1975. Zinc deficiency in California rice. California Agric.
29(9): 8-9.
Mohammad, A., Hamad, R.J., Khan, E.A. and Mohammad, R. 2005.Comparative response of
diverse rice varieties to green manuring (Sesbania aculeata). J Res (Sci).16(1): 39-43.
Mortvedt, J.J. 1979. Crop response to zinc sources applied alone or with suspensions. Fertil Sol.
23: 64-79.
Naik, S.K. and Das, D.K. 2008.Relative performance of chelated zinc and zinc sulphate for
lowland rice (Oryza sativa L.). Nutr Cycl Agroecosystems. 81(3): 219-227.

19
 ACCEPTED MANUSCRIPT

Nayyar, V.K., Takkar, P.N., Bansal, R.L., Singh, S.P., Kaur, N.P. and Sadana, U.S. 1990.
Micronutrients in soils and crops of Punjab. Research Bulletin, Department of Soils,
Punjab Agricultural University (PAU), Ludhiana, Punjab, India. 146p.
Nene, Y.L. 1966. Symptoms, causes and control of khaira disease of paddy. Bull Indian
Phytopathology Soc.3: 97-101.
Norman, R.J., Wilson, C.E.Jr. and Salton, N.A. 2003. Soil fertilization and mineral nutrition in
US mechanized rice culture. In: Rice origin, history, technology and production. Smith,
C.W. and Hilday, R.H. (Eds). John Wiley, New Jersey, USA. pp. 331-412.
Olsen, R., Cole, C.V., Watanabe, F.S. and Dean, L.A. 1954. Estimation of available phosphorus
in soils by extraction with sodium bicarbonate. Circular-939, United States Department
of Agriculture, Washington DC, USA.
Ortiz, E.M.E. and Garcia, O.A. 1998. Process of sorption of zinc in three soils of Colombia

t
ip
cultivated with rice (Oryza sativa) of the dry land variety Llano-5. In: Proceedings of the
VIII Congress of Soil Sciences, Santa Marta, Colombia. Suelos-Ecuatoriales. 28: 112-
117.

cr
Pooniya, V., Shivay, Y.S., Rana, A., Nain, L. and Prasanna, R. 2012. Enhancing soil nutrient
dynamics and productivity of Basmati rice through residue incorporation and zinc

us
fertilization. European J Agron. 41: 28-37.
Prasad, R. 2005. Rice-wheat cropping systems. Adv Agron. 86: 255-339.
Prasad, R. 2006. Zinc in soils and in plants, human & animal nutrition. Indian J Fertil. 2(9): 103-
an
119.
Prasad, R., Prasad, B.L. and Sakal, R. 1991. Effect of submergence on native DTPA-extractable
Zn, Fe, Cu and Mn in some old alluvial rice soils of South Bihar. J Indian Soc Soil
M
Sci.39: 773-776.
Prasad, R., Shivay, Y.S. and Kumar, D. 2014. Agronomic biofortification of cereal grains with iron and
zinc. In Donald L. Sparks, editor: Adv Agron. 125: 55-91.
d

Prasad, R., Shivay, Y.S., Kumar, D. and Sharma, S.N. 2006. Learning by Doing Exercises in Soil
Fertility (A practical manual for soil fertility), Division of Agronomy, Indian
e

Agricultural Research Institute, New Delhi, India. 68p.

pt

Rani, N.S., Prasad, G.S.V. and Viraktamath, B.C. 2009. National system for evaluation of
Basmati rices for yield and quality traits. Indian Far. 59 (1): 7-11.
Rattan, R.K., Datta, S.P., Saharan, N. and Katyal, J.C. 1997. Zinc in Indian Agriculture. Fertil
ce

News. 42(12): 75-89.

Ray, S.S. and Gupta, R.P. 2001. Effect of green manuring and tillage practices on physical
properties of puddled loam soil under rice-wheat cropping system. J Indian Soc Soil Sci.
Ac

49(4): 670-678.
Reddy, C.N. and Patrick, W.H. Jr. 1977. Effect of redox potential on stability of zinc and copper
chelates in flooded soil. Soil Sci Soc America J. 41: 729-732.
Robertson, G.P., Paul, E.A. and Harwood, R.R. 2000. Greenhouse gases in intensive agriculture:
Contributions of individual gases to the radioactive forcing of the atmosphere. Sci. 289:
1922-1925.
Sharma, S.N. and Prasad, R. 1999. Effect of Sesbania green-manuring and mungbean residue
incorporation on productivity and nitrogen uptake of a rice-wheat cropping system.
Bioresource Techn. 87: 171-175.
Shaver, T.M., Westfall, D.G. and Ronaghi, M. 2007. Zinc fertilizer solubility and its effects on
zinc bioailability over time. J Plant Nutr. 30(1): 123-133.

20
 ACCEPTED MANUSCRIPT

Shihua, T. and Wenqiang, F. 2000. Nutrient management in the rice-wheat cropping system in
the Yangtze river flood plain. In: Soil and crop management practices for enhanced
productivity of the rice-wheat cropping system in the Sichuan province of China. Hobbs,
P.R. and Gupta, R.K. (Eds), Rice-Wheat Consortium for the Indo-Gangetic Plains, New
Delhi, India. pp. 24-34.
Shivay, Y. S., Prasad, R. and Pal, M. 2014. Effect of conditioning zinc sulfate heptahydrate
(ZnSHH) with zinc oxide (ZnO) and neem oil on growth, productivity, zinc
biofortification of grain and zinc uptake by Basmati rice. J Plant Nutr. 37(11): 1873-
1884.
Shivay, Y. S., Prasad, R. and Pal, M. 2015a. Effects of source and method of zinc application on
yield, zinc biofortification of grain, and Zn uptake and use efficiency in chickpea (Cicer
arietinum L.). Commu Soil Sci Plant Anal. 46(17): 2191-2200.

t
ip
Shivay, Y. S., Prasad, R., Kaur, R. and Pal, M. 2015b. Relative efficiency of zinc sulphate and
chelated zinc on zinc biofortification of rice grains and zinc use-efficiency in Basmati
rice. Proc Natl Acad Sci. (India) Sect B (Biol Sci). DOI 10.1007/s40011-015-0544-7.

cr
Shivay, Y.S. and Prasad, R.2012. Zinc-coated urea improves productivity and quality of Basmati rice
(Oryza sativa L.) under zinc stress condition. J Plant Nutr. 35: 928–951.

us
http://dx.doi.org/10.1080/01904167.2012.663444
Shivay, Y.S., Kumar, D. and Prasad, R. 2007a. Effect of zinc fertilization on physical grain
quality of basmati rice. Interl Rice Res Notes. 32(1): 41-42.
an
Shivay, Y.S., Kumar, D. and Prasad, R. 2008a. Relative efficiency of zinc sulfate and zinc oxide-
coated urea in rice-wheat cropping system. Commu Soil Sci Plant Anal. 39(7&8): 1154-
1167.
M
Shivay, Y.S., Kumar, D. and Prasad, R. 2008b. Effect of zinc-enriched urea on productivity, zinc
uptake and efficiency of an aromatic rice-wheat cropping system. Nutr Cyc
Agroecosystems. 81(3): 229-243.
d

Shivay, Y.S., Kumar, D., Ahlawat, I.P.S. and Prasad, R. 2007b. Relative efficiency of zinc oxide
and zinc sulphate coated urea for rice. Indian J Fertil. 3(2): 51-56.
e

Shivay, Y.S., Prasad, R. and Rahal, A. 2010. Genotypic variation for productivity, zinc
utilization efficiencies, and kernel quality in aromatic rices under low available zinc
pt

conditions. J Plant Nutr. 33: 1835-1848.

Shivay, Y.S., Prasad, R. and Rahal, A. 2010. Genotypic variation for productivity, zinc
ce

utilization efficiencies, and kernel quality in aromatic rices under low available zinc
conditions. J Plant Nutr. 33: 1835-1848.
Siddiq, E.A. 2006. Genetic improvement of rice in perspective. Indian Far. 56(7): 10-16.
Ac

Siddiq, E.A., Vemireddy, L.R. and Nagraju, J. 2012. Basmati rices: Genetics, breeding and trade.
Agric Res. 1(1): 25-36.
Singh, A.K., Khan, S.K. and Nongkynrih, P. 1999. Transformation of zinc in wetland rice soils
in relation to nutrition of crop. J Indian Soc Soil Sci.47: 248-253.
Singh, V.P. and Singh, A.K. 2009. History of Basmati rice research and development in India.
Indian Far. 59(1): 4-6.
Slaton, N.A., Gbur, E.E., Wilson, C.E. and Norman, R.J. 2005a. Rice response to granular
zinc sources varying in water-soluble zinc. Soil Sci Soc America J. 69: 443-452.
Slaton, N.A., Norman, R.J. and Wilson, C.E. 2005b. Effect of zinc source and application
time on zinc uptake and grain yield of flooded-irrigated rice. Agron J. 97: 272-278.
Srivastava, P.C., Ghosh, D. and Singh, V.P. 1999. Evaluation of different zinc sources for
lowland rice production. Biol Fertil Soils. 30: 168-172.

21
 ACCEPTED MANUSCRIPT

Srivastava, P.C., Singh, S.K. and Mishra, B. 2006. Crop response and profitability to applied
secondary and micronutrients. Indian J Fertil. 2(8): 45-51.
Subbiah, B.V. and Asija, G.L. 1956. A rapid procedure for the estimation of available nitrogen in
soils. Curr Sci. 25(8): 259-260.
Takkar, P.N., Singh, M.V. and Ganeshmurthy, A.N. 1997. A critical review of plant nutrient
supply needs, efficiency and policy issues for Indian agriculture for the year 2000:
Micronutrients and Trace elements. In: Kanwar, J.S. and Katyal, J.C. (Eds.) Plant
Nutrient, Supply Efficiency and Policy Issues: 2000-2025. National Academy of
Agricultural Sciences, New Delhi, India. pp. 238-264.
Tejada, M., Gonzalez, J.L., Martinez, A.M.G. and Parrado, J. 2008a. Application of a green
manure and green manure composted with beet vinasse on soil restoration: Effects on soil
properties. Bioresource Techn. 99: 4949-4957.

t
ip
Tejada, M., Gonzalez, J.L., Martinez, A.M.G. and Parrado, J. 2008b. Effects of different green
manures on soil biological properties and maize yield. Bioresource Techn. 99: 1758-
1767.

cr
Walkley, A.J. and Black, I.A. 1934. An examination of the Degtjareff method for determination
of soil organic matter and a proposed modification of the chromic acid titration method.

us
Soil Sci. 37: 29-38.
Whitbread, A.M., Blair, G.J. and Lefroy, R.D.B. 2000. Managing legume leys, residues and
fertilisers to enhance the sustainability of wheat cropping systems in Australia 1. The
an
effects on wheat yields and nutrient balances. Soil Tillage Res. 54(1&2): 63-75.
WHO. 2002. The World Health Report 2002. Geneva: World Health Organization. pp. 1-14.
Yadvinder-Singh, Khind, C.S. and Bijay-Singh. 1991. Efficient management of leguminous
M
green manures in wetland rice. Adv Agron. 45: 135-189.
e d
pt
ce
Ac

22
 ACCEPTED MANUSCRIPT

TABLE I
Effect of summer green manuring crops and Zn fertilizer sources on Zn concentration of rice
grain and straw and DTPA-extractable Zn in soil after harvest of Basmati rice

DTPA-extractable
Zn concentration Zn concentration in
Zn in soil after rice
in grain straw
Treatment harvest
(mg kg-1 grain) (mg kg-1 straw)
(mg kg-1)

2009 2010 2009 2010 2009 2010

Summer-green manuring crops

t
Sesbania aculeata (Dhaincha) 31.5 32.4 167.5 172.1 0.96 1.05

ip
Crotalaria juncea (Sunhemp) 29.8 29.6 162.1 162.4 0.91 0.93

cr
Vigna unguiculata (Cowpea) 27.2 26.7 155.6 156.2 0.83 0.85
Summer fallow 24.2 23.7 150.2 151.0 0.76 0.75

us
SEm± 0.47 0.55 2.05 2.37 0.007 0.011
LSD (P=0.05) 1.61an 1.90 7.08 8.16 0.023 0.039
Zn sources
Control 21.1 19.6 146.4 146.8 0.69 0.70
M
ZnSO4.7H2O (21 % Zn) 30.3 30.3 164.4 165.9 0.93 0.97
ZnSO4.H2O (33 % Zn) 29.3 29.1 159.6 161.3 0.86 0.86
d

ZnO (82 % Zn) 26.5 25.6 152.5 153.2 0.75 0.74

ZnSO4.7H2O + ZnO (50 % + 50 %) 28.1 28.1 156.7 158.1 0.81 0.82
e

EDTA-chelated Zn (12 % Zn) 33.9 35.7 173.5 177.4 1.14 1.27

pt

SEm± 0.29 0.35 0.68 0.94 0.011 0.011

ce

LSD (P=0.05) 0.84 0.99 1.96 2.70 0.030 0.032

LSD = Least significant difference
SEm± = Standard error of mean (±)
Ac

23
 ACCEPTED MANUSCRIPT

TABLE II
Correlation and regression coefficients between Zn concentration in Basmati rice dry matter and
DTPA-extractable Zn concentration in soil

S. Zn content in rice 2009 2010

No. dry matter (mg kg-1) R 2
Regression R 2
Regression
1) At 15 DAT 0.923** Y = 8.791X + 130.8 0.924** Y = 9.544X + 132.9

2) At 30 DAT 0.928** Y = 10.65X + 128.9 0.941** Y = 9.589X + 138.2

3) At 45 DAT 0.973** Y = 12.60X + 125.0 0.963** Y = 11.10X + 134.2

t
ip
4) At 60 DAT 0.972** Y = 15.95X + 121.9 0.955** Y = 14.21X + 131.1

5) At 75 DAT 0.974** Y = 16.12X + 124.5 0.955** Y = 14.89X + 130.1

cr
6) At 90 DAT 0.954** Y = 18.40X + 118.7 0.939** Y = 17.03X + 124.4

us
Y= DTPA- Zn and X= explanatory variable
**= Significant at p ≤0.05
an
Figures: 1 to 5 (C)
M
Sesbania aculeata (Dhaincha) Crotalaria juncea (Sunhemp)
d

Vigna unguiculata (Cowpea) Summer fallow

6.5
e

5.5
pt
DTPA- Zn (mg kg-1)

4.5
ce

3.5
Ac

2.5

1.5

0.5
15 DAT 30 DAT 45 DAT 60 DAT 75 DAT 90 DAT

Days after transplanting 2009

24
 ACCEPTED MANUSCRIPT

Control ZnSO4.7H2O (21% Zn)

ZnSO4.H2O (33% Zn) ZnO (82% Zn)
ZnSO4.7H2O + ZnO (50% + 50%) EDTA-chelated Zn ( 12% Zn)
8.5

7.5
DTPA- Zn (mg kg-1)

6.5

5.5

4.5

t
ip
3.5

cr
2.5

1.5

us
0.5
15 DAT 30 DAT 45 DAT 60 DAT 75 DAT 90 DAT
an
Days after transplanting 2009

Fig. 1 Effect of summer green manuring crops and Zn fertilizer sources on DTPA-extractable Zn
M
concentrations (mg kg-1) in soil with time after transplanting of Basmati rice during 2009. The
vertical bars represent LSD0.05 values.
e d
pt
ce
Ac

25
 ACCEPTED MANUSCRIPT

Sesbania aculeata (Dhaincha) Crotalaria juncea (Sunhemp)

Vigna unguiculata (Cowpea) Summer fallow
6.5

5.5
DTPA- Zn (mg kg-1)

4.5

3.5

2.5

t
ip
1.5

cr
0.5
15 DAT 30 DAT 45 DAT 60 DAT 75 DAT 90 DAT

us
Days after transplanting 2010

Control an ZnSO4.7H2O (21% Zn)

ZnSO4.H2O (33% Zn) ZnO (82% Zn)
ZnSO4.7H2O + ZnO (50% + 50%) EDTA-chelated Zn ( 12% Zn)
M
8.5

7.5
d

6.5
DTPA- Zn (mg kg-1)

5.5
pt

4.5
ce

3.5

2.5
Ac

1.5

0.5
15 DAT 30 DAT 45 DAT 60 DAT 75 DAT 90 DAT

Days after transplanting 2010

Fig. 2 Effect of summer green manuring crops and Zn fertilizer sources on DTPA-extractable Zn
concentrations (mg kg-1) in soil with time after transplanting of Basmati rice during 2010. The
vertical bars represent LSD 0.05 values.

26
 ACCEPTED MANUSCRIPT

Sesbania aculeata (Dhaincha) Crotalaria juncea (Sunhemp)

Vigna unguiculata (Cowpea) Summer fallow
215
Zn content of dry matter (mg kg-1)

205
195
185
175
165

t
ip
155
145

cr
135
15 DAT 30 DAT 45 DAT 60 DAT 75 DAT 90 DAT

us
Days after transplanting 2009

Control
an ZnSO4.7H2O (21% Zn)
ZnSO4.H2O (33% Zn) ZnO (82% Zn)
M
ZnSO4.7H2O + ZnO (50% + 50%) EDTA-chelated Zn ( 12% Zn)
220
Zn content of dry matter (mg kg-1)

210
d

200
e

190
pt

180
ce

170
160
Ac

150
140
130
15 DAT 30 DAT 45 DAT 60 DAT 75 DAT 90 DAT
Days after transplanting 2009

Fig. 3 Effect of summer green manuring crops and Zn fertilizer sources on the periodic Zn
concentrations (mg kg-1) in Basmati rice dry matter during 2009. The vertical bars represent
LSD0.05 values.

27
 ACCEPTED MANUSCRIPT

Sesbania aculeata (Dhaincha) Crotalaria juncea (Sunhemp)

Vigna unguiculata (Cowpea) Summer fallow
220
Zn content of dry matter (mg kg-1)

210
200
190
180
170

t
160

ip
150

cr
140
15 DAT 30 DAT 45 DAT 60 DAT 75 DAT 90 DAT

us
Days after transplanting 2010

Control an ZnSO4.7H2O (21% Zn)

ZnSO4.H2O (33% Zn) ZnO (82% Zn)
ZnSO4.7H2O + ZnO (50% + 50%) EDTA-chelated Zn ( 12% Zn)
M
220
Zn content of dry matter (mg kg-1)

210
d

200
e

190
pt

180
170
ce

160
150
Ac

140
130
15 DAT 30 DAT 45 DAT 60 DAT 75 DAT 90 DAT
Days after transplanting 2010

Fig. 4 Effect of summer green manuring crops and Zn fertilizer sources on the periodic Zn
concentrations (mg kg-1) in Basmati rice dry matter during 2010. The vertical bars represent
LSD0.05 values.

28
 ACCEPTED MANUSCRIPT

2009 2010
220 220
Y = 8.791X + 130.8 Y = 9.544X + 132.9
R² = 0.923 R² = 0.924
Zn content in rice dry matter (mg kg-1)

Zn content in rice dry matter (mg kg-1)

200 200

180 180

160 160

t
ip
140 140

cr
120 120

us
15 DAT 15 DAT
100 100
0 2 4 6 8 an 0 2 4 6 8

DTPA- Zn (mg kg-1) DTPA- Zn (mg kg-1)

M
240 220
Y = 10.65X + 128.9 Y = 9.589X + 138.2
R² = 0.928 R² = 0.941
d

220
Zn content in rice dry matter (mg kg-1)

200
Zn content in rice dry matter (mg kg-1)
e

200
pt

180

180
ce

160
160
Ac

140
140

120 120

30 DAT 30 DAT
100 100
0 2 4 6 8 0 2 4 6 8

DTPA- Zn (mg kg-1) DTPA- Zn (mg kg-1)

Fig. 5 (A) Relationship between Zn concentration in rice dry matter and DTPA-extractable Zn
concentration in soil with time after Basmati rice transplanting.

29
 ACCEPTED MANUSCRIPT

2009 2010
220 220
Y = 12.60X + 125.0 Y = 11.10X + 134.2
R² = 0.973 R² = 0.963

Zn content in rice dry matter (mg kg-1)

200 200
Zn content in rice dry matter (mg kg-1)

180 180

160 160

t
ip
140 140

cr
120 120

us
45 DAT 45 DAT
100 100
0 2 4 6 8
an 0 2 4 6 8

DTPA- Zn (mg kg-1) DTPA- Zn (mg kg-1)

M
220 220
Y = 15.95X + 121.9 Y = 14.21X + 131.1
R² = 0.972 R² = 0.955
d

200
Zn content in rice dry matter (mg kg-1)

200
Zn content in rice dry matter (mg kg-1)

e
pt

180 180
ce

160 160
Ac

140 140

120 120

60 DAT 60 DAT
100 100
0 2 4 6 0 2 4 6
DTPA- Zn (mg kg-1) DTPA- Zn (mg kg-1)

Fig. 5 (B) Relationship between Zn concentration in rice dry matter and DTPA-extractable Zn
concentration in soil with time after Basmati rice transplanting.

30
 ACCEPTED MANUSCRIPT

2009 2010
220 220
Y = 16.12X + 124.5 Y = 14.89X + 130.1
R² = 0.974 R² = 0.955
Zn content in rice dry matter (mg kg-1)

Zn content in rice dry matter (mg kg-1)

200 200

180 180

160 160

t
ip
140 140

cr
120 120

us
75 DAT 75 DAT
100 an 100
0 1 2 3 4 5 0 1 2 3 4 5

DTPA- Zn (mg kg-1) DTPA- Zn (mg kg-1)

M
220
220 Y = 17.03X + 124.4
Y = 18.40X + 118.7 R² = 0.939
d

R² = 0.954
Zn content in rice dry matter (mg kg-1)

200
Zn content in rice dry matter (mg kg-1)

200
e
pt

180 180
ce

160 160
Ac

140 140

120 120

90 DAT 90 DAT
100 100
0 1 2 3 4 0 1 2 3 4

DTPA- Zn (mg kg-1) DTPA- Zn (mg kg-1)

31
 ACCEPTED MANUSCRIPT

Fig. 5 (C) Relationship between Zn concentration in rice dry matter [above ground plant parts including leaf, stem
and panilces (at later stages of crop growth)] and DTPA-extractable Zn concentration in soil with time after Basmati
rice transplanting.

t
ip
cr
us
an
M
e d
pt
ce
Ac

32