Scientia Horticulturae, 44 ( 1990 ) 191-199 191

Elsevier Science Publishers B.V., A m s t e r d a m

Physiological changes in asparagus spears

immediately after harvest

R.E. Lill, G.A. King and E.M. O'Donoghue

Horticultural Research Centre, MAFTech, Ministry ofAgriculture and Fisheries, PrivateBag, Levin
(New Zealand)
(Accepted for publication 12 February 1990)

ABSTRACT

Lill, R.E., King, G.A. and O'Donoghue, E.M., 1990. Physiological changes in asparagus spears im-
mediately after harvest. Scientia Hortic., 44:191-199.

Respiratory activity of asparagus spears measured in the field showed a 4-fold increase between the
butt and the tip of 190-ram spears. After harvest, there was an immediate rise in the respiration rate,
followed by a rapid drop during 24 h at 16 ° C to a constant level, ~ 30% of the peak respiration rate.
Strong gradients of sugars and proteins were measured along the spears with low levels of sugars and
high levels of proteins present in the spear tips. Sugars declined markedly in the first 24 h after har-
vest, particularly in spear tips. Proteins in spear tips were unchanged 24 h after harvest, but had
decreased 25% by 72 h. Total free amino acids remained steady for the first 24 h after harvest, but
increased by 75% at 48 h. Asparagine/aspartic acid increased, whereas glutamine/glutamic acid and
proline decreased in concentration substantially during the first 24 h after harvest. Tips of taller spears
had a lower sugar content and more protein than tips of short spears.

Keywords: amino acids; Asparagus officinalis L; protein; respiration; sugars.

INTRODUCTION

Asparagus spears are very active metabolically and highly perishable dur-
ing handling and storage (Lill, 1980; King et al., 1988 ). A shelf life of 4-5
days after storage for 3-4 weeks is desired by the New Zealand industry to
enable sea freight of asparagus to Northern Hemisphere markets, but is not
yet reliably attainable due to tissue deterioration during shelf life (Lill, 1980;
King et al., 1986, 1988 ). A more complete understanding of the deteriorative
process is required before significant progress can be made in extending the
shelf life of asparagus after storage.
Perishability relates in part to the tissue type of the organ. In asparagus, the
pattern of deterioration is influenced by the heterogeneity of the tissues: the
tip comprises actively dividing meristematic cells grading into a zone of cel-
lular elongation, whereas the butt comprises more mature tissue where cell

0304-4238/90/$03.50 © 1990 - - Elsevier Science Publishers B.V.

192 R.E. LILL ET AL.

elongation has ceased and the vascular tissue is lignifying. Highly metabolic
tissues generally have poor storage potential (Burton, 1982).
Tissue heterogeneity is reflected in physiological gradients in the spear. Tip
sections of detached spears produce CO2 at four times the rate of butt sections
1 day after harvest (Saltveit and Kasmire, 1985 ). Tips have high concentra-
tions of total nitrogen (Culpepper and Moon, 1939), protein (Hsaio et al.,
1981; Saltveit and Kasmire, 1985; King et al., 1988 ), free amino acids ( Salt-
veit and Kasmire, 1985 ) and dry matter (Culpepper and Moon, 1939; Scott
and Kramer, 1949), but are low in soluble carbohydrate (Culpepper and
Moon, 1939; Hsaio et al., 1981; King et al., 1988) in comparison with the
butt.
Some of these gradients appear to change as the spear grows. Culpepper and
Moon (1939) report that total sugars in the tip of 200-mm spears are one-
tenth the level in the tip of 100-mm spears, whereas levels in the butt are
similar in both heights of spear. Nitrogen levels appear to be closely related
to the zone of elongation, with levels increasing rapidly above this zone to-
ward the tip (Culpepper and Moon, 1939). The nitrogen concentrations in
the tip do not alter in taller spears, but because the amount of fully elongated
tissue increases, taller spears have more tissue with low nitrogen. A reduction
in free amino acids has been observed in the tips as spears grow, with proline
decreasing most (Sagisaka and Araki, 1983 ).
To better understand the physiological response of spears to harvesting, we
have determined the respiratory rates and composition of unharvested spears
at a range of heights, followed by a study of changes in composition after
harvest.

MATERIALS AND METHODS

Plant material. - Asparagus spears (Asparagus officinalis L. cultivar 'Lim-

bras 10' ) were obtained from production trials at the Levin Horticultural Re-
search Centre.

Respiration rate measurements. - C O 2 production was measured on intact

spears using a portable infrared gas analyser (Analytical Development Com-
pany Ltd., Hoddesdon, U.K. ). A cuvette ( 10 ml total volume) was sealed to
appropriate spear sections using closed-cell foam plastic, and a constant flow
of air ( 150-300 ml m i n - i, depending on temperature ) was pumped through
the cuvette to the analyser. Regression curves relating spear diameter to weight
of the spear section (r2---0.90 tip, n = 8 ; r2=0.98 mid and butt, n = 16) were
used to calculate tissue weight for non-destructive analysis of respiration.
The respiration rate of spears growing in the field was measured on five
spears each of 40, 110 or 190 m m height. Respiration rates of tip (0-30 m m ) ,
mid (75-105 m m ) and butt ( 150-180 m m ) sections were measured. Spear
CHANGES IN HARVESTED ASPARAGUS SPEARS 19 3

temperature in the field at the time of measurement was 33 ° C. Weather con-

ditions were still and sunny. Five spears in each of three replicates (blocked
on butt diameter) at the different heights were then harvested and divided
into tip, mid and butt sections prior to freeze-drying for compositional
analyses.
Respiration rate changes after harvest were determined by measuring tip
(0-30 m m ), mid ( 60-90 m m ) and butt ( 120-150 m m ) sections of five spears
in the field at 16 ° C. Insufficient 180-mm long spears were available for this
experiment so shorter 150-mm long spears were used. These spears were cut,
trimmed to 150 m m and transferred to a temperature-controlled room at
16 °C. Respiration rate was measured at intervals over 72 h storage. Addi-
tional spears were harvested, trimmed and held at 16 °C for compositional
analyses. Five spears in each of three replicates blocked on butt diameter were
sampled at 0, 24 and 72 h, divided into tip, mid and butt sections, and freeze-
dried prior to compositional analysis.

Sugar and acid analyses. - Triplicate samples of 20 mg freeze-dried tissue

were extracted with 80% methanol at 70°C for 2 min. Acids were separated
on Sephadex G-50. Sugars and acids (released from the Sephadex with 4%
formic acid) were dried, silylated with Trisil-Z (Pierce) and analysed sepa-
rately by a gas liquid chromatograph (GLC) fitted with a 5-m methyl silicone
capillary column (530/zm diameter) and a flame ionization detector. The
temperature programme for sugars was 165 ° C for 1 min, rising at 20 ° C m i n - 1
to 220 ° C, hold for 0.5 min, then rising at 25 ° C m i n - 1to 270 oC. The temper-
ature programme for acids was 115 °C for 2 min, rising at 15 °C rain-1 to
160 oC, hold for 1 min, then rising at 20 ° C m i n - ~ to 225 ° C. Quantification
was by external standard. This method was adapted from that reported by
King et al. (1988).

Protein analyses. - Total protein was determined from duplicate samples of

10 mg freeze-dried tissue. Chlorophyll was removed as previously described
(King et al., 1988 ) prior to solution of protein by boiling for 10 min in 0.1 M
NaOH. Cell debris was removed by centrifugation and the pellet re-extracted
as above. Protein was estimated using a modified Bradford method (Scopes,
1982 ) with bovine serum albumin as the standard. Absorbence readings were
taken at 750 nm.

- Free amino acids were extracted from 500 mg freeze-

F r e e a m i n o acids.
dried tip tissue by homogenization with 5 ml 80% ethanol. Cell debris was
removed by centrifugation (Ordonez and Burgos, 1980) and the pellet was
re-extracted twice as above. Extracts were pooled and dried under vacuum.
The residue was derivatised with 7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole prior
194 R.E. LILLET AL.

to analysis by high performance liquid chromatography (Watanabe and Imai,

1982).

RESULTS

Respiration and composition during spear growth. - The respiration rate of

spears growing at different heights in the field showed a large gradient, declin-
ing from tip to butt (Table 1 ). No trend was observed between tips or mid
sections of spears of different heights.
Compositional analyses of tip sections of spears at different heights showed
that soluble sugars were less in taller spears, with the strongest gradient occur-
ring for glucose (Table 1 ). Fructose and sucrose had similar gradients, with
concentrations in tip sections of 190-mm spears ~ 50% the concentration in
40-ram spears. Malic acid tended to be less in shorter spears, while citric acid
was present in constant amounts amongst spears of different heights. Protein
content tended to be higher in tips of longer spears. There was little difference
in composition in mid sections between spears of different heights (Table 1 ).

Post-harvest respiration rate and changes in composition. - By 2 h after har-

vest, the respiration rate had increased markedly in all sections of the spear
(Fig. 1 ). Subsequently, respiration decreased rapidly until 24 h after harvest
when the rates approached constant values at ~ 30% of the respiratory peak
of each section.
Large changes in composition occurred over the storage period (Table 2 ).
Soluble sugars declined substantially during the first 24 h after harvest. This
decline was greatest (in percentage terms) in the upper regions of the spear
and glucose was more affected than fructose. Levels of glucose, fructose and
sucrose were similar in spear tips, but while sucrose was present at similar
TABLE 1

Respiration rate (mg CO2 kg- ~h - a, (standard error) ) and compositional analysis (all data mg g- 1 dry
weight, (standard error) ) of growing spears of different heights at 33 ° C. Tip sections were 0-30 mm,
mid sections 75-105 mm and butt sections 150-180 mm

Section Spear Respiration Glucose Fructose Sucrose Malic Citric Protein

height rate acid acid
(mm)
Tip 190 2939 (306) 7.7 (1.2) 21.7 (4.4) 5.6 (0.1) 5.9 (0.1) 7.0 (2.5) 346(1)
110 3482(194) 16.7(1.2) 31.3(1.1) 7.4(1.3) 5.4(0.8) 7.2 (1.2) 335 (7)
40 2665 (185) 25.8 (6.6) 43.7 (5.8) 10.2 (1.3) 3.6 (0.5) 6.9 (0.7) 312 (6)
Mid 190 1054 (274) 64.0 (4.6) 111.6 (7.7) 3.0 (0.3) 7.4 (1.0) 5.7 (1.0) 174 (3)
110 1080(65) 82.0(9.0) 104.9(12.3) 4.2(0.3) 6.7(0.7) 5.6 (0.6) 167 (13)
Butt 190 571 (55) 115.4 (10.6) 149.8 (20.8) 12.4 (1.7) 8.4 (0.5) 4.6(1.3) 110 (1)
CHANGESIN HARVESTEDASPARAGUSSPEARS 195

1400"

1200"

~= 1000" /
i "

d
o 800 ",q
E~
E

600 .1\.
'\ k ""--..
i ........
TIP . . . . . . . . o

d~
re
400 - ~...
x. %.

........ - .... ..._~_.__MJD__ . . . . . .

2001 "------..,.
.....................................
/IIIIIT
/TIT
I I I I
..B.,U..TT
........................ i

0 24 4"8 7'2
Storage time (h)

Fig. 1. Respiration rate of asparagus spear sections during 72 h storage at 16 ° C. Bars = standard
error.

TABLE 2

Composition of asparagus spear sections during 72 h storage at 16 °C (all data mg g - ~dry weight,
(standard error) ). Tip sections were 0-30 ram, mid sections 60-90 mm and butt sections 120-
150 mm from 150-mm long spears

Section Time Glucose Fructose Sucrose Malicacid Citric Protein

(h) acid
Tip 0 17.3 (2.8) 26.5 (3.2) 12.3 (2.8) 3.8 (0.3) 7.0 (0.3) 243 (8)
24 4.7 (2.0) 8.5 (1.9) 2.9 (0.9) 3.9 (0.6) 9.4 (5.2) 257 (5)
72 4.7 (1.5) 12.7 (0.8) 7.8 (2.1) 7.6 (1.1) 15.0 (2.6) 185 (15)
Mid 0 111.7 (18.8) 157.7 (18.8) 12.0 (1.9) 4.2 (1.6) 7.8 (1.4) 135 (6)
24 26.0 (6.4) 77.9 (7.3) 1.9 (0.6) 3.7 (0.5) 4.8 (0.5) 141 (4)
72 56.3 (16.1) 124.2 (13.0) 15.2 (0.9) 10.4 (1.5) 3.7 (0.4) 126 (5)
Butt 0 173.3 (5.2) 200.7 (15.3) 22.7 (4.9) 5.1 (0.7) 4.2 (0.7) 43 (2)
24 68.2 (4.0) 100.9 (3.6) 8.7 (5.8) 5.2 (0.3) 5.3 (2.3) 42 (1)
72 86.3 (8.9) 142.5 (12.4) 11.7 (1.3) 12.5 (0.9) 3.3 (0.3) 45 (2)
196 R.E. HLLETAL.

concentrations throughout the spear, glucose and fructose concentrations in-

creased substantially toward the spear butt. Sugar levels increased from 24 to
72 h after harvest, b u t remained m u c h lower than at harvest except for su-
crose in the m i d sections.
Malic and citric acid concentrations remained constant over the first 24 h
after harvest, b u t almost d o u b l e d in the tip by 72 h (Table 2). This change
occurred for malic acid in the m i d and butt sections. Citric acid did not in-
crease in m i d and butt sections during 72 h after harvest.
Protein content did not change over the first 24 h, b u t declined substan-
tially in the tip by 72 h after harvest (Table 2 ). This change was not apparent
in other sections o f the spear where protein concentrations were lower.
Total free amino acids increased significantly between 24 and 48 h (Table
3 ), although alanine, arginine and hydroxyproline did not change over 72 h.
Asparagine/aspartic acid d o u b l e d during the first 24 h to b e c o m e the m a j o r
amino acids, whereas glutamine/glutamic acid halved from its position as a
m a j o r c o m p o n e n t at harvest. Collectively, asparagine/aspartic acid and glu-
t a m i n e / g l u t a m i c acid comprised > 50% of the free amino acids at harvest.

TABLE 3

Free amino acid content (#mol g-~ dry weight, (standard error) ) of asparagus tips (0-30 mm
of 150-mm long spears) during 72 h storage at 16 °C

Amino acid Time after harvest (h)

0 24 48 72
Alanine 30.0 (2.6) 24.3 (3,0) 37.3 (1.8) 33.7 (2.7)
Arginine 20.0 (2.6) 20.7 (2.8) 31.0 (5.8) 30.0 (3.1)
Asparagine/aspartic acid 68.3 (2.8) 163.3 (3.3) 306.7 (13.3) 303.3 (46.3)
Glutamine/glutamic acid 180.0(20.8) 89.7 (11.3) 67.3 (11.1) 106.0 (7.0)
Glycine 9.4 (0.2) 9.7 (0.7) 18.7 (0.9) 15.7 (0.9)
Hydroxyproline 0.7 (0.1) 1.1 (0.1) 1.1 (0.2) 1.4 (0.3)
Isoleucine 2.2 (0.1) 7.2 (0.9) 24.3 (0.3) 37.3 (3.8)
Leucine 8.1 (1.2) 12.0 (0.6) 23.3 (0.9) 27.3 (1.9)
Lysine 1.7 (0.1) 7.4 (1.9) 16.7 (0.7) 21.7 (3.2)
Methionine 3.6 (0.2) 3.9 (0.5) 6.1 (0.5) 6.3 (1.0)
Phenylalanine 2.8 (0.2) 7.3 (1.0) 18.7 (0.9) 29.0 (3.5)
Proline 19.0 (3.1) 4.0 (0.1) 5.7 (0.4) 5.3 (O.7)
Serine 30.3 (2.3) 44.3 (2.3) 66.7 (2.2) 75.0 (2.9)
Threonine 8.9 (0.6) 16.3 (0.9) 20.7 (0.3) t8.0 (o.o)
Tryptophan 0.3 (0.0) 1.3 (0.2) 3.4 (0.1) 4.9 (0.1)
Tyrosine 1.4 (0.2) 2.9 (0.5) 3.4 (0.4) 2.9 (0.2)
Valine 15.7 (1.2) 21.3 (1.9) 50.3 (2.3) 74.3 (3.8)
Total 402 (17) 436 (15) 701 (33) 792 (71)
CHANGES IN HARVESTED ASPARAGUS SPEARS 197

Proline was the only other amino acid to decline, dropping by 75% over 24 h,
and remaining stable thereafter.

DISCUSSION

The asparagus spear tip is unusual from a post-harvest viewpoint because

it is a shoot, characterized by intense metabolic activity, and is of a highly
perishable nature. Our data contribute to the knowledge on three major issues
of asparagus physiology: respiration whilst plants are growing in the field,
changes in the composition of individual soluble carbohydrates during stor-
age and changes in amino acid composition during storage.
Respiratory activity of asparagus spears has not been measured in the field
before, nor has activity in different sections of intact spears been followed
after harvest. In the field, the respiration rate of spear tips was over 5 times
greater than the butt sections. This reflects a major gradient in metabolic ac-
tivity along the spear (Saltveit and Kasmire, 1985), with apical tissue (in-
volved in active cell division and the initial stages of cell elongation) showing
the highest respiration rate.
A major rise in respiration rate was measured 2 h after harvest. Spear tem-
perature would have had to increase~ 5 °C ( Q l o = 2 ) to explain this effect.
The temperature of these spears in the field was 16 °C and they were moved
into a 16 ° C cool room within 15 min of cutting. A more probable explanation
is that a respiratory surge occurred in response to harvesting and handling.
This should be investigated further.
The pattern of change in respiration rate in all sections showed a rapid de-
cline during the first day after harvest to reach a stable constant rate. How-
ever, even at this stable rate the respiratory gradien t from tip to butt re-
mained. This pattern of respiration rate change is very similar to that reported
in whole spears (Platenius, 1942; King et al., 1988) and in a range of other
vegetables (Platenius, 1942 ). The changes are slower and less pronounced at
lower temperatures (Platenius, 1942; Saltveit and Kasmire, 1985; King et al.,
1988 ). Trippi et al. ( 1988 ) associated a decline in respiratory activity with
senescence in cut flowers and suggested it is related to nucleotide metabolism.
It seems probable that the supply and control of metabolic energy is vulnera-
ble to disruption, which quickly leads to loss of normal respiratory function.
This change in response to harvest may be similar in a wide range of plant
tissue.
The gradients in sugars in freshly harvested spears generally reflect those
observed by Hsaio et al. ( 1981 ), although we observed lower levels of sugars
in the spear tip, and rather higher levels of fructose in mid and butt sections.
The observation that the sugar concentration in the tip section is influenced
by spear height confirms earlier work (Culpepper and Moon, 1939) and sug-
gests that there is a very high turnover of these simple sugars with the pool
198 R.E. LILL ET AL.

size influenced markedly by the rate of import. It is likely that imports from
sugars mobilized in the crown and storage roots will become more restricted
as the spear grows.
Gradients in organic acids were not as clear cut. There was a tendency for
malic acid to be higher in the tips of longer spears and to be at lower concen-
trations in tips than butts. Between 24 and 72 h, the acid pools altered, pre-
sumably reflecting changing metabolism in response to substrate depletion
after harvest.
Protein concentrations more clearly indicated a disruption in metabolism,
particularly in the tip where significant loss of protein occurred between 24
and 72 h. It is in this region that a massive decline in soluble carbohydrate
pools during the first 24-h period must force the tissue to use alternative res-
piratory substrates.
Calculation of the respiratory requirement for carbohydrate in spear tips
over the 72-h period indicated that, after accounting for the loss of soluble
carbohydrate, there was a deficit of 46 mg carbohydrate per tip. The calcula-
tion for m i d sections indicated a deficit there of 20 mg. Butt sections however
lost 29 mg more carbohydrate than required for respiration. These calcula-
tions suggest some translocation of soluble carbohydrate from spear butts to
the upper parts of the spear, but still far short of the respiratory d e m a n d in
tip and m i d sections. This supports the hypothesis that asparagus spears use
an alternative substrate to carbohydrate to support their high post-harvest
respiration rate.
Protein is the most likely candidate as asparagus spears are thought to use
protein in normal respiratory metabolism (Platenius, 1942, 1943; King et al.,
1988) and the large increase in free amino acids after 24 h supports this
hypothesis.
Asparagine/aspartic acid and glutamine/glutamic acid are the major com-
ponents of the amino acid pool and, together with proline, appear to be key
elements in asparagus metabolism because all changed appreciably during the
24-h period prior to substantial protein loss.
The high level of protein in tips compared to butts correlates well with
smaller cells in active division in this tissue, whereas cells at the base of the
spear are fully expanded and are in the first stages of secondary thickening. It
is less clear why tips of longer spears should have higher concentrations of
proteins than tips of shorter spears. We think it could relate to activity in
axillary buds which are more actively developing in 190-ram spears than in
40-mm spears.
We conclude that asparagus spears are very active metabolically and that
the control of that metabolism is vulnerable to disruption. In future studies,
we aim to better characterize this disruption.
CHANGES IN HARVESTED ASPARAGUS SPEARS 199

ACKNOWLEDGEMENTS

We thank Wilhelmina Martin and Gus van der Mespel for technical assis-
tance with this project. The amino acid analyses were carded out by Dr. David
Woollard, MAFQual Auckland Dairy Laboratory, Lynfield Agricultural
Centre.

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