Journal of Tea Science Research. 2016, Vol. 6, No.

4, 1-11
http://jtsr.biopublisher.ca

Research Article

Open Access

Drought Induced Physiological and Biochemical Changes in Leaves of

Developing Seedlings of Tea [ Camellia sinensis (L) O Kuntze ] Cultivars
Upadhyaya H.1, , Dutta B.K.2, Panda S.K.3
1 Department of Botany and Biotechnology, Karimganj College , Karimganj-788710, Assam, India
2 Microbial and Agricultural Ecology Laboratory, Department of Ecology and Environmental Sciences, Assam (Central) University , Silchar, 788011, India
3 Plant Biochemistry and Molecular Biology Laboratory, School of Life sciences, Assam ( Central ) University ,Silchar, 788011, India
Corresponding author email: hkupbl_au@rediffmail.com
Journal of Tea Science Research, 2016, Vol.6, No.4 doi: 10.5376/jtsr.2016.06.0004
Received: 28 Oct., 2015
Accepted: 15 Dec., 2015
Published: 28 Jan., 2016
2016 Upadhyaya et al., This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Upadhyaya H., Dutta B.K., and Panda S.K., 2016, Drought induced physiological and biochemical changes in leaves of developing seedlings of tea [Camellia
sinensis (L) O Kuntze] cultivars, Journal of Tea Science Research, 6(4), 1-11 (doi: 10.5376/jtsr.2016.06.0004)

Abstract Drought is one of the important environmental stress affecting agricultural productivity around the world. In this study, an
attempt has been made to understand drought induced biochemical alterations in different clones of Camellia sinensis [TV-1, TV-20,
TV-29 and TV-30]. Drought stress induced decrease in total chlorophyll and carotenoid, phenolics concentration and increases in
proline concentration, lipid peroxidation and polyphenols oxidase activity as a consequent of decrease in leaf relative water content
(RWC). Decreased Na+ and K+ concentration caused osmotic stress in leaves decreasing NR activity, and ultimately reducing leaf
relative growth rate. Thus, drought induced a range of physiological and biochemical alterations causing membrane damage and loss
in cellular functions ultimately leading to reduction in growth of one of the most important economic crop like tea. In comparison ,
TV-1 showed better drought tolerance by maintaining higher endogenous K+ and proline content and a balance Na+/K+ ratio in leaves.
Keywords Drought. relative growth rate(RGR); chlorophyll; phenols.proline; lipid peroxidation; Camellia sinensis.

1 Introduction

stress results in stomatal closure and reduced

transpirations rate, a decrease in water potential of
plant tissues, decrease in photosysthesis and growth
inhibition (Tahi et al. 2007), accumulation of abscisic
acid (ABA), proline, mannitol, sorbitol, formation of
radical scavenging compounds (ascorbate, glutathione,
tocopherol etc) and systhesis of proteins. Decrease in
photosysthesis is due to the limitted CO2 assimilation
caused by stomatal closure and decrease is total
chlorophyll content. Osmotic adjustment has been
considered as beneficial to drought tolerant
mechanism in field crop species. Na+ and K+ content
of leaf affect and regulates the osmotic potential of the
plant during stress conditions and hence Na+/ K+ ratio
should be adequate for stress acclimatization by plant
through osmotic adjustment . However, there is no
report on the changes of these ions in response to
drought in Camellia sinensis. The lowering of
osmotic potential by adjustment also minimizes the
opportunity for significant water loss to occur from
leaf tissue. This helps cells of higher plants to

Drought is one of the more important environmental

stresses affecting agricultural productivity around the
world and may result in considerable yield reduction
(Boyer 1982). Drought is a meteorological term that
denotes a period without rains during which soil water
content is reduced and plants suffer from lack of water.
Drought affects on the morphology anatomy and
physiology of the plants. Physiological, biochemical,
and anatomical responses however occur much earlier
than the usual symptoms of wilting, which may be
permanent or temporary depending on the availability
of soil moisture. The physiological, biochemical
and molecular mechanism involved in cellular and
whole plant responses to drought therefore generate
considerable interest and are frequently reviewed
(Ingram and Bartel, 1996; Chakraborty et al.
2002; Kar 2002; Shinozaki et al. 2002; Yordanav et al.
2003; Francois Tardieu 2003; Upadhyaya and Panda
2004; Reddy et al. 2004; Jeyaramraja et al. 2005;
Sakuma et al. 2006; Ohashi et al. 2006). Water
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withstand water deficit by maintaining sufficient
turgor to proceed (Grima and Krieg 1992 a,b).
Water stress induced pigment degradation, gross
decline in protein level, increased proline content,
carbohydrate status, lipid peroxidation, in plants
have also been reviewed ( Kar 2002 ) . Tea is second
only to water as the most consumed beverage in the
world. It has been used medicinally for centuries in
India and China. Green tea is comparatively healthier
than black and olong tea. The active constituents in
green tea are powerful antioxidants called polyphenols
(catechins) and flavonols. Research shows that tea
consumption is healthy and help fighting various
health problems which has also been reviewed by
many authors ( Kabir 2002; Higdon and Frei, 2003;
Cabrera et al. 2006). Tea is one of the most important
economic crops in Barak/Brahmaputra valley/ Dooars
and other hilly terrains of India (i.e. Darjeeling,
Himachal, Nilgiri and Uttaranchal). Tea plant being
perennial crops is subjected to different environmental
stress, drought being one of the important amongst
them. In N.E. India, generally tea suffers from drought
during November to April. In this region irrigation is
increasingly used as an insurance against drought to
maintain the productivity of tea during this period.
The influence of irrigation on the potential yield of tea
in this region has also been studied (Panda et al. 2003).
Thus the present investigation was undertaken for
understanding the mechanism of drought stress
induced physiological and biochemical alterations in
selected clone of Camellia sinensis L (O) Kuntze.
Drought tolerance in tea can be assessed through some
physiological and biochemical parameters under
moisture stress and these parameters can be used as
selection criteria for drought tolerance in the selection
and breeding programs of tea (Handique and
Manivel 1990). In the present experiment, the
field soil was used in the pot. The aim was to test the
responses of selected clones to drought stress imposed
by withholding water in the pot. Such pot experiments
have been used to asses the drought tolerance in plants
(Chakraborty et al., 2002; Sharma and Kumar, 2005,
Xu and Zhou, 2007, etc). However, the field
conditions were different from pot as the tea is a deep
rooted plant and soil area is not limited it can
penetrate the soil deep and it is obvious that it can

withstand more days of dehydration stress in the

field condition. During the period of natural drought
plant encounters long period without rain as it is
grown in rainfed ecosystem. Thus it is quite relevant
to evaluate drought responses of plant like tea in
potted conditions and correlate its responses in
relation to field performance as because all the plants
were grown in the same size pots and under same
environmental conditions.

2 Materials and Methods

Four commonly growing clonal varieties of Camellia
sinensis L. (O) Kuntze ( viz. TV-1, TV-20, TV-29 &
TV-30 ) seedlings of uniform age, one and half year
old were procured from. Tocklai Tea Research Station,
Silcoori, Silchar.
The seedlings grown in field soil in polyethene
sleeves were procured from the nursery of near by tea
Garden of Durgakona and brought to the laboratory.
The seedlings were potted after removing polyethene
sleeves and adding field soil. The plants were
acclimatized for 10-15 days in laboratory conditions
and were grown under natural light with well irrigation.
As tea is a shade loving plant, the seedlings were
grown in a shed where the intensity of light ranges
from 150-300 mol m- 2 s- 1 and 200-400 mol m- 2 s- 1
inside and outside the shed respectively. All control
and treated plants were kept in similar growth
conditions during acclimatization and treatment
imposition.
After 10-15 days of acclimatization, drought is
imposed by withholding water for 20 days. Well
watered plant is considered as control. The level
of stress was quantified by measuring changes in soil
moisture and RWC of leaf in stressed plant relative to
control. After dehydration, soil moisture content
decreased to (12.88 1.34)% and (3.55 0.28 )%
after 10 and 20d of stress imposition respectively
relative to control (23.46 1.62)%. The average
temperature range during experimental period was
noted as 25.1 32.3C and 12.5 24.7C max/min
respectively. The average relative humidity during the
experiment period was 88-96% and 38-67%
morning/afternoon respectively. All the leaf samplings were
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reagent and boiled in a water bath at 100C for 30 min.
After cooling the reaction mixture, 6.0 ml of toluene
were added and after cyclomixing it, absorbance was
read at 570 nm. Total phenolics were extracted from
tea leaves in 80 % (v/v) ethanol and were estimated as
per the method of Mahadevan and Sridhar (1982)
using Follin Ciocalteau reagent and Na2CO3.

done during morning hours between 8 am to 9 am.

Fresh mass of leaf was measured in three replicates
using five leaves and expressed as g leaf -1 . For
dry mass measurement same leaves were oven dried at
80C for 48 h and expressed as g leaf -1. The relative
gain in leaf total fresh mass was determined as relative
growth rate (RGR, [g d-1] of leaf or the increase in
total leaf fresh mass per unit of existing mass per unit
time), and calculated according to the formula:

Lipid peroxidation was measured as the amount of

TBARS determined by the thiobarbituric acid (TBA)
reaction as described by Heath and Packer (1968). The
leaf tissues (0.2 g) were homogenised in 2.0 ml of
0.1% ( w/v trichloroacetic acid (TCA). The homogenate
was centrifuged at 10,000g for 20 min. To 1.0 ml of the
resulting supernatent, 1.0 ml of TCA (20%) containing
10.5% (w/v) of TBA and 10L (4% in ethanol) BHT
(butylated hydroxytolune) were added. The mixture
was heated at 950C for 30min in a water bath and then
cooled in rice. The contents were centrifuged at
10,000 g for 15 min and the absorbancy was measured
at 532 nm and corrected for 600 nm. The concentration of
MDA were calculated using extinction coefficient of
155mM-1cm-1.

RGR=(InM2-InM1)/(t2-t1)
where M2 was the final total fresh mass (g), M1 was
the initial total fresh mass (g), t2 was the number of
days since initiation of experiment and t1 was day 0.
Relative water content (RWC) was measured by
following the methods of Barrs and Weatherly (1962).
Tea leaves were sampled, oven dried and digested in a
HNO 3-HCl (3:1,v/v) mixture and Na + and K +
concentrations were determined by Flame Photometer
(Systronics, India) (Jackson, 1973)
The stability of leaf membranes, was assessed by
determining leakage of electrolytes from leaf discs
placed in 20 ml of deionised water for 24 h at room
temperature and measuring the electrical conductivity
before and after autoclaving the samples and
Electrolytic leakage was determined as described by
Dionisio Sese and Tobita (1998) and calculated using
formula.

Leaf tissues were homogenized with potassium

phosphate buffer pH 6.8 (0.1M) containing 0.1 mM
EDTA, 1% PVP and 0.1 mM PMSF in prechilled
mortar pestle. The extract was centrifuged at 40C for
15 min at 17000 g in a refrigerated cooling centrifuge.
The supernatant was used for the assay of polyphenol
oxidase (PPO). PPO were assayed using pyrogallol as
substrate in 1.0 ml of enzyme extract. according to
Kar and Mishra (1976).After incubations at 250C for 5
min, the reaction was stopped with additions of 1.0 ml
of 10 % H2SO4. The purpurogallin formed was read at
430 nm. 1 unit of enzyme activity is defined as
that amount of enzyme which forms 1 mole of
purpurogallin formed per minute under the assay
conditions. Total soluble protein content was
estimated as per the method of Bradford (1976) using
BSA as standard.

EL = EC1/EC2 X 100
Where - EC1 is the initial conductivity of the sample
before autoclaving and EC2 is the final conductivity
measured after autoclaving the sample.
Leaves were extracted in cold with 80 % acetone. The
chlorophyll and carotenoid contents were estimated as
per the methods of Arnon (1949).
Proline concentration in tea leaves was determined
following the method of Bates et al. (1973). Leaf
sample (0.5 g)was homogenized with 5 ml of
sulfosalicylic acid (3%) using mortar and pestle and
filtered through Whatman No. 1 filter paper. The
volume of filtrate was made upto 10 ml with
sulfosalicylic acid and 2.0 ml of filtrate was incubated
with 2.0 ml glacial acetic acid and 2.0 ml ninhydrin

Nitrate Reductase (NR) activity in tea leaves was

extracted and estimated as per the methods of Singh
and Mallik (1980). The enzyme extracted in 0.1M
phosphate buffer (pH7.2) and the homogenate was
centrifuged in cooling centrifuge at 10,000 g at -4C.
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The resulting supernatant was used for the assay of
enzyme activity. The assay mixture comprised of 1.0
ml phosphate buffer (0.1M) (pH7.2), 0.5 ml NADH
(1.5 mM), 0.5 ml distilled water and 1.0 ml enzyme
extract. It is then incubated at 25C. The enzyme
reaction was initiated by adding 0.5 ml KNO3 (0.1 M)
and incubated for 30 min. Then 0.8 ml zinc acetate
(0.1 M) was added and centrifuged and 3.0 ml of the
supernatant was retained. To this 2.0 ml [1 %
sulfanilamide in 1.5 N HCl and 0.2 % N-naphthalene
diamine hydrochloride (NEDH)], mixed in equal
volume was added. After 10 min absorbency was read
at 540 nm. NR activity was expressed as moles NO2
min-1.g-1 FW.
Each experiment was done in triplicate and repeated
thrice and data presented are mean standard error
(SE). The results were subjected to ANOVA using
GLM factorial model on all the parameters. Tukey test
was used for comparision between pairs of treatments.
For relationship between relative water content and
proline accumulation, K+ content of leaf and its RWC,
lipid peroxidation and solute leakage of leaf tissue,
lipid peroxidation and decrease in RWC of leaf ,
relative growth rate of leaf and changes in total
chlorophyll content a linear regression was performed.
The data analysis were carried out using statistical
package SPSS 7.5.

Figure 1 Changes in solute leakage, relative growth rate(RGR)

of leaf and relative water content (RWC) in four clonal
varieties of Camellia sinensis ( TV-1, TV-20, TV-29 & TV-30 )
subjected to drought. Control (white); 10 days of drought
(gray) ; 20 days of drought (black) imposition. Data presented
are mean SE (n=3). Different lower case letters indicates
differences between drought treatment in the same variety ,
while * indicates differences within the varieties at P< .05
according to multiple comparision by Tukey test.

3 Results
Relative growth rate (RGR) of leaf showed uniformly
decline trend with progressive soil moisture stress in
all the tested clones (Figure 1B). Changes in RGR of
leaf due to10d of stress imposition was minimum in
TV-30 and TV-1 but after 20d of stress RGR changes
among the clones was not significant though decrease
growth was about 95%.The amount of solute
leakage is an index of membrane damage. Drought
induced increase in solute leakage in all the tested
clones of Camellia sinensis (Figure 1A). After 20d of
stress, in comparison with control plants solute
leakage increases to 130.84, 89.75, 150 and 153.92%
in TV-1, TV-17, TV-20, TV-29 and TV-30 respectively.
However, TV-1 and TV-20 showed comparatively
lesser amount of solute leakage during stress
conditions. Relative water content (RWC) of leaf
uniformly decreased with decreasing soil moisture

content imposed by 20 d of water withholding (Figure

1C). After 20 d of drought, RWC of leaf was
found to be 56.261.21, 52.871.08, 44.990.43 and
52.661.03% relative to control with 92.151.24,
90.212.83, 87.57 6.11 and 91.64 4.92 % in TV-1,
TV-20, TV-29 and TV-30.
The contents of Na+ ion increased with the progress of
water stress imposition, apparently showing highest
Na+ content in TV-29(41.73%) with lowest in TV-1 as
a result of 20 d of drought stress in comparision with
control plants (Figure 2A). On the other hand,
water stress induced decrease in K accumulation
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was observed in leaves of Camellia sinensis , but
highest K+ content was maintained by TV-1 (27.16%)
and TV-29 (13.85%) even after 20 d of drought
imposition as depicted in Fig 2B. There was increase
in Na+/ K+ ratio with
progressive days of water
withholding in TV-20 (105.80%),TV-29 (64.58%)
& TV-30 (283.25%), but with exception TV-1
(37.09%) showed decrease in Na+/ K+ ratio with the
increasing intensity of drought stress (Figure 2C).
Photosynthetic pigments (chlorophyll and carotenoid)
content was found to be decreased with increasing
days of stress imposition. Drought induced
degradation of chlorophyll and carotenoid was
maximum in TV-29 (57.91 & 82.38%) and TV-20
(55.10 & 86.13%) respectively as depicted in Fig.3A
& B. Phenolic compounds are widely distributed in
plants and are mainly produced to protect plants from
stress, ROS, wounds, UV light, disease and herbivores
(Dixon and Paiva 1995). Total phenolics content in tea
includes catechins and polyphenols, which was found
to be decreased with the imposition of drought (Figure
3C). Maximum decrease in phenolics content was
observed in TV-20(37.63%) and TV-2928.15%).
Increase in compatible solutes like proline, Glycine
betain etc., is the characteristic feature of plants
acclimatizing stress conditions. In this study proline
accumulation during water stress condition was
maximum in TV-1 (Figure 4A). Drought stress
induced significant increase in PPO activity was
observed in all the tested tea cultivars, maximum
increase being shown by TV-29 (Figure 4B). Highest
PPO activity was shown by TV-29(458.82%) and
TV-1 (424.91%). MDA content was maximum in
TV-29 (420.41%) after 20 d of stress imposition,
whereas TV-30(58.95%) showed minimum increase as
compared to control plant (Figure 4C).

Figure 2 Changes in Na+, K+ content and Na+/ K+ ratio in four

clonal varieties of Camellia sinensis ( TV-1, TV-20, TV-29 &
TV-30 ) subjected to drought . Control (white); 10 days of
drought (gray); 20 days of drought (black) imposition. Data
presented are mean SE (n=3) and significant differences are
shown as in (Figure 1)

The interesting aspect in this study is that there was no

significant correlation between relative water content
and changes in proline content of leaf (r2 = 0.15, ns)
(Figure 6 A). In our study, increase in Na+ content is
followed by significant decrease in K+ content of leaf
which is significantly correlated with decrease in
RWC of leaf (r2=0.50, p<0.01) as depicted in Figure 6
B. Concomitant with the increase in MDA content of
leaf amount of solute leakage also increased which
was evident from the significant correlation (r2=0.66,
p<0.001) between increase in lipid peroxidation and

As indicated in Figure 5A, NR activity decreased due

to drought in four clones of tea, where minimum
decrease was observed in TV-1(62.59%) and TV-30
(72.03%), that could be correlated with stress induced
decrease in total soluble protein content in tested
clones of Camellia sinensis (Figure 5B).
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Figure 4 Changes in proline content, polyphenol oxidase (PPO)

activity and MDA content in four clonal varieties of Camellia
sinensis (TV-1, TV-20, TV-29 & TV-30) subjected to drought.
Control (white ); 10 days of drought (gray) ; 20 days of drought
(black ) imposition . Data presented are mean SE (n=3) and
significant differences are shown as in Figure 1

Figure 3 Changes in total chlorophyll, carotenoid and phenolic

contents in four clonal varieties of Camellia sinensis (TV-1,
TV-20, TV-29 & TV-30) subjected to drought. Control (white) ;
10 days of drought (gray); 20 days of drought (black)
imposition. Data presented are mean SE (n=3) and significant
differences are shown as in Figure 1

comparision with control plants. On the other hand,

water stress induced decrease in K accumulation
was observed in leaves of Camellia sinensis, but
highest K + content was maintained by TV-1
even after 20 d of drought imposition. This is in
contradiction with the findings of Osmond et al.
(1980) and Martinez et al. (2003) who reported
involvement of Na + and K + accumulation in the
osmotic adjustment of leaf tissues to low external
water potential in Artiplex species during water stress.
Sodium contribution to osmotic adjustment may be
indirectly by triggering osmolyte synthesis independent of
osmotic stress was however demonstrated by Subbarao
et al. (2001). In our study, increase in Na+ content is

amount of solute leakage as shown in Figure 6C.

Increase in MDA content is significantly correlated
with decrease in RWC of leaf (r2 = 0.39, p<0.05) as
depicted in Figure 6D. There was significant
correlation between chlorophyll degradation and
RGR of leaf (r2 = 0.40, p<0.05) (Figure 6 E).

4 Discussion
Drought imposition resulted in significant changes of
growth and biochemical responses in various clones of
Camellia sinensis. The contents of Na+ ion increased
with the progress of water stress imposition,
apparently showing highest Na+ content in TV-29 with
lowest in TV-1 as a result of 20 d of drought stress in
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followed by significant decrease in K+ content of leaf
which is significantly correlated with decrease in
RWC of leaf (r2 = 0.50, p< .01) as depicted in Fig. 6 B.
K+ nutrition in plants is known to enhanced drought
resistance , water use efficiency and growth under

the present study, relative growth rate (RGR) of leaf

showed uniformly decline trend with increasing
drought stress. The growth of leaf is directly or
indirectly related with the various metabolism
dependent on its osmotic potential which is regulated
by potassium content of leaf. The amount of solute
leakage is an index of membrane damage. Drought
induced increase in solute leakage in all the tested
clones of Camellia sinensis. However, TV-1 and TV-30
showed comparatively lesser amount of solute leakage
during stress conditions. Drought induced membrane
damage could be due to enhanced membrane lipid
peroxidation which was evident from increased MDA
content with the progress of stress imposition,
observed in four clones of tea. MDA content was
maximum in TV-29 after 20 d of stress imposition,
whereas TV-30 showed minimum increase as compared to
control plant. Concomitant with the increase in MDA
content of leaf amount of solute leakage also increased
which was evident from the significant correlation (r2
= 0.66, p< .001) between increase in lipi d
peroxidation and amount of solute leakage as shown
in Fig. 6 C. Thus, it apparently suggests that increase
in leakage of solute could be due to drought induced
lipid peroxidation of membrane disturbing the
structural and functional integrity of cell causing
ultimate reduction in growth of tea plant.
Photosynthetic pigments (chlorophyll and carotenoid )
content was found to be decreased with increasing
days of stress imposition.Drought induced
degradation of chlorophyll and carotenoid was
maximum in TV-29 and TV-20 respectively. There
was significant correlation between chlorophyll
degradation and RGR of leaf (r2 = 0.40, p< .05).
Decrease in chlorophyll and carotenoid in response to
water stress was reported earlier (Upadhyaya and
Panda, 2004). Such degradation of chlorophyll
pigments may eventually decrease photosynthetic
efficiency in plants which might be one of the potent
causes of reduction in growth of plant. However, a
very recent study also provides evidence for a
significant positive relationship between transpiration
efficiency and leaf chlorophyll concentration in plant
(Sheshshayee et al. 2006). During prolonged periods
of drought, the decrease in water availability for

Figure 5 Changes in nitrate reductase (NR) activity and total

soluble protein content in four clonal varieties of Camellia
sinensis ( TV-1, TV-20, TV-29 & TV-30 ) subjected to drought .
Control (white) ; 10 days of drought (gray) ; 20 days of drought
(black) imposition . Data presented are mean SE (n=3) and
significant differences are shown as in Figure 1.

drought condition (Egilla et al., 2001 ). Thus it

suggested adequate K + fertilizer application of tea
plant may facilitate osmotic adjustment improving its
drought tolerance potential. There was increase in
Na + / K + ratio with progressive days of water
withholding in TV-20, TV-29 & TV-30, but with
exception TV-1 showed decrease in Na+/ K+ ratio with
the increasing intensity of drought stress (Figure 2C).
As K + is the main ion regulating osmotic
adjust ment in leaves , incr ease in Na + /K + ra tio
suggested occurrence of osmotic stress in three tea
cultivars (TV-20, TV-29 & TV-30) but exceptional
decreased Na+/ K+ ratio in TV-1 is the indication of its
comparatively better drought accl imatizing
potential. Water deficit stress, often called drought
affects the growth and productivity of plants
growing in natural or agricultural field conditions. In
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transport-associated processes leads to changes in the
concentrations of many metabolites, followed by
disturbances in amino acid and carbohydrate

metabolism. For example, there is an increase in the

synthesis of compatible solutes such as special amino
acids (e.g. Proline), sugars and sugar-alcohols, and

Figure 6 Relationship between relative water content and proline accumulation (A), K+ content of leaf and its RWC (B), lipid
peroxidation and solute leakage of leaf tissue (C), lipid peroxidation and decrease in RWC of leaf (D), relative growth rate of leaf and
changes in total chlorophyll content in four clonal varieties of Camellia sinensis subjected to drought . The open square, closed
triangle, open circle and closed circle denotes TV-1, TV-20, TV-29 and TV-30 variety of tea subjected to drought treatment
respectively. *, ** and *** indicates significant correlation at P<.05,.01 and .001 respectively.

Gly betaine.Acclimation to drought requires responses

that allow essential reactions of primary metabolism
to continue and enable the plant to tolerate water
deficits. In the complex interplay of natural conditions,
simple water deficits are unlikely to occur, since they
intrinsically affect the acquisition of essential nutrients
such as N etc. The reduction of NO 3 to NO 2
catalyzed by NR is considered to be the rate-limiting
step of N assimilation. NR activity is coordinated with

the rate of photosynthesis Indeed, drought-induced N

deficiency was found to limit recovery of
photosynthesis in plants. In situations of water
deprivation, maximal foliar extractable NR activity
has been found to decrease in some cases (Foyer et al.,
1998). In the present study, NR activity decreased
due to drought in four clones of tea , where
minimum decrease was observed in TV-1 and TV-30,
that could be correlated with stress induced decrease
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in total soluble protein content in tested clones of
Camellia sinensis. Similar results were already
reported in other plants (Foyer et al., 1998; Panda,
2002; Xu and Zhou, 2006). Increase in compatible
solutes like proline, Glycine betain etc., is the
characteristic feature of plants acclimatizing stress
conditions. In this study proline accumulation during
water stress condition was maximum in TV-1. Proline
acts as an osmoprotectant and greater accumulation of
proline in TV-1 suggested genotypic tolerance of tea
to water deficit stress, as proline accumulation helps
in maintaining the water relations, prevents membrane
distortion and acts as a hydroxyl radical scavenger.
Drought induced biochemical modifications and
proline metabolism has also been studied in other
plants (Sanker et al., 2007). The molecular mechanism
of quenching of reactive oxygen species proline has
also been well reviewed ( Matysik et al., 2001).
However, Wu et al., (2007) reported osmotic
adjustment in Citrus seedling colonized by Glomus
versiforme subjected to drought stress did not
correlate with proline but with ions like K+, Ca2+ , etc
and carbohydrates. Thus, similarly K+ might play
critical role in providing osmotic adjustment in
response to drought stress in tea cultivars as evidenced
by significant correlation between changes in
endogenous K+ content and RWC of leaves in stress
imposed tea cultivars (r2 = 0.50, p< .01). Also higher
K + content of TV-1 in stressed condition showed
better drought tolerance in comparision with other
clones. Relative water content (RWC) of leaf
uniformly decreased with decreasing soil moisture
content imposed by 20 d of water withholding (Fig.
1C). The interesting aspect in this study is that there
was no significant correlation between relative water
content and changes in proline content of leaf (r2 =
0.15, ns) ( Figure 6A).

good health drink. Polyphenol oxidases (PPO) are

widely distributed in the plant and play a role in
oxygen scavenging and defense against stress (Esen,
1993; Shivishankar, 1988). The antioxidant activity of
phenolic compounds is mainly due to their redox
properties. The medicinal value of phenolics in human
health is well documented (Caia et al., 2004; Tona et
al., 2004 ). In this study, drought stress induced
significant increase in PPO activity was observed in
all the tested tea cultivars, maximum increase being
shown by TV-29. PPO catalyses the O2-dependent
oxidation of mono- and o diphenols to o-diquinones,
where secondary reactions may be responsible for the
defense reaction and hypersensitive response (Mayer
and Harel, 1991) .Moreover, it is proposed that PPO
activity may regulate the redox state of phenolic
compounds and become involved in the
phenylpropanoid pathway (Kojima and Takeuchi 1989;
Nakayama et al., 2000).
In conclusion, it can be said that drought induced a
range of physiological and biochemical alterations
causing membrane damage and loss in cellular
functions ultimately leading to reduction in growth of
one of the most important economic crop like tea. In
comparision TV-1 showed better drought tolerance by
maintaining higher endogenous K+ and proline content
and a balance Na+/K+ ratio in leaves. Hence, the
present result may provide insight into underlying
mechanism of tea plant in response to soil drought at
the juvenile stage. However, more investigation is
required in the field at different plant growth stages in
the future. It can be further confirmed by studying
various physiological and biochemical responses of
different tea cultivars in field conditions and
conducting experiments to study the responses of
those cultivars to rehydration and post stress
manipulation of soil and leaf nutrients which are
already initiated at our laboratory.

Phenolic compounds are widely distributed in plants

and are mainly produced to protect plants from stress,
ROS, wounds, UV light, disease and herbivores
(Dixon and Paiva, 1995). Total phenolics content in
tea includes catechins and polyphenols, which was
found to be decreased with the imposition of drought
(Fig.3C). Tea polyphenols are mostly catechins which
have potential antioxidant properties that makes tea a

Acknowledgement
The authors thank Mr S.M. Bhati, General Manager,
Tocklai Tea Estate, Silcoorie, Silchar for providing Tea
seedlings throughout the experimental work.
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