Chapter IV - Shodhgangashodhganga.inflibnet.ac.in/bitstream/10603/5496/9/09...Chapter IV 91 4.2...

Chapter IV

Real-time qPCR Monitoring of Genes Involved in Water Stress Tolerance in Tea

Chapter IV

89

41 Introduction

Water stress is one of the most important environmental factor that limits

crop productivity and quality throughout the world resulting in substantial

loss every year (Tan et al 2008) Efficient irrigation is one of the key factor

for sustainable agriculture as pointed out by many researchers from time to

time (Hillel and Vlek 2005 Khan et al 2006 Hsiao et al 2007 Pannunzio

et al 2008 Pannunzio 2008) However due to continuous depletion and

uneven distribution of water resources in many regions of the world it has

become necessary to genetically improve the performance of crop plants in

such water stress environments for a sustainable and economically viable

solution

Successful cultivation of tea is highly dependent on availability of rain water

There are reports of yield losses every year due to recurring prolong dry

spell in the tea growing areas (Barua 1989 Jain 1999) of India and other

countries like Kenya and Sri Lanka Keeping in view the continuous increase

of drought hit regions due to decreased rainfall the present need of the tea

industry is a good quality high yielding tea genotype that can withstand and

perform well during the prolonged dry spells of the year One of the ways to

achieve this goal is to study the drought responsive mechanism in tea at

molecular level to identify the different drought responsive pathways and

their corresponding genes which would potentially contribute towards

development of drought tolerant tea plants However the development of

drought tolerant tea genotypes has been hindered by lack of knowledge of

more precise physiological and molecular parameters that reflect genetic

potential for improved productivity under water stress environment

Comparative study of the transcriptome of a drought tolerant tea genotype

with a susceptible one is one of the potential ways to decipher the

mechanism of drought tolerance in tea plant We have identified a large

numbers of differentially expressed transcripts in a drought tolerant cultivar

TV23 by comparative transcriptome analysis with a susceptible cultivar

S3A3 (Chapter III) from an induced water stress experiment by SSH

approach Some of these differentially expressed transcripts are highly

represented in TV23 compared to S3A3 which may potentially be

Chapter IV

90

contributing for contrasting drought tolerance behaviour of TV23 However

the expression pattern of these transcripts has to be studied at various

degrees of water stress as well as in unstressed conditions to correlate their

potential role that they might play during water stress We have identified

(Chapter III) a total of 49 drought responsive transcripts represented by 3 or

more than 3 ESTs in the drought tolerant cultivar TV23 In the present study

we have selected 29 transcripts that are highly represented in TV23

compared to S3A3 and studied their expression pattern at different stages

of drought in both the cultivars using qRT-PCR

Water stress response in plants involves expression of a large number of

genes encoding proteins with an adaptive role These complex set of genes

are thought to function not only in protecting cells from water deficit by

production of important metabolic proteins but also in regulation of genes for

signal transduction in water stress response (Shinozaki et al 2003) The

products of these genes can be classified into two groups The first group

includes genes encoding proteins that function directly in the protection of

plant cells against stresses such as heat stress proteins LEA proteins

osmoprotectants detoxification enzymes and free radical scavengers

whereas the second group includes those that control gene expression and

signal transduction (Wang et al 2003) Monitoring the expression of these

genes at mRNA level in different tissues or in tissues under different degrees

of stress by qRT-PCR is a well established technique for study of differential

expression pattern in real time during water stress environment Real-time

RT-PCR is the most sensitive method for detection of low abundance mRNA

(Bustin 2000) Here we have used qRT-PCR to monitor the expression

pattern of selected genes at three stages of drought induction The qRT-

PCR analysis of these genes at BWS will further validate our SSH library

constructed in Chapter III Therefore the present study was carried out with

the objective to validate the SSH library by monitoring the expression pattern

of highly induced drought responsive genes from library BWE3E4 at BWS

and also to monitor their expression pattern at WS and AWS in both

cultivars

Chapter IV

91

42 Materials and Methods

421 Plant materials and RNA Isolation

The plant material used in the present study for isolation of RNA was a bud

along with 1st and 2nd leaf collected during induced water stress experiment

(Chapter II) Total RNA was isolated from tissues of TV23 and S3A3

collected at three stages of drought induction ie before wilting stage

(TV23BWS and S3A3BWS) wilting stage (TV23WS and S3A3WS) and

after wilting stage (TV23AWS and S3A3AWS) using RNAqueous kit

(Ambion USA Cat No Am1912) following the manufacturers protocol Total

RNA was also isolated from TV23 grown under well watered condition

(TV23C) The purity and concentration of RNA was checked using a

spectrophotometer (BioPhotometer Eppendorf Germany) and integrity was

ensured by running a 1 denaturing agarose gel

422 Selection of Internal Control Genes for Normalization

For internal standard we have selected four housekeeping genes viz 26S

rRNA 18S rRNA Camellia tubulin and ribulose-1 5-bisphosphate

carboxylaseoxygenase and tested their variability of expression at three

stages of drought induction (BWS WS and AWS) in both the cultivars under

consideration The gene sequences were retrieved from public database and

primer was designed using Primer3 [(httpfrodowimiteduprimer3)

(Rozen et al 2000)] The details of gene accession numbers and designed

primers are given in Table 41

Table 41 Housekeeping gene and primers used for normalization

Genes Gene Bank acc number

Primer sequence (5ʹ - 3ʹ) Length (nt)

26S rRNA AY283368 TCAAATTCCGAAGGTCTAAAG CGGAAACGGCAAAAGTG

21 17

18S rRNA AY563528 GGCCGGCTCCGTTACTTTG GTTTCAGCCTTGCGACCATACTC

19 23

Camellia tubulin DQ444294 AGCGTGCGGTTTGCATGA GCCCAAAGGTTTGGCATCA

18 19

Ribulose-1 5-bisphosphate carboxylaseoxygenase

EF0110751 AAGCACAATTGGGAAAAGAAG AAAGTGAAAATGAAAAGCGACAAT

21 24

Chapter IV

92

423 Selection of Drought Induced Genes for qRT-PCR Assays

Among highly represented drought induced ESTs in BWE3E4 library (Table

32 Chapter III) we have selected 29 genes (Table 42) for expression

studies based on their existing report of induction under drought and other

environmental stresses and also on the role they might play in giving drought

tolerance to plants (Table 43)

4231 Primer Design

ESTs with high quality value representing the above genes were considered

for primer designing using Primer3 A total of 29 primer pairs were designed

whose length varies from 20 nt (nucleotide) to 22 nt and Tm from 60oC to

64oCThe details of selected genes and corresponding primer sequence and

product size are given in Table 42

424 Standardization of PCR Parameters and Product size Verification

The PCR parameters for all the housekeeping genes and the selected

drought induced genes were standardized in a gradient thermal cycler

(Mastercycler gradient Eppendorf Germany) before going in to the qRT-

PCR analysis The product sizes were verified by running an agarose gel

These standardised PCR profiles were used during qRT-PCR analysis

425 Two Step qRT-PCR

4251 Reverse Transcription

1 microg total RNA was reverse transcribed using Transcriptor First Strand

cDNA Synthesis kit (Roche Germany Cat No 04379012001) following the

manufacturers protocol Briefly 1 microg of total RNA and 1 microl of anchored-

oligo(dT)18 primer and required volume of nuclease free water to make the

final volume of 13 microl were gently mixed in a 02 ml nuclease free PCR tube

and incubated at 65ordmC for 10 minutes in a thermal cycler (ABI 2720) to

denature the template-primer mixture The tube was immediately chilled on

Table 42 Details of drought induced genes selected for expression studies

EST ID dbEST acc no

BlastX homology with E-value Designed primer sequence 5ʹ - 3ʹ (20 ndash 22 nt)

Tm (ordmC) Product size (bp)

N23E3E4_A176 HS393998 Abscisic stress ripening protein (ASR1) 3e-19

ACCACCAACGCCTATGGAAG CTGCAGCCACAGCTGCTATC

6220 6224

268

N23E3E4_A482 HS394253 Ascorbate peroxidase 6e-109

CACGATGCTGGGACGTATGA CTGCAACAACACCAGCAAGC

6306 6298

120

N23E3E4_A682 HS395169 APE1 2e-49 GGGAAATGTGCCACTCCAAT TCGCGAATCATTTGCAGAAC

6205 6223

164

SSH2_269 HS396211 Copper-containing amine oxidase 5e-21 GCTTCGATCTAAAACCGGTCA AGCTGCAAACAACACCCAAG

6147 6126

211

N3E4E3_A388 HS393556 Clp G3PD 5e-28 TGGTCAAGGTAGTGGCATGG TCCTTCTCAGCAGGGCTTGT

6192 6241

157

N23E3E4_A691 HS394527 Glutathione S-transferase 2e-06 GGTCCTCCAACTACCGACTCC AAAAGGGCCAGCATTCAGAC

6190 6000

191

N23E3E4_A261 HS394072 Loss of the timing of ET and JA biosynthesis 2

1e-26 AATGGGCAAGTGGTTGGAGA TTGATTTCGCTAGCACGTTTCA

6276 6297

206

N23E3E4_A552 HS394303 ACC oxidase 1e-15

TATGCCACCGTGAAGTTCCA TGACCCACTCTTAAGCTGTTGC

6243 6171

106

N23E3E4_A233 HS394047 Lipase 2e-20

CGTGCAGAATTGCAATGGAT AGACAAACCGCAACCCAACT

6198 6186

158

23E3E4_A139 HS393777 Dehydrin 4e-14 GAGGCCATCATCAGCAGCAT CTGGTCTTTGTGCCCACCTG

6319 6289

155

SSH1_226 HS395745 DNA J protein 4e-38 CAGTGCAAGGCTCTTGAAGG ACCACCACCGTGCATATCAT

6110 6108

180

N23E3E4_A147 HS393974 Ethylene responsive transcription factor 4e-46

TGACTTTGGGTGGGGAGAAT TCATTTGGGACTCGAAAGCA

6165 6169

206

N23E3E4_A680 HS394396 Aquaporin 1 MIP family 6e-59 GAGTGCTGCACCGATGAATG GTGCCAGAGACTCCCACGTC

6282 6328

203

N23E3E4_A157 HS393983 Ehylene- induced esterase 8e-35 GTCTCATGGCCACAAGGTCA TTTTGTGGGGGAACTTGTCC

6213 6204

209

N23E3E4_A244 HS394057 Hydrogen peroxide induced protein 1 5e-20 ACCTGGACGAACAGGTCCAT AAGGCCTTCTGCTCCAACAC

6177 6172

184

Chapter IV

94

Table 42 Contd

EST ID dbEST acc no


Tm (ordmC) Product Size (bp)

N23E3E4_A169 HS393992 NO SIMILARITY -- CCAGGAGGCTAAGGAGCAACTA GCCACCATCAAGGTTAAGGCTA

6206 6250

235

N23E3E4_A139 HS393967 Hydroxyproline-rich glycoprotein 3e-59 AATGCCGAAGCTTGCTGTGT3 TTCAGGAAAGCCCTCAGCAG

6319 6289

162

3E4_A16 HS394695 Glutathion peroxidase 5e-50 ATCAGTTTGGCGAGCAGGAG GCTGTCCCCAAGAAGTCCAC

6279 6202

170

N23E3E4_612 HS394342 Elicitor responsive gene 3 8e-33 TCCATTCCAATACCGTCGAA GCATGGCGACAGCTTCATAC

6123 6176

217

N23E3E4_A17 HS393845 Proline-rich protein 7e-53

TACCCTGTTGGCGTTCCATC CGCCATCTCCAGGTCAAGTC

6314 6313

217

SSH1_415 HS395830 Cinnamoyl-CoA reductase 3e-50 ACTGCTATGGGCCCAATTCT AAGTGCCTTCCAGTTGCTGA

6085 6098

179

N23E3E4_A164 HS393988 Zn finger protein 2e-72

TGGAAGCTTTGCCACCACTT CGATGGTTGGCACAGTCAAA

6303 6305

222

SSH2_304 HS396243 Diaminopimelate decarboxylase 2e-29 ATAGGGGGCTTCCAACTCCA TCTCTGCATGACGGATCTTGG

6296 6362

159

N23E3E4_A182 HS394003 TRAPP complex 2e-29 TCCCATGTGGAATCATACGG AGCATGGTTCATTCAGGGAGA

6153 6194

299

N23E3E4_A301 HS394100 Jasmonate ZIM domain protein 6e-06

AAAAAGCGTTTCGGGTGAGA CCCCAAAAACAAACCAAACA

6201 6060

280

N23E3E4_A338 HS394126 Eukaryotic translation initiation factor 5A4 6e-32 AGGATGGATTCGGTGAAGGA AGATTGCTGCACACCCACAT

6176 6156

196

N23E3E4_A317 HS394113 TCP transcription factor 1e-28 AAAGATGGGCACAGCAAGGT CGCTCAATTGCTGAGGAGTG

6482 6403

220

N23E3E4_A460 HS394234 Drm3-like protein 3e-32

ATCAGCTCTGGGACGACACC GTCGGCGGTGGTTTCAGTAT

6264 6222

169

N23E3E4_A693 HS395085 Hexose transporter 7e-33

TTCCCACTCGAAATCCGTTC GGTCATCACCACCACGAAGA

6228 6198

153

Chapter IV

95

Table 43 Functions of selected genes reported in the literature

Selected genes showing homology with

Stress regulation or putative function References

Abscisic stress ripening protein (ASR1)

Induced under water stress enhance drought tolerance ABA signalling

Silhavy et al 1995 Yang et al 2005

Ascorbate peroxidase Drought and salt tolerance Badawi et al 2004

APE1 Involved in acclimatization of photosynthesis during stress Drought responsive

Walters et al 2003

Copper-containing amine oxidase ABA induced stomatal closure during stress An et al 2008

Clp G3PD Heat shock responsive ABA mediated growth and stomatal closure

Yang et al 1993 Munoz-Bertomeu et al 2011

Glutathione S-transferase Improved drought and salt tolerance Ji et al 2010

Loss of the timing of ET and JA biosynthesis 2

Function unknown in plants Kushwaha et al 2009

ACC oxidase Ethylene biosynthesis during stress

Dehydrin Drought cold and salinity responsive drought tolerance

Battaglia et al 2008 Rinne et al 1998 Zhu et al2000 Nylander et al 2001 Kimura et al 2003 kim et al 2010

DNA J protein Induced under drought Alamillo et al 1995 Coca et al 1996 Campalans et al 2001

Ethylene responsive transcription factor

Transcription factors that mediate exoression of defers related ethylene responsive genes

Ecker 1995

Aquaporin 1 MIP family Key gene for plant water balance and water use efficiency

Knepper 1994 Tyerman et al 2002 Aharon et al 2003 Kaldenhoff and Fischer 2006 Maurel 2007

Hydrogen peroxide induced protein 1

functions in plant development growth and stress responses

Neill et al 2002 Foyer and Noctor 2005

Chapter IV

96

Table 43 Contd

Selected proteins Stress regulation or putative function References

LEA Drought tolerance Imai et al 1996 Xu et al 1996 Swire-Clark and Marcotte 1999 Zhang et al 2000

Hydroxyproline-rich glycoprotein

functions in defense against plant pathogen Lamport 1970

Glutathion peroxidase Drought tolerance Miao et al 2006

Elicitor responsive gene 3 Defence related Day et al 2002

Proline-rich protein Drought responsive Harrak et al 1999

Cinnamoyl-CoA reductase Defense signalling Umemura et al 2006

Zn finger protein Functions in growth differentiation transcription signal transduction and oncogenesis

Reddy et al 1992

Diaminopimelate decarboxylase

Lysine metabolism during stres Less and Galali 2008

TRAPP complex Functions in vesicular transport Sacher et al 2001

Jasmonate ZIM domain protein

Functions in Jasmonate (JA) signalling during plant growth development and defense

Chini et al 2007 Melotto et al 2008

Eukaryotic translation initiation factor 5A

Functions in plant growth and development by regulating cell division cell growth and cell death

Feng et al 2007

TCP transcription factor Involved in flower development regulation of growth and cell division shoot branching

Broholm et al 2008 Li et al 2005 Poza-Carrion et al 2007

Drm3-like protein Dehydration responsive Zhang et al 2009

Hexose transporter Induced during leaf senescence Ehness et al 1997

ice To the above mix 4 microl of 5X transcriptor reverse transcriptase reaction

buffer 05 microl protector RNase inhibitor (40Umicrol) 2 microl deoxynucleotide mix

(10 mM each) and 2 microl transcriptor reverse transcriptase (20Umicrol) were

added mixed briefly centrifuged and incubated at 50ordmC for 1 hour The

reverse transcriptase enzyme was deactivated by heating the tube at 85ordmC

for 5 min The first strand was stored at -20ordmC for downstream application

4252 Quantitative Real Time PCR (qRT-PCR)

LightCycler 480 SYBR green I Master kit (Cat No 04707516001 Roche

Germany) was used to carry out the expression assays in a LightCycler 480

II real time machine (Roche Germany) The reverse transcribed first strand

cDNA was used as template All the reactions were carried out in a 20 microl

volume prepared according to the protocol mentioned in the kit manual

Three technical replicates of each reaction were used to reduce the error

rate The following amplification programme was used pre incubation at

95ordmC for 5 min followed by 45 cycles of amplification with 10 sec

denaturation at 95ordmC annealing for 5 - 20 sec (primer dependent) and

extension at 72ordmC for 30 sec This was followed by one cycle of melting

curve analysis to check specificity of amplified product The temperature for

melting curve analysis was 95ordmC for 5 sec 65ordmC for 1 min and 97ordmC for

continuous acquisition This was followed by one cycle of final cooling at

40ordmC

4253 Data Acquisition

Data on the expressions were obtained in the form of crossing point (Cp) the

point where the samplersquos fluorescence curve turns sharply upward The data

acquisition was done employing the 2nd derivative maximum method

(Tichopad et al 2003 2004) as computed by the software of LightCycler

(Roche Diagnostics) Carousel-based system

426 Computation of Normalization Factor

We have used geNorm (httpmedgenugentbe~jvdesompgenorm)

software to find out the most stable genes among the set of housekeeping

Chapter IV

98

genes at three stages of drought geNorm calculates a gene expression

normalization factor for each tissue sample based on the geometric mean of

a user defined number of reference genes (Vandesompele et al 2002) It

eliminates the most unstable genes from the group of housekeeping genes

step by step and ends up with two most stable reference genes In the actual

process the raw Cp values of all the housekeeping genes are transformed

into relative quantities using delta Ct method These values are than used for

computation of normalization factor in geNorm for all the biological

replicates

427 Relative Quantification

In relative quantification assays the expression of the target gene is

expressed as ratio of target-to-reference gene in the same sample Here the

mean CP values (obtained from three technical replicates) of all the target

genes of all the biological replicates (seven biological replicates in our case)

under consideration were transformed into relative quantities using delta Ct

method The normalized expression level was calculated by dividing the

quantities by respective normalization factor This normalised expression

value of a gene in different biological replicates was compared to know the

relative expression level

428 Hierarchical Clustering of Drought Induced Genes

The hierarchical clustering was performed to study the expression profile of

genes across the seven biological replicates to identify groups of genes that

share similar expression profile This was performed using GenePattern

(Reich et al 2006) on line server

(httpgenepatternbroadinstituteorggppagesindexjsf)

43 Results

431 Expression of Housekeeping Genes

Four genes were tested using geNorm software in order to find out the most

stable housekeeping genegenes under water stress in the seven biological

replicates of the two cultivars Step wise exclusion of most unstable genes

Chapter IV

99

by geNorm ended up with two most stable housekeeping genes viz

ribulose-1 5-bisphosphate carboxylaseoxygenase and 18S rRNA The

normalization factors thus calculated for these stable housekeeping genes in

different samples are given in Table 44

432 Relative Expression of Drought Induced Genes

The relative expression studies of selected genes by qRT-PCR in both the

tolerant and susceptible cultivars have shown diverse expression pattern

(Appendix 41) Expression of some genes was found to be continuously up-

regulated in both the cultivars while some were found to be down-regulated

Some of the genes were found to be either up-regulated or down-regulated

transiently in either of the cultivars in three stages of drought However the

expression levels of these genes were different in both the cultivars To get a

clear picture we compared the expression pattern of these genes in both the

cultivars separately at three stages of drought ie BWS WS and AWS

(Figure 41 and 42) The relative expression values of all the genes

considered at three stages of drought in both the cultivar is given in

Appendix 41

4321 Relative Expression in TV23

The relative expression of these genes in tolerant cultivar TV23 has shown

four distinct patterns (Figure 41) The group A (12 genes) have been found

to be continuously up-regulated while group B (8 genes) continuously down

Table 44 Normalization factor of housekeeping genes calculated using geNorm

Biological replicates 18S Rubisco Normalization Factor

TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100

regulated in three stages with progression of drought Two genes in group B

(elicitor-responsive gene 3 and cinnamoyl-CoA reductase) with very high

expression values (110821 and 142724) at BWS are not shown in Figure

41 as their inclusion in the graph drastically makes the drowning of other

bars The genes in group C (6 genes) have shown up-regulation at WS

followed by down-regulation at AWS The group D consists of only one gene

showing down-regulation at WS followed by up-regulation at AWS

D

0 50 100 150 200 250

Copper-containing amine oxidase

Loss of the timing of ET and JA hellip


Aquaporin 1 PIP subfamily

Proline-rich protein


Zinc finger

LEA

TRAPP complex

TCP24 transcription factor

Ascorbate peroxidase

Jasmonate ZIM-domain protein

DNA J protein

clp G3PD

Glutathion peroxidase

Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin


Abscisic stress ripening protein

ACC oxidase

Ethylene-responsive transcription factor

Glutathione-S transferase theta

Ethylene-induced esterase

Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C

Figure 41 Expression pattern of drought induced genes in TV23 at three stages

of drought

Chapter IV

101

4322 Relative Expression in S3A3

The expression pattern of the genes in S3A3 at three stages of drought can

be divided into three distinct groups as shown in figure 42 Group A

represents five genes with continuous up-regulation at three stages of

drought while group B represent 20 genes with up-regulation at WS followed

by down-regulation at AWS The group C represent only four genes that are

down-regulated at WS followed by up-regulation at AWS A total of 19 genes

out of 29 considered for study have shown up-regulation at BWS

A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein

Elicitor responsive protein

Cinnamoyl CoA reductase



Aquaporin MIP family

Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)

Figure 42 Expression pattern of drought induced genes in S3A3 at three stages of drought

Chapter IV

102

433 Comparative Expression Pattern of Drought Induced Genes

The water stress experiment (Chapter II) has indicated that the drought

tolerance of TV23 is much higher compared to S3A3 therefore we were

more interested to find out the genes that are up-regulated in TV23 at three

stages of drought in comparison to S3A3 We have divided the genes into

three categories based on their highest expression level in TV23 at each

stage of drought and compared them with the S3A3 (Figure 43 44 and

45) at that stage There are 11 genes (elicitor responsive gene 3 and

cinnamoyl-CoA reductase were not shown to avoid drowning of other bars in

the graph) whose expression was found to be highest at BWS of TV23

compared to WS and AWS We compared the expression of these genes

with S3A3 at that stage (Figure 43) However the expression of

choloroplast glyceraldehydes-3 phosphate dehydrogenase in this group was

found to be down-regulated when compared to control plant There were 6

genes highly expressed at WS in TV23 as compared to other two stages of

drought Of these only three were found to be highly expressed compared

to WS of S3A3 (Figure 44) These three genes are glutathione S-

transferase ethylene-responsive transcription factor and ACC oxidase

Likewise there were 12 genes that showed highest expression in TV23 at

AWS compared to WS and BWS Nine genes out of these 12 were found to

be highly expressed at AWS of TV23 when compared with the AWS of

S3A3 (Figure 45) Therefore of these 29 genes 21 (9 in BWS 3 in WS

and 9 in AWS) have shown higher relative expression in TV23 compared to

S3A3 in three stages of drought There were two genes viz loss of the

timing of ET and JA biosynthesis 2 and hydrogen peroxide induced protein 1

which showed similar pattern of expression with continuous up-regulation

with the progression of drought in both the cultivars at three stages of

drought (relative expression values are represented by bold numbers in

Appendix 41 Other two genes having similar expression pattern with up-

regulation at WS followed by down-regulation at AWS in both the cultivars

are ethylene-responsive transcription factor and glutathione-S transferase

whose relative expression values are indicated as shaded box in Appendix

41 Genes that showed completely opposite pattern of relative expression in

Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)

Figure 43 Comparison of expression of genes having highest expression at

BWS in TV23 with BWS of S3A3

0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase


Glutathione-S transferase

Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)


WS in TV23 with WS of S3A3

Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex

TCP transcription factor



DNA J protein

ACC oxidase

Clp G3PD




Glutathione S-transferase

Hexose transporter

Lipase

APE1

Eukaryotic translation initiation factor 5A4

Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)

Figure 46 Comparison of relative expression of drought induced genes in TV23 and S3A3 at BWS

0 100 200 300

Coper-containing amine oxidase

Loss of the timing of ET and hellip

Hydrogen peroxide induced hellip




Zinc f inger protein

LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)

Figure 45 Comparison of expression of genes having highest expression at AWS in

TV23 with AWS of S3A3

Chapter IV

105

both the cultivars at three stages of drought are the DNA J elicitor-

responsive gene 3 and cinnamoyl-CoA reductase The expression of these

three genes (their relative expression values are indicated by bold and

underline in Appendix 41) were found to be decreased in TV23 with the

increase of drought while in S3A3 they showed a completely opposite

pattern of expression with continuous increase There is another group

represented by 9 genes (their relative expression values are indicated as

bold italics and underlined in Appendix 41) whose expression followed a

similar pattern in both the cultivars with continuous up-regulation at BWS

and WS However all these genes were found to be further up-regulated in

TV23 at AWS while they are down-regulated at AWS in S3A3 The gene

chloroplast glyceraldehyde-3 phosphate dehydrogenase (indicated as bold

italics in Appendix 41) was found to be down-regulated in TV23 compared

to control The comparison of expression at BWS (column indicated by super

scribing with in Appendix 41 Figure 46) of both the cultivar has shown 19

genes that are up-regulated in TV23 compared to S3A3 Two highly

expressed genes (elicitor responsive gene 3 and cinnamoyl-CoA reductase)

are not shown in Figure 46 to avoid drowning of others Among these the

maximum expression was observed for cinnamoyl-CoA reductase and

elicitor responsive gene 3 (their expression values are indicted by super

scribing with yen in Appendix 41)

434 Hierarchical Cluster Analysis of Drought Induced Genes

The hierarchical cluster analysis of the expression profiles of the drought

induced genes showed differences between drought tolerant and susceptible

cultivar (Figure 47) Clustering separated the seven biological replicates into

two groups (A and B Figure 47) indicating tolerant and susceptible cultivars

clustered separately However the biological replicate TV23 (AWS) is

included in susceptible group B The results also showed that there were

four distinct expression patterns (cluster I to IV Figure 47) of the genes in

three stages of drought in both the cultivars The cluster I is represented by

11 genes whose expression is higher at WS and AWS of TV23 and WS of

S3A3 among all three stages considered The gene for ethylene-

responsive transcription factor is however very weakly induced compared to

Chapter IV

106

other genes in that group although its expression is slightly higher than

TV23C in all the biological replicates The cluster II includes 5 genes whose

expression is higher in two stages (WS and AWS) of S3A3 and also in

AWS of TV23 The cluster III includes two genes whose expression is down-

regulated in comparison to TV23C in both the cultivars with the progression

of drought The cluster IV is represented by 11 genes that are highly

Figure 47 Hierarchical cluster analysis of drought induced genes in TV23 and

S3A3 at three stages of drought


Aquaporin 1 MIP family




LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter

Glutathione peroxidase



DNA J protein

Lipase


Elicitor responsive gene 3

Drm3 like protein

A B

I

II

III

IV

Chapter IV

107

expressed in BWS and WS of TV23 among seven biological replicates Of

these 9 genes were found to be highly expressed in BWS and remaining 2

genes in WS Among these 29 drought induced genes three most highly

expressed genes in all the biological replicates are cinnamoyl-CoA

reductase followed by elicitor-responsive gene 3 and lipase (class 3) with

relative expression values of 1427244 110821 and 171732 respectively

435 Validation of Subtractive Library

To validate our subtractive library we have compared the relative expression

of drought induced genes mostly selected from the library BWE3E4

Nineteen genes (65) out of 29 selected were found to be highly expressed

(Figure 46) in TV23 compared to S3A3 at BWS This clearly showed that

the efficiency of subtraction was as high as 65

44 Discussion

When plants experience environmental stresses such as drought salinity

etc they activate a diverse set of physiological metabolic and defence

processes to survive and to sustain growth Tolerance and susceptibility to

these stresses are very complex Drought tolerance or susceptibility is a

multigenic trait and therefore difficult to control Transcriptomics proteomics

and gene expression approaches are commonly used to identify the

activation and regulation of several stress-related transcripts and proteins

which are generally classified into two major groups One group is involved

in signalling cascades and in transcriptional control whereas members of

the other group functions in membrane protection as osmoprotectants as

antioxidants and as ROS scavengers (Shinozaki and Yamaguchi-Shinozaki

1997) Manipulation of genes that protects and maintains cellular functions

or that maintains the structure of cellular components has been the major

target of attempts to produce plants having enhanced stress tolerance The

study of expression pattern of such genes at different level of water stress

will pave the way to understand the drought responsive mechanisms in tea

In the present study we have studied the expression pattern of 29 genes

under induced drought condition in two tea cultivars with contrasting drought

tolerance behaviour in field as well as in controlled condition Among these

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there are genes that codes for proteins involved in membrane protection

eg LEA protein which has been reported to play roles in membrane and

protein stabilization and cellular homeostasis in several species (Amiard et

al 2003 Wang et al 2004 Houston et al 2005 Boudet et al 2006

Cameron et al 2006) aquaporin that facilitates transport of water across

cell membranes (Agre et al 1993 Chrispeels et al 1994a Chrispeels et

al 1994b) dehydrins involved in osmotic adjustment (Taiz and Zeiger

2002 Brini et al 2007) antioxidant enzymes such as ascorbate peroxidase

(Yamaguchi et al 1995) glutathione transferase (Noctor et al 1998)

glutathione peroxidase (Miao et al 2006) involved in scavenging ROS The

other group includes genes encoding hydrogen peroxide induced protein

that functions in plant development growth and stress responses

particularly those induced by environmental stimuli (Neill et al 2002 Foyer

and Noctor 2005) transcription factors like Zn finger protein (Leon and

Roth 2000) abscisic acid stress ripening protein (Cakir et al 2003) and

TCP transcription factor Besides we have monitored the expression of

other genes encoding proteins as listed in Table 43 that are reported to be

induced during water and other environmental stresses in planta It has been

found that the expression pattern of all these genes (as evident from the

Figure 48a-c) are either continuously or transiently up-regulated or down-

regulated during progressive drought which also implies that these genes

are not constitutive but are drought responsive

We monitored the expression of some of the transcription factors that are

found to be up-regulated or down-regulated during the induced drought

experiment The Zinc finger protein is a C3HC4-type RING finger whose

expression was found to be induced by drought (Figure 48a) This protein

has been reported to have crucial roles in the growth differentiation

transcription signal transduction and oncogenesis (Reddy et al 1992) and

also involved in the ubiquitin-mediated protein degradation pathway (Lorick

et al 1999) The transgenic Arabidopsis over-expressing the gene has been

found to have increased cellular ABA and drought tolerance compared to

wild type plant (Ko et al 2006) We have observed higher induction of this

gene in S3A3 compared to TV23 at BWS and WS However its expression

Chapter IV

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Figure 48a Relative Expression of drought induced gene at three stages of

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Chapter IV

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Chapter IV

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is down-regulated at AWS in S3A3 where as in TV23 it continues to

increase with progressive drought

Another gene whose expression was induced by drought is ethylene-

responsive transcription factor which is a well known transcription factor

having DNA binding domain that mediates the expression of defense related

ethylene responsive genes (Ecker 1995) by binding with the 11 bp

consensus sequence (TAAGAGCCGCC) called GCC box (Ohme-Takagi

and Shinshi 1995 Shinshi et al 1995) of ethylene responsive element

(ERE) in the promoter region Over-expression of ethylene-responsive

transcription factor gene (JERF3) in tobacco has been reported to increase

adaptation to drought salt and freezing by activating the expression of

oxidative stress related genes such as superoxide dismutase and by

decreasing the accumulation of ROS Therefore higher expression of this

gene in TV23 may be playing a positive role for giving drought tolerance by

reducing the accumulation of ROS and enhancing the production of ROS

scavengers like ascorbate peroxidase (Figure 48a) and glutathione

peroxidase (Figure 48c) Another transcription factor gene the eukaryotic

translation initiation factor 5A4 (eIF5A4) was also highly induced by drought

in TV23 compared to S3A3 This gene has been widely reported to be

involved in plant growth and development by regulating cell division cell

growth and cell death (Feng et al 2007) The partial inactivation of eIF 5A

gene by over-expressing antisense DHS (Deoxyhypusine synthase) cDNA

gene in transgenic Arabidopsis has been reported to delay senescence and

resistance to drought stress (Wang et al 2003) But in our study we have

observed a high expression of the gene in the tolerant tea cultivar TV23

(Figure 48c) which contradicts the above result Moreover its expression

level decreased further at WS and AWS at which we observed maximum

senescence of lower mature leaves during the induced drought experiment

Therefore this contrasting expression pattern of eIF5A in tea needs to be

considered for further investigation that will help to identify its role in plant

adaptation to stress This may be due to interaction of (genotype x

environment) factor as because drought is a multigenic trait and we did our

Chapter IV

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drought induced experiment in pot condition which seems to be different

from natural field condition

TCP transcription factor whose up-regulation was found to be low in TV23

compared to S3A3 during the induced water stress experiment It is a

member of a large group of transcription factor family called TCP family

which in Arabidopsis is represented by 24 genes This family has been

reported to be involved in flower development (Broholm et al 2008)

regulation of growth and cell division (Li et al 2005) shoot branching (Poza-

Carrion et al 2007) etc There is no report of induction of this transcription

factor under water stress

The LEA gene found to be highly expressed in the present study (Figure

48a) belongs to group 3 or D-29 family (classification adapted by Dure et

al 1989 Dure 1993) The role of LEA proteins towards improving drought

tolerance in plants and yeast has been reported in several studies (Imai et

al 1996 Xu et al 1996 Swire-Clark and Marcotte 1999 Zhang et al

2000) The expression profile of LEA gene under stress supports its role as

protective molecule that enables cells to survive protoplasmic water

depletion (Ingram and Bartels 1996) Studies on over-expression of LEA

gene also support its protective roles by improving the stress tolerance of

plants For example the expression of Barley gene HVA1 in wheat and rice

increases drought tolerance (Sivamani et al 2000) and also over-

expression of LEA genes PMA80 and PMA1959 (Cheng et al 1995) and

OsLEA3-1 (Xiao et al 2007) increases dehydration tolerance in transgenic

rice These results are consistent with our findings that the expression of

LEA in drought tolerant cultivar TV23 is much higher compared to

susceptible cultivar (Figure 48a) at BWS The expression was found to

increase further in TV23 with increase of water stress in WS and AWS

compared to S3A3 This contrasting expression pattern of the gene might

be contributing towards higher drought tolerance of TV23

Another gene whose expression was found to be highly induced is

aquaporin 1 that code for protein of MIP (Membrane Intrinsic protein) family

and PIP (Plasma membrane intrinsic protein) subfamily which is well

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documented to play a key role in plant water balance and water use

efficiency (Knepper 1994 Tyerman et al 2002 Aharon et al 2003

Kaldenhoff and Fischer 2006 Maurel 2007) by mediating movements of

small solutes and water across membranes (Maurel et al 2009) However

the role of aquaporin in leaf water transport under drought conditions

remains unclear (Heinen et al 2009) It has been reported that transgenic

tomato over-expressing tobacco aquaporin 1 showed higher stomatal

conductance transpiration rate and net photosynthesis (Sade et al 2010)

which are very much essential for a plant to maintain growth In present

study we have found higher expression of the gene in TV23 compared to

S3A3 (Figure 48a) This higher expression of the gene may be responsible

for efficient movement of water to growing tissues in TV23 compared to

S3A3 to maintain a water balance under induced drought condition The

higher drought tolerance of TV23 has also been supported by its higher

photosynthesis and transpiration rate observed during the induced drought

experiment compared to S3A3 at a given level of water stress (Chapter II)

Moreover there are reports of reduction in drought tolerance and hydraulic

conductivity in Arabidopsis with reduced aquaporin 1 gene expression

(Siefritz et al 2002) This finding is also consistent with our findings that

expression of aquaporin 1 gene in susceptible is less compared to the

tolerant cultivar

Dehydrin is another gene that was found to be highly induced in the present

study in both the cultivars However its expression goes down as drought

became more severe at AWS (Figure 48b) The higher expression of

dehydrin in our experiment is consistent with the findings that it accumulates

under drought and other stresses like low temperature and or salt-stress in

vegetative tissues of plant (Ingram et al 1996rsquo Allagulova et al 2003

Close 1997) Dehydrin is a type of LEA group protein originally designated

as D-11(Battaglia et al 2008) Studies have suggested many functions of

dehydrin in buffering water sequestering ions stabilizing membranes or

acting as chaperones or molecular shields (Rorat et al 2006 Tunnacliffe et

al 2007 Koag et al 2003 Goyal et al 2005 Kovacs et al 2008 Kruger

et al 2002) Although constitutively expressed the accumulation of dehyrin

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was found to be increased in many studies involving stresses like drought

cold salinity and ABA (Battaglia et al 2008 Rinne et al 1998 Houde et

al 1992 Wood and Goldsbrough 1997 Ismail et al 1999 Zhu et al2000

Nylander et al 2001 Kimura et al 2003 Kim et al 2010 Borovskii et al

2002) which also corroborates our findings Higher expression of this gene

was observed in TV23 compared to susceptible cultivar S3A3 This higher

expression level of dehydrin in both the cultivars compared to control plants

seems to be involved in the dehydrationosmotic stress response in tea and

might contribute to the drought tolerance behaviour This assumption is

supported by data showing that under conditions of water deficit

transcription of dehydrin gene is significantly higher in drought-tolerant than

in drought-sensitive species A correlation between plant drought tolerance

and dehydrin accumulation was also found in Sorghum and in Sunflower

(Wood et al 1997 Cellier et al 1997) Therefore the higher expression of

dehydrin may be responsible for high drought tolerance behaviour of TV23

compared to S3A3

DNA J a heat shock protein reported to involve in a variety of cellular

processes including protein folding protein transport across membranes

regulation of protein degradation modulation of protein activity and

prevention of irreversible protein aggregation (Sun et al 2001 Lee et al

1995 Mohd et al 2006) Induction of this protein under water stress has

been well reported in plants (Alamillo et al 1995 Coca et al 1996

Campalans et al 2001) The higher expression of this gene in TV23 at BWS

(Figure 48b) may be attributed for higher drought tolerance of TV23 Our

observation is consistent with the findings that the higher expression of heat

shock proteins enhance drought tolerance in tobacco and other plants

(Cascardo et al 2000 Eun and Choo 2006) However its expression

sharply decreased at WS and AWS in both the cultivars (Figure 48b)

A secondary effect of dehydration in plant is the production of ROS like

singlet oxygen superoxide anion radicals hydroxyl radicals and hydrogen

peroxide (Smirnoff 1998 Bartels 2001 Apel and Hirt 2004) in chloroplast

as well as in mitochondria Higher accumulation of ROS is found to be toxic

as it can oxidize proteins lipids and DNA (Halliwell and Gutteridge 1999)

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leading to cell death (Asada 1987 Dat et al 2000 Hammond-Kosack et al

1996) Therefore production of ROS has to be fine tuned to prevent

oxidative damage In fact plant has developed mechanisms to keep the

concentrations of ROS under tight control by producing antioxidants which

may be enzyme or non-enzyme proteins Therefore alleviation of oxidative

damage and increase resistance is correlated with an efficient antioxidative

system (Smirnoff 1998 Shalata et al 2001 Kranner et al 2002) In the

present study we have monitored the expression of such genes coding for

antioxidative proteins like ascorbate peroxidase glutathione peroxidase and

glutathione S-transferase The expression of these genes has been found to

be highly up-regulated in TV23 compared to S3A3 at BWS (Figure 48a-c)

The higher expression of these genes indicates the accumulation of ROS

during drought stress For example the accumulation of ROS like H2O2 is

substantiated by higher expression of H2O2 induced protein (Figure 48a) in

the present study Hydrogen peroxide is an interesting molecule which can

play dual role during stress it induces a defense response at lower

concentration while at higher concentration it results into programmed cell

death (PCD) (Vandenabeele et al 2003) which is essential for a number of

developmental processes and environmental responses including aleurone

cell death the hypersensitive response to pathogens and allelopathic

plantndashplant interactions (Bethke and Jones 2001 Bais et al 2003 Apel and

Hirt 2004) We have observed starting of senescence browning and drying

of lower leaves (cell death) in both the cultivars at WS and AWS during the

water stress experiment This may be due to higher accumulation of H2O2 as

indicated by higher expression of hydrogen peroxide induced protein in WS

and AWS The higher accumulation of H2O2 is also supported by higher

expression of ascorbate peroxidase a scavenging enzyme for H2O2 during

the experiment (Figure 48a) This molecule has also been reported to

increase the level of glutathione S-transferase (Levine et al 1994 Chen et

al 1996 Polidoros and Scandalios 1999) an enzyme responsible for the

detoxification of highly reactive lipid peroxidation products generated from

oxidative stress damaged membranes (Polidoros and Scandalios 1999)

during drought In fact the expression of glutathione S-transferase was

found to be highly up-regulated in both the cultivars during the experiment

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(Figure 48b) Transgenic plants over-expressing these antioxidant enzymes

have been reported to have improved drought tolerance characters For

example transgenic Tobacco and Arabidopsis over-expressing ascorbate

peroxidase (Badawi et al 2004) and glutathione peroxidase (Miao et al

2006) respectively were found to have higher drought tolerance compared to

wild type plant Likewise over-expression of glutathione S-transferase gene

was found to improve drought and salt tolerance in transgenic Tobacco (Ji et

al 2010) Therefore the higher expression of these ROS scavenging

enzymes in our study might be responsible for higher drought tolerance of

TV23 as reported in other plants (Li et al 1998 Bowler et al 1992)

The gene encoding Copper-containing amine oxidase was found to be

induced during progressive drought in both the cultivars to varying degrees

(Figure 48a) This enzyme catalyses the catabolism of polyamine putrescine

to produce H2O2 ammonia and ∆1-pyrroline The H2O2 thus produced is

reported to be involved in ABA induced stomatal closure during stress (An et

al 2008) ∆1-pyrroline is further reduced by pyrroline-5-carboxylate

reductase to produce proline a well known osmoprotectant involved in

protection of cells from damage due to water stress (Shinozaki et al 1997)

in plants The high expression of this gene in S3A3 (15 days) compared to

TV23 (22 days) at BWS may be correlated with the earlier closing of

stomata in S3A3 than TV23 which was actually observed during the

induced drought experiment

During severe drought stress plant has to depend on reserve food for

carbon source for survival When drought becomes so severe that it results

in drought induced leaf senescence the reserved food in the form of

triacylglycerol (TAG) from dying leaves and other tissues are converted to

fatty acid and glycerol by lipase class 3 enzyme (Trigacylglycerol lipase or

class 3 lipase) which are further converted to sugars to support growth

Although seeds are the main storage for TAG its synthesis and

accumulation has also been reported in chloroplast as a means for

sequestering free fatty acids that are rapidly de-esterified from galactolipids

in the event of stress (Sakaki et al 1990a and b Navari-Izzo et al 1990)

and during natural senescence (Kaup et al 2002) In our study we have

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observed a very high up-regulation of lipase (Class 3) in TV23 compared to

S3A3 at BWS followed by sharp decrease in WS and AWS (Figure 48c)

This higher expression of lipase may be contributing for higher efficiency of

TV23 to convert the stored and senescence induced TAG into fatty acids

and glycerol These two compounds are further metabolised to produce

sugars required to maintain metabolic processes to sustain during water

stress period Therefore the high expression of Lipase in TV23 might be

responsible for supplying carbon source for maintaining metabolic processes

to sustain during water stress This source of energy might be contributing in

TV23 to withstand prolonged drought up to 25 days (WS) compared to 18

days in S3A3 in the present study

The gene encoding cinnamoyl-CoA reductase that catalyzes the synthesis of

lignin via phenyl propanoid pathway (Piquemal et al 1998 Jones et al

2001 Goujon et al 2003 Kawasaki et al 2006 Leple et al 2007

Wadenback et al 2008 Zhou et al 2010) have been found to be highly

expressed in the tolerant TV23 at BWS This high induction of the gene

under drought is consistent with the findings of So et al (2010) A strong

correlation between leaf lignin content and drought tolerance has been

reported in maize that can be used as an index for evaluation of drought

tolerance (Hu et al 2009) Assuming that high expression of cinnamoyl-CoA

reductase observed (Figure 48b) will eventually result in to higher

accumulation of lignin in TV23 which may be contributing for its higher

drought tolerance

Diaminopimelate decarboxylase is an enzyme that catalyzes the

decarboxylation of diaminopimelate to produce lysine We have observed

up-regulation of this gene under drought in both the cultivars (Figure 48a)

However its expression was found higher in TV23 and continues to increase

at WS and AWS Probably this is the first report of induction of this gene

under drought stress in plants

APE1 (Acclimation of photosynthesis to the environment) is another gene

highly expressed in tolerant cultivar in the present study at BWS (Figure

48c) The differential expression of the gene under drought has been

Chapter IV

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reported in ryegrass (Liu and Jiang 2010) Plants subjected to variation in

environmental temperature has the capacity to acclimate the process of

photosynthesis (for review see Berry and Bjorkman 1980 and Down ton et

al 1984) that improves the efficiency with which light energy is used in

photosynthesis (Chow et al 1990 Walters and Horton 1995) Study on

ape1 gene in Arabidopsis mutants showed acclimation-defective phenotype

(Walters et al 2003) The higher expression of this gene in TV23 under

induced drought (Figure 48c) may therefore be helping the plant to

acclimatize their photosynthetic apparatus to survive under drought

condition However the expression of the gene was found to be decreased

at WS

Chloroplast glyceraldehyde-3-phosphate dehydrogenase is the gene among

all the genes considered in present study whose expression has been found

to be continuously down-regulated compared to the control plant with the

progression of drought (Figure 48b) A similar result of down regulation of

the gene has also been reported in Arabidopsis under heat shock treatment

(Yang et al 1993) and in water-deceit tolerance in C4 perennial grass

species (Zhao et al 2011) Recently the mutant studies in Arabidopsis has

also indicated that it play a positive role in ABA mediated growth and

stomatal closure (Munoz-Bertomeu et al 2011) Therefore down regulation

of the gene in the present study might be associated with the down

regulation of carbon metabolism and ABA signalling during drought stress

One of the responses of plants to environmental stress is establishing a

temporary growth arrest that allows adaptation to adverse conditions

Inhibited or reduced growth is considered to be an adaptive feature for

survival allowing plants to employ multiple resources to combat stress

(Xiong and Zhu 2001) This was also observed in our study by down-

regulation of Drm3 or Auxin repressed gene (Figure 48c) This down

regulation of the gene during drought stress may be associated in arresting

the growth of the plant to overcome the drought situation

Elicitors are molecules that can trigger accumulation of phytoalexin in plants

They are often pathogen derived and can trigger the biosynthesis of one or a

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combination of the hormone-like compounds jasmonic acid (JA) salicylic

acid and ethylene (Reymond and Farmer 1998) These hormones are

widely reported to involve in different signal transduction pathways in plants

under stress condition Therefore the higher expression of elicitor

responsive gene 3 in the present study may be associated in stress related

signal transduction pathways under drought The hydroxyproline-rich

glycoprotein is a structural cell wall protein (Lamport 1970) that functions in

defense against plant pathogen The expression of gene encoding this

protein was found to be up-regulated due to drought in the present study at

BWS Its expression pattern in the two cultivars under consideration was

found opposite to each other (Figure 48c) The down-regulation of the

protein under drought has been reported for example repression of cell wall

extensions (a hydroxyproline-rich glycoprotein) whose function is to cross

link cell walls after elongation (Carpita and Gibeau 1993) was found to

reduce cross linking of cell walls in drought-stressed P acutifolius compared

to wild type non-stressed plant (Micheletto et al 2007) Down-regulation of

this gene in the present study therefore may be involved in suppression of

growth by suppressing the cell wall cross linking under drought stress

Jasmonate ZIM-domain protein (JAZ) whose expression was found to be

down-regulated or nil at BWS in the present study (Figure 48c) has been

reported to act as a repressors (Chini et al 2007 Melotto et al 2008) of

Jasmonate (JA) signalling during plant growth development and defense

signalling However expression of the gene was found to increase

continuously with the advancement of drought after BWS (Figure 48c) LEJ2

(Loss of the timing of ET and JA biosynthesis 2) whose expression was

found to be continuously up-regulated at all the stages of drought (Figure

48c) in the present study encodes proteins that belongs to a group

containing CBS (cystathionine β-synthase) domain whose function are

largely unknown in plants (Kushwaha et al 2009) Some of these proteins

have been reported to enhance oxidative stress tolerance in transgenic

Arabidopsis (Luhua et al 2008) Therefore high expression of the gene

may also play a role during drought stress which needs further investigation

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The Asr1 gene belongs to the Asr gene family (named after abscisic acid

stress and ripening) classified as a new group of LEA (Caramelo et al

2008 Battaglia et al 2008) has been reported to be induced under water

stress (Silhavy et al 1995 Chang et al 1995) and involved in adaptation to

dry climates (Frankel et al 2003) It is also reported to involve in abscisic

acid signalling and used to develop transgenic Arabidopsis with enhance

drought and salt tolerance (Yang et al 2005) It can also act as a

transcription factor targeting the enhancer of a hexose transporter gene

(Cakir et al 2003) It was proposed that Asr1 is a link between sugar and

ABA metabolism and signalling Therefore Asr1 might act as non-histone

chromosomal protein involved in protection against a range of stress signals

by modulating cell sugar traffic The localization of this protein has been

reported in both nucleus and cytoplasm (Wang et al 2005) Its function in

the nucleus has been proposed in protecting DNA structure during water

loss and in gene regulation upon stress by changing DNA topology (Iusem et

al 1993 Silhavy et al 1995) The role of Asr1 as transcription factor for

Hexose transporter gene is also consistent with our findings that the hexose

transporter gene under water stress is highly induced During the drought

induced experiment we have observed senescence at WS in the lower

mature leaves This senescence may results in the coordinated

degradation of macromolecules and the mobilization of regained

nutrients like nitrogen carbon and minerals from senescing tissues into

other growing tissue (in our case to the buds along with 1st and 2nd leaf)

parts of the plant (Buchanan 1997 Nooden 1988) The induction of

hexose transporter has also been reported during leaf senescence (Ehness

et al 1997) This mobilization is further facilitated by higher expression of

hexose transporter gene as observed in the present study (Figure 48c) The

over expression of hexose transporter in TV23 is further supported by the

over expression of ASr1 gene that act as a transcription factor of the former

Therefore the higher induction of hexose transporter and Asr1 gene in TV23

may be responsible for its higher drought tolerance Its higher expression in

TV23 at wilting stage may be contributing towards efficient mobilization of

carbon source from the senescence leaf to the growing leaves to cope with

the drought stress The low level expression of the gene in the susceptible

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cultivar at BWS might have resulted in inefficient mobilization of the carbon

source to the growing parts of the plants which is further supported by the

low level expression of Asr1 gene at BWS of S3A3

Drought avoidance in plants include closing of stomata to minimize water

loss adjustment of sinksource allocation by increasing root growth and

decrease canopy by reducing growth and shedding of older leaves (Fischer

et al 1978) Accelerated leaf senescence and leaf abscission are

associated with drought in nature as a means to decrease canopy size In

perennial plant like tea this strategy may contribute to the survival of the

plant under drought stress In the present induced drought experiment we

have observed that the senescence of lower mature leaves in case of

susceptible cultivar started much earlier (18 days) compared to the tolerant

cultivar (25 days) This delayed leaf senescence in TV23 might be

responsible for its higher drought tolerance compared to S3A3 which is

consistent with the findings that delayed leaf senescence induces extreme

drought tolerance in Sunflower (Rivero et al 2007) Transgenic Tobacco

plants were generated expressing an isopentenyltransferase (enzyme that

catalyzes the rate-limiting step in cytokinin synthesis) gene from

Agrobacterium driven by promoter of senescence associated receptor

protein kinase (SARK) of bean Remarkably the suppression of drought-

induced leaf senescence resulted in outstanding drought tolerance as shown

by vigorous growth after a long drought period that killed the control plants

Such transgenic plants were also found to maintain high water contents and

retained photosynthetic activity during drought

ACC oxidase is the enzyme that catalyses one of the rate-limiting steps for

ethylene biosynthesis in plants (Wang et al 2003) Ethylene is a plant

hormone that involved in a wide range o f physiological processes in plants

including germination (Gallardo et al 1994) abscission (Abeles 1973

Jacobs1968 Osborne 1973) senescence (Grbic et al 1995) flowering

and fruit ripening (Theologis 1993) Ethylene is also involved in plant

responses to external signals for example responses to wounding

pathogens and stress caused by environmental pollutants such as ozone

(Zarembinski et al 1994) In the present study we have observed higher

Chapter IV

122

expression of of ACC oxidase gene in TV23 at BWS compared to S3A3

Therefore we can speculate that this higher expression of the gene also

resulted in to the higher production of ethylene at BWS Although increased

production of ethylene has been reported to induce leaf senescence in

plants (Smart 1994) in our experiment we did not observe any leaf

senescence in both the cultivars at BWS At this stage both the plants have

shown wilting phenotype but there was no visible symptoms of leaf

senescence Therefore higher production of ethylene in the present study

may not be related to leaf senescence Moreover similar results has also

been obtained in different studies of ethylene-insensitive mutants where it

was shown that ethylene is neither necessary nor on its own sufficient

to induce leaf senescence Leaves should reach a certain age to be

susceptible to the ethylene signal (Buchanan-Wollaston et al 2003)

Ethylenersquos involvement in senescence appears to be related to the timing of

the process (Nam 1997 Grbic and Bleecker 1995) Ethylene has been

reported to inhibit the ABA mediated leaf stomatal closure (Tanaka et al

2005) which is a very important mechanism for a plant to survive in a water

stress environment Assuming that the high expression of ACC oxidase

gene in drought tolerant cultivar also resulted in higher production of

ethylene compared to susceptible cultivar present expression studies have

shown an opposite picture with higher ethylene production in TV23

compared to S3A3 It has been reported (Yang et al 2005) that stomata

gets closed much later in tolerant Arabidopsis thaliana developed by over

expressing LLA23 gene (an ortholog of Asr1 from Lily) compared to wild

type at same level of drought The higher production of ethylene in the

tolerant cultivar may therefore be attributed to inhibition of stomatal closure

mediated by ABA during water stress to maintain the normal physiological

events Moreover we have observed that closing of stomata started much

earlier in S3A3 compared to TV23 (data not shown) Therefore higher

expression of ACC oxidase might be responsible for giving drought tolerance

in TV23 by inhibiting the ABA mediated stomatal closure through higher

production of ethylene However there are reports of both decrease

(Morgan et al 1990 Feng and Barker 1992 Narayana et al 1991 Eklund

et al 1992) and increase (Bergner and Teichmann 1993 Huberman et al

Chapter IV

123

1993 Michelozzi et al 1995 Tudela and Primo Millo 1992) of ethylene

production during water stress Therefore role of ethylene during water

stress has to be further investigated Ethylene has been reported to induce

the expression of a complex set of genes for example ethylene

biosynthesis genes (Whittaker et al 1997) ethylene receptor genes (Hua et

al 1998) hydrolase genes (Burns et al 1998) and pathogenesis-related

genes (Koiwa et al 1994) Among these we have observed high expression

of an ethylene induced esterase gene in TV23 compared to S3A3 at BWS

during the drought experiment (Figure 48b)

45 Conclusion

In the present study we have monitored the expression of 29 genes at three

stages of drought in two tea cultivars with contrasting drought tolerance

character Transient or continuous up- or down-regulation of these genes

clearly indicates that these genes are true drought responsive and not

constitutive Out of 29 genes selected for expression studies from library

BWE3E4 19 genes were found to be up-regulated in TV23 compared to

S3A3 at BWS which accounts for a subtraction efficiency as high as 65

The relative expression of all the genes (except chloroplast glyceraldehydes-

3-phosphate dehydrogenase) considered in the present study have shown

up-regulation in both the cultivars compared to control plant However the

expression pattern varies between the two cultivars at different level of

induced drought stress We were more interested in those genes whose

relative expression at BWS was higher in tolerant cultivar TV23 compared to

susceptible cultivar S3A3 Because we presume that these are the

potential group of genes (early responsive) that could be responsible for

extending drought tolerance period of TV23 to 22 days (BWS) as compared

to 15 days (BWS) for S3A3 during the induced drought experiment

Therefore high expression of these genes in TV23 might contribute for its

higher drought tolerance However there are few genes (as discussed

above) whose relative expression was found higher in S3A3 The down-

regulation of these genes in TV23 may also play a key role that may

probably contribute for higher drought tolerance We have identified two

Chapter IV

124

genes (elicitor-responsive gene 3 and cinnamoyl-CoA reductase) whose

expression was exceptionally higher in the tolerant cultivar compared to

susceptible (Appendix 41 Figure 46) at BWS It is interesting to note that

the expression patterns of these two genes are completely opposite

(continuously increasing in S3A3 and continuously decreasing in TV23 with

the progressive drought Appendix 41) in the two cultivars at all the stages

of drought (Figure 48b) These genes therefore may involve in higher

drought tolerance behaviour of TV23 Further studies are required to explore

the role of these two genes along with others during drought stress in tea

We have studied the expression of these genes under controlled

environmental variables in pot condition and there is always a genotype and

environment interaction factor (Genotype x Environment) that affects the

gene expression pattern in plants Therefore the expression patterns in the

present study may differ from field drought condition One has to consider

these factors to further substantiate the findings of present study that might

deviate in field condition Therefore from the present study we can conclude

that we have identified a set of gene whose expression was highly affected

by drought and these might be the potential group of genes responsible for

contrasting drought tolerance in tea Further studies are required to know the

functional role of these genes during drought stress that may pave the way

in understanding the drought tolerance mechanism in tea and help to devise

future strategies for improvement of drought tolerance There are several

ways to further substantiate the role of these genes in drought tolerance

mechanism for example full length cloning and heterologous expression to

study the structure (which is the subject matter of next chapter of this thesis)

and putative functions of these proteins study of drought tolerance of

transgenic plants over-expressing these proteins

References

1 Abeles F B 1973 Ethylene in Plant Biology Academic Press New-

York pp178 ndash 190

Chapter IV

125

2 Agre P Sasaki S Chrispeels M J 1993 Aquaporins a family of

water channel proteins Am J Physiol 261 F461

3 Aharon R Shahak Y Wininger S Bendov R Kapulnik Y Galili

G 2003 Over-expression of a plasma membrane aquaporin in

transgenic tobacco improves plant vigor under favorable growth

conditions but not under drought or salt stress Plant Cell 15 439 ndash

447

4 Alamillo J Almogura C Bartels D Jordano J 1995 Constitutive

expression of small heat shock proteins in vegetative tissues of the

resurrection plant Craterostigma planatgenium Plant Mol Biol 29 1093

ndash 1099

5 Allagulova C R Gimalor F R Shakirova F M Vakhitov V A

2003 The plant dehydrins Structure and putative functions

Biochemistry (Mosc) 68 945 ndash 951

6 Amiard T Guenzi A C Martin B Cushman J C 2003 Tolerance

of mannitol-accumulating transgenic wheat to water stress and salinity

Plant Physiol 131 1748 ndash 1755

7 An Z Jing W Liu Y Zhang W 2008 Hydrogen peroxide generated

by copper amine oxidase is involved in abscisic acid-induced stomatal

closure in Vicia faba J Expt Bot 59(4) 815 ndash 825

8 Apel K and Hirt H 2004 Reactive oxygen species metabolism

oxidative stress and signal transduction Annu Rev Plant Biol 55 373

ndash 399

9 Asada K and Takahashi M 1987 Production and scavenging of

active oxygen in photosynthesis In Photoinhibition (Kyle DJ et al

eds) pp 227 ndash 287 Elsevier

10 Badawi G H Kawanoa N Yamauchi Y Shimada E Sasaki R

Kubo A Tanaka K 2004 Over-expression of ascorbate peroxidase in

tobacco chloroplasts enhances the tolerance to salt stress and water

deficit Physiologia Plantarum 121 231 ndash 238

11 Bais H P Vepachedu R Gilroy S Callaway R M Vivanco J M

2003 Allelopathy and exotic plant invasion from molecules and genes

to species interactions Science 3011377 ndash 1380

Chapter IV

126

12 Bartels D 2001 Targeting detoxification pathways an efficient

approach to obtain plants with multiple stress tolerence Trends Plant

Sci 6 284 ndash286

13 Barua D N 1989 Science and practice in Tea culture (Calcutta Tea

Research Association) pp 509

14 Battaglia M Olvera-Carrillo Y Garciarrubio A Campos F

Covarrubias A A 2008 The enigmatic LEA proteins and other

hydrophilins Plant Physiol 148(1) 6 ndash 24

15 Bergner G and Teichmann G 1993 A role for ethylene in plants

responding to soil water shortage J Plant Growth Regul 12 67 ndash 72

16 Berry J A and Bojorkman O 1980 Photosynthetic response and

adaptation to temperature in higher plants Annu Rev Plant Physiol 31

491 ndash 543

17 Bethke P C and Jones R L 2001 Cell death of barley aleurone

protoplasts is mediated by reactive oxygen species Plant J 25 19 ndash

29

18 Borovskii G B Stupnikova I V Antipina A I Vladimirova S V

Voinikov V K 2002 Accumulation of dehydrin-like proteins in the

mitochondria of cereals in response to cold freezing drought and ABA

treatment BMC Plant Biol 2 5

19 Boudet J Buitink J Hoekstra F A Rogniaux H Larre C Satour

P Leprince O 2006 Comparative analysis of the heat stable

proteome of radicles of Medicago truncatula seeds during germination

identifies late embryogenesis abundant proteins associated with

desiccation tolerance Plant Physiol 140 1418 ndash 1436

20 Bowler C Montagu M V Inze D 1992 Superoxide dismutase and

stress tolerance Ann Rev Plant Physiol Plant Mol Biol 43 83 ndash 116

21 Brini F Hanin M Lumbreras V Irar S Pages M Masmoudi K

2007 Over-expression of wheat dehydrin DHN-5 enhances tolerance to

salt and osmotic stress in Arabidopsis thaliana Plant Cell Rep 26

2017 ndash 2026

22 Broholm S K Tahtiharju S Laitinen R A E Albert V A Teeri T

H Elomaa P 2008 A TCP domain transcription factor controls flower

Chapter IV

127

type specification along the radial axis of the Gerbera (Asteraceae)

inflorescence PNAS USA 26 9117 ndash 9122

23 Buchanan-Wollaston V 1997 The molecular biology of leaf

senescence J Expt Bot 48 181 ndash 199

24 Buchanan-Wollaston V Earl S Harrison E Mathas E Navabpour

S Page T and Pink D 2003 The molecular analysis of leaf

senescence ndash a genomics approach Plant Biotech J 1 3 ndash 22

25 Burns J K Lewandowski D J Nairn C J Brown G E 1998

Endo-1 4-beta-glucanase gene expression and cell wall hydrolase

activities during abscission in Valencia orange Physiol Plant 102 217

ndash 225

26 Bustin S A 2000 Absolute quantification of mRNA using real-time

reverse transcription polymerase chain reaction assays J Mol

Endocrinology 25 169 ndash 193

27 Cakir B Agasse A Gaillard C Saumonneau A Delrot S

Atanassova R 2003 A grape ASR protein involved in sugar and

abscisic acid signalling Plant Cell 15(9) 2165 ndash 2180

28 Cameron K D Teece M A Smart L B 2006 Increased

accumulation of cuticular wax and expression of lipid transfer protein in

response to periodic drying events in leaves of tree tobacco Plant

Physiol 140 176 ndash 183

29 Campalans A Pages M Messeguer R 2001 Identification of

differentially expressed genes by the cDNA-AFLP technique during

dehydration of almond (Prunus amygdalus) Tree Physiol 21 633 ndash

643

30 Caramelo J J and Iusem N D 2008 When cells lose water lessons

from biophysics and molecular biology Prog Biophys Mol Biol 99(1) 1

ndash 6

31 Carpita N C and Gibeau D M 1993 Structural models of primary

cell walls in flowering plants consistency of molecular structure with the

physical properties of the walls during growth Plant J 3 1 ndash 30

32 Cascardo J Almeida R Buzeli R Carolino S Otoni W Fontes

E 2000 The phosphorylation state and expression of soybean BiP

Chapter IV

128

isoforms are differentially regulated following abiotic stresses J Biol

Chem 275 14494 ndash 14500

33 Cellier F G Conejero J Breitler C Casse F 1997 Molecular and

physiological responses to water deficit in drought-tolerant and drought-

sensitive lines of sunflower1 Accumulation of dehydrin transcripts

correlates with tolerance Plant Physiol 116 319 ndash 328

34 Chang S Puryear J D Dias M A D L Funkhouser E A Newton

R J Cairney J 1995 Gene expression under water deficit in loblolly

pine (Pinus taeda L) isolation and characterization of cDNA clones

Physiol Plant 95 1 ndash 10

35 Chen W Chao G Singh K B 1996 The promoter of a H2O2-

inducible Arabidopsis glutathione S-transferase gene contains closely

linked OBF- and OBP1-binding sites Plant J 10 955 ndash 966

36 Cheng Z Targolli J Huang X Wu R 1995 Wheat LEA genes

PMA80 and PMA enhance dehydration tolerance of transgenic rice

(Oryza sativa L) Mol Breeding 10 71 ndash 82

37 Chini A Fonseca S Fernandez G Adie B Chico J M Lorenzo

O Garcia-Casado G Lopez-Vidriero I Lozano F M Ponce M R

Micol J L Solano R 2007 The JAZ family of repressors is the

missing link in jasmonate signalling Nature 448 666 ndash 671

38 Chow W S Melis A Anderson J M 1990 Adjustments of

photosystem stoichiometry in chloroplasts improve the quantum

efficiency of photosynthesis PNAS USA 87 7502 ndash 7506

39 Chrispeels M J and Agre P 1994a Aquaporins water channel

proteins of plant and animal cells Trends Biochem Sci 19 421 ndash 425

40 Chrispeels M J and Maurel C 1994b Aquaporins the molecular

basis of facilitated water movement through living plant cells Plant


41 Close T J 1997 Dehydrins a commonality in the response of plants

to dehydration and low temperature Physiol Plant 100 291 ndash 296

42 Coca M Almoguera C Thomas T Jordano J 1996 Differential

regulation of small heat-shock genes in plants Analysis of a water

stress-inducible and developmentally activated sunflower promoter

Plant Mol Biol 31 863 ndash 876

Chapter IV

129

43 Dat J Vandenabeele S Vranova E Van Montagu M Inze D Van

Breusegem F 2000 Dual action of the active oxygen species during

plant stress responses Cell Mol Life Sci 57 779 ndash 795

44 Downton W J S Berry J A Seemann J R 1984 Tolerance of

photosynthesis to high temperature in desert plants Plant Physiol 74

786 ndash 790

45 Dure L 1993 Structural motifs in LEA proteins In T J Close E A

Bray Eds Plant responses to cellular dehydration during

Environmental Stress American Society of Plant Physiologists

Rockville MD pp 91 ndash 103

46 Dure L Crouch M Harada J J Ho T Mundy J Quatrano R S

Thomas T L Sung Z R 1989 Common amino acid sequence

domains among the LEA proteins of higher plants Plant Mol Biol 12

475 ndash 486

47 Ecker J R 1995 The ethylene signal transduction pathway in plants

Science 268 667 ndash 675

48 Ehness R and Roitsch T 1997 Co-ordinated induction of mRNAs for

extracellular invertase and a glucose transporter in Chenopodium

rubrum by cytokinins Plant J 11 539 ndash 548

49 Eklund L Gieociala E and Hallgren J E 1992 No relation between

drought stress and ethylene production in Norway spruce Physiol

Plant 86 297 ndash 300

50 Eun K and Choo B 2006 Over-expression of tobacco NtHSP70-1

contributes to drought-stress tolerance in plants Plant Cell Rep 25

349 ndash 358

51 Feng H Chen Q Feng J Zhang J Yang X Zuo J 2007

Functional characterization of the Arabidopsis eukaryotic translation

initiation factor 5A-2 that plays a crucial role in plant growth and

development by regulating cell division cell growth and cell death


52 Feng J and Barker A V 1992 Ethylene evolution and ammonium

accumulation by tomato plants under water and salinity stresses Part-

II J Plant Nutr 15 2471 ndash 2490

Chapter IV

130

53 Fischer R A and Tuner N C 1978 Plant productivity in arid and

semiarid zones Ann Rev of Plant Physiol 29 277 ndash 317

54 Foyer C H and Noctor G 2005 Redox homeostasis and antioxidant

signalling A metabolic interface between stress perception and

physiological responses Plant Cell 17 1866 ndash 1875

55 Frankel N Hasson E Iusem N D Rossi M S 2003 Adaptive

evolution of the water stress-induced gene Asr2 in Lycopersicon

species dwelling in arid habitats Mol Biol Evol 20(12) 1955 ndash 1962

56 Gallardo M Gallardo M E Matilla A J De Rueda P M Sanchez-

Calle I M 1994 Inhibition of polyamine synthesis by cyclohexylamine

stimulates the ethylene pathway and accelerates the germination of

Cicer arietinum seeds Physiologia Plantarum 91 9 ndash 16

57 Goujon T Ferret V Mila I Pollet B Ruel K Burlat V Joseleau

J P Barriere Y Lapierre C Jouanin L 2003 Down-regulation of

the AtCCR1 gene in Arabidopsis thaliana effects on phenotype lignins

and cell wall degradability Planta 217 218 ndash 228

58 Goyal K Walton L J Tunnacliffe A 2005 LEA proteins prevent

protein aggregation due to water stress Biochem J 388 151 ndash 157

59 Grbic V and Bleecker A B 1995 Ethylene regulates the timing of

leaf senescence in Arabidopsis Plant J 8 595 ndash 602

60 Halliwell B and Gutteridge J M C 1999 Free Radicals in Biology

and Medicine 3rd Edn Oxford University Press New York

61 Hammond-Kosack K E and Jones J D G 1996 Resistance gene-

dependent plant defense responses Plant Cell 8 1773 ndash 1791

62 Heinen R B Ye Q Chaumont F 2009 Role of aquaporins in leaf

physiology J Expt Bot 60 2971 ndash 2985

63 Hillel D and Vlek P 2005 The sustainability of irrigation Adv Agron

87 55 ndash 84

64 Houde M Danyluk J Laliberte J F Rassart E Dhindsa R S

Sarhan F 1992 Cloning characterization and expression of a cDNA

encoding a 50-kilodalton protein specifically induced by cold

acclimation in wheat Plant Physiol 99(4) 1381 ndash 1387

65 Houston N L Fan C Xiang J Q Schulze J M Jung R Boston

R S 2005 Phylogenetic analyses identify 10 classes of the protein

Chapter IV

131

disulphide isomerase family in plants including single-domain protein

disulphide isomerase related proteins Plant Physiol 137 762 ndash 778

66 Hsiao T C Steduto P Fereres E 2007 A systematic and

quantitative approach to improve water use efficiency in agriculture

Irrig Sci 25 209 ndash 231

67 Hu Y Li W-Ch Xu Y-Q Li G-J Liao Y Fu F-L 2009 Differential

expression of candidate genes for lignin biosynthesis under drought

stress in maize leaves J Appl Genet 50(3) 213 ndash 223

68 Hua J Sakai H Nourizadeh S Chen Q G Bleecker A B Ecker

J R Meyerowitz E M 1998 EIN4 and ERS2 are members of the

putative ethylene receptor gene family in Arabidopsis Plant Cell 10

1321 ndash 1332

69 Huberman M Pressman E Jaffe M J 1993 Pith autolysis in plants

IV The activity of polygalacturonase and cellulase during drought

stress-induced pith autolysis Plant Cell Physiol 34 795 ndash 801

70 Imai R Chang L Ohta A Bray E A Takagi M 1996 A Lea-class

gene of tomato confers salt and freezing tolerance when expressed in

Saccharomyces cerevisiae Gene 170 243 ndash 248

71 Ingram J and Bartels D 1996 The molecular basis of dehydration

tolerance in plants Annu Rev Plant Physiol Plant Mol Biol 47 377 ndash

403

72 Ismail A M Hall A E Close T J 1999 Allelic variation of a

dehydrin gene cosegregates with chilling tolerance during seedling

emergence PNAS USA 96 13566 ndash 13570

73 Iusem N D Bartholomew D M Hitz W D Scolnik P A 1993

Tomato (Lycopersicon esculentum) transcript induced by water deficit

and ripening Plant Physiol 102 1353 ndash 1364

74 Jacobs W P 1968 Hormonal regulation of leaf abscission Plant


75 Jain N K 1999 Global advances in tea science (New Delhi Aravali

Books International) pp 882

76 Ji W Zhu Y Li Y Yang L Zhao X Cai H Bai X 2010 Over-

expressing of a glutathione S-transferase gene GsGST from wild type

Chapter IV

132

Soybean (Glycine soja) enhances drought and salt tolerance in

transgenic tobacco Biotechnol Lett 32(8) 1173 ndash 1179

77 Jones L Ennos A R Turner S R 2001 Cloning and

characterization of irregular xylem4 (irx4) a severely lignin-deficient

mutant of Arabidopsis The Plant J 26 205 ndash 216

78 Kaldenhoff R and Fischer M 2006 Aquaporins in plants Acta Physiol

(Oxf) 187 169 ndash 176

79 Kaup M T Froese C D Thompson J E 2002 A role for

diacylglycerol acyltransferase during leaf senescence Plant Physiol

129 1616 ndash 1626

80 Kawasaki T Koita H Nakatsubo T Hasegawa K Wakabayashi

K Takahashi H Umemura K Umezawa T Shimamoto K 2006

cinnamoyl-CoA reductase a key enzyme in lignin biosynthesis is an

effector of small GTPase Rac in defense signalling in rice PNAS USA

103 230 ndash 235

81 Khan S Tariq R Yuanlai C Blackwell J 2006 Can irrigation be

sustainable Agric Water Manage 80 87 ndash 99

82 Kim S Y and Nam K H 2010 Physological roles of ERD10 in abiotic

stresses and seed germination in Arabidopsis Plant Cell Rep 29 203

ndash 209

83 Kimura M Yamamoto Y Y Seki M Sakurai T Sato M Abe

T Yoshida S Manabe K Shinozaki K Matsui M 2003

Identification of Arabidopsis genes regulated by high light-stress using

cDNA microarray Photochem Photobiol 77(2) 226 ndash 233

84 Knepper M A 1994 The aquaporin family of molecular water

channels PNAS USA 91 6255 ndash 6258

85 Ko J H Yang S H Han K H 2006 Upregulation of an Arabidopsis

RING-H2 gene XERICO confers drought tolerance through increased

abscisic acid biosynthesis Plant J 47 343 ndash 355

86 Koag M C Fenton D R Wilkens S Close T J 2003 The binding

of maize DHN1 to lipid vesicles gain of structure and lipid specificity


Chapter IV

133

87 Koiwa H Sato F Yamada Y 1994 Characterization of accumulation

of tobacco PR-5 proteins by IEF-immunoblot analysis Plant Cell


88 Kovacs D Kalmar E Torok Z Tompa P 2008 Chaperone activity

of ERD10 and ERD14 two disordered stress-related plant proteins


89 Kranner I Beckett R P Wornik S Zorn M Pfeifhofer H W 2002

Revival of resurrection plant correlates with its antioxidant status Plant

J 31 13 ndash 24

90 Kruger C Berkowitz O Stephan U W Hell R 2002 A metal-

binding member of the late embryogenesis abundant protein family

transports iron in the phloem of Ricinus communis L J Biol Chem

277(28) 25062 ndash 25069

91 Kushwaha H R Singh A K Sopory S K Singla-Pareek S L

Pareek A 2009 Genome wide expression analysis of CBS domain

containing proteins in Arabidopsis thaliana (L) Heynh and Oryza sativa

L reveals their developmental and stress regulation BMC Genomics

10 200

92 Lamport D T A 1970 Cell wall metabolism Annu Rev Plant Physiol

21 235 ndash 270

93 Lee J Hubel A Schoffl F 1995 Derepression of the activity of

genetically engineered heat shock factor causes constitutive

synthesis of heat shock proteins and increased thermotolerance in

transgenic Arabidopsis Plant J 8 603 ndash 612

94 Leon O and Roth M 2000 Zinc fngers DNA binding and protein-

protein interactions Biol Res 33 21 ndash 30

95 Leple J C Dauwe R Morreel K Storme V et al 2007

Downregulation of cinnamoyl-coenzyme A reductase in poplar

multiple-level phenotyping reveals effects on cell wall polymer

metabolism and structure The Plant Cell 19 3669 ndash 3691

96 Levine A Tenhaken R Dixon R Lamb C 1994 H2O2 from the

oxidative burst orchestrates the plant hypersensitive disease resistance

response Cell 79 583 ndash 593

Chapter IV

134

97 Li C Potuschak T Colon-Carmona A Gutierrez R A Doerner P

2005 Arabidopsis TCP20 links regulation of growth and cell division

control pathways PNAS USA 102(36) 12978 ndash 12983

98 Li L Staden J V Jager A K 1998 Effects of plant growth

regulators on the antioxidant system in seedlings of two maize cultivars

subjected to water stress Plant Growth Regul 25 81 ndash 87

99 Liu S and Jiang Y 2010 Identification of differentially expressed

genes under drought stress in perennial ryegrass Physiologia

Plantarum 139 375 ndash 387

100 Lorick K L Jensen J P Fang S Ong A M Hatakeyama S

Weissman A M 1999 RING fingers mediate ubiquitin-conjugating

enzyme (E2)-dependent ubiquitination PNAS USA 96(20) 11364 ndash

11369

101 Luhua S Ciftci-Yilmaz S Harper J Cushman J Mittler R 2008

Enhanced tolerance to oxidative stress in transgenic Arabidopsis plants

expressing proteins of unknown function Plant Physiol 148 280 ndash 92

102 Maurel C 2007 Plant aquaporins novel functions and regulation

properties FEBS Lett 581 2227 ndash 2236

103 Maurel C Santoni V Luu D-T Wudick M M Verdoucq L 2009

The cellular dynamics of plant aquaporin expression and functions

Plant Biol 12 690 ndash 698

104 Melotto M Mecey C Niu Y Chung H S Katsir L Yao J Zeng

W Thines B Staswick P E Browse J Howe G A He S Y

2008 A critical role of two positively charged amino acids in the Jas

motif of Arabidopsis JAZ proteins in mediating coronatine- and

jasmonoyl isoleucine-dependent interactions with the COI1 F-box

protein Plant J 55 979 ndash 988

105 Miao Y Lv D Wang P Wang Xue-Chen Chen J Miao C

Songa Chun-Peng 2006 An Arabidopsis glutathione peroxidase

functions as both a redox transducer and a scavenger in abscisic acid

and drought stress responses The Plant Cell 18 2749 ndash 2766

106 Micheletto S Rodriguez-Uribe L Hernandez R Richins R D

Curry J OrsquoConnell M A 2007 Comparative transcript profiling in

Chapter IV

135

roots of Phaseolus acutifolius and P vulgaris under water deficit stress

Plant Science 173 510 ndash 520

107 Michelozzi M Johnson J D Warrag E I 1995 Responses of

ethylene and chlorophyll in two eucalyptus clones during drought New

For 9(3) 197 ndash 204

108 Mohd N Anju B Dhananjay S 2006 Immunogenicity and

protective efficacy of DnaJ (hsp40) of Streptococcus pneumoniae

against lethal infection in mice Vaccine 24 6225 ndash 6231

109 Morgan P W 1990 Effects of abiotic stresses on plant hormone

systems ndash In stress responses In plants adaptation mechanisms (R

Alscher and J Gumming eds) pp 313 ndash 314 Wiley-Liss Inc Publ

New York ISBN0-471-56810-4

110 Munoz-Bertomeu J Bermudez M A Segura J Ros R 2011

Arabidopsis plants deficient in plastidial glyceraldehyde-3-phosphate

dehydrogenase show alterations in abscisic acid (ABA) signal

transduction interaction between ABA and primary metabolism J Expt

Bot 62(3) 1229 ndash 1239

111 Nam H G 1997 The molecular genetic analysis of leaf senescence

Curr Opin Biotech 8 200 ndash 207

112 Narayana L Lalonde S Saini H S 1991 Water-stress-induced

ethylene production in wheat a fact or artefact Plant Physiol 96 406

ndash 410

113 Navari-Izzo F Vangioni N Quartacci M F 1990 Lipids of soybean

and sunflower seedlings grown under drought conditions

Phytochemistry 29(7) 2119 ndash 2123

114 Neill N Desikan R Hancock J 2002 Hydrogen peroxide signalling

Curr Opin Plant Biol 5 388 ndash 395

115 Noctor G and Foyer C H 1998 Ascorbate and glutathione keeping

active oxygen under control Annu Rev Plant Physiol Plant Mol Biol 49

249 ndash 279

116 Nooden L D 1988 Whole plant senescence In Senescence and

aging in plants (Nooden L D and Leopold A C Eds) Academic

Press San Diego 391 ndash 439

Chapter IV

136

117 Nylander M Svensson J Palva E T Welin B V 2001 Stress-

induced accumulation and tissue-specific localization of dehydrins in

Arabidopsis thaliana Plant Mol Biol 45(3) 263 ndash 279

118 Ohme-Takagi M and Shinshi H 1995 Ethylene-inducible DNA

binding proteins that interact with an ethylene-responsive element

Plant Cell 7 173 ndash 182

119 Osborne D J 1973 Shedding of Plant Parts Academic Press New

York pp 125 ndash 147

120 Pannunzio A 2008 Efectos de sustentabilidad de los sistemas de

riego por goteo en arandanos 113 p Tesis MgSc Universidad de

Buenos Aires Facultad de Ciencias Veterinarias Buenos Aires

Argentina

121 Pannunzio A Texeira P Perez D Sbarra G Grondona y A 2008

Efectos de sustentabilidad de los sistemas de riego de araacutendanos en la

Pampa Huacutemeda p 24-25 Libro de Resuacutemenes del I Congreso

Latinoamericano de Arandanos y otros berries Agosto 2008

Universidad de Buenos Aires Facultad de Agronomiacutea (FAUBA)

Buenos Aires Argentina

122 Piquemal J Lapierre C Myton K OrsquoConnell A Schuch W

Grima-Pettenati J Boudet A M 1998 Down-regulation of cinnamoyl-

CoA reductase induces significant changes of lignin profiles in

transgenic tobacco plants The Plant J 13 71 ndash 83

123 Polidoros A N and Scandalios J G 1999 Role of hydrogen

peroxide and different classes of antioxidants in the regulation of

catalase and glutathione S-transferase gene expression in maize (Zea

mays L) Physiol Plant 106 112 ndash 120

124 Poza-Carrion C Aguilar-Martinez J A Cubas P 2007 Role of TCP

gene BRANCHED1 in the control of shoot branching in Arabidopsis

Plant Signalling amp Behavior 2(6) 551 ndash 552

125 Reddy B A Etkin L D Freemont P S 1992 A novel zinc finger

coiled-coil domain in a family of nuclear proteins Trends Biochem Sci

17(9) 344 ndash 345

126 Reich M Liefeld T Gould J Lerner J Tamayo P Mesirov J P

2006 GenePattern 20 Nature Genetics 38 no 5 pp 500 ndash 501

Chapter IV

137

127 Reymond P and Farmer E E 1998 Jasmonate and salicylate as

global signals for defense gene expression Curr Opin Plant Biol 1 404

ndash 411

128 Rinne P Willing A Kaikuranta P 1998 Onset of freezing tolerance

in birch (Betula pubescens Ehrh) involves LEA proteins and

osmoregulation and is impaired in an ABA-deficient genotype Plant

Cell Environ 21 601 ndash 11

129 Rivero R M Kojima M Gepstei A Sakakibara H Milter R

Gepstein S Blumwald E 2007 Delayed leaf senescence induces

extreme drought tolerance in a flowering plant PNAS USA 104(49)

19631 ndash 19636

130 Rorat T 2006 Plant dehydrins-tissue location structure and function

Cell Mol Biol Lett 11 536 ndash 556

131 Rozen S and Skaletsky H J 2000 Primer3 on the WWW for general

users and for biologist programmers Plant Mol Biol 5 69 ndash 76

132 Sade N Gebretsadik M Seligmann R Schwartz A Wallach R

Moshelion M 2010 The role of Tobacco aquaporin1 in improving

water use efficiency hydraulic conductivity and yield production under

salt stress Plant Physiol 152 245 ndash 254

133 Sakaki T Kondo N Yamada M 1990a Pathway for the synthesis of

triacylglycerols from monogalactosyldiacylglycerols in ozone-fumigated

spinach leaves Plant Physiol 94 773 ndash 780

134 Sakaki T Saito K Kawaguchi A Kondo N Yamada M 1990b

Conversion of monogalactosyldiacylglycerols to triacylglycerols in

ozone-fumigated spinach leaves Plant Physiol 94 766 ndash 772

135 Shalata A Mittova V Volokita M Guy M Tal M 2001 Response

of the cultivated tomato and its wild salt-tolerant relative Lycopersicon

pennellii to salt-dependent oxidative stress The root antioxidative

system Physiologia Plantarum 112 487 ndash 494

136 Shinozaki K and Yamaguchi-Shinozaki K 1997 Gene expression

and signal transduction in water stress response Plant Physiol 115

327 ndash 334

Chapter IV

138

137 Shinozaki K Yamaguchi- Shinozaki K Seki M 2003 Regulatory

network of gene expression in the drought and cold stress responses


138 Shinshi H Usami S Ohme-Takagi M 1995 Identification of an

ethylene-responsive region in the promoter of a tobacco class I

chitinase gene Plant Mol Biol 27 923 ndash 932

139 Siefritz F Tyree M T Lovisolo C Schubert A Kaldenhoff R

2002 PIP1 plasma membrane aquaporins in tobacco From cellular

effects to function in plants Plant Cell 14 869 ndash 876

140 Silhavy D Hutvagner G Barta E Banfalvi Z 1995 Isolation and

characterization of a water-tress-inducible cDNA clone from Solanum

chacoense Plant Mol Biol 27 587 ndash 595

141 Sivamani E Bahieldin A Wraith J M Al-Niemi T Dyer W E Ho

T-HD Qu R 2000 Improved biomass productivity and water use

efficiency under water deficit conditions in transgenic wheat

constitutively expressing the barley HVA1 gene Plant Sci 155 1 ndash 9

142 Smart C M 1994 Gene expression during leaf senescence New

Phytol 126 419-448

143 Smirnoff N 1998 Plant resistance to environmental stress Curr Opin


144 So Hyun-Ah Chung E Cho Chang-Woo Kim Kee-Young Lee

Jai-Heon 2010 Molecular cloning and characterization of soybean

cinnamoyl-CoA reductase induced by abiotic stresses Plant Pathol J

26(4) 380 ndash 385

145 Sun W Bernard C Van D Van M Verbruggen N 2001 At-

HSP17 6A encoding a small heat-shock protein in Arabidopsis can

enhance osmotolerance upon over-expression Plant J 27 407 ndash 415

146 Swire-Clark G A and Marcotte W R Jr 1999 The wheat LEA

protein Em functions as an osmoprotective molecule in Saccharomyces

cerevisiae Plant Mol Biol 39 117 ndash 128

147 Taiz L and Zeiger E 2002 Plant physiology 3rd edn Sunderland M

A Sinauer Associates Publishers

148 Tan J Zhao H Hong J Han Y Li H Zhao W 2008 Effects of

exogenous nitrick oxide on photosynthesis antioxidant capacity and

Chapter IV

139

proline accumulation in wheat seedlings subjected to osmotic stress

World J Agri Sci 4(3) 307 ndash 313

149 Tanaka Y Sano T Tamaoki M Nakajima N Kondo N

Hasezawa S 2005 Ethylene inhibits abscisic acid-Induced stomatal

closure in Arabidopsis Plant Physiol 138 2337 ndash 2343

150 Theologis A 1993 One rotten apple spoils the whole bushel the role

of ethylene in fruit ripening Cell 70 181 ndash 184

151 Tichopad A Didier A Pfaffl M W 2004 Inhibition of real-time RT-

PCR quantification due to tissue-specific contaminants Mol Cell

Probes 18 45 ndash 50

152 Tichopad A Dilger M Schwarz G Pfaffl M W 2003 Standardized

determination of real-time PCR efficiency from a single reaction set-up

Nucl Acids Res 31(20) e122

153 Tudela D and Primo Millo E 1992 1-Aminocyclopropane-1-

carboxylic acid transported from roots to shoots promotes leaf

abscission in Cleopatra mandrin (Citrus rashni Hort ex tan) seedlings

rehydrated after water stress Plant Physiol 100 131 ndash 137

154 Tunnacliffe A and Wise M J 2007 The continuing conundrum of the

LEA proteins Naturwissenschaften 94(10) 791 ndash 812

155 Tyerman S D Niemietz C M Bramley H 2002 Plant aquaporins

multifunctional water and solute channels with expanding roles Plant


156 Vandenabeele S Kelen K V D Dat J Gadjev I Boonefaes T

Morsa S Rottiers P Slooten L Montagu M V Zabeau M Inze

D Breusegem F V 2003 A comprehensive analysis of hydrogen

peroxide- induced gene expression in tobacco PNAS USA 100(26)

16113 ndash 16118

157 Vandesompele J De Preter K Pattyn F Poppe B Van Roy N

De Paepe A Speleman F 2002 Accurate normalization of real-time

quantitative RT-PCR data by geometric averaging of multiple internal

control genes Genome Biology 3 341 ndash 3411

158 Wadenback J Von Arnold S Egertsdotter U Walter M H Grima-

Pettenati J Goffner D Gellerstedt G Gullion T Clapham D 2008

Lignin biosynthesis in transgenic Norway spruce plants harbouring an

Chapter IV

140

antisense construct for cinnamoyl- CoA reductase (CCR) Transgenic

Research 17 379 ndash 392

159 Walters R G and Horton P 1995 Acclimation of Arabidopsis thaliana

to the light environment changes in photosynthetic function Planta

197 306 ndash 312

160 Walters R G Shephard F Rogers J J Rolfe S A Horton P

2003 Identification of mutants of Arabidopsis defective in acclimation of

photosynthesis to the light environment Plant Physiol 131 472 ndash 481

161 Wang H J Hsu C M Jauh G Y Wang C S 2005 A Lilly pollen

ASR protein localizes to both cytoplasm and nuclei requiring a nuclear

localization signal Physiol Plant 123 314 ndash 320

162 Wang W Vinocur B Altman A 2003 Plant responses to drought

salinity and extreme temperatures towards genetic engineering for

stress tolerance Planta 218 1 ndash 14

163 Wang W Vinocur B Shoseyov O Altman A 2004 Role of plant

heat-shock proteins and molecular chaperones in the abiotic stress

response Trends in Plant Sci 9 244 ndash 252

164 Whittaker D J Smith G S Gardner R C 1997 Expression of

ethylene biosynthetic genes in Actinidia chinensis fruit Plant Mol Biol

34 45 ndash 55

165 Wood A J and Goldsbrough P B 1997 Characterization and

expression of dehydrins in water stressed Sorghum bicolor Physiol


166 Xiao B Huang Y Tang N Xiong L 2007 Over-expression of a

LEA gene in rice improves drought resistance under the field

conditions Theor Appl Genet 115 35 ndash 46

167 Xiong L and Zhu J K 2001 Abiotic stress signal transduction in

plants molecular and genetic perspectives Physiol Plant 112 152 ndash

166

168 Xu D Duan X Wang B Hong B Ho T Wu R 1996 Expression

of a late embryogenesis abundant protein gene HVA1 from barley

confers tolerance to water deficit and salt stress in transgenic rice


Chapter IV

141

169 Yamaguchi K Mori H Nishimura M 1995 A novel isoenzyme of

ascorbate peroxidase localized on glyoxysomal and leaf peroxisomal

membranes in pumpkin Plant Cell Physiol 36(6) 1157 ndash 1162

170 Yang Chin-Ying Chen Yu-Chuan Jauh Guang Yuh Wang Co-

Shine 2005 A Lily ASR protein involves abscisic acid signalling and

confers drought and salt resistance in Arabidopsis Plant Physiol 139

836 ndash 846

171 Yang Y Kwon Hawk-Bin Peng Hsiao-Ping Shih Ming-Che 1993

Stress response and metabolic regulation of glyceraldehyde-3-

phosphate dehydrogenase genes in Arabidopsis Plant Physiol 101

209 ndash 216

172 Zarembinski T I and Theologis A 1994 Ethylene biosynthesis and

action a case of observation Plant Mol Biol 26 579 ndash 597

173 Zhang L Ohta A Takagi M Imai R 2000 Expression of plant

group 2 and group 3 lea genes in Saccharomyces cerevisiae revealed

functional divergence among LEA proteins J Biochem (Tokyo) 127

611 ndash 616

174 Zhao Y Du H Wang Z Huang B 2011 Identification of proteins

associated with water-deficit tolerance in C4 perennial grass species

Cynodon dactylon times Cynodon transvaalensis and Cynodon dactylon

Physiologia Plantarum 141 40 ndash 55

175 Zhou R Jackson L Shadle G Nakashima J Temple S Chen F

Dixon R A 2010 Distinct cinnamoyl-CoA reductases involved in

parallel routes to lignin in Medicago truncatula PNAS USA 107

17803 ndash 17808

176 Zhu B Choi D W Fenton R Close T J 2000 Expression of the

barley dehydrin multigene family and the development of freezing

tolerance Mol Gen Genet 264 145 ndash 153

Chapter IV

89

41 Introduction

Water stress is one of the most important environmental factor that limits

crop productivity and quality throughout the world resulting in substantial

loss every year (Tan et al 2008) Efficient irrigation is one of the key factor

for sustainable agriculture as pointed out by many researchers from time to

time (Hillel and Vlek 2005 Khan et al 2006 Hsiao et al 2007 Pannunzio

et al 2008 Pannunzio 2008) However due to continuous depletion and

uneven distribution of water resources in many regions of the world it has

become necessary to genetically improve the performance of crop plants in

such water stress environments for a sustainable and economically viable

solution

Successful cultivation of tea is highly dependent on availability of rain water

There are reports of yield losses every year due to recurring prolong dry

spell in the tea growing areas (Barua 1989 Jain 1999) of India and other

countries like Kenya and Sri Lanka Keeping in view the continuous increase

of drought hit regions due to decreased rainfall the present need of the tea

industry is a good quality high yielding tea genotype that can withstand and

perform well during the prolonged dry spells of the year One of the ways to

achieve this goal is to study the drought responsive mechanism in tea at

molecular level to identify the different drought responsive pathways and

their corresponding genes which would potentially contribute towards

development of drought tolerant tea plants However the development of

drought tolerant tea genotypes has been hindered by lack of knowledge of

more precise physiological and molecular parameters that reflect genetic

potential for improved productivity under water stress environment

Comparative study of the transcriptome of a drought tolerant tea genotype

with a susceptible one is one of the potential ways to decipher the

mechanism of drought tolerance in tea plant We have identified a large

numbers of differentially expressed transcripts in a drought tolerant cultivar

TV23 by comparative transcriptome analysis with a susceptible cultivar

S3A3 (Chapter III) from an induced water stress experiment by SSH

approach Some of these differentially expressed transcripts are highly

represented in TV23 compared to S3A3 which may potentially be

Chapter IV

90
































cultivars

Chapter IV

91



























21 17


19 23


18 19



21 24

Chapter IV

92







4231 Primer Design






















EST ID dbEST acc no





6220 6224

268



6306 6298

120


6205 6223

164


6147 6126

211


6192 6241

157


6190 6000

191



6276 6297

206



6243 6171

106

N23E3E4_A233 HS394047 Lipase 2e-20


6198 6186

158


6319 6289

155


6110 6108

180



6165 6169

206


6282 6328

203


6213 6204

209


6177 6172

184

Chapter IV

94

Table 42 Contd

EST ID dbEST acc no




6206 6250

235


6319 6289

162


6279 6202

170


6123 6176

217



6314 6313

217


6085 6098

179



6303 6305

222


6296 6362

159


6153 6194

299



6201 6060

280


6176 6156

196


6482 6403

220



6264 6222

169



6228 6198

153

Chapter IV

95









Walters et al 2003













Ecker 1995






Chapter IV

96

Table 43 Contd










Reddy et al 1992









Feng et al 2007

























40ordmC










Chapter IV

98









replicates

















43 Results





Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

90
































cultivars

Chapter IV

91



























21 17


19 23


18 19



21 24

Chapter IV

92







4231 Primer Design






















EST ID dbEST acc no





6220 6224

268



6306 6298

120


6205 6223

164


6147 6126

211


6192 6241

157


6190 6000

191



6276 6297

206



6243 6171

106

N23E3E4_A233 HS394047 Lipase 2e-20


6198 6186

158


6319 6289

155


6110 6108

180



6165 6169

206


6282 6328

203


6213 6204

209


6177 6172

184

Chapter IV

94

Table 42 Contd

EST ID dbEST acc no




6206 6250

235


6319 6289

162


6279 6202

170


6123 6176

217



6314 6313

217


6085 6098

179



6303 6305

222


6296 6362

159


6153 6194

299



6201 6060

280


6176 6156

196


6482 6403

220



6264 6222

169



6228 6198

153

Chapter IV

95









Walters et al 2003













Ecker 1995






Chapter IV

96

Table 43 Contd










Reddy et al 1992









Feng et al 2007

























40ordmC










Chapter IV

98









replicates

















43 Results





Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

91



























21 17


19 23


18 19



21 24

Chapter IV

92







4231 Primer Design






















EST ID dbEST acc no





6220 6224

268



6306 6298

120


6205 6223

164


6147 6126

211


6192 6241

157


6190 6000

191



6276 6297

206



6243 6171

106

N23E3E4_A233 HS394047 Lipase 2e-20


6198 6186

158


6319 6289

155


6110 6108

180



6165 6169

206


6282 6328

203


6213 6204

209


6177 6172

184

Chapter IV

94

Table 42 Contd

EST ID dbEST acc no




6206 6250

235


6319 6289

162


6279 6202

170


6123 6176

217



6314 6313

217


6085 6098

179



6303 6305

222


6296 6362

159


6153 6194

299



6201 6060

280


6176 6156

196


6482 6403

220



6264 6222

169



6228 6198

153

Chapter IV

95









Walters et al 2003













Ecker 1995






Chapter IV

96

Table 43 Contd










Reddy et al 1992









Feng et al 2007

























40ordmC










Chapter IV

98









replicates

















43 Results





Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

92







4231 Primer Design






















EST ID dbEST acc no





6220 6224

268



6306 6298

120


6205 6223

164


6147 6126

211


6192 6241

157


6190 6000

191



6276 6297

206



6243 6171

106

N23E3E4_A233 HS394047 Lipase 2e-20


6198 6186

158


6319 6289

155


6110 6108

180



6165 6169

206


6282 6328

203


6213 6204

209


6177 6172

184

Chapter IV

94

Table 42 Contd

EST ID dbEST acc no




6206 6250

235


6319 6289

162


6279 6202

170


6123 6176

217



6314 6313

217


6085 6098

179



6303 6305

222


6296 6362

159


6153 6194

299



6201 6060

280


6176 6156

196


6482 6403

220



6264 6222

169



6228 6198

153

Chapter IV

95









Walters et al 2003













Ecker 1995






Chapter IV

96

Table 43 Contd










Reddy et al 1992









Feng et al 2007

























40ordmC










Chapter IV

98









replicates

















43 Results





Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808





EST ID dbEST acc no





6220 6224

268



6306 6298

120


6205 6223

164


6147 6126

211


6192 6241

157


6190 6000

191



6276 6297

206



6243 6171

106

N23E3E4_A233 HS394047 Lipase 2e-20


6198 6186

158


6319 6289

155


6110 6108

180



6165 6169

206


6282 6328

203


6213 6204

209


6177 6172

184

Chapter IV

94

Table 42 Contd

EST ID dbEST acc no




6206 6250

235


6319 6289

162


6279 6202

170


6123 6176

217



6314 6313

217


6085 6098

179



6303 6305

222


6296 6362

159


6153 6194

299



6201 6060

280


6176 6156

196


6482 6403

220



6264 6222

169



6228 6198

153

Chapter IV

95









Walters et al 2003













Ecker 1995






Chapter IV

96

Table 43 Contd










Reddy et al 1992









Feng et al 2007

























40ordmC










Chapter IV

98









replicates

















43 Results





Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

94

Table 42 Contd

EST ID dbEST acc no




6206 6250

235


6319 6289

162


6279 6202

170


6123 6176

217



6314 6313

217


6085 6098

179



6303 6305

222


6296 6362

159


6153 6194

299



6201 6060

280


6176 6156

196


6482 6403

220



6264 6222

169



6228 6198

153

Chapter IV

95









Walters et al 2003













Ecker 1995






Chapter IV

96

Table 43 Contd










Reddy et al 1992









Feng et al 2007

























40ordmC










Chapter IV

98









replicates

















43 Results





Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

95









Walters et al 2003













Ecker 1995






Chapter IV

96

Table 43 Contd










Reddy et al 1992









Feng et al 2007

























40ordmC










Chapter IV

98









replicates

















43 Results





Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

96

Table 43 Contd










Reddy et al 1992









Feng et al 2007

























40ordmC










Chapter IV

98









replicates

















43 Results





Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808
























40ordmC










Chapter IV

98









replicates

















43 Results





Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

98









replicates

















43 Results





Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

99

















Appendix 41







TV23(BWS) 206E-01 216E-01 05655

S3A3(BWS) 847E-01 735E-01 21140

TV23(WS) 296E-01 193E-01 06406

S3A3(WS) 818E-01 339E-01 14118

TV23(AWS) 415E-02 791E-02 01536

S3A3(AWS) 963E-01 730E-01 22462

TV23C 100E+00 100E+00 26805

Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

100








D

0 50 100 150 200 250







Zinc finger

LEA

TRAPP complex




DNA J protein

clp G3PD


Hexose transporter

Lipase

APE1


Drm3-like protein

Dehydrin



ACC oxidase




Expression level

Gen

es

TV23 (AWS)

TV23 (WS)

TV23(BWS)

A

B

C


of drought

Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

101









A

B

C

0 20 40 60 80 100 120 140 160



DNA J protein






Dehydrin



Zinc f inger


LEA

TRAPP complex



ACC oxidase





Hexose transporter

Lipase



APE1

Drm3-like protein

Clp G3PD

Expression level

Gen

es

S3A3 (AWS)

S3A3(WS)

S3A3(BWS)


Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

102


































Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

103

0 50 100 150 200

DNA J protein



Hexose transporter

Lipase

APE1


Drm3-like protein

Clp G3PD

Expression level

Ge

ne

s

TV23C

S3A3(BWS)

TV23(BWS)



0 10 20 30 40 50 60 70 80 90

Dehydrin



ACC oxidase



Expression level

Ge

ne

s

TV23C

S3A3(WS)

TV23(WS)



Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

104

0 20 40 60 80 100 120 140 160 180





Dehydrin



Zinc finger



LEA

TRAPP complex




DNA J protein

ACC oxidase

Clp G3PD





Hexose transporter

Lipase

APE1


Drm3-like protein

Expression level

Ge

ne

s

S3A3(BWS)

TV23(BWS)


0 100 200 300








LEA

TRAPP complex




Expression level

Gen

es

TV23C

S3A3(AWS)

TV23(AWS)



Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

105


































Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





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Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

106














LEA

Zinc finger



Dehydrin






TRAPP complex

Clp G3PD

APE1

ACC oxidase


Hexose transporter




DNA J protein

Lipase



Drm3 like protein

A B

I

II

III

IV

Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

107













44 Discussion




















Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

108


































Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

109


drought

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

160

180

BWS WS AWS

Rel

ativ

e E

xp

ress

ion


protein1

TV23S3A3

0

50

100

150

200

250

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Zinc finger protein

TV23S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xp

ress

ion

LEA

TV23S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

TRAPP complex

TV23

S3A3

0

10

20

30

40

50

60

70

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

20

40

60

80

100

120

140

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3Ahellip

Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

110

0

2

4

6

8

10

12

14

16

18

20

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n

Ethylene-responsive


TV23

S3A30

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

DNA J protein

TV23

S3A3

0

200

400

600

800

1000

1200

BWS WS AWS

Re

lati

ve

Ex

pre

ssio

n


TV23

S3A3

0

5

10

15

20

25

30

35

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

200

400

600

800

1000

1200

1400

1600

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

Dehydrin

TV23

S3A3

0

10

20

30

40

50

60

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23

S3A3

0

10

20

30

40

50

60

70

80

90

BWS WS AWS

Re

lati

ve E

xpre

ssio

n

ACC oxidase

TV23

S3A3

0

5

10

15

20

25

30

35

40

45

50

BWS WS AWS

Re

lati

ve E

xpre

ssio

n


TV23S3A3

0

2

4

6

8

10

12

14

BWS WS AWS

Re

lati

ve

Exp

ress

ion

Clp G3PD

TV23

S3A3


drought

Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

111

































Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

112

































Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

113





















tolerant cultivar













Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

114
















compared to S3A3


















Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

115



































Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

116


































Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

117























drought tolerance










Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

118











at WS






















Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

119

































Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

120



































Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

121


































Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

122



































Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

123










45 Conclusion























Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

124




























References



Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

125







447




ndash 1099












ndash 399











Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

126



Sci 6 284 ndash286










491 ndash 543



29















2017 ndash 2026



Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

127











ndash 225














643



ndash 6






Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

128


Chem 275 14494 ndash 14500

































Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

129






786 ndash 790








475 ndash 486











349 ndash 358









Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

130




























87 55 ndash 84







Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

131












1321 ndash 1332









403













Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

132







(Oxf) 187 169 ndash 176



129 1616 ndash 1626





103 230 ndash 235





ndash 209













Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

133









J 31 13 ndash 24




277(28) 25062 ndash 25069





10 200


21 235 ndash 270














Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

134













11369





















Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

135





For 9(3) 197 ndash 204












Bot 62(3) 1229 ndash 1239





ndash 410








249 ndash 279




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

136












Argentina




















17(9) 344 ndash 345



Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

137



ndash 411








19631 ndash 19636





















327 ndash 334

Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

138


















Phytol 126 419-448






26(4) 380 ndash 385











Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

139



























16113 ndash 16118








Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

140





197 306 ndash 312















34 45 ndash 55









166





Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV

141







836 ndash 846




209 ndash 216






611 ndash 616








17803 ndash 17808




Chapter IV - Shodhgangashodhganga.inflibnet.ac.in/bitstream/10603/5496/9/09...Chapter IV 91 4.2...

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Transcript of Chapter IV - Shodhgangashodhganga.inflibnet.ac.in/bitstream/10603/5496/9/09...Chapter IV 91 4.2...