Abiotic Stress and Plant Response

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Abiotic stress and plant responses from the whole vine

to the genes_058 86..93

G.R. CRAMER

Department of Biochemistry and Molecular Biology, University of Nevada, Reno, NV 89557, USA

Corresponding author: Dr Grant R. Cramer, fax +1 775 784 1650, email [email protected]

Abstract

Drought, salinity and extreme temperatures significantly limit the distribution of grapes around the

world. In this review, the literature of grape responses to abiotic stress with particular reference to whole

plant and molecular responses observed in recent studies is discussed. A number of short-term and

long-term studies on grapevine shoots and berries have been conducted using a systems biology

approach. Transcripts, proteins and metabolites were profiled. Water deficit, salinity and chilling altered

the steady-state abundance of a large number of transcripts. Common responses to these stresses includedchanges in hormone metabolism, particularly abscisic acid (ABA), photosynthesis, growth, transcription,

protein synthesis, signalling and cellular defences. Some of the transcriptional changes induced by stress

were confirmed by proteomic and metabolomic analyses. More than 2000 genes were identified whose

transcript abundance was altered by both water deficit and ABA. Different gene sets were used to map

molecular pathways regulated by ABA, water deficit, salinity and chilling in grapevine. This work

supports the hypothesis that ABA is a central regulator of abiotic stress tolerance mechanisms. ABA

affects signalling pathways that trigger important molecular activities involving metabolism, transcrip-

tion, protein synthesis, and cellular defence and also regulates important physiological responses such as

stomatal conductance, photoprotection and growth. Systems biology approaches are providing more

comprehensive understanding of the complex plant responses to abiotic stress. The molecular sets

generated from mapping the ABA-inducible stress responses provide numerous targets for genetic and

cultural manipulation for improved plant protection and grape quality.

Keywords: abscisic acid, salinity, water deficit

Introduction

Levitt (1980) defined stress as an environmental factor

that is potentially unfavourable to an organism and resis-

tance as the ability of the organism to survive such an

environmental factor. Survival is an important issue, but

when one considers crops, one is also concerned about

production of the harvestable portion of the plant. So

there are at least two variables that one must differentiate

when one is discussing stress resistance or tolerance. The

mechanisms for these types of resistances can be quite

different.

Common environmental factors or abiotic stresses

around the world include, drought (water deficit), salin-

ity, temperature, and acid soils (Tester and Bacic 2005).

Rarely is there a single abiotic stress affecting a plant;

almost always there are interacting factors. For example,

plants exposed to the direct sun in the field will most

likely be exposed to levels of light that can cause photo-

inhibition. If the plant is sufficiently stressed that its pho-

tosynthesis is reduced, then the plant will almost

certainly be experiencing the secondary effects of photo-inhibition as well as the effects of the primary stress (e.g.

drought, salinity, high or low temperatures). Thus, while

we may study such stresses in the lab in an artificial

environment and elucidate important mechanisms of

resistance, one must consider the whole environment

that a plant grows in to fully comprehend the stress

resistance mechanisms needed in the field.

A common response to many stresses (e.g. water

deficit, salinity, high and low temperatures, herbivory,

and pathogens) is an osmotic response (Tester and Bacic

2005, Lopez et al. 2008). Either the whole plant experi-

ences osmotic stress or specific cells under attack expe-

rience the osmotic stress. The osmotic stress is caused by

water loss, which is a daily problem for any land plant

that is photosynthesising and transpiring.

There are many abiotic stresses that significantly limit

the distribution of grapes around the world. These

stresses reduce crop yields, but only water deficit has been

used in a positive way to enhance flavour and quality

characteristics of the berries (Roby et al. 2004, Chapman

et al. 2005). In part, this effect is because of reduced shoot

vigour and competition for carbon resources (a change

in source to sink relationship). Berry size can also be

reduced, concentrating flavours and colour by increasingthe skin surface: berry mass ratio (the skin being a sig-

nificant tissue for producing flavours, tannins and

colour). In addition, there are fundamental biochemical

86 Abiotic stress and plant responses Australian Journal of Grape and Wine Research 16, 8693, 2010

doi: 10.1111/j.1755-0238.2009.00058.x

2009 Australian Society of Viticulture and Oenology Inc.


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changes in berries under water deficit that cause impor-

tant metabolic changes that influence berry flavour and

quality (Castellarin et al. 2007a, Deluc et al. 2009).

Semi-arid regions where grapes are grown not only

suffer from water deficit but also are prone to salinisation.

Extreme cold can limit distribution in the northern- and

southern-most latitudes and at high elevation. In addi-tion, frost and chilling damage occurs during the spring

and can limit yields in more moderate climates. Heat and

light stress often interact with water deficit to accelerate

water loss and plant strain.

This paper reviews the literature of grape responses to

abiotic stress with particular emphasis on whole plant

and molecular responses observed in investigations per-

formed in the authors research group. It will not be

possible to discuss all abiotic stresses in this review. Our

group has focused on the effects of water deficit, salinity

and chilling stress on grapes. Experiments were con-

ducted at many levels: in the field (water deficit only),

greenhouse and the laboratory, and at the whole plant(growth and photosynthetic processes) and molecular

levels (transcripts, proteins and metabolites). A systems

biology approach was used to integrate the data in an

attempt to better understand the plants responses to

abiotic stresses (Cramer et al. 2005). This review is

divided into two sections: the first part will discuss the

effects of stress on growing shoot tips and a smaller

second part will address briefly the effect of water deficit

on berry metabolism.

Shoot responses to water deficit, salinity

and chillingVitis vinifera grapevines grow well in arid and semi-arid

environments. V. vinifera has relatively high drought

tolerance (McKersie and Leshem 1994, Grimplet et al.

2007a) and genetic variability within the species exists

(Gaudillere et al. 2002). Once established in a deep soil

with adequate water retention characteristics, grapevines

produce root systems many meters deep enabling the vines

to survive severe water deficits. However, global warming

is causing significant climate change, making it necessary

for future crop adaptation (Howden et al. 2007). Increased

temperatures along with changing rainfall will require

improved crop performance under water deficits.

Abscisic acid (ABA) is a hormone that is central to

regulating the plant response to osmotic stress (Seki et al.

2007). It operates at the whole plant level regulating

important processes such as shoot and root growth,

thereby affecting the root to shoot ratio (Munns and

Cramer 1996). It also regulates transpiration and water

loss via stomatal closure. ABA also operates at the

molecular level by regulating gene transcription, protein

synthesis, signalling pathways, ion transport (and the

transport of other organic molecules) and the production

of important protectants against dehydration and photo-

inhibition (Yang et al. 2006, Seki et al. 2007).

ABA in the xylem sap is a key root to shoot signal indrought-stressed plants (Schachtman and Goodger 2008).

Other chemical signals that may influence stomatal con-

ductance during water deficits include malate, protons,

cytokinins, and ABA conjugates (Schachtman and

Goodger 2008). Peptides and proteins may also act as

signals in the xylem (Aki et al. 2008). Metabolites, pep-

tides and proteins have been analysed in the xylem sap of

maize (Alvarez et al. 2008) and while protein concentra-

tions are affected by water deficit, ABA was considered

the most important signal in the xylem sap that regulatestranspiration. Small RNAs (e.g. mRNAs) play significant

roles in abiotic stress resistance (Sunkar et al. 2007, Li

et al. 2008), including drought resistance (Li et al. 2008).

However, the role of peptides, proteins and mRNAs in the

xylem sap remains unclear and requires further investi-

gation (Schachtman and Goodger 2008).

It is a very interesting coincidence that cells closely

associated with the xylem (e.g. xylem parenchyma) are

located where hydraulic conductance to the epidermis

(expanding cells) is most restricted for water flow and

growth (Tang and Boyer 2002), where the expression of

nine-cis-epoxycarotenoid dioxygenase (NCED), the rate

limiting enzyme for ABA biosynthesis, first increases inresponse to water deficit (Endo et al. 2008), and where

the aquaporins VvPIP1;1 and VvPIP2;2 are most abun-

dantly expressed in grapevine roots (Vandeleur et al.

2008). It is intriguing that VvPIP1;1 expression appears to

be quite responsive to changes in water potentials influ-

encing cell and root hydraulic conductivity (Vandeleur

et al. 2008).

There is a wide range of water-use efficiencies

between grape cultivars (Bota et al. 2001). These differ-

ences are largely attributed to variation in stomatal con-

ductance in response to water deficits (Bota et al. 2001,

Schultz 2003, Soar et al. 2006), but also can be related todifferences in the change of root hydraulic conductance

and aquaporin (water channel) expression in response to

water deficit (Vandeleur et al. 2008). Water deficit

increases ABA concentrations in the xylem sap and leaves

of grapevine and changes in stomatal conductance are

well correlated with ABA concentrations of the xylem sap

(Okamoto et al. 2004, Soar et al. 2004, Pou et al. 2008).

ABA also influences hydraulic conductance (Hose et al.

2000), aquaporin gene expression (Tyerman et al. 2002,

Kaldenhoff et al. 2008) and embolism repair (Lovisolo

et al. 2008) in grapevines. Furthermore, there is signifi-

cant variation in ABA concentrations between rootstocks

originating from different Vitis species and which have an

influence on scion (V. vinifera L. cv. Shiraz) photosynthe-

sis and stomatal conductance (Soar et al. 2006). Thus, it

appears that ABA plays a vital role in grapevine water

relations during osmotic stress.

In addition to reductions in stomatal conductance,

water deficit, salinity and low temperatures can reduce

photosynthesis by photoinhibition of photosystem II (Lia-

kopoulos et al. 2006, Cramer et al. 2007, Tattersall et al.

2007). In grapevines, the main protective mechanisms

for photoinhibition appear to be related to non-

photochemical quenching involving xanthophylls and

the D1 repair rate (Hendrickson et al. 2004). In addition,photorespiration was able to dissipate up to 20% of the

energy flux contributing to electron transport. In Arabi-

dopsis, ABA can protect the plant against photoinhibition

Cramer Abiotic stress and plant responses 87



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by affecting fibrillin concentrations (Yang et al. 2006).

Fibrillins are lipid-binding proteins that enhance photo-

system II tolerance to light stress.

The ABA metabolic pathway is displayed in Figure 1.

NCED (nine-cis-epoxycarotenoid dioxygenase) repre-

sents the first committed step in ABA biosynthesis

(Schwartz et al. 2003) and its transcript abundance is

often correlated with ABA concentrations. In grapevine,

stomatal conductance is negatively correlated with ABA

concentrations in the xylem sap and ABA concentrations

in the leaves are correlated with the transcript abundance

of VvNCED1 (Soar et al. 2004).

In one study (access to the original data can be found at

http://www.plexdb.org in the experiment entitled VV2),

the impacts of long-term water deficit and salinity on

grapevine shoot tips was investigated (Cramer et al. 2007).

A unique stress experiment was performed with Cabernet

Sauvignon comparing the gradual effects of a decreasing

water deficit with equivalent salinity. Transcript and

metabolite profiling provided consistent and corroborativeresults with growth assays. At equivalent water potentials,

water deficit had a more severe effect than salinity on

growth, gene expression and specific metabolites.

Stress broadly affected many gene transcripts involved

with metabolism, protein fate, transport, transcription,

cellular defence and communication/signalling (Cramer

et al. 2007). Gene expression of the ABA and ethylene

pathways was particularly increased by stress compared

with other hormone pathways and was negatively corre-

lated with stem water potentials. Energy metabolism was

strongly increased by stress. There were higher concen-

trations of glucose, malate and proline in water deficit-

treated plants as compared with salinised plants. These

differences were linked to differences in gene expression

and other metabolites of the photosynthetic, gluconeo-

genic and photorespiratory pathways. Not only are these

solutes likely to aid plants in osmotic adjustment, but also

may help plants cope with reactive oxygen species detoxi-

fication and photoinhibition.

The simultaneous monitoring of the abundance of

both transcripts and metabolites has reinforced the obser-

vations made with each metric alone. These observations

demonstrate the information synergy derived from theuse of integrative functional genomic approaches. The

changes in the individual transcript abundance of many

genes were similar to changes in our short-term study

Figure 1. The ABA metabolicpathway. BHASE, beta-carotenehydroxylase; ABA1, zeaxanthinepoxidase; NPQ1, violaxanthinde-epoxidase; CCD1,(9,10[9,10]carotenoidcleavage dioxygenase); NCED,nine-cis-epoxycarotenoiddioxygenase; ABA2, xanthoxindehydrogenase; ABAHASE,(+)-abscisic acid8-hydroxylase; UGT,

UDP-glucoseglucosyltransferase; BGL1,b-glucosidase. Shaded boxesrepresent different cellularcompartments and have acompartment name inside ofthem, except for the grey boxaround BGL1, which representsendomembranes just as forABAHASE.

-Carotene

BHASEChloroplast

C13-norisoprenoid

&-Cryptoxanthin

Zeaxanthin

BHASE

CCD1

C14

-dialdehyde

Antheraxanthin

ABA1

ABA1

NPQ1

High

Light

Low

LightnthophyllCycle

nthophyllCycle

Violaxanthin

9 cis neoxanthin

NPQ1g

Xan

Xan

9-cis-neoxanthin

Dihydroxy

phaseic acid

NCED

Cytoplasm

ABA-aldehyde

Phaseic acidABA2

AAO3

Abscisic acid8-Hydroxy abscisic

acidABAHASE

EndomembranesABA-GE

UGT

BGL1

Xanthoxin




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(Tattersall et al. 2007); the original data can be found in

the experiment entitled VV1 in http://www.plexdb.org),

however, there were indications that a larger and more

complex response in the acclimation process occurred

with a gradual long-term stress. In another study (Vincent

et al. 2007), proteomic analysis indicated that the

decrease in growth and stem water potentials of the vinewas correlated with decreased amounts of proteins

involved in photosynthesis and protein synthesis.

Common transcript responses in shoot tips to

drought, salinity and chilling (VV1 and VV2)

In the short-term stress experiment (Tattersall et al. 2007),

we found that grapevine shoot tips had distinct differences

in response to chilling as compared with water deficit or

salinity. Nevertheless, there were also some common

attributes for all three stresses. Microarray data sets for

both the long-term (water deficit and salinity, VV2) and

short-term (water deficit, salinity and chilling, VV1)

experiments were surveyed to determine the set of tran-scripts in shoot tips that had common responses to these

stresses (Table 1). There were 15 transcripts that were

positively up-regulated in all conditions. Two of them were

related to the biosynthesis of sugars (raffinose and treha-

lose), which putatively contribute to improved tolerance

to osmotic stress (Taji et al. 2002, Downie et al. 2003).

About half (8 of 15) were transcription factors and the rest

were involved in signalling or stress responses (ABA, Ca 2+

or H2O2). The catalase was up-regulated only after severe

stress over time. The 9-cis-epoxycarotenoid dioxygenase 1

is NCED1, the rate-limiting step in ABA biosynthesis. The

proteins in Table 1 most likely operate together in acommon stress response pathway in grapevine.

Particularly interesting was the up-regulation of a

serine/threonine protein kinase that has high homology

with ATSR1 in Arabidopsis (Table 1). ATSR1 influences the

TOR gene in yeast, the central regulating kinase that

stimulates and integrates metabolism for growth (Baena-

Gonzalez and Sheen 2008). Two transcription factors are

worth noting. ATHB-12 in Arabidopsis is a homeodomain

leucine zipper class I protein that responds to ABA

through ABI1, a protein phosphatase. One of the func-tions of ATHB-12 is to inhibit growth (Olsson et al. 2004,

Moes et al. 2008). The bZIP (basic-leucine zipper) tran-

scription factor listed in Table 1 is very similar to a family

of ABF transcription factors in Arabidopsis (Choi et al.

2000). They respond to ABA and bind to the abscisic acid

responsive element motif in the promoter region of ABA-

inducible genes. Many of the genes are involved in stress

tolerance mechanisms.

It is apparent from the data in Table 1 and the previous

discussion of the literature that ABA plays a central role in

the response of grapevine to osmotic stress. Data from two

recent studies in Arabidopsis were utilised (Huang et al.

2007, Matsui et al. 2008) in order to gain a wider viewof ABAs role during osmotic stress. Both studies deter-

mined the response of the Arabidopsis transcriptome to

ABA treatment using two different technologies and

approaches. The set of transcripts affected significantly

by ABA from each experiment were downloaded and their

sequences BLASTed against the grape transcript sequences

that were affected significantly by water deficit. Note that

this set of genes is a subset of all the water deficit-

responsive genes, indicating that there are factors other

than ABA that contribute to water deficit responses as

well. The assumption made here is that these transcripts in

this subset are also affected by ABA (this hypothesisremains to be validated by grapevine experiments with

ABA applications or ABA mutants). The corresponding

grape gene sets were placed into functional categories and

Table 1. Common responses for all stresses and both experiments (VV1 and VV2).

Affy probe Set ID Gene Protein MIPS 2.1 functional category

1608995_at GSVIVG00025569001 Raffinose synthase 01.05 C-metabolism

1617699_at GSVIVG00010921001 Trehalose phosphatase 01.05 C-metabolism

1609107_at GSVIVG00014947001 C2H2 zinc finger 11.02.03.04.01 transcription activation

1609172_at GSVIVG00027622001 NAC domain protein 11.02.03.04.01 transcription activation1610064_at GSVIVG00023994001 WRKY DNA-binding protein 11.02.03.04.01 transcription activation

1617931_at GSVIVG00000517001 ATHB-12 11.02.03.04.01 transcription activation

1618998_at GSVIVG00030292001 RAV transcription factor 11.02.03.04.01 transcription activation

1619029_at GSVIVG00036604001 bZIP transcription factor 11.02.03.04.01 transcription activation

1620621_at GSVIVG00015416001 NAC domain protein 11.02.03.04.01 transcription activation

1614779_s_at GSVIVG00025566001 Remorin-like protein 16.03.01 DNA binding

1620564_at GSVIVG00023957001 Serine/threonine protein kinase 30.01.05.01 protein kinase cascades

1608587_at GSVIVG00036780001 Calcium dependent protein kinase 30.01.09.03 Ca2+ mediated signal transduction

1610871_s_at GSVIVG00002880001 Catalase 2 32.07.07.01 catalase reaction

1608022_at GSVIVG00000988001 9-cis-epoxycarotenoid dioxygenase 1 36.20.18.05 abscisic acid response

1614892_at No hit Protein phosphatase 2C ABI1 (PP2C) 36.20.18.05 abscisic acid response

Gene names were obtained from a BLAST of the probe set sequence against the 8x assembly of the grape genome. These data were collected from the grape annotation

page of http://www.plexdb.org.

The probe set identification number for the Affymetrix grape genome array (version 1.0).




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these grape gene sets separated into those that had an

increased or decreased abundance during water deficit.

The results from this analysis (Figure 2) are quite

intriguing. Although the data sets are represented by quite

large differences in the numberof transcripts (2095 vs 644)

that are matched because of the different technologies

used (tiling vs genome array, respectively), there are some

interesting commonalities that stand out. For example, the

transcript abundance was decreased for nearly all of the

grapevine transcripts affected by water deficit that are

involved in protein synthesis. The differences in the %

values between these two studies (5 vs 30% in Figure 2)

was affected largely by the number of total transcripts

involved in either set (2095 vs 644). The large decrease in

protein synthesis transcripts was a rather striking result

and is consistent with our earlier proteomic study (Vincent

et al. 2007), in which a decrease in proteins involved in

protein synthesis was highly correlated with the inhibition

of growth. It is not clear whether the inhibition of protein

synthesis by ABA is the cause of the growth reduction or

protein synthesis is merely responding to feedback regu-

lation in response to inhibited growth (less demand or

need for proteins). This hypothesis requires further testing

before any firm conclusions can be made.In both transcript sets from the two technologies

(Figure 2), there was an increased proportion of tran-

scripts with increased abundance that were involved in

transcription and cell rescue, defence and virulence.

This is consistent with ABA playing a role in stress toler-

ance. The increased down-regulation of genes involved in

biogenesis of the cell is consistent with the action of ABA

and ATHB-12 described previously, however, these pro-

portional differences are rather small. This data analysis

serves only in the development of new hypotheses in

order to stimulate new ideas and areas of research.

Caution should be used in the interpretation of these

results until additional experiments are performed. Other

differences between other functional categories were not

consistent between the studies and definitely require

further testing.

Transcript responding to osmotic stress only

There are several transcripts that responded in all osmotic

stress treatments, but not in chilling. They include NF-YA

TF (1613912_at), a transcription factor responsive to ABA

that affects approximately 130 Arabidopsis genes (Li et al.

2008). The transcript abundance of this transcription

factor is regulated by a mRNA (miR169) that is affected

by osmotic stress. Other transcription factors affected by

osmotic stress include a member of the DREB family

(1608315_at), which bind to a drought-responsiveelement in the promoter of drought-induced genes (Liu

et al. 1998), ERF1 (1609559_at), a member of a family of

ethylene response factors that bind to a cis-element in

Figure 2. Functional categorisation of a subset of grape transcripts responsive to water deficit and also linked with Arabidopsisgenes thatare affected by ABA. This subset grape transcripts is referred to as WD-ABA transcripts. ABA responsive genes in Arabidopsiswere identifiedusing the Arabidopsistiling array data set from Matsui et al. (2008) and the Arabidopsisgenome array data set from Huang et al. (2007). Theinitial subset of genes in grapes responsive to water deficit and which were also identified as ABA-responsive genes in Arabidopsis werefunctionally categorised (black bars) and then separated into those genes with increased (grey bars) or decreased (white bars) transcriptabundance during water deficit. Each subset is displayed separately with the subset of 2095 grape transcripts based on the tiling array dataon the left and the subset of 644 grape transcripts based upon the genome array data on the right. Functional categories are organised alongthe y-axis from the highest to the lowest number of transcripts in a functional category (bottom to top). The % of transcript set refers to the% of total transcripts in that particular set of transcripts that were placed into that functional category. The tiling array data are from Matsuiet al. (2008) and the genome array data are from Huang et al. (2007).




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ethylene inducible genes (Hao et al. 1998), and KNAT3

(1615625_at), a homeodomain protein involved in regu-

lating cell patterning and organ development (Truernit

et al. 2006). In addition, osmotic stress affected CRY2

(1610235_at), a blue-light receptor that is a positive

regulator of flowering (Liu et al. 2008), thioredoxin

(1615681_at), a chloroplast protein involved in redoxregulation of photosynthesis and stress responses (Foyer

et al. 2009), LTP (1616990_s_at), a lipid transfer protein

that may be involved in signalling, and a flavonol-o-

glucosyl transferase (1618155_at).

Berry responses to water deficit

Using transcript and metabolite profiling, our group

showed that water deficit had significant impacts on the

metabolism of grape berries (Deluc et al. 2009). Metabolic

responses appear to be dependent on the cultivar and

the colour of the grape. Water deficit particularly affected

ABA, carotenoid, amino acid, fatty acid and phenylpro-

panoid metabolism in two cultivars, Cabernet Sauvignonand Chardonnay, but in different ways. Water deficit

increased ABA concentrations in Cabernet Sauvignon

berries, but not Chardonnay berries. ABA is known to

enhance proline, sugar and anthocyanin accumulation in

plants and the increased ABA concentration in Cabernet

Sauvignon by water deficit was consistent with this

hypothesis resulting in increased accumulation of proline,

sugar and anthocyanins relative to well-watered controls.

In Chardonnay, water deficit did not increase ABA con-

centration above that of well-watered berries. Likewise,

sugar and proline concentration were not significantly

different from the well-watered controls. The responseof anthocyanins to water deficit was unaffected and is

irrelevant because Chardonnay berries cannot produce

anthocyanins. Water deficit increased the transcript

abundance of genes (lipoxygenase and hydroperoxide

lyase) involved in fatty metabolism, a pathway known to

affect berry and wine aromas. In Chardonnay, water

deficit activated parts of the phenylpropanoid, energy,

carotenoid and isoprenoid metabolic pathways that con-

tributed to increased concentrations of antheraxanthin

and flavonols. The effects of water deficit on metabolism

had important impacts on berry constituents that influ-

enced flavour and quality characteristics in both grapes

and wine and might contribute to increased antioxidants

and human-health benefits.

Regulated-deficit irrigation has been shown by several

research groups to improve berry and wine quality (Roby

et al. 2004, Chapman et al. 2005, Castellarin et al.

2007a,b, Deluc et al. 2009). Application of water deficit

early in the season before vraison resulted in greater

concentrations of anthocyanins and phenolics (Matthews

and Anderson 1988, Matthews et al. 1990). Colour dif-

ferences were the result of increased anthocyanin synthe-

sis caused by water deficit applied either early or late in

the season (Matthews and Anderson 1988, Castellarin

et al. 2007a, Deluc et al. 2009). It was suggested that bothABA and sugar signalling might affect accelerated antho-

cyanin development. Additions of ABA and rhamnose to

grape berry skins have induced anthocyanin biosynthesis

in a synergistic manner (Hiratsuka et al. 2001) and ABA

applications to Crimson Seedless grapes increased their

red colour and the transcript abundance of a UDP-

glucose: flavonoid 3-O-glucosyltransferase (UFGT) gene,

the rate limiting step in anthocyanin biosynthesis (Peppi

et al. 2008). The increase of anthocyanin concentration is

a common response in stressed plants and probably func-tions to provide greater photoprotection (Merzlyak et al.

2008a,b). In red wines, of course, it serves to provide

deeper and darker colour, which is often perceived as an

indicator of quality by the consumer.

A berry tissue analysis using global gene expression

techniques indicated that water deficit affected the mRNA

abundance of 13% of genes at grape maturity within the

three tissues of the berry (skin, pulp and seeds), with the

greatest changes located in the pulp and skin (Grimplet

et al. 2007b). While the function of many of the genes

differentially expressed within the seed and pulp remain

to be elucidated, other genes over-represented in the skin

were clearly associated with phenylpropanoid metabo-lism, ethylene, pathogenesis-related responses, energy

metabolism and stress responses.

Proteomic and metabolomic analyses of the berry

tissues indicated significant changes because of water

deficit (Grimplet et al. 2009). Water deficit altered the

abundance of approximately 7% of proteins in the peri-

carp, but had little effect on the expression of proteins in

the seeds. Comparison of the protein and transcript

expression profiles indicated that 32% of the pericarp and

69% of the seed proteins had similar quantitative expres-

sion patterns. This indicated that proteins might be influ-

enced by post-transcriptional processes. There wereincreases in the relative abundance of proteolytic enzymes

in the skin, and the enzyme, cytosolic ascorbate peroxi-

dase. There were increases in isoflavone reductase,

glutamate decarboxylase andan endochitinase in the pulp.

About half of the 32 metabolites measured had differences

in abundance between tissues with water deficit, including

some sugars, amino acids and organic acids.

Conclusion

Osmotic stress is a common feature of many abiotic stresses

that afflict grapevines. A systems biology approach has

enabled a broad picture of grapevine responses to abiotic

stress to be drawn. By comparing and contrasting

responses of grapevine to different stress treat-

ments, common and distinct stress-response pathways in

grapevine were elucidated. Many of the grapevine

responses to osmotic stress appear to be transcriptionally

regulated, but proteomic studies indicate that there are

post-translational controls as well. ABA acts as a central

regulator of many osmotic responses in grapevines.

NCED1 appears to be the rate-limiting step for the bulk of

ABA biosynthesis, because it is highly correlated with

ABA concentrations in both shoots and berries. ABA acts

to reduce water loss and increase stress tolerance in grape-

vines. It reduces stomatal conductance and limits leaf areaexpansion. It affects the abundance of many transcripts

and proteins involved in many different plant processes

including osmo- and photoprotection, photosynthesis,




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and growth. One of the most striking features from these

analyses is the potential inhibitory effect of ABA on the

transcript abundance of nearly all genes involved in

protein synthesis that respond to water deficit. Modern

biology is going through a revolution and grapevine

research is no exception. It is expected that we

will make much more rapid progress in the developmentof stress tolerance genotypes in grapevines in the near

future, because there are large genetic resources

for grapevine and there are a large number of high-

throughput genomic tools available to conduct functional

genomic analyses.

Acknowledgements

I would like to thank all my grapevine colleagues who

have contributed to such an interesting field of science. I

would especially like to thank my lab personnel and

colleagues at UNR for their valuable help in the work cited

in this review. I would also like to acknowledge that much

of the research from my lab that was cited in this review

was supported by a grant (DBI-0217653) from the

National Science Foundation (NSF) Plant Genome

program.

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Manuscript received: 3 February 2009Revised manuscript received: 20 April 2009

Accepted: 17 May 2009



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Abiotic Stress and Plant Response