ORIGINAL PAPER

The expression pattern of b-glucosidase genes (VvBGs)during grape berry maturation and dehydration stress

Guojun Zhang • Chaorui Duan • Ya Wang • Yanping Wang •

Kai Ji • Haiying Xu • Shengjie Dai • Qian Li • Pei Chen •

Yufei Sun • Yan Wu • Hao Luo • Ping Leng

Received: 20 September 2012 / Accepted: 21 December 2012 / Published online: 28 December 2012

� Springer Science+Business Media Dordrecht 2012

Abstract In order to understand more about the role of

b-glucosidase gene expression in modulating ABA level,

the expression pattern of three cDNAs (VvBG1, VvBG2 and

VvBG3) which encode b-glucosidase in ripening grape

berries was analyzed in the presence or absence of dehy-

dration stress. The results show that expression of these

three VvBG genes was markedly different. Expression of

VvBG1 and VvBG2 increased rapidly from veraison to

reach a maximum at harvest or several days immediately

before harvest, and coincident with ABA accumulation

during berry development and ripening. However, expres-

sion of VvBG3 differed from VvBG1 and VvBG2 in that

transcript levels declined from the early young fruit stage

through veraison after which there was no further change.

At 10 days before harvest, dehydration treatment of

detached grape berries up-regulated the expression of

VvBG1 and enhanced ABA accumulation whereas the

expression of VvBG2 was down-regulated, VvBG3 was

unaffected by dehydration stress. However, in the leaves,

dehydration treatment up-regulated the expression of

VvBG1 and stimulated the accumulation of ABA but down-

regulated expression of VvBG2 and VvBG3. Based on the

results obtained, it is concluded that the expression pattern

of the three VvBGs is both temporal and tissue specific.

Furthermore, expression of the VvBGs might play a role in

the regulation of ABA content during berry ripening and in

the response of berries to dehydration stress.

Keywords Grape � ABA � VvBGs � Berry ripening �Dehydration

Introduction

Fruit ripening is a complex physiological and biochemical

process regulated by genes and is also influenced by the

environment (Giovannoni 2001). To date, our understanding

of the regulatory and signalling pathways of climacteric fruit

ripening at the molecular level has increased substantially

(Yen et al. 1995; Zegzouti et al. 1999; Alexander and Grierson

2002; Vrebalov et al. 2002; Tieman et al. 2000, 2001; Kevany

et al. 2007; Itkin et al. 2009). However, little is known about

the molecular mechanisms regulating non-climacteric fruit

ripening. Generally speaking, non-climacteric fruits do not

display either an ethylene or respiratory climacteric, and for

many, ripening is regulated by ABA. NCED is a key gene in

the biosynthesis pathway of ABA (Iuchi et al. 2001; Chernys

and Zeevaart 2007; Huang et al. 2001). Even so, Wheeler et al.

(2009) showed that expression of NCED is not consistent with

the level of ABA, suggesting that a more complex regulation

mechanism is involved in mediating accumulation of ABA

which includes both synthesis and catabolism (Nambara and

Marion-poll 2005; Seo and Koshiba 2002;Taylor et al. 2000;

Ren et al. 2010, 2011;Sun et al. 2011, 2012). In general, active

ABA is rapidly catabolized to inactive structures in higher

plants through two main routes (Zeevaart and Creelman 1998,

Zeevaart 1999; Barthe et al. 2000) i.e. hydroxylation to pha-

seic and dihydrophaseic acid; and conjugation to glucose

esters. The former involves conversion of ABA to 80-hydroxy

ABA (HOABA) while in the latter is ABA is glucosylated to

G. Zhang � C. Duan � Y. Wang � Y. Wang � K. Ji � S. Dai �Q. Li � P. Chen � Y. Sun � Y. Wu � H. Luo � P. Leng (&)

College of Agronomy and Biotechnology,

China Agricultural University, Beijing 100193, China

e-mail: pleng@cau.edu.cn

G. Zhang � H. Xu

Research Institute of Pomology, Forestry Academy of Sciences,

Beijing 100093, China

123

Plant Growth Regul (2013) 70:105–114

DOI 10.1007/s10725-012-9782-3

either ABA-glucosyl ester (–GE) or ABA-glucosyl ether

(-GS).

Glucosyltransferases (GTase) are thought to play an

important role in the biosynthesis of many plant secondary

metabolites including plant hormones like ABA (Cowan

2001; Oritani and Kiyota 2003; Lee et al. 2006) and do so by

transferring nucleoside diphosphate-activated sugars to recep-

tors of low molecular weight substrates. ABA-GE could be

synthesized from ABA and UDP-D-Glc (UDPG) by a GTase

(Lehmann and Schutte 1980; Schwarzkopf and Miersch 1992;

Lee et al. 2006). ABA-GE plays an important role in the

regulation of ABA (Xu et al. 2002; Li et al. 2012; Sun et al.

2010). The b-glucosidases could catalyze the the release of

ABA-GE back into active ABA to rapidly adjust ABA level

(Lee et al. 2006). Thus, b-glucosidases have been implicated

in key developmental processes, such as growth, pathogen

defense, and hormone hydrolysis (Esen 1993; Kleczkowski

et al. 1995). Recently, considerable progress has been made in

elucidating the functions of b-glucosidases for activation of

plant hormone groups, including ABA (Lee et al. 2006).

In this study, the key genes involved in ABA catabolism,

VvBG1, VvBG2 and VvBG3 were isolated and expression

pattern of these genes during grape fruit development and

ripening was investigated in the presence and absence of

dehydration stress.

Materials and methods

Genes silico and phylogenetic analysis

The VvBG1, VvBG2 and VvBG3 full-length cDNA sequen-

ces of grape were obtained from the nucleotide database of

NCBI (http://www.ncbi.nlm.nih.gov/nucleotide/). The open

reading frames (ORF) were determined by using NCBI ORF

Finder (http://www.ncbi.nlm.nih.gov/gorf/gorf.html). Deduced

amino acid sequences of VvBGs were aligned with the

homologous proteins in Hordeum vulgare, Lotus japonicus,

Medicago truncatula, and Zea mays using ClustalX 2.0.12

software in the default setting. The alignment results

were edited and marked using BOXSHADE 3.21 software

(http://www.ch.embnet.org/software/BOX_form.html). The phy-

logenetic tree was constructed by using the neighbor-joining

(N-J) method in MEGA 4.0.2 software with the bootstrap

analysis setting at 1,000 replicates for evaluating the reli-

ability of different phylogenetic groups. Tree files were

viewed and edited using MEGA 4.0.2 software.

Berry sampling

Fourteen-year-old grapevines (Vitis vinifera L. cv.Muscat

Hamburg) growing on the campus of China Agricultural

University, Beijing, China were used as a source of plant

material. The experimentation was performed in 2011, full

bloom of grapevines occurred in mid-May and veraison

occurred approximately 60 days after full bloom (DAFB).

Samples were taken every few days from 20 DAFB and

immediately frozen in liquid nitrogen and stored at -80 �C

until required. To define the stage of berry development, 30

randomly selected berries were weighed and an average

berry weight was calculated. The level of total soluble

sugar and titratable acid content were measured.

Dehydration stress treatments

Both wilting leaves and normal leaves were sampled from

grapevines, and immediately frozen with liquid nitrogen,

powdered, mixed and stored at -808C for further use. For

testing the effects of dehydration treatment on grape ber-

ries, the fruit harvested at 10 days before harvest stage

were divided into the control (group 1) and dehydration

treatment (group 2). Fruit water loss was measured from 20

fruits every day, the same fruit being weighted at each

sampling point. Water loss at harvest was set as 0 and was

expressed as a percentage of original fruit weight for the

further days. The entire experiment was repeated twice.

RNA extraction, reverse transcription-PCR (RT-PCR),

and sequencing

Total RNA was extracted from 1 g of flesh using the hot borate

method (Wan and Wilkins 1994). Synthesis of the first-strand

cDNA from 2 lg of total RNA was conducted using a

Moloney murine leukemia virus reverse transcriptase (Takara,

Dalian, China) and used as a template for amplifying NCEDs

with degenerate primers (forward: 50-TTYGAYGGIGAYGG

IATGGTICA-30; reverse: 50-TCCCAIGCRTTCCAIARRT-

GRAA-30) designed from the conserved sequences of plant

NCEDs (AF224671, Z97215, DQ028471, DQ028472,

AY337613). To obtain the 30 nucleotide sequences, RACE-

PCR was performed using the 50/30RACE System for Rapid

Amplification of cDNA Ends, invitrogenTM according to the

manufacturer’s instructions. The PCR products were

sequenced by Invitrogen (Shanghai, China).

Real-time quantitative PCR

First-strand cDNA synthesis was conducted using the

PrimeScriptTM RT reagent kit (TaKaRa, Dalian, China)

from 1.0 lg total RNA. The sequences of the primer pairs

used for each gene are shown in Table 1. Real-time PCR

was performed using the SYBR Premix Ex TaqTM kit

(TaKaRa). Reactions contained 1 ll of primer mix, 2 ll

cDNA template, 10 ll SYBR Premix Ex TaqTM (29) mix

and 7 ll water for a total volume of 20 ll. Reactions were

carried out under the following conditions: 95 �C/30 s

106 Plant Growth Regul (2013) 70:105–114

123

(1 cycle); 95 �C/15 s, 58 �C/20 s; 72 �C/15 s (40 cycles),

using a Rotor-Gene 3000 system (Corbett Research, Aus-

tralia). The PCR product of each gene was confirmed by

agarose gel electrophoresis and double-strand sequencing.

The amplified fragment of each gene was subcloned and

used to generate efficiency curves. Relative fold expression

for each gene was calculated by the Rotor-Gene 6.1.81

software. The transcript of SAND was used to standardize

each reaction run with respect to RNA integrity, sample

loading and inter-PCR variations.

Determination of ABA content

For ABA extraction, 1 g of flesh was ground in a mortar and

homogenized in extraction solution (80 % methanol, v/v).

Extracts were centrifuged at 10,000 g for 20 min. The

supernatant liquid was eluted through a Sep-Pak C18 car-

tridge (Waters, Milford, MA, USA) to remove non-polar

compounds, and then stored at -20 �C for enzyme-linked

immunosorbent assay (ELISA). The ELISA procedures were

conducted according to the instructions provided by the

manufacturer (China Agricultural University, Beijing,

China). The stepwise procedure for the indirect ELISA of

ABA was as follows. Each well of a micro titer plate had been

pre-coated with ABA-BSA conjugate diluted in coating

buffer by the manufacturer (China Agricultural University,

Beijing, China). To each well were added 50 ll of standard

or sample in assay buffer (8.0 g NaCl, 0.2 g KH2PO4, 2.96 g

Na2HPO4�12H2O, 1.0 ml Tween-20, 1.0 g gelatin, added to

1,000 ml H2O) followed by 50 ll of ABA antibody diluted

1:2,000 in assay buffer. The plates were incubated for 0.5 h

at 37 �C and then washed four times with scrubbing buffer

(the same was done with assay buffer but there was no gel-

atin). Anti-mouse IgG coupled to alkaline phosphatase

(100 ll of a 1:1,000 dilution) was added to each well and the

plates incubated for 0.5 h at 37 �C. The plates were washed

as above, and then 100 ll of a 1–2 mg ml-1 solution of

o-phenylenediamine substrate and 0.04 % of 30 % H2O2 in

substrate buffer (5.10 g C6H8O7�H2O, 18.43 g Na2H-

PO4�12H2O, 1.0 ml Tween-20, added 1,000 ml H2O) were

added to each well. After several minutes, 50 ll of

2.0 mol l-1 H2SO4 were added to each well to terminate the

reaction. The absorbance was read at 490 nm with a Thermo

Electron (labsystems) Multiskan MK3 (PIONEER Co.,

China). The concentration of ABA in a sample was calcu-

lated from Logit B/B0-transformed standard curve data

where B and B0 are the absorbance values in the presence and

absence of the competing antigen, respectively.

Determination of soluble sugar content and titratable

acidity

Ten grams of sample was ground into powder in a mortar and

pestle with liquid nitrogen. A total of 0.5 g of powder and

10 mL of 80 % (v/v) ethanol were mixed and incubated for

3 min at 80 �C, centrifuged at 10,000 g for 10 min, and the

supernatant was collected into a 100 mL triangular flask. Ten

milliliters of 80 % (v/v) ethanol was added to the residues, and

the extraction was repeated twice. The supernatants were

pooled, and the remaining residue was washed and filtered

with 1 mL of 80 % (v/v) ethanol. The filtrate was moved into a

10 mL test tube, and two drops of 5 % N-naphthol was added

and mixed. Then, concentrated sulfuric acid was added slowly

along the wall of the tube, until there was no purple ring in

the layer, indicating that the sugar was completely extracted

from the sample. The combined supernatants were evaporated

in a boiling-water bath and washed twice with 20 mL of

ultrapure water, made up to a volume of 50 mL, and 2 mL was

removed for LC-18 solidphase extraction. The soluble sugar

content was determined by the anthrone colorimetric method

(Laurentin and Edwards 2003) using a SP-1900UV spectro-

photometer (Ledon Technologies). Standard D-(?)-Glc

(G8270-100G) was purchased from Sigma. The assays were

repeated three times.

Table 1 Specific primers used

for the real-time quantitative

PCR

Primer names Sequences GenBank number

VvBG1-F 50-TGAACCTTACATAGTTGCCCACCAT-30 GU480917

VvBG1-R 50-AATCCCCATACATCAGAGGGTCAAT-30

VvBG2-F 50-ATAGTGAAGAAGAGGGCAGGCACG-30 XM_003632325

VvBG2-R 50-GCGGCCATATCTGCAAGAAAGTC-30

VvBG3-F 50-GCCGCAGAATAGTAGAAGACTTTGC-30 XM_002272377

VvBG3-R 50-GCAATATAAGGCTCGGTTGATGAGT-30

VvNCED1-F 50-GGTGGTGAGCCTCTGTTCCT-30 AY337613

VvNCED1-R 50-CTGTAAATTCGTGGCGTTCACT-30

VvCYP707A1-F 50-GGTCACTTGGAGGGTAATTAC-30 XM_002282197

VvCYP1707A1-R 50-TGTTGTCGGCGATTTGATCCT-30

SAND-F 50-GCGACGAGTGAGTCTGGGATTG -30 XM_002285134

SAND-R 50-TGAAATAGAAGCATCGTCCTCATCG-30

Plant Growth Regul (2013) 70:105–114 107

123

The titratable acidity was determined by repeated titra-

tion with 4.00 mol/L NaOH (2 drops of 1 % phenol-

phthalein added) to a faint pink (Long and He 2002). Based

on the volume of NaOH solution used for titration calcu-

lates the titratable acidity expressed as tartaric acid.

Results

Gene isolation and analysis

The three VvBGs full-length cDNA sequences from grape

were isolated and designated as VvBG1, VvBG2 and

VvBG3, respectively. A phylogenetic tree shows the

genetic relationship of VvBGs with BGs reported in other

species (Fig. 1). As shown on the phylogenetic tree,

VvBG1 was closely related to AtBG40 (NM_102418.3)

(Theologis et al. 2000) in Arabidopsis and VvBG2 was

closely related to MaBG (JN833573.1) in banana. And

VvBG3 was closely related to HvBG (L41869.1) (Leah

et al. 1995) in barley.

The deduced amino acid sequence lengths of VvBG1,

VvBG2 and VvBG3 were 505aa, 512aa and 518aa, respec-

tively while similarities in the deduced amino acid

sequences were 40 to 50 %. Based on multiple alignments

of these protein sequences (Fig. 2), most of the functional

residues or domains were observed to be well-conserved

within this gene family. These catalytic amino acids were

surrounded by a characteristic consensus of conserved

amino acids, already recognized in known, active b-gluco-

sidases. The catalytic acid/base was found in the conserved

sequence TFNEP in b-glucosidases. The nucleophilic glu-

tamate was always found in the consensus sequence

I/VTENG (Opassiri et al. 2003; Withers et al. 1990; Wang

et al. 1995).

Development of grape berries and changes in ABA

concentration

The dynamic process of Muscat Hamburg grape berry

development is obviously divided into three major phases

spanning a period of 11 weeks after full bloom (Xu et al.

1995):1–3 weeks for the first rapid berry growth phase

(Phase I), 4–8 weeks for the lag phase (Phase II)

and 9–11 weeks for the second rapid berry growth phase

(Phase III). Morphologically berry color changes from dark

green to light green, and further to red green 50-70 DAFB.

As shown in Fig 3, soluble sugar content (mainly glucose

and fructose) increases rapidly 54 DAFB while acid con-

tent decreases sharply, which indicates the onset of grape

berry ripening (veraison). ABA concentration declined to a

minimum 20–46 DAFB during the immature green stage.

At veraison, the level of ABA increased markedly and

reached a maximum 60 DAFB, followed by a gradual

decline in peel, pulp and seed tissues (Fig. 4).

The expression of VvNCED1, VvCYP707A1 and VvBGs

genes during grape berry development and ripening

Transcript levels of VvNCED1, VvCYP707A1, and VvBGs

genes were analyzed using real-time quantitative PCR. The

expression patterns of these genes during grape berry

development are shown in Figs. 4 and 5. The VvNCED1

gene was expressed strongly 24 DAFB, and then decreased

gradually with berry development. Expression of this gene

increased with the onset of ripening and reached a maxi-

mum 85 DAFB concomitant with accumulation of ABA.

The expression pattern of VvCYP707A1 was negatively

correlated with VvNCED1 transcripts and ABA content.

Expression of VvBG1 was upregulated whereas expression

of VvBG2 displayed either one or two peak values during

development and ripening of grape berries. Unlike VvBG1

and VvBG2, the expression of VvBG3 decreased gradually

throughout grape berry development (Fig. 5).

Fig. 1 Phylogenetic tree of the BG genes subfamily. A phylogenetic

tree of Neighbor-Joining was constructed with ClustalX software

using the deduced protein sequences. Genes studied in this experi-

ment are marked by black filled circle. Branch lengths are propor-

tional to the number of amino acid substitutions. Sequence data

mentioned in this article can be found in the GenBank data libraries

under the following accession numbers: Grape (Vv)-VvBG1 (GU480

917.1), VvBG2 (XM_003632325.1) and VvBG3 (XM_002272377.2);

Arabidopsis (At)-AtBG29 (NM_001125052.1), AtBG40 (NM_1024

18.3); Alfalfa (Mt)-MtBG (EU078903.1); Barley (Hv)-HvBG(L41869.1); Brazil Rubber Tree (Hb)-HbBG (EF100816.1); Lotus

japonicus (Lj)-LjBG (EU710845.1); Lodgepole Pine (Pc)-PcBG(AF072736.1); Maize (Zm)-ZmBG (U33816.1); Musa acuminata

AAA Group (Ma)-MaBG (JN833573.1); Rice (Os)-OsBG (AY1292

94.1); Shamrock (Tr)-TrBG (X56733.1); Sweet cherry (Pa)-PaBG(U39228.1); Watermelon (Cl)-ClBG (HQ681280.1); Wheat (Ta)-TaBG(AB236422.1)

108 Plant Growth Regul (2013) 70:105–114

123

Expression of VvBG1, VvBG2 and VvBG3 genes

in response to dehydration

To examine whether expressions of the VvBG1, VvBG2 and

VvBG3 genes was induced by dehydration stress and any

relationship to changes in ABA content in vegetative

tissues, expressions of these genes in leaves (Fig. 6) and

fruit tissues (Figs. 7, 8) was analyzed after dehydration

stress treatment. A comparative analysis of transcript

accumulation and ABA content in leaves and fruit tissues

treated for 7 days under low RH (45 %) was performed.

After exposure to dehydration stress, a marked increase in

Fig. 2 Amino acid sequence alignment of grape VvBGs with related

b-glucosidases. The grape cDNA derived sequences are labeled as

VvBG1 (GenBank ID:ADD17684.1), VvBG2 (GenBank ID:XP_0036

32373.1), VvBG3 (GenBank ID:XP_002272413.1); HvBG is Horde-um vulgare b-glucosidases (GenBank ID: ACF07998.1); LjBG is

Lotus japonicus b-glucosidases (GenBank ID: ACD65510.1); MtBG

is Medicago truncatula b-glucosidases (GenBank ID: ABW76288.1);

ZmBG is Zea mays b-glucosidases (GenBank ID: AAD10503.1); The

catalytic acid/base and nucleophile residues are marked by black filledcircle, while those conserved residues making contacts with the sugar

are marked by triangle

Plant Growth Regul (2013) 70:105–114 109

123

ABA and VvBG1 transcript accumulation was detected

(Figs. 6,7,8). During the dehydration experiment, fruits lost

20–23 % of their water content 4 days after dehydration.

The ABA content of berries increased significantly after

dehydration stress, and after 3 days was 2 times the initial

level, and in leaves, ABA content increased by a similar

amount. Real time RT-PCR revealed that the expression

patterns of the VvBGs genes under dehydration stress were

relatively complex (Figs. 6,8). Expression of VvBG1 was

significantly up-regulated in the fruits and leaves. In con-

trast to VvBG1, expression of VvBG2 was either marginally

up-regulated or down-regulated by dehydration stress in

fruit tissues and leaves. The expression of VvBG3 was

down-regulated in leaves but no significant difference was

observed in fruit tissues (Figs. 6,8).

Discussion

Our data indicate that the expression of b-glucosidase

genes plays an important but as yet unidentified role in

modulation of ABA concentration of grape berry. The

expression of VvNCED1 coincided with ABA accumula-

tion and fruit development, and it might be the major gene

involved in ABA biosynthesis in grape berry. The

expression of VvCYP707A1 was confirmed as a negative

regulatory factor of ABA, corroborating previous reports

(Krochko et al. 1998; Saito et al. 2004; Nambara and

Marion-Poll 2005; Setha et al. 2005). In addition to

expression of VvNCED1 and VvCYP707A1 in grape, the

transcription of the b-glucosidase genes (BGs) could also

impact levels of ABA. The three cDNAs were expressed in

Fig. 3 Changes in soluble sugar content and titratable acidity of

berries of Muscat Hamburg during grape development and ripening.

The experiments were repeated three times and the bars represent

standard error (SE)

Fig. 4 Change in ABA content and expression of VvNCED1,

VvCYP707A1 in peel, pulp and seed of Muscat Hamburg during

grape development and ripening. SAND was used to standardize each

reaction run with respect to RNA integrity, sample loading, and inter-

PCR variations. Values presented are mean ± SE (n C 3)

110 Plant Growth Regul (2013) 70:105–114

123

Fig. 5 Expression of VvBG1, VvBG2 and VvBG3 in the peel, pulp

and seed of Muscat Hamburg during grape development and ripening.

SAND rRNA was used to standardize each reaction run with respect to

RNA integrity, sample loading and inter-PCR variations. Values

presented are mean ± SE (n C 3)

Fig. 6 Expression of VvBG1, VvBG2 and VvBG3 in dehydration

stressed leaves of grape and changes in ABA content. SAND rRNA

was used to standardize each reaction run with respect to RNA

integrity, sample loading and inter-PCR variations. Expression of

each gene is presented as relative fold change. Values presented are

mean ± SE (n C 3)

Fig. 7 ABA content of control and dehydration stress-treated

detached grape berries. Fruits were detached from vines and

dehydrated under ambient conditions. After determination of water

loss the endogenous ABA content was determined as described in

‘‘Materials and methods’’. Results are the mean ± SD of the three

measurements

Plant Growth Regul (2013) 70:105–114 111

123

the fruit pulp, peel and seed during development and

maturation. For example, transcript levels of VvBG1

and VvBG2 were relatively low during early berry growth

and development, but increased during the period from

coloration to fruit ripening and peaked 60 DAFB coinci-

dent with ABA accumulation, which indicates that VvBG1

and VvBG2 might play a role in regulating the level of

ABA during the ripening stage (Lee et al. 2006; Sun et al.

2010). VvBG3 transcript levels were higher during early

young berry growth, and then decreased with fruit devel-

opment and ripening. These results indicate that ABA level

of grape berries might be impacted by all three VvBGs

during the fruit maturation stage, and by VvBG1 and

VvBG2 during the late ripening stage, while VvBG3

appears to play a role in early fruit development. Confir-

mation however, awaits investigation of the expression

pattern in response to exogenous (?)-ABA and/or by

investigating ABA catabolism per se via measurement of

endogenous 80 OH-ABA, PA and DPA in future work.

ABA accumulation in fruit has been shown to correlate

with abiotic stresses (Cakir et al. 2003; Deluc et al. 2007,

2009; Zhang et al. 2009a, b; Leng et al. 2009; Ji et al. 2012).

Three or four days after dehydration stress treatment,

significant increases were found in ABA content and in

transcript levels of VvBG1 and VvBG2 in the berries com-

pared with those in the control tissue samples (Figs. 7, 8).

ABA activates its own inactivation to DPA via the tran-

scriptional activation of CYP707A1 gene expression (Saito

et al. 2004; Umezawa et al. 2006; Okamoto et al. 2009). This

suggess that endogenous ABA content is modulated by a

dynamic balance between biosynthesis and catabolism.

Based on the results above, potential function of the VvBGs

and their putative role in modulating ABA content exists

during grape berry growth and development and response to

abiotic stress, and further research on the VvBGs enzyme

activity and substrate specificity is required in future work.

In conclusion, the expression of VvBGs might play a

role in the modulation of ABA level during grape berries

development and respond to dehydration stress.

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Fig. 8 Expression of VvBG1,VvBG2 and VvBG3 in

dehydration stressed grape

berries. SAND rRNA was used

to standardize each reaction run

with respect to RNA integrity,

sample loading and inter-PCR

variations. Expression of each

gene is presented as relative fold

change. Values presented are

mean ± SE (n C 3)

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