Transcriptome changes in grapevine (Vitis viniferaL.) cv. Malbec leaves induced by ultraviolet-B radiation
© Pontin et al; licensee BioMed Central Ltd. 2010
Received: 29 June 2010
Accepted: 20 October 2010
Published: 20 October 2010
Ultraviolet-B radiation (UV-B, 280-315 nm) is a natural component of sunlight, which has numerous regulatory effects on plant physiology. The nature of the response to UV-B is dependent on fluence rate, dose, duration and wavelength of the UV-B treatment. Some reports have analyzed the changes in gene expression caused by UV-B light on several plant species using microarray technology. However, there is no information on the transcriptome response triggered by UV-B in grapevine. In this paper we investigate the gene expression responses of leaves from in vitro cultured Vitis vinifera cv. Malbec plants subjected to the same dose of biologically effective UV-B radiation (4.75 kJ m-2 d-1) administered at two different fluence rates (16 h at ≅ 8.25 μW cm-2, 4 h at ≅ 33 μW cm-2) using a new custom made GrapeGen Affymetrix GeneChip®.
The number of genes modulated by high fluence rate UV-B doubled the number of genes modulated by low fluence UV-B. Their functional analyses revealed several functional categories commonly regulated by both UV-B treatments as well as categories more specifically modulated depending on UV-B fluence rate. General protective responses, namely the induction of pathways regulating synthesis of UV-B absorbing compounds such as the Phenylpropanoid pathway, the induction of different antioxidant defense systems and the activation of pathways commonly associated with pathogen defense and abiotic stress responses seem to play critical roles in grapevine responses against UV-B radiation. Furthermore, high fluence rate UV-B seemed to specifically modulate additional pathways and processes in order to protect grapevine plantlets against UV-B-induced oxidative stress, stop the cell cycle progression, and control protein degradation. On the other hand, low fluence rate UV-B regulated the expression of specific responses in the metabolism of auxin and abscisic acid as well as in the modification of cell walls that could be involved in UV-B acclimation-like processes.
Our results show the UV-B radiation effects on the leaf transcriptome of grapevine (Vitis vinifera cv. Malbec) plantlets. Functional categories commonly modulated under both UV-B treatments as well as transcripts specifically regulated in an UV-B-intensity dependent way were identified. While high fluence rate UV-B had regulatory effects mainly on defense or general multiple-stress responses pathways, low fluence rate UV-B promoted the expression of genes that could be involved in UV-B protection or the amelioration of the UV-B-induced damage. This study also provides an extensive list of genes regulating multiple metabolic pathways involved in the response of grapevine to UV-B that can be used for future researches.
Ultraviolet-B radiation (UV-B, wavelength range 280 to 315 nm) is a natural component of solar radiation. Most of the UV-B solar radiation is absorbed by the stratospheric ozone layer and other atmospheric gases and therefore only a minor proportion reaches the Earth's surface. The level of UV-B is dependent on several factors such as latitude, season, time of day, cloud cover, and altitude .
The effects of UV-B have been analyzed on diverse plants species and vary depending on UV-B fluence rates, duration and wavelength of the UV-B treatment [2–7]. Exposure to UV-B amounts much higher than those found in nature causes tissue necrosis and induces the expression of many genes normally involved in defense, wounding, or general stress responses. That is, several studies have reported damage to DNA, proteins and membranes and the inhibition of protein synthesis and photosynthetic reactions [4, 8, 9].
Ultraviolet-B radiation is not necessarily a damage-inducing source of stress but instead can act as an important environmental cue in higher plants, regulating several key developmental plant responses. At ambient UV-B levels, crosstalk between wounding and UV-B signaling pathways seem to modify plant-insect interactions . Moreover, exposure to such low non-damaging levels of UV-B has numerous regulatory effects on plant morphology, physiology, and biochemistry [3, 5, 11]. These low fluence rates of UV-B promote the expression in a range of genes that are known to be involved in UV-B protection or amelioration of UV-B damage [3, 5, 12, 13]. Among the most important protective mechanisms in higher plants are the accumulation of UV-absorbing phenolic compounds in epidermal tissues [9, 14, 15] and the enhancement of cellular antioxidant systems [3, 8]. Similar responses have also been shown in grapevine plants (Vitis vinifera L. cv. Malbec) [16, 17].
Over the last years efforts have been focused on the study of the molecular responses of plants to UV-B radiation. Several reports have analyzed the changes in gene expression caused by UV-B in many plant species [12, 13, 18–20] using microarray analysis. Interestingly, these studies have shown that some UV-B response pathways are shared with other environmental cues, while additional pathways may account for UV-B-specific responses , pointing out the existence of common as well as stress-specific gene expression profiles .
In grapevine (Vitis vinifera L.) plants there is no available information on the changes in gene expression triggered by UV-B radiation. However, different microarray platforms have been used in this species to analyze transcriptomic variation during berry development [22–24], water and salinity stresses , virus infection  and fungal pathogen attack .
Malbec is a grapevine cultivar well adapted to the growing conditions of Argentine, were it became the emblematic icon of the country's wine industry. In the province of Mendoza, the major wine-producing region in the country, vineyards are located at heights ranging from 500 to 1500 m a.s.l. Such variations in altitude account for different fluence rates and dosages of UV-B reaching the vineyards . Thus, Malbec plants growing at different altitude develop different UV-B responsive mechanisms that are relevant for the final composition of grape berries and affect the quality of wine [16, 17].
In the present study we used a new custom made Affymetrix GrapeGen GeneChip™ (Lijavetzky et al., In preparation) to investigate gene expression responses of leaf tissues from in vitro cultured grapevine cv. Malbec plants to the environmental level of UV-B radiation found at 1000 m a.s.l. (4.75 kJ m-2 d-1, calculated for the range 310-315 nm) , administered at two different fluence rates: "low UV-B" (16 h at 8.25 μW cm-2) or "high UV-B" (4 h at 33 μW cm-2 UV-B). The minimum and maximum UV-B fluence rate of solar radiation perceived by vineyards at 1,450 m a.s.l. at early morning and noon for the Mendoza's region is of c.a 9 and 35 μW cm-2, respectively .
The functional classification of the differentially expressed genes and the pictorial representation of the significantly modulated classes were accomplished by means of the MapMan software  using adapted files specifically designed for the custom made Grapegen GeneChip™. Our results showed that transcription in grapevine leaves is differentially affected by UV-B fluence rate. We found that the number of modulated probe sets (up- and down-regulated) under high fluence rate UV-B was two-fold higher than the number of probe sets modulated under low intensity UV-B. Transcripts commonly modulated under both UV-B treatments as well as transcripts specifically regulated in an UV-B-fluence rate dependent way were identified. The functional analysis of the differentially expressed genes showed that while high fluence rate UV-B had regulatory effects on defense or general stress response pathways, low fluence rate UV-B promoted the expression of genes involved in UV-B protection or the alleviation of the UV-B damage, likely contributing to the acclimation of plants to UV-B exposure.
Results and Discussion
High UV-B has a stronger effect on gene expression than low UV-B
Functional analyses of transcriptome changes in response to UV-B light treatments
Significantly modulated processes under High UVB
Cellular response overview.Biotic stress
Carbohydrate metabolism.Sugar binding proteins
Protein metabolism and modification.Molecular chaperone.HSP
Regulation overview.Nucleic acid metabolism.TFs.NAC
Regulation overview.Nucleic acid metabolism.TFs.Pathogenesis-related
Regulation overview.Nucleic acid metabolism.TFs.WRKY
Secondary metabolism.Phenylpropanoids.General pathway
Cellular response overview.Cell growth and death.Cell cycle
Signalling.Light signalling.Blue light signalling
General responses induced by UV-B radiation
Nine functional categories were found to be significantly up-regulated by both UV-B treatments, whereas no functional category was found down-regulated by both UV-B treatments. Interestingly, different functional categories were specifically modulated (either induced or repressed) depending on UV-B fluence rate (Tables 1 and 2). These results are in agreement with previous reports indicating that different UV-B fluence rates can elicit different responses on diverse plant species [3, 5–7]. For example, in maize certain genes are only induced above certain thresholds of UV-B irradiance intensity, suggesting the operation of diverse signaling pathways at different fluence rates within the same species .
In grapevine, chalcone synthase (CHS) substrates can also be used by stilbene synthase (STS) enzymes for the production of stilbenes, which are classified as phytoalexins because of their role in plant defense mechanisms against fungal pathogens [31, 32]. Among stilbenes, resveratrol (trans-3,5,4'-trihydroxystilbene) is the most prominent compound . The accumulation of resveratrol in grapevine berries and leaves is induced by several stresses such as fungal infection, injury and UV-B light exposure [16, 31]. As shown in Figure 3, many genes assigned to the functional category "Phytoalexins" (BIN 19.4.2) were induced under both UV-B treatments. Among them, genes mainly encoding STS and resveratrol synthases gave significance to this functional category  (Additional files 6 and 7). The enhanced representation of "Phenylpropanoid general pathway" and "Phytoalexins" categories show that UV-B promoted the expression of genes involved in the induction of different antioxidant defense systems. This is in agreement with the observation that in grapevine the synthesis of anthocyanins, flavonols, quercetin and kaempferol is promoted by exposure of plants to environmental doses of UV-B light .
Plant hormones play important roles in diverse growth and developmental processes as well as in various plant responses to biotic and abiotic stresses. In particular, ethylene, auxins, abscisic acid (ABA) and brassinosteroids have been associated with the UV-acclimated phenotypes [13, 17, 20, 35]. As shown in Tables 1 and 2 and Additional file 8a, the "Ethylene metabolism" (BIN16.1.2) and "Ethylene responsive TFs" (TFs: Transcription Factor/s, BIN 22.214.171.124) were functional categories significantly over represented under both UV-B treatments. It has been established that ethylene as well as ethylene response factor (ERF) proteins play important regulatory roles in plant pathogen resistance and abiotic stress response [36, 37]. ERFs belong to a large family of APETALA2-domain-containing TFs that bind to a GCC-box present in the promoters of many ethylene inducible defense-related genes . We found that several genes, putatively annotated as ERFs and ethylene responsive proteins, were commonly over expressed under both UV-B treatments (Additional files 6 and 7). These results are in agreement with a previous study carried out by Ulm et al.  in Arabidopsis, showing that activation of the ethylene pathway is a general response to UV-B exposition. In grapevine, ethylene plays an important role in berry development and ripening processes including the regulation of gene expression for anthocyanin biosynthesis and accumulation . As shown here, ethylene also seems to participate in the modulation of leaf responses to UV-B radiation (Tables 1 and 2, Additional file 8a).
The WRKY proteins share a DNA binding domain which contains an invariant WRKYGQK sequence (after which the domain was named, ). The WRKY TF super family is involved in a diverse set of biological functions including pathogen defense, abiotic stress responses and plant development [43, 44]. As shown in Figure 4, several genes encoding WRKY factors showed an increased expression in response to both UV-B treatments (Additional files 6 and 7), suggesting a role for these factors as components of grapevine protection mechanisms against potentially damaging UV-B radiation.
Responses specifically regulated by high fluence rate UV-B radiation
High fluence rate UV-B significantly modulated gene expression in additional functional categories. Among the up-regulated processes three functional categories were significantly overrepresented: BIN 5.1 corresponding to "Sugar binding proteins", BIN 11.4.1 "Heat shock proteins" (Hsps) and BIN 126.96.36.199 "Pathogenesis related TFs" (Table 1).
The functional category of "Sugar binding proteins" (Table 1, Additional file 8c) includes genes mainly encoding different kind of lectins (curculin-like, -Mannose-binding- lectin, D-galactoside L-rhamnose binding lectin, and lectin 2) and glycoproteins (Additional file 6). Lectins comprise a miscellaneous group of proteins able to bind carbohydrate residues of different chemical nature in a specific and reversible way. Although protection against pathogens is the major function attributed to lectins, their accumulation in plant tissues have also been described under different abiotic stresses or as a result of growth and developmental processes [45–47]. These proteins could play a role in the protection of plant cell against damaging effects of free radicals as hydrogen peroxide . Thus, in the high fluence UV-B response, the induction of lectins could be interpreted as a mechanism to shorten the duration of the "oxidative burst" and to protect grapevine leaves against high UV-B induced-ROS.
Hsps and their regulatory transcription factors (Hsfs) play a broad role as molecular chaperones in the tolerance to multiple environmental stresses apart from heat stress . Some Hsps act as molecular chaperones counteracting protein denaturation and aggregation while other Hsps including ubiquitin and certain proteases, target nonnative proteins for degradation . Hsps function may extend beyond their chaperone activity, limiting the damage that results from ROS accumulation. In fact, exposure to UV-B radiation, which increases cellular concentrations of hydrogen peroxide, activates Hsfs expression . In agreement with those reports our results show that genes encoding several Hsps (with mitochondrial, chloroplastic, and cytoplasmic localization) as well as Hsfs are differentially up-regulated under high fluence rate UV-B (Additional file 6).
The induction of the expression of genes in the functional category of "Pathogenesis related TFs" detected under high UV-B but not under low UV-B (Table 1, Figure 4) could be part of the same general response to biotic stress (BIN 4.2) observed at both UV-B fluence rates mentioned above. Given the existence of a slight difference in the number of genes induced under each UV-B treatment this category is significantly represented only under high UV-B. However, this category must be relevant under both conditions.
Down regulation of gene expression is also an important effect of UV-B light exposure although as mentioned above fluence rates have specific effects on the functional categories that are down regulated. The strong down-regulation of the category "Cell cycle" (BIN 4.13) is a specific feature of high UV-B (Table 1, Figure 2). Indeed, our results revealed that the activities of several cell cycle-related genes are temporally paused in the leaves of grapevine plantlets exposed to high UV-B (Additional file 9). High UV-B specifically down-regulates the expression of genes encoding cyclins, cyclin-dependent kinase inhibitors, kinesins and cell division control proteins (Additional file 9). It is likely advantageous for grapevine cells to temporally halt cell cycle, to allow for repairing DNA damage and to prevent the introduction of mutations into the DNA of daughter cells. It is well documented that UV-B photons may cause cellular damage by generating photoproducts of DNA . DNA damage is also known to induce cell cycle arrest and rapid protein turnover via the proteasome [15, 53]. In fact, it has been proposed that UV-B could delay cell division by arresting cell cycle in the G1/S transition phase .
Finally, among the functional categories specifically regulated by high UV-B, "Blue light signaling" (BIN 8.2.1) is also found down-regulated (Table 1, Additional file 8e). Genes encoding TFs acting downstream of phototropin photoreceptors are differentially repressed under high UV-B (Additional file 9). Phototropins (phot1 and phot2) are blue-light (BL)-sensitive receptor kinases, involved in the control of BL-activated stomatal opening and chloroplast relocation in addition to the phototropism response [55, 56]. It is known that these phototropins use signal transducers for different photoinduced movement responses in different tissue. Thus, in the phototropic response of hypocotyls, nonphototropic hypocotyl 3 (NPH3) is a common regulator in the phot1-and phot2-signaling pathways . Also, it was shown that NPH3-mediated phototropin signaling optimizes the efficiency of BL-perception by inducing both optimal leaf positioning and leaf flattening . Our results show a significant down-regulation of the NHP3 transcription factor (Additional file 9) suggesting that high UV-B might trigger a negative phototropic response of grapevine leaves as an escape response to protect leaf tissues against potentially damaging UV-B radiation.
Differential responses regulated by low UV-B radiation
Significantly modulated processes under Low UVB
Cellular response overview.Abiotic stress.Anoxia
Cellular response overview.Biotic stress
ATPase family associated with various cellular activities
Regulation overview.Nucleic acid metabolism.TFs
Regulation overview.Nucleic acid metabolism.TFs.NAC
Regulation overview.Nucleic acid metabolism.TFs.WRKY
Secondary metabolism.Phenylpropanoids.General pathway
Cell wall metabolism
Cell wall metabolism.Cell wall modification
Cellular response overview.Stress Miscellaneous
Exposure of grapevine plantlets to low UV-B specifically down-regulates genes in the functional category "Stress miscellaneous" (BIN 4.10, Table 2, Figure 2). An overview of the genes which gave significance to this category shows that low UV-B affected components commonly involved in stress responses such as putative Thaumatin-like proteins, Pathogenesis-related protein PR-1 precursor, putative senescence-associated proteins, Allergen V5 Tpx-1 and chitinase-like proteins (Additional file 10) [62, 63]. These genes have been associated with plant defense to pathogens attack or environmental stress but they are also developmentally regulated . In our experiment, their expression could be more related to growth processes and secondary metabolic changes induced by low UV-B, than to responses triggered by UV-B stress.
Another functional category whose expression was specifically enhanced by low fluence rate UV-B is the "ATPase family" (BIN 15, Table 2, Additional file 8d). Genes encoding a particular class of ATPases, named AAA ATPase is specifically up-regulated under this UV-B treatment (Additional file 7). ATPase Associated with various cellular Activities (AAA) proteins are characterized by the presence of one or several conserved motives including the Walker A and Walker B required for ATP binding and hydrolysis, respectively, and a highly conserved amino acid sequence termed the second region of homology (SRH, ). This protein family is commonly distributed among eukaryotes and is involved in several cellular functions. Information on the role of AAA ATPases is very limited in plants. Some studies have related them with degradation of thylakoid proteins by the 26S proteasome . Our results might suggest an effect of low fluence rate UV-B on chloroplast proteins turnover.
Finally, exposure of grapevine plantlets to low UV-B has a repressive effect on the category "Auxin metabolism" (BIN 16.1.3, Table 2, Additional file 8a). Our results show that genes belonging to auxin responsive SAUR and Aux/IAA family, auxin response factors and auxin transporter-like proteins are down-regulated in the grapevine leaves exposed to low UV-B (Additional file 10). Thus, down-regulation of auxin signaling components or auxin transport supports a role for auxins in the response to low UV-B fluence light. Similar results were found in the study of pathogen resistance responses, where a repression of a number of auxin responsive genes (including genes SAUR, Aux/IAA, auxin importer AUX1, auxin exporter PIN7) were significantly repressed ; supporting the idea that down-regulation of auxin signaling contributes to induce immune responses in plants .
The leaf transcriptome of grapevine was differentially affected by UV-B fluence rate. The number of genes modulated (up- and down-regulated) by UV-B light exposure was two-fold higher under high fluence rate UV-B than under low fluence rate UV-B. Analysis of the functional categories differentially represented under high and low UV-B revealed that the existence of common and specific responses regulated at different UV-B intensity. General protective responses such as the induction of pathways regulating the synthesis of UV-B absorbing phenolic compounds, the induction of antioxidant defense systems, and the activation of signal transduction pathways associated with biotic and abiotic stress responses played critical roles independently of fluence rate. Furthermore, increased fluence rates triggered additional stress responses involving the induction of heat shock and sugar binding proteins and causing a rapid decline in the expression of genes involved in cell cycle. Then again, low fluence rate UV-B promoted additional transcriptional responses that could be interpreted as acclimation processes such as the induction of ABA catabolism and chloroplast protein turnover associated ATPases or the repression of genes involved in cell wall modification and auxin metabolism and responses. These findings provide a preliminary functional genomics framework to understand the complexity of grapevine UV-B radiation responses.
Plant material and in vitro culture
Wood cuttings corresponding to three-nodal lignified segments derived from virus-free plants of Vitis vinifera L. cv. Malbec were planted in 2.5 l plastic pots containing hydrated-perlite and grown under greenhouse conditions. One-node cuttings coming from three-month old plants were collected and used as explants for in vitro culture. Explants were surface sterilized with 75% ethanol for 3 min and then with 15% sodium hypochlorite for 15 min. The cuttings were rinsed three times in sterile dH2O and cultivated on solid medium composed of full MS medium salts and vitamins , 30 g l-1 sucrose, 1 mg l-1 6-benzylaminopurine (BAP), and 7.5 g l-1 agar for shoot initiation and proliferation of grapevine shoot-tip explants. In vitro shoot tips were subcultured (five times) at 35-40-day intervals into half-strength MS micro- and macro-nutrients, excluding FeEDTA (0.1 mM Na2EDTA + 0.1 mM FeSO4) and supplemented with full-strength MS vitamins, 30 g l-1 sucrose, 0.5 μM 1-naphthaleneacetic acid and 7.5 g l-1 agar. Only one explant per bottle (12 cm height × 6.5 cm diameter) was planted. Cultures were maintained in a growth chamber at 25 ± 2 ºC under 16 h photoperiod provided by cool-white fluorescents tubes at a photosynthetic photon flux density of 80 μmol m-2 s-1. Bottle tops were covered with low-density polyethylene, which does not absorb any wavelength of the photosynthetic light.
UV-B exposure conditions
In vitro grown plants (45-48 d old, having 6 full expanded leaves) were exposed to UV-B radiation in the same controlled growth chambers described above. Throughout the UV-B treatment, all plantlets were exposed to a background intensity of photosynthetically active radiation (PAR, 80 μmol m-2 s-1) provided by cool-white fluorescent lamps. For different light treatments, supplemental UV-B was given using a Philips TL 100W/01 tube (311 and 313 nm spectrum peaking, Philips, Eindhoven, The Netherlands) suspended 40 cm above the pots. A total effective dose of 311 nm normalized UV-B corresponding to 4.75 kJ m-2 d-1 was provided in two different treatments: 1) "low UV-B" (16 h at ≅8.25 μW cm-2 irradiance) and "high UV-B" (4 h at ≅33 μW cm-2 irradiance) supplied at the end of the 16h-day photoperiod). As a control of no UV-B radiation, plantlets were exposed for 16 h under the same UV-B tube covered with a polyester filter (100 μm clear polyester plastic, Oeste Aislante, Buenos Aires, Argentina), which absorbed more than 95% of UV-B without affecting PAR. It is worth mentioning that UV-B irradiance registered at ca. 1,500 m a.s.l., the altitude where the most reputed vineyards are located in the Mendoza's region, is about 35 μW cm-2 solar noon. A LI-250A light meter with a LI-190 quantum sensor (LI-COR Inc., Lincoln, NE, USA) and a PMA2200 radiometer with a PMA2102 UV-B detector (Solar Light Company Inc., Glenside, USA) were used to measure PPFD and UV-B, respectively.
As a control for circadian effects, only the fully expanded-upper leaves from two plantlets collected from each experimental treatment were harvested and pooled immediately at the end of the light treatments. Samples were immediately frozen in liquid nitrogen and conserved at -80 ºC until RNA extraction. Three independently grown, harvested, and extracted sets of samples corresponding to each experimental treatment were prepared as biological replicates.
RNA isolation and GeneChip®hybridization
Total RNA was extracted from leaf tissues using the TRIZOL reagent (Invitrogen) according to standard procedures. DNase digestion of contaminating DNA in the RNA samples was carried out with the RNase-Free DNase Set (QIAGEN). Final RNA purification was performed using the RNeasy Mini Kit (QIAGEN) according to standard protocols. Samples were analyzed at the Genomics Unit of the Spanish National Centre for Biotechnology (CNB-CSIC, Madrid). RNA integrity analyses were done with an Agilent's Bioanalyzer 2100. Probe synthesis, microarrays hybridization, washing, staining and scanning with the GeneChip™ Scanner 3000 were performed according to the Affymetrix GeneChip® Expression Analysis Technical Manual.
Differential expression and functional analyses
The mRNA expression profiles of the different treatments described above were compared using the Affymetrix GrapeGen custom GeneChip™ (Lijavetzky et al., In preparation). This new GeneChip generated for grapevine (genome size about 475 Mb) contains 23096 probe sets. Probe sets design was based on the publicly available Unigen information at the National Center for Biotechnology Information  by July 2006 (342576 grapevine ESTs). The probe sets represent Vitis vinifera consensus sequences (when overlapped) from Cabernet Sauvignon, Muscat Hamburg, Pinot Noir, Chardonnay and Shiraz cultivars. In order to determine the number of non-redundant genes represented in the GeneChip we mapped the original 23096 probe sets to the recently released 12X grapevine genomic sequence  and we determined that the GeneChip contains probes sets for ca. 15800 different annotated genes. Differential expression analyses were performed using the entire probe set information. In order to carry out the functional analyses with unique probe sets we processed the corresponding produced gene list using a Microsoft Excel spreadsheet containing both the original and "12X" probe set information. Non-redundant probe sets codes were replaced with their unique gene identifier at the "12X" while redundant probe sets were merged and we assigned their median value to the corresponding unique identifier at the 12X.
Data analysis for each treatment was performed using biological triplicates. Normalization was performed with the help of the normalization tool within the GEPAS 4.0 suite [72, 73]. For this procedure we used all the default Affymetrix methods defined by the GEPAS 4.0 system which were implemented in the "affy" package from Bioconductor : a) background correction by Robust Multi-array Average (RMA) expression measure , b) between array standardization based upon quantiles , and c) PM-MM adjustment using "pmonly" (just PM values are used). Differential expression analysis (P < 0.05) was carried out using the Clear Test method  implemented at the GEPAS 4.0 suite. This method tries to avoid the problem caused by t-tests when declaring genes with relatively small expression changes as differentially expressed if they show low observed variances. The Clear Test proposes to use the z-test as an alternative and combines that test with a χ2 test to evaluate variability  what helps to prevent the identification of genes with high observed variances as differentially expressed. Functional analyses of the differentially expressed genes were done with the MapMan software based on the Wilcoxon Rank Sum Test .
Classification of the GrapeGen GeneChip™ probesets and MapMan files preparation
Original Vitis vinifera sequences used for the GrapeGen GeneChip™ probe design and annotation were classified into a MapMan "mapping file" after performing automated and manual database searches. Sequence were blasted against the UniProtKB/Swiss-Prot plant proteins sequence database  and classified into a modified and adapted "mapping file" using as a reference model the original Arabidopsis file  as well as the related information present in other databases such as GO , KEGG , Swiss-Prot  and TAIR . Out of the 23046 grapevine sequences, 15783 were actually sorted in 36 BINs and several subBINs (Additional file 3). A total of 22 pictorial "pathway" representations were also mainly adapted from Arabidopsis while some specific files were originally developed (Figures 2, 3, 4 and 5, Additional file 8).
Real time quantitative RT-PCR (qRT-PCR)
Reactions and cDNA synthesis were performed according to Lijavetzky et al. . Transcript levels were determined using a 7300 Real-Time PCR System (Applied Biosystems) and SYBR Green dye (Applied Biosystems). Gene specific primers (Additional file 3) were designed using the Oligo Explorer 1.2 software (Gene Link) based on the corresponding probe set sequence from the custom GrapeGen GeneChip™. No-template controls were included for each primer pair, and each PCR reaction was completed in triplicate. Data were analyzed using the 7300 SDS software 1.3 (Applied Biosystems). Dissociation curves for each amplicon were then analyzed to verify the specificity of each amplification reaction. Transcript level was calculated using the standard curve method and normalized against grapevine EFα1 gene (UniGene Vvi.1750) used as reference control.
All microarray expression data produced in our study are publically available at the ArrayExpress database  under the accession number E-MEXP-2541.
The authors thank S. Gómez Talquenca (Estación Experimental INTA Mendoza) for providing us with the virus-free V. vinifera cv. Malbec segments and Jerome Grimplet (Instituto de Ciencias de la Vid y del Vino, Logroño, Spain) for providing additional unpublished information on GrapeGen GeneChip annotation and probe set redundancy. This study was supported by the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET, PID 5028 to RB), Secretaría de Ciencia y Técnica de la Nación (SECyT, PICT 08-12398 and PICT 20-20093 to RB) and Genoma España (Project GrapeGen to JMMZ). M. Pontin was recipient of a CONICET post-doctoral scholarship. We also thank Cheo Machín (Genoma España) for careful editing of the manuscript and David Montaner and Ximo Dopazo (Centro de Investigación Principe Felipe) for the statistical advice.
- Madronich S, McKenzie RL, Caldwell MM, Björn LO: Changes in ultraviolet radiation reaching the Earth's surface. AMBIO. 1995, 24: 143-152.Google Scholar
- Blanding CR, Simmons SJ, Casati P, Walbot V, Stapleton AE: Coordinated regulation of maize genes during increasing exposure to ultraviolet radiation: identification of ultraviolet-responsive genes, functional processes and associated potential promoter motifs. Plant Biotechnol J. 2007, 5 (6): 677-695. 10.1111/j.1467-7652.2007.00282.x.PubMedView ArticleGoogle Scholar
- Brosché M, Strid Å: Molecular events following perception of ultraviolet-B radiation by plants. Physiol Plant. 2003, 117 (1): 1-10. 10.1034/j.1399-3054.2003.1170101.x.View ArticleGoogle Scholar
- Casati P, Walbot V: Rapid transcriptome responses of maize (Zea mays) to UV-B in irradiated and shielded tissues. Genome Biol. 2004, 5 (3): R16-10.1186/gb-2004-5-3-r16.PubMedPubMed CentralView ArticleGoogle Scholar
- Frohnmeyer H, Staiger D: Ultraviolet-B radiation-mediated responses in plants. Balancing damage and protection. Plant Physiol. 2003, 133: 1420-1428. 10.1104/pp.103.030049.PubMedPubMed CentralView ArticleGoogle Scholar
- Jenkins GI: Signal transduction in responses to UV-B radiation. Annu Rev Plant Biol. 2009, 60: 407-431. 10.1146/annurev.arplant.59.032607.092953.PubMedView ArticleGoogle Scholar
- Ulm R, Nagy F: Signalling and gene regulation in response to ultraviolet light. Curr Opin Plant Biol. 2005, 8 (5): 477-482. 10.1016/j.pbi.2005.07.004.PubMedView ArticleGoogle Scholar
- Jansen MAK, Gaba V, Greenberg BM: Higher plants and UV-B radiation: balancing damage, repair and acclimation. Trends Plant Sci. 1998, 3 (4): 131-135. 10.1016/S1360-1385(98)01215-1.View ArticleGoogle Scholar
- Rozema J, van de Staaij J, Björn LO, Caldwell M: UV-B as an environmental factor in plant life: stress and regulation. Trends Ecol Evol. 1997, 12 (1): 22-28. 10.1016/S0169-5347(96)10062-8.PubMedView ArticleGoogle Scholar
- Stratmann J: Ultraviolet-B radiation co-opts defense signaling pathways. Trends Plant Sci. 2003, 8 (11): 526-533. 10.1016/j.tplants.2003.09.011.PubMedView ArticleGoogle Scholar
- Caldwell MM, Ballaré CL, Bornman JF, Flint SD, Björn LO, Teramura AH, Kulandaivelu G, Tevini M: Terrestrial ecosystems, increased solar ultraviolet radiation and interactions with other climatic change factors. Photochem Photobiol Sci. 2003, 2 (1): 29-38. 10.1039/b211159b.PubMedView ArticleGoogle Scholar
- Casati P, Walbot V: Gene expression profiling in response to ultraviolet radiation in maize genotypes with varying flavonoid content. Plant Physiol. 2003, 132 (4): 1739-1754. 10.1104/pp.103.022871.PubMedPubMed CentralView ArticleGoogle Scholar
- Ulm R, Baumann A, Oravecz A, Máté Z, Ádám É, Oakeley EJ, Schafer E, Nagy F: Genome-wide analysis of gene expression reveals function of the bZIP transcription factor HY5 in the UV-B response of Arabidopsis. Proc Natl Acad Sci USA. 2004, 101 (5): 1397-1402. 10.1073/pnas.0308044100.PubMedPubMed CentralView ArticleGoogle Scholar
- Bornman JF, Reuber S, Cen YP, Weissenböck G: Ultraviolet radiation as a stress factor and the role of protective pigments. Plants and UV-B: Responses to Environmental Change. Edited by: Lumsden PJ. 1997, Cambridge: Cambridge University Press, 157-168. full_text.View ArticleGoogle Scholar
- Logemann E, Wu SC, Schroder J, Schmelzer E, Somssich IE, Hahlbrock K: Gene activation by UV light, fungal elicitor or fungal infection in Petroselinum crispum is correlated with repression of cell cycle-related genes. Plant J. 1995, 8 (6): 865-876. 10.1046/j.1365-313X.1995.8060865.x.PubMedView ArticleGoogle Scholar
- Berli F, D'Angelo J, Cavagnaro B, Bottini R, Wuilloud R, Silva MF: Phenolic Composition in Grape (Vitis vinifera L. cv. Malbec) Ripened with Different Solar UV-B Radiation Levels by Capillary Zone Electrophoresis. J Agric Food Chem. 2008, 56 (9): 2892-2898. 10.1021/jf073421+.PubMedView ArticleGoogle Scholar
- Berli F, Moreno D, Piccoli P, Hespanhol-Viana L, Silva MF, Bressan-Smith R, Cavagnaro B, Bottini R: Abscisic acid is involved in the response of grape (Vitis vinifera L.) cv. Malbec leaf tissues to ultraviolet-B radiation by enhancing ultraviolet-absorbing compounds, antioxidant enzymes and membrane sterols. Plant Cell Environ. 2009, 33 (1): 1-10.PubMedGoogle Scholar
- Brosché M, Schuler MA, Kalbina I, Connor L, Strid Å: Gene regulation by low level UV-B radiation: identification by DNA array analysis. Photochem Photobiol Sci. 2002, 1 (9): 656-664. 10.1039/b202659g.PubMedView ArticleGoogle Scholar
- Casati P, Stapleton AE, Blum JE, Walbot V: Genome-wide analysis of high-altitude maize and gene knockdown stocks implicates chromatin remodeling proteins in response to UV-B. Plant J. 2006, 46 (4): 613-627. 10.1111/j.1365-313X.2006.02721.x.PubMedView ArticleGoogle Scholar
- Hectors K, Prinsen E, De Coen W, Jansen MAK, Guisez Y: Arabidopsis thaliana plants acclimated to low dose rates of ultraviolet B radiation show specific changes in morphology and gene expression in the absence of stress symptoms. New Phytol. 2007, 175 (2): 255-270. 10.1111/j.1469-8137.2007.02092.x.PubMedView ArticleGoogle Scholar
- Kilian J, Whitehead D, Horak J, Wanke D, Weinl S, Batistic O, D'Angelo C, Bornberg-Bauer E, Kudla J, Harter K: The AtGenExpress global stress expression data set: protocols, evaluation and model data analysis of UV-B light, drought and cold stress responses. Plant J. 2007, 50 (2): 347-363. 10.1111/j.1365-313X.2007.03052.x.PubMedView ArticleGoogle Scholar
- Deluc LG, Grimplet J, Wheatley MD, Tillett RL, Quilici DR, Osborne C, Schooley DA, Schlauch KA, Cushman JC, Cramer GR: Transcriptomic and metabolite analyses of Cabernet Sauvignon grape berry development. BMC Genomics. 2007, 8: 429-10.1186/1471-2164-8-429.PubMedPubMed CentralView ArticleGoogle Scholar
- Grimplet J, Deluc LG, Tillett RL, Wheatley MD, Schlauch KA, Cramer GR, Cushman JC: Tissue-specific mRNA expression profiling in grape berry tissues. BMC Genomics. 2007, 8: 187-10.1186/1471-2164-8-187.PubMedPubMed CentralView ArticleGoogle Scholar
- Terrier N, Glissant D, Grimplet J, Barrieu F, Abbal P, Couture C, Ageorges A, Atanassova R, Léon C, Renaudin JP, et al: Isogene specific oligo arrays reveal multifaceted changes in gene expression during grape berry (Vitis vinifera L.) development. Planta. 2005, 222 (5): 832-847. 10.1007/s00425-005-0017-y.PubMedView ArticleGoogle Scholar
- Cramer GR, Ergül A, Grimplet J, Tillett RL, Tattersall EAR, Bohlman MC, Vincent D, Sonderegger J, Evans J, Osborne C, et al: Water and salinity stress in grapevines: early and late changes in transcript and metabolite profiles. Funct Integr Genomics. 2007, 7 (2): 111-134. 10.1007/s10142-006-0039-y.PubMedView ArticleGoogle Scholar
- Espinoza C, Vega A, Medina C, Schlauch K, Cramer G, Arce-Johnson P: Gene expression associated with compatible viral diseases in grapevine cultivars. Funct Integr Genomics. 2007, 7 (2): 95-110. 10.1007/s10142-006-0031-6.PubMedView ArticleGoogle Scholar
- Figueiredo A, Fortes AM, Ferreira S, Sebastiana M, Choi YH, Sousa L, Acioli-Santos B, Pessoa F, Verpoorte R, Pais MS: Transcriptional and metabolic profiling of grape (Vitis vinifera L.) leaves unravel possible innate resistance against pathogenic fungi. J Exp Bot. 2008, 59 (12): 3371-3381. 10.1093/jxb/ern187.PubMedView ArticleGoogle Scholar
- Thimm O, Blasing O, Gibon Y, Nagel A, Meyer S, Kruger P, Selbig J, Muller LA, Rhee SY, Stitt M: MAPMAN: a user-driven tool to display genomics data sets onto diagrams of metabolic pathways and other biological processes. Plant J. 2004, 37 (6): 914-939. 10.1111/j.1365-313X.2004.02016.x.PubMedView ArticleGoogle Scholar
- Green R, Fluhr R: UV-B-induced PR-1 accumulation is mediated by active oxygen species. Plant Cell. 1995, 7 (2): 203-212. 10.1105/tpc.7.2.203.PubMedPubMed CentralView ArticleGoogle Scholar
- Ballaré CL, Rousseaux MC, Searles PS, Zaller JG, Giordano CV, Robson TM, Caldwell MM, Sala OE, Scopel AL: Impacts of solar ultraviolet-B radiation on terrestrial ecosystems of Tierra del Fuego (southern Argentina). An overview of recent progress. J Photochem Photobiol B: Biol. 2001, 62: 67-77. 10.1016/S1011-1344(01)00152-X.View ArticleGoogle Scholar
- Jeandet P, Douillet-Breuil AC, Bessis R, Debord S, Sbaghi M, Adrian M: Phytoalexins from the Vitaceae: biosynthesis, phytoalexin gene expression in transgenic plants, antifungal activity, and metabolism. J Agric Food Chem. 2002, 50 (10): 2731-2741. 10.1021/jf011429s.PubMedView ArticleGoogle Scholar
- Langcake P, Pryce RJ: A new class of phytoalexins from grapevines. Experientia. 1977, 33 (2): 151-152. 10.1007/BF02124034.PubMedView ArticleGoogle Scholar
- Lijavetzky D, Almagro L, Belchi-Navarro S, Martínez-Zapater JM, Bru R, Pedreno MA: Synergistic effect of methyljasmonate and cyclodextrin on stilbene biosynthesis pathway gene expression and resveratrol production in Monastrell grapevine cell cultures. BMC Res Notes. 2008, 1: 132-10.1186/1756-0500-1-132.PubMedPubMed CentralView ArticleGoogle Scholar
- Tang K, Zhan JC, Yang HR, Huang WD: Changes of resveratrol and antioxidant enzymes during UV-induced plant defense response in peanut seedlings. J Plant Physiol. 2010, 167 (2): 95-102. 10.1016/j.jplph.2009.07.011.PubMedView ArticleGoogle Scholar
- Jansen MAK: Ultraviolet-B radiation effects on plants: induction of morphogenic responses. Physiol Plant. 2002, 116 (3): 423-429. 10.1034/j.1399-3054.2002.1160319.x.View ArticleGoogle Scholar
- Ludwig AA, Saitoh H, Felix G, Freymark G, Miersch O, Wasternack C, Boller T, Jones JDG, Romeis T: Ethylene-mediated cross-talk between calcium-dependent protein kinase and MAPK signaling controls stress responses in plants. Proc Natl Acad Sci USA. 2005, 102 (30): 10736-10741. 10.1073/pnas.0502954102.PubMedPubMed CentralView ArticleGoogle Scholar
- Park JM, Park CJ, Lee SB, Ham BK, Shin R, Paek KH: Overexpression of the tobacco Tsi1 gene encoding an EREBP/AP2-type transcription factor enhances resistance against pathogen attack and osmotic stress in tobacco. Plant Cell. 2001, 13 (5): 1035-1046. 10.1105/tpc.13.5.1035.PubMedPubMed CentralView ArticleGoogle Scholar
- Hao D, Ohme-Takagi M, Sarai A: Unique mode of GCC box recognition by the DNA-binding domain of ethylene-responsive element-binding factor (ERF domain) in plants. J Biol Chem. 1998, 273 (41): 26857-26861. 10.1074/jbc.273.41.26857.PubMedView ArticleGoogle Scholar
- Chervin C, El-Kereamy A, Roustan JP, Latché A, Lamon J, Bouzayen M: Ethylene seems required for the berry development and ripening in grape, a non-climacteric fruit. Plant Sci. 2004, 167 (6): 1301-1305. 10.1016/j.plantsci.2004.06.026.View ArticleGoogle Scholar
- Olsen AN, Ernst HA, Leggio LL, Skriver K: DNA-binding specificity and molecular functions of NAC transcription factors. Plant Sci. 2005, 169 (4): 785-797. 10.1016/j.plantsci.2005.05.035.View ArticleGoogle Scholar
- Olsen AN, Ernst HA, Leggio LL, Skriver K: NAC transcription factors: structurally distinct, functionally diverse. Trends Plant Sci. 2005, 10: 79-87. 10.1016/j.tplants.2004.12.010.PubMedView ArticleGoogle Scholar
- Eulgem T, Rushton PJ, Robatzek S, Somssich IE: The WRKY superfamily of plant transcription factors. Trends Plant Sci. 2000, 5 (5): 199-206. 10.1016/S1360-1385(00)01600-9.PubMedView ArticleGoogle Scholar
- Eulgem T, Somssich IE: Networks of WRKY transcription factors in defense signaling. Curr Opin Plant Biol. 2007, 10 (4): 366-371. 10.1016/j.pbi.2007.04.020.PubMedView ArticleGoogle Scholar
- Seki M, Narusaka M, Ishida J, Nanjo T, Fujita M, Oono Y, Kamiya A, Nakajima M, Enju A, Sakurai T, et al: Monitoring the expression profiles of 7000 Arabidopsis genes under drought, cold and high-salinity stresses using a full-length cDNA microarray. Plant J. 2002, 31 (3): 279-292. 10.1046/j.1365-313X.2002.01359.x.PubMedView ArticleGoogle Scholar
- Babosha AV: Inducible lectins and plant resistance to pathogens and abiotic stress. Biochem (Moscow). 2008, 73 (7): 812-825. 10.1134/S0006297908070109.View ArticleGoogle Scholar
- Mishkind M, Keegstra K, Palevitz BA: Distribution of wheat germ agglutinin in young wheat plants. Plant Physiol. 1980, 66: 950-955. 10.1104/pp.66.5.950.PubMedPubMed CentralView ArticleGoogle Scholar
- Hoson T, Masuda Y: Concanavalin A inhibits auxin-induced elongation and breakdown of (1-3), (1-4) beta-D-glucans in segments of rice coleoptiles. Plant Cell Physiol. 1995, 36: 517-523.Google Scholar
- Bhaglal P, Singh P, Bhullar SS, Kumar S: Drought stress induced accumulation of wheat germ agglutinin in the developing embryos of wheat (Triticum aestivum L.) and its likely role in vivo. J Plant Physiol. 1998, 153: 163-166.View ArticleGoogle Scholar
- Swindell WR, Huebner M, Weber AP: Transcriptional profiling of Arabidopsis heat shock proteins and transcription factors reveals extensive overlap between heat and non-heat stress response pathways. BMC genomics. 2007, 8 (1): 125-10.1186/1471-2164-8-125.PubMedPubMed CentralView ArticleGoogle Scholar
- Gurley WB: HSP101: a key component for the acquisition of thermotolerance in plants. Plant Cell. 2000, 12 (4): 457-460. 10.1105/tpc.12.4.457.PubMedPubMed CentralView ArticleGoogle Scholar
- Miller G, Mittler R: Could heat shock transcription factors function as hydrogen peroxide sensors in plants?. Ann Bot. 2006, 98 (2): 279-288. 10.1093/aob/mcl107.PubMedPubMed CentralView ArticleGoogle Scholar
- Britt AB: DNA damage and repair in plants. Annu Rev Plant Biol. 1996, 47 (1): 75-100. 10.1146/annurev.arplant.47.1.75.View ArticleGoogle Scholar
- Molinier J, Oakeley EJ, Niederhauser O, Kovalchuk I, Hohn B: Dynamic response of plant genome to ultraviolet radiation and other genotoxic stresses. Mutat Res. 2005, 571 (1-2): 235-247.PubMedView ArticleGoogle Scholar
- Hirt H, Mink M, Pfosser M, Bogre L, Gyorgyey J, Jonak C, Gartner A, Dudits D, Heberle-Bors E: Alfalfa cyclins: differential expression during the cell cycle and in plant organs. Plant Cell. 1992, 4 (12): 1531-1538. 10.1105/tpc.4.12.1531.PubMedPubMed CentralView ArticleGoogle Scholar
- Kinoshita T, Doi M, Suetsugu N, Kagawa T, Wada M, Shimazaki K: Phot1 and phot2 mediate blue light regulation of stomatal opening. Nature. 2001, 414: 656-660. 10.1038/414656a.PubMedView ArticleGoogle Scholar
- Sakai T, Kagawa T, Kasahara M, Swartz TE, Christie JM, Briggs WR, Wada M, Okada K: Arabidopsis nph1 and npl1: blue light receptors that mediate both phototropism and chloroplast relocation. Proc Natl Acad Sci USA. 2001, 98: 6969-6974. 10.1073/pnas.101137598.PubMedPubMed CentralView ArticleGoogle Scholar
- Inada S, Ohgishi M, Mayama T, Okada K, Sakai T: RPT2 Is a signal transducer involved in phototropic response and stomatal opening by association with phototropin 1 in Arabidopsis thaliana. Plant Cell. 2004, 16: 887-896. 10.1105/tpc.019901.PubMedPubMed CentralView ArticleGoogle Scholar
- Inouea SI, Kinoshitaa T, Takemiyaa A, Doib M, Shimazakia KI: Leaf positioning of Arabidopsis in response to blue Light. Mol Plant. 2008, 1 (1): 15-26. 10.1093/mp/ssm001.View ArticleGoogle Scholar
- de Torres-Zabala M, Truman W, Bennett MH, Lafforgue G, Mansfield JW, Egea PR, Bögre L, Grant M: Pseudomonas syringae pv. tomato hijacks the Arabidopsis abscisic acid signalling pathway to cause disease. EMBO J. 2007, 26 (5): 1434-10.1038/sj.emboj.7601575.PubMedPubMed CentralView ArticleGoogle Scholar
- Zhu JK: Salt and drought stress signal transduction in plants. Annu Rev Plant Biol. 2002, 53 (1): 247-273. 10.1146/annurev.arplant.53.091401.143329.PubMedPubMed CentralView ArticleGoogle Scholar
- Liu Y, Zhong ZC: Interactive effects of a-NAA and UV-B radiation on the endogenous hormone contents and growth of Trichosanthes kirilowii Maxim seedlings. Acta Ecologica Sinica. 2009, 29: 244-248. 10.1016/j.chnaes.2009.08.007.View ArticleGoogle Scholar
- Pocock KF, Hayasaka Y, McCarthy M, Waters EJ: Thaumatin-like proteins and chitinases, the haze-forming proteins of wine, accumulate during ripening of grape (Vitis vinifera) berries and drought stress does not affect the final levels per berry at maturity. J Agric Food Chem. 2000, 48: 1637-1643. 10.1021/jf9905626.PubMedView ArticleGoogle Scholar
- Salzman RA, Tikhonova I, Bordelon BP, Hasegawa PM, Bressan RA: Coordinate accumulation of antifungal proteins and hexoses constitutes a developmentally controlled defense response during fruit ripening in grape. Plant Physiol. 1998, 117: 465-472. 10.1104/pp.117.2.465.PubMedPubMed CentralView ArticleGoogle Scholar
- Goes da Silva F, Iandolino A, Al-Kayal F, Bohlmann MC, Cushman MA, Lim H, Ergul A, Figueroa R, Kabuloglu EK, Osborne C, et al: Characterizing the Grape Transcriptome. Analysis of Expressed Sequence Tags from Multiple Vitis Species and Development of a Compendium of Gene Expression during Berry Development. Plant Physiol. 2005, 139 (2): 574-597. 10.1104/pp.105.065748.View ArticleGoogle Scholar
- Beyer A: Sequence analysis of the AAA protein family. Protein Sci. 1997, 6 (10): 2043-2058. 10.1002/pro.5560061001.PubMedPubMed CentralView ArticleGoogle Scholar
- Ogura T, Wilkinson AJ: AAA+ superfamily ATPases: common structure--diverse function. Genes to Cells. 2001, 6 (7): 575-597. 10.1046/j.1365-2443.2001.00447.x.PubMedView ArticleGoogle Scholar
- Wang D, Pajerowska-Mukhtar K, Culler AH, Dong X: Salicylic acid inhibits pathogen growth in plants through repression of the auxin signaling pathway. Curr Biol. 2007, 17: 1784-1790. 10.1016/j.cub.2007.09.025.PubMedView ArticleGoogle Scholar
- Bari R, Jones JDG: Role of plant hormones in plant defence responses. Plant Mol Biol. 2009, 69 (4): 473-488. 10.1007/s11103-008-9435-0.PubMedView ArticleGoogle Scholar
- Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant. 2000, 15: 473-497. 10.1111/j.1399-3054.1962.tb08052.x.View ArticleGoogle Scholar
- National Center for Biotechnology Information. [http://www.ncbi.nlm.nih.gov/]
- Grape Genome Browser (12X). [http://www.genoscope.cns.fr/externe/GenomeBrowser/Vitis/]
- Gene Expression Profile Analysis Suite (GEPAS). [http://gepas.bioinfo.cipf.es/]
- Tarraga J, Medina I, Carbonell J, Huerta-Cepas J, Minguez P, Alloza E, Al-Shahrour F, Vegas-Azcarate S, Goetz S, Escobar P, et al: GEPAS, a web-based tool for microarray data analysis and interpretation. Nucleic Acids Res. 2008, W308-314. 10.1093/nar/gkn303. 36 Web ServerGoogle Scholar
- BIOCONDUCTOR, an open source software for bioinformatics. [http://www.bioconductor.org/]
- Irizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, Scherf U, Speed TP: Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics. 2003, 4 (2): 249-264. 10.1093/biostatistics/4.2.249.PubMedView ArticleGoogle Scholar
- Bolstad BM, Irizarry RA, Astrand M, Speed TP: A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics. 2003, 19 (2): 185-193. 10.1093/bioinformatics/19.2.185.PubMedView ArticleGoogle Scholar
- Valls J, Grau M, Sole X, Hernandez P, Montaner D, Dopazo J, Peinado MA, Capella G, Moreno V, Pujana MA: CLEAR-test: combining inference for differential expression and variability in microarray data analysis. J Biomed Inform. 2008, 41 (1): 33-45. 10.1016/j.jbi.2007.05.005.PubMedView ArticleGoogle Scholar
- Schneider M, Lane L, Boutet E, Lieberherr D, Tognolli M, Bougueleret L, Bairoch A: The UniProtKB/Swiss-Prot knowledgebase and its Plant Proteome Annotation Program. J Proteomics. 2009, 72 (3): 567-573. 10.1016/j.jprot.2008.11.010.PubMedPubMed CentralView ArticleGoogle Scholar
- Ashburner M, Ball CA, Blake JA, Botstein D, Butler H, Cherry JM, Davis AP, Dolinski K, Dwight SS, Eppig JT, et al: Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. Nat Genet. 2000, 25 (1): 25-29. 10.1038/75556.PubMedPubMed CentralView ArticleGoogle Scholar
- Kanehisa M, Goto S, Kawashima S, Okuno Y, Hattori M: The KEGG resource for deciphering the genome. Nucleic acids research. 2004, D277-10.1093/nar/gkh063. 32 DatabaseGoogle Scholar
- Cosgrove DJ: Relaxation in a high-stress environment: The molecular bases of extensible cell walls and cell enlargement. Plant Cell. 1997, 9: 1031-1041. 10.1105/tpc.9.7.1031.PubMedPubMed CentralView ArticleGoogle Scholar
- Lijavetzky D, Almagro L, Belchi-Navarro S, Martinez-Zapater JM, Bru R, Pedreno MA: Synergistic effect of methyljasmonate and cyclodextrin on stilbene biosynthesis pathway gene expression and resveratrol production in Monastrell grapevine cell cultures. BMC Res Notes. 2008, 1: 132-10.1186/1756-0500-1-132.PubMedPubMed CentralView ArticleGoogle Scholar
- ArrayExpress database. [http://www.ebi.ac.uk/microarray-as/ae/]
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.