Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

A Vitis vinifera WRKY transcription factor, VvWRKY1, enhances Plasmopara viticola resistance in transgenic grapevines through activation of jasmonic acid signalling pathway

ABSTRACT: WRKY genes are transcription factors involved in plant response to pathogen attacks in many plant species. These proteins have been shown to activate expression of defence genes in a salicylic acid- and/or jasmonic acid-dependent signalling pathway. To understand the molecular mechanisms involved in grapevine defence, we previously identified a WRKY gene, VvWRKY1, which was able to enhance tolerance to fungal pathogens when overexpressed in tobacco. To elucidate its role in grapevine, we generated transgenic grapevines that overexpress VvWRKY1. Microarray analyses were performed to compare global gene expression in leaves of the transgenic and wild-type lines. Results showed that expression of genes encoding defence-related proteins was enhanced in the transgenic 35S::VvWRKY1 line. Quantitative RT-PCR analysis confirmed that three genes putatively involved in jasmonic acid signalling pathway, two genes encoding JASMONATE ZIM-domain (JAZ) proteins and one lipoxygenase, are over-expressed. The ability of VvWRKY1 to trans-activate their corresponding promoters was confirmed by transient expression assay in grape protoplasts. After challenging with the downy mildew pathogen Plasmopara viticola, resistance was enhanced in the transgenic line compared to the wild-type line. These results suggest that VvWRKY1 transcription factor is able to control plant disease resistance to one of the main grapevine pathogen by activating jasmonic acid signalling pathway in grapevine.

ORGANISM(S): Vitis vinifera

SUBMITTER: Christian Kappel

PROVIDER: E-MTAB-1077 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Over-expression of VvWRKY1 in grapevines induces expression of jasmonic acid pathway-related genes and confers higher tolerance to the downy mildew.

Marchive Chloé C Léon Céline C Kappel Christian C Coutos-Thévenot Pierre P Corio-Costet Marie-France MF Delrot Serge S Lauvergeat Virginie V

PloS one 20130114 1

Most WRKY transcription factors activate expression of defence genes in a salicylic acid- and/or jasmonic acid-dependent signalling pathway. We previously identified a WRKY gene, VvWRKY1, which is able to enhance tolerance to fungal pathogens when it is overexpressed in tobacco. The present work analyzes the effects of VvWRKY1 overexpression in grapevine. Microarray analysis showed that genes encoding defence-related proteins were up-regulated in the leaves of transgenic 35S::VvWRKY1 grapevines. ...[more]

PMID: 23342101

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Project description:The experiment is aiming to analyze the effect of overexpression of the Arabidopsis At1g06160 AP2/ERF-domain transcription factor on gene expression and the involvement of the At1g06160-regulated genes in the jasmonic acid or jasmonic acid/ethylene signal transduction pathways. The experiment was performed with 12 hybridizations, using 24 samples of species [Arabidopsis thaliana], using 12 arrays of array design [Agilent Arabidopsis 3 Oligo Microarray [G4142A]], producing 12 raw data files and 0 transformed and/or normalized data files. In the first two hybridizations (#1 and #2), two independent transgenic Arabidopsis lines containing a construct carrying the Arabidopsis gene At1g06160 fused to an inducible promoter were used. This promoter is activated after the addition of the chemical estradiol to the plant medium. Addition of estradiol results in overexpression of At1g06160. For each independent line, total RNA from estradiol-treated and -untreated plants were collected and labeled with Cy3 and Cy5, respectively, and hybridization was performed for each At1g06160-inducible line. Two other hybridizations (#3 and #4) were performed using two independent transgenic Arabidopsis control lines containing a construct carrying the GUS gene fused to an inducible promoter. For each independent line, total RNA from estradiol-treated and -untreated plants were collected and labeled with Cy3 and Cy5, respectively and hybridization was performed for each GUS-inducible line. The last 8 hybridizations were performed using wild-type Arabidopsis plants. The aim was analyze the effect of the jasmonic acid hormone or the effect of a combination of the jasmonic acid and ethylene hormones on gene expression in wild-type. Hybridizations #5 and #6: wild-type Arabidopsis seedlings were treated with jasmonic acid (JA) or with the control DMSO and total RNA was collected after 8 hours. Total RNA from JA-treated and DMSO-treated plants were collected and labeled with Cy3 and Cy5, respectively, and hybridization was performed. A second hybridization was performed using independent biological samples that were treated and labeled similarly. Hybridizations #7 and #8: wild-type Arabidopsis seedlings were treated with jasmonic acid (JA) or with the control DMSO and total RNA was collected after 24 hours. Total RNA from JA-treated and DMSO-treated plants were collected and labeled with Cy3 and Cy5, respectively, and hybridization was performed. A second hybridization was performed using independent biological samples that were treated and labeled similarly. Hybridizations #9 and #10: wild-type Arabidopsis seedlings were treated with jasmonic acid (JA) and ethylene or with the control DMSO/Na-phosphate and total RNA was collected after 8 hours. Total RNA from JA/ethylene-treated and DMSO/Na-phosphate-treated plants were collected and labeled with Cy3 and Cy5, respectively, and hybridization was performed. A second hybridization was performed using independent biological samples that were treated and labeled similarly. Hybridizations #11 and #12: wild-type Arabidopsis seedlings were treated with jasmonic acid (JA) and ethylene or with the control DMSO/Na-phosphate and total RNA was collected after 24 hours. Total RNA from JA/ethylene-treated and DMSO/Na-phosphate-treated plants were collected and labeled with Cy3 and Cy5, respectively, and hybridization was performed. A second hybridization was performed using independent biological samples that were treated and labeled similarly.

Project description:Background: Global climate change, in particular the entailed predicted temperature increase, will noticeably affect plants vegetative and reproductive development. High temperatures alter the composition of the grapevine fruit, one of the most important fruits produced worldwide. This is leading to variable yield and quality, already observed in many growing regions in recent years. However, physiological processes underlying temperature response and tolerance of the grapevine fruit have hardly been investigated. Currently, all studies on fleshy fruits investigating their abiotic stress response on a molecular level were conducted during the day but possible night-specific variations were overlooked. The present study explores the grapevine fruit transcriptomic response at different developmental stages upon heat stress at day and night. Methodology/Principal Results: Short heat stresses (2 h) were applied at day and night to vines bearing clusters sequentially ordered according to the developmental stages along their vertical axis. The recently proposed microvine model was grown in climatic chambers in order to circumvent common constraints and biases introduced in field experiments with perennial vines. Post-véraison berry heterogeneity inside clusters was evaded upon constituting homogenous batches following organic acid and sugar measurements on individual berries. A whole genome transcriptomic approach was subsequently conducted using NimbelGen® 090918 12X microarrays (30K). Results revealed important differences in heat stress responsive pathways according to day or night treatment, in particular regarding genes within phenylpropanoid metabolism. Precise distinction of post-véraison stages led to a stage-specific detection of anthocyanin-related transcripts repressed by heat. Important modifications in cell wall-related processes as well as indications for a heat-induced delay of ripening and sugar accumulation were observed at véraison and reversed in later stages. Conclusion: This first day - night study on heat stress adaption of the grapevine berry shows that the transcriptome of fleshy fruits is differentially affected by abiotic stress at night. The present results emphasize the necessity to include different developmental stages and especially different time points in transcriptomic studies. A total of 12 samples were analyzed representing three berry developmental stages (two after the onset of ripening, one during green growth). At each stage, heat stress was applied at day and night. Controls and treated berry samples were drawn in triplicates (two in duplicates) at day and at night on the microvine dwarf (Dwarf Rapid Cycling and Continous Flowering; DRCF) gibberellin-insensitive (GAI) mutant.

Dataset Information

A Vitis vinifera WRKY transcription factor, VvWRKY1, enhances Plasmopara viticola resistance in transgenic grapevines through activation of jasmonic acid signalling pathway

Publications

Over-expression of VvWRKY1 in grapevines induces expression of jasmonic acid pathway-related genes and confers higher tolerance to the downy mildew.

Similar Datasets

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