Project description:The specificity and synergisms of methyl jasmonate and cyclodextrin elicitor effects in grapevine were analysed at the transcrptomic level. To this end, 12-14 day old cell cultures established from Vitis vinifera L. cv Monastrell calli were elicited with 50 mM cyclodextrin (CD), 100 µM methyl jasmonate (MJ) or the combination of both elicitros (MJ+CD). The experiment was carried out in triplicate and untreated cell cultures were followed in parallel as a control. Cells of each culture were isolated just prior and 24h after the treatment to obtain RNA. GrapeGen GeneChip microarray hybridization was performed from each sample. ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Pablo Carbonell-Bejerano. The equivalent experiment is VV44 at PLEXdb.]
Project description:Plants regenerated from tissue culture frequently show somaclonal variation. In this study we compared the transcriptomic and epigenetic state of embryogenic callus of grapevine with leaves from mature grapevine plants. In particular, we focussed on the expression of transposable elements and changes in siRNA abundance and genome-wide methylation in these tissues.
Project description:Plants regenerated from tissue culture frequently show somaclonal variation. In this study we compared the transcriptomic and epigenetic state of embryogenic callus of grapevine with leaves from mature grapevine plants. In particular, we focussed on the expression of transposable elements and changes in siRNA abundance and genome-wide methylation in these tissues.
Project description:Plants regenerated from tissue culture frequently show somaclonal variation. In this study we compared the transcriptomic and epigenetic state of embryogenic callus of grapevine with leaves from mature grapevine plants. In particular, we focussed on the expression of transposable elements and changes in siRNA abundance and genome-wide methylation in these tissues.
Project description:Somatic variation is a valuable source of trait diversity in clonally propagated crops. In grapevine, which has been clonally propagated worldwide for centuries, important phenotypes such as white berry colour are the result of genetic changes caused by transposable elements. Additionally, epiallele formation may play a role in determining geo-specific (‘terroir’) differences in grapes and thus ultimately in wine. This genomic plasticity might be co-opted for crop improvement via somatic embryogenesis, but that depends on a species-specific understanding of the epigenetic regulation of transposable element (TE) expression and silencing in these cultures. For this reason, we used whole-genome bisulphite sequencing, mRNA sequencing and small RNA sequencing to study the epigenetic status and expression of TEs in embryogenic callus, in comparison with leaf tissue.
Project description: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.
Project description:Mature grapevine berries at the harvesting stage (MB) are very susceptible to the gray mold fungus Botrytis cinerea while veraison berries (VB) are not. We conducted simultaneous microscopic and transcriptomic analyses of the pathogen and the host to investigate the infectious process developed by B. cinerea on MB versus VB, and the plant defense mechanisms deployed to stop the fungus development. On the pathogen side, our genome-wide transcriptomic data revealed that B. cinerea genes up-regulated during infection of MB are enriched in functional categories related to necrotrophy such as degradation of plant cell wall, proteolysis, membrane transport, reactive oxygen species generation and detoxification. Quantitative-PCR on a set of representative genes related to virulence and microscopic observations further demonstrated that the infection is also initiated on VB but stops at the penetration stage. On the plant side, genome-wide transcriptomic analysis and metabolic data revealed a defense pathways switch during berry ripening. In response to B. cinerea infection, VB activated a burst of reactive oxygen species (ROS), the salicylate (SA)-dependent defense pathway, the synthesis of the resveratrol phytoalexin and cell-wall strengthening. In opposite, infected MB activated the jasmonate (JA)-dependent pathway which does not stop the fungal necrotrophic process. Grapevine berries at veraison (VB) and harvesting stages (MB) were inoculated with Botrytis cinerea B05-10 and samples were taken at 24h and 48h post-inoculation. An additional uninfected control sample taken at 0h post-inoculation was included in the experimental design. 3 replicates per sample were performed. The total-RNA samples were labeled and used for hybridization on NimbleGen 12plex Vitis vinifera gene expression array.