The stilbene resveratrol is a stress metabolite produced by Vitis vinifera grapevines during fungal infection, wounding or UV radiation. Resveratrol is synthesised particularly in the skins of grape berries and only trace amounts are present in the fruit flesh. Red wine contains a much higher resveratrol concentration than white wine, due to skin contact during fermentation. Apart from its antifungal characteristics, resveratrol has also been shown to have cancer chemopreventive activity and to reduce the risk of coronary heart disease. It acts as an antioxidant and anti-mutagen and has the ability to induce specific enzymes that metabolise carcinogenic substances. The objective of this pilot study was to investigate the feasibility of developing wine yeasts with the ability to produce resveratrol during fermentation in both red and white wines, thereby increasing the wholesomeness of the product. To achieve this goal, the phenylpropanoid pathway in Saccharomyces cerevisiae would have to be introduced to produce p-coumaroyl-CoA, one of the substrates required for resveratrol synthesis. The other substrate for resveratrol synthase, malonyl-CoA, is already found in yeast and is involved in de novo fatty-acid biosynthesis. We hypothesised that production of p-coumaroyl-CoA and resveratrol can be achieved by co-expressing the coenzyme-A ligase-encoding gene (4CL216) from a hybrid poplar and the grapevine resveratrol synthase gene (vst1) in laboratory strains of S. cerevisiae. This yeast has the ability to metabolise p-coumaric acid, a substance already present in grape must. This compound was therefore added to the synthetic media used for the growth of laboratory cultures. Transformants expressing both the 4CL216 and vst1 genes were obtained and tested for production of resveratrol. Following beta-glucosidase treatment of organic extracts for removal of glucose moieties that are typically bound to resveratrol, the results showed that the yeast transformants had produced the resveratrol beta-glucoside, piceid. This is the first report of the reconstruction of a biochemical pathway in a heterologous host to produce resveratrol.

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BackgroundPolygalacturonase-inhibiting proteins (PGIPs) directly limit the effective ingress of fungal pathogens by inhibiting cell wall-degrading endopolygalacturonases (ePGs). Transgenic tobacco plants over-expressing grapevine (Vitis vinifera) Vvpgip1 have previously been shown to be resistant to Botrytis infection. In this study we characterized two of these PGIP over-expressing lines with known resistance phenotypes by gene expression and hormone profiling in the absence of pathogen infection.ResultsGlobal gene expression was performed by a cross-species microarray approach using a potato cDNA microarray. The degree of potential cross-hybridization between probes was modeled by a novel computational workflow designed in-house. Probe annotations were updated by predicting probe-to-transcript hybridizations and combining information derived from other plant species. Comparing uninfected Vvpgip1-overexpressing lines to wild-type (WT), 318 probes showed significant change in expression. Functional groups of genes involved in metabolism and associated to the cell wall were identified and consequent cell wall analysis revealed increased lignin-levels in the transgenic lines, but no major differences in cell wall-derived polysaccharides. GO enrichment analysis also identified genes responsive to auxin, which was supported by elevated indole-acetic acid (IAA) levels in the transgenic lines. Finally, a down-regulation of xyloglucan endotransglycosylase/hydrolases (XTHs), which are important in cell wall remodeling, was linked to a decrease in total XTH activity.ConclusionsThis evaluation of PGIP over-expressing plants performed under pathogen-free conditions to exclude the classical PGIP-ePG inhibition interaction indicates additional roles for PGIPs beyond the inhibition of ePGs.

Sorghum is particularly drought tolerant compared with other cereal crops and is favoured for subsistence farming in water scarce regions of the world. This study was conducted to identify South African sorghum landraces with superior drought tolerance compared with a drought tolerant breeding line (P898012). Seedlings of 14 South African sorghum landrace accessions were initially screened for drought tolerance by assessing percentage leaf water content (LWC) during progressive water deficit. Four landraces (designated LR5, LR6, LR35 and LR36) recorded higher LWC than P898012.These were subsequently evaluated with P898012 during the reproductive growth stage, for their physiological responses to mild (four days) and severe (six days) water stress treatments and a moderate re-watered treatment on day seven. Plant height, soil moisture and LWC were measured during harvests. Chlorophyll, carotenoid and proline contents were quantified. All five genotypes maintained LWC above 80% during mild and severe stress treatments. For LR35 and LR36, LWC were recorded within 8% less in comparison to their well-watered controls following the moderate re-watered treatment.Significantly higher chlorophyll and carotenoid contents were recorded for both LR6 and LR35 in comparison to P898012 during severe stress. When LWC was reduced in LR36 (to 73.68%) and LR35 (to 73.51%), their proline content significantly increased by 14-and 16-fold, respectively. In this study, we have identified four previously uncharacterised sorghum genotypes exhibiting drought tolerance and described their physiological responses during water deficit and moderate re-watering. Aside from their application to breeding, these landraces are valuable resources to elucidate genetic mechanisms that enable drought tolerance in South African sorghum.3