Elsevier

Plant Science

Volume 171, Issue 1, July 2006, Pages 132-138
Plant Science

The ripening wine grape berry skin transcriptome

https://doi.org/10.1016/j.plantsci.2006.03.002Get rights and content

Abstract

Ripening of the grape berry immediately precedes harvesting and is the key phase which determines the composition of wine and table grapes. Although it is apparent that changes in gene expression levels play a role in initiating ripening, it is unknown to what extent gene expression levels differ between cultivars during ripening. We have undertaken a comparison of a selection of wine grape cultivars using a 9200 feature cDNA microarray to gauge the extent to which transcript levels differ between wine grape cultivars in the berry skin during ripening. Clones for microarray slide fabrication were sourced from a cDNA library constructed from mRNA derived from randomly sampled berries. The berries were collected at weekly intervals over four weeks of berry development including véraison. The microarray slides were hybridised with Cy3 and Cy5 labelled cDNA derived from the skin of ripening berries of the cultivars Cabernet Sauvignon, Pinot Noir, Shiraz, Chardonnay, Riesling, Sauvignon Blanc and Semillon. Only a small proportion of the genes within the berry skin showed a three-fold or greater difference in expression level after ripening commenced. Most of the differences appear to arise from environmental signals rather than genome differences.

Introduction

Whilst environmental, agronomic and processing factors influence the character of wine, each wine grape cultivar produces its own distinctive wine [1]. The biochemical basis of cultivar differences have been identified, many of which are manifested in the grape berry and therefore impact on the character of wine. Parameters which have been measured and compared in both the grape berry and wine include protein, amino acid, anthocyanin and tannin content [2]. The amino acid content and profile when measured at similar levels of maturity has been shown to vary in a cultivar dependent manner [3], and the key wine quality determinants of anthocyanin and proanthocyanin content and profile are also cultivar dependent [4], [5].

Ripening of the grape berry is a key growth phase which determines the nature of the material used in the production of both wine and table grapes since it immediately precedes harvesting. Although the initiation of ripening is heavily dependent on gene expression changes [6], [7], hybridisation based technologies suggest once ripening commences, gene expression levels within the berry skin are relatively stable within the first few weeks of this phase [7], [8]. However, it is unknown to what extent gene expression levels differ between cultivars during ripening.

Several comparisons of wine grape cultivars at the genotypic and phenotypic level, including gene expression studies of selected genes and groups of genes, have been undertaken. Until recently, however, no studies had attempted to compare transcript levels of grape cultivars on a genome-wide basis. Burger and Botha [9] used cDNA-AFLP to compare gene expression in Cabernet Sauvignon and Clairette blanche berries early in the ripening phase of berry development, one-week post-véraison, with gene expression late in berry development immediately prior to harvest. Their work identified candidate genes that changed their level of expression over the study period within each of these cultivars and may have identified genes differentially expressed when comparing the two cultivars. However, cDNA-AFLP is based on restriction fragment length differences and consequently sequence banding patterns which suggest differential gene expression between cultivars may simply reflect polymorphisms in selective restriction enzyme sites [9].

In order to determine the extent to which transcript levels differ between wine grape cultivars, we have undertaken a comparison of a selection of cultivars using a 9200 feature cDNA microarray. The cultivars chosen for this study were Cabernet Sauvignon, Pinot Noir, Shiraz, Chardonnay, Riesling, Sauvignon Blanc and Semillon. All are important wine grape cultivars that produce their own distinctive wine in addition to being represented by substantial plantings throughout the world. The work presented here reports upon an attempt to ascertain the degree to which these cultivars differ at the transcript level during the ripening phase of berry development and if the observed differences have implications for functional differences between the cultivars.

Section snippets

Plant materials

Cy3 and Cy5 labelled cDNA hybridised to the microarray slides was derived from the berry skins of Vitis vinifera cultivars Shiraz, Cabernet Sauvignon, Pinot Noir, Chardonnay, Riesling, Sauvignon Blanc and Semillon. With the exception of the cultivar Shiraz, the berries were collected from the Queensland Department of Primary Industries vineyard situated at Stanthorpe, Queensland, Australia, at midday on the same day, 13 weeks post Shiraz flowering during the 1999–2000 growing season. Berry

Results

The berry flesh of all cultivars had soluble solid concentrations of approximately 10% or more indicating all were in the ripening phase of their development (Table 1). The anthocyanin data support this assertion since all cultivars that produce coloured berries were also producing anthocyanins at the time of RNA sampling (Table 2). None of the white grape cultivars (Chardonnay, Riesling, Sauvignon Blanc, Semillon) had detectable levels of anthocyanins.

The threshold of significance for

Discussion

The objective of the work presented here was to determine the degree to which the cultivars under scrutiny differed at the transcript level during the ripening phase of berry development. The first point to note is that for each of the cultivars, only a relatively small number of genes had expression levels that differed from week 13 Shiraz berry skin. Specifically, five of the six cultivars fell within the range of 1.3% (Sauvignon Blanc) to 2.35% (Chardonnay) of array features that were either

Acknowledgements

The authors wish to thank the following: Angelo Puglisi (Ballandean Estate Wines, Ballandean, Qld, Australia) for generous provision of plant material and David Oag (Queensland Horticulture Institute, Applethorpe, Qld, Australia) for general viticultural advice.

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    Current address: Queensland Agricultural Biotechnology Centre, Department of Primary Industries, University of Queensland, St. Lucia, Qld 4072, Australia.

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