Effect of Culture Conditions on Conidia Formation by Elsinoë ampelina, the Causal Organism of Grapevine Anthracnose.

Elsinoë ampelina, the causal organism of grapevine anthracnose, can be easily grown in culture, yet its sporulation is poor and unstable in culture. In this study, we sought the optimum conditions for a simple method to stably generate conidia. We first examined the optimum period of incubation for young colonies grown on potato dextrose agar (PDA) in water. The resultant number of conidia showed a logarithmic increase, which slowed at about 8 to 10 h. This suggests that 8 to 10 h of preculture would provide a sufficient number of conidia under the culture conditions used. A high negative correlation between colony density on PDA and the number of resultant conidia existed: colonies grown at >2.5 colonies per cm2 produced few or no conidia, whereas those grown at <1.0 colony per cm2 stably produced as many as 2.9 × 106 conidia. The optimum condition for preculture was to incubate colonies grown for 6 days on PDA at a density of <1.0 colony per cm2. The conidia obtained by our method were pathogenic on the grape cultivar Rizamat.

Genomic and genetic analysis of Myb-related genes that regulate anthocyanin biosynthesis in grape berry skin.

As a result of natural hybridization and human selection over millennia, the skin colors of grapes have become greatly diversified. The color is determined by the quantity and composition of anthocyanins. Color-skinned cultivars accumulate anthocyanins in their skins, whereas white-skinned cultivars do not. Myb-related transcription-factor genes such as VvmybA1 regulate anthocyanin biosynthesis. VvMYBA2r, VlmybA1-1, VlmybA1-2, and VlmybA2, which are homologs of VvmybA1, also regulate anthocyanin biosynthesis. In this study, we isolated a novel Myb-related sequence, VlmybA1-3, from cultivars of Vitis labruscana (Vitis vinifera x Vitis labrusca) by means of inverse PCR, and confirmed by means of transient gene expression assay that the gene regulates anthocyanin biosynthesis in grape berry skin. Seedlings of V. labruscana with two functional haplotypes at a region of berry color loci accumulated more anthocyanins than seedlings with a single functional haplotype. In addition, we investigated the haplotypes at the region in 35 cultivars (both V. vinifera and V. labruscana), and found certain typical characteristics. These findings will contribute to the selection of seedlings with high anthocyanin quantities in breeding programs for wine and table grapes, and will help elucidate the origin and evolution of Vitis species.