Botrytis cinerea Infection of Grape Flowers: Light and Electron Microscopical Studies of Infection Sites.

ABSTRACT Specific floral organs including the calyptra, stigma, and receptacle area of glasshouse-grown grapevines (Vitis vinifera cv. Cabernet Sauvignon) were inoculated with aqueous suspensions of Botrytis cinerea conidia, and the initial steps involved in colonization and infection of the host tissues were studied for several days postinoculation using light microscopy as well as scanning and transmission electron microscopy. Conidia germinated on all floral organs examined and became attached to the host surface within 48 h after inoculation. However, in all cases the vast majority of conidia accumulated in a channel-like gap between the ovary and the calyx that extended in a narrowing fashion into the flower interior where the ovary joined the receptacle. Very few conidial germ tubes were detected in the style following inoculation of the stigma, and no evidence for their growth toward the ovaries could be found. In contrast, hyphae were more abundant in the receptacle area, regardless of the site of inoculation. Tips of the calyx became necrotic and mycelium formed in the gap between ovary and calyx within 72 h following inoculation, providing a major point of colonization and infection. B. cinerea colonized dehiscent calyptras within 72 h of inoculation, providing a potential source of inoculum from calyptras that remained stuck in the cluster. The results suggest that the grape flower's receptacle area is the predominant site of infection for B. cinerea, although a minor portion of infections may also occur through the stigma and style.

Botrytis cinerea Infection in Grape Flowers: Defense Reaction, Latency, and Disease Expression.

ABSTRACT Inflorescences of field-grown grapevines (Vitis vinifera L. cv. Gamay) were inoculated with a Botrytis cinerea conidia suspension or dried conidia at different stages during bloom in moist weather. Approximately 10% of the conidia germinated within 72 h, resulting in two to three times more latent infections than uninoculated controls in pea-size (7 mm in diameter) berries. In surface-sterilized pea-size berries, latent B. cinerea was present predominantly in the receptacle area. After veraison, latent B. cinerea also was found in the style and, in mature berries, latent colonies were distributed throughout the pulp. Inoculation at full bloom led to the highest disease severity (66%) at harvest, compared with 38% in controls. Stilbene stress metabolites in the flowers were measured by high-performance liquid chromatography. Resveratrol accumulated mainly after pre-bloom and full-bloom inoculation, but did not prevent infection. Piceid levels did not change following inoculation, while epsilon-viniferin was found in necrotic tissues only, and pterostilbene and alphaviniferin were not detected at all. B. cinerea conidia suspensions also were applied to various locations on flowers of pot-grown cvs. Pinot Noir and Chardonnay. Inoculation of the receptacle area, but not that of the stigma and ovary, resulted in latent infections. Stilbene synthesis was similar to the field results, with resveratrol accumulating mainly in the calyptra and receptacle area. Constitutive soluble phenolic compounds (mainly derivatives of quercetin and hydroxy-cinnamic acid) were present at high concentrations in the calyptra but at low levels in the receptacle area. These experiments confirmed bloom as a critical time for B. cinerea infection in grapes and suggest that the most likely site of infection is the receptacle area or cap scar exposed at anthesis. Stilbenes may have a limited role in inhibition of flower infection and latency in susceptible grape cultivars, and epsilon-viniferin may be a by-product rather than a deterrent of infection.