Surface colonization, penetration, and lesion formation on grapes inoculated fresh or after cold storage with single airborne conidia of Botrytis cinerea
Infection of grapes by different densities of airborne conidia of Botrytis cinerea was investigated on table grapes (cultivar Dauphine) harvested ripe (16°Brix) and inoculated fresh, or after SO2 treatment and 8-week storage at -0.5°C. Berries were detached at each inoculation and dusted with dry conidia in a settling tower. Following inoculation, the fresh berries were incubated for 24 h at high relative humidity (≥93%), or were overlaid with wet sterile paper towels. Cold-stored berries were incubated at high relative humidity. The effect of conidial density on surface colonization, penetration, and lesion formation was determined by surface sterilization, isolation, and freezing studies on fresh berries. Only symptom expression was determined on cold-stored berries. Fluorescence microscopy of skin segments showed that conidia were consistently deposited as single cells, and not in pairs or groups, on berry surfaces. Individual conidia, at all densities tested, readily infected the cold-stored berries and formed separate lesions after 2 days. Although the cold-stored berries were highly susceptible, lesion numbers were not related to conidial density at low inoculum dosages (0.67 to 2.60 conidia per mm2 berry surface). Lesion numbers tended to increase exponentially at higher dosages (3.24 to 3.88 conidia per mm2 berry surface). Individual conidia, however, did not induce any disease symptoms on fresh berries. Removal of the pathogen after 24-h incubation from the surface of fresh berries by ethanol, and subsequent incubation of excised skin segments revealed that, irrespective of the conidial density or the wetness regime, less than 2% of skin segments were penetrated. Furthermore, increasing densities of conidia did not lead to higher rates of surface colonization and skin penetration. The low incidence of disease caused on fresh berries and high disease incidence induced after prolonged cold storage indicated that infection was not governed by conidial density on berry surfaces, but by the level of host resistance.