Table grapes (cv. Dauphine) at different phenological stages were dusted in a settling tower with dry conidia of Botrytis cinerea. The berries were incubated for periods of 3 to 96 h at high relative humidity (RH; ±93% RH, moist berries), or were covered with a film of water (wet berries). Germination of the solitary conidia, appressorium formation, stilbene and suberin induction by germlings, and germling viability were examined by fluorescence microscopy after each incubation period. Isolation and freezing studies were conducted to determine surface colonization (berries left unsterile) and penetration (surface-disinfested berries). Symptoms were determined on berries incubated at a specific wetness regime, kept dry for 10 days, and then incubated for 4 days at high RH. Microscopic observations indicated that germination was delayed on immature berries, but proceeded at a high rate on mature berries. Growth was invariably restricted on moist berries. Attempted penetration was always direct. Stilbene and suberin were generally induced early and were intense on berries at the pea-size and bunch closure stages. Dieback of conidia and germlings occurred at a significantly higher rate on wet than moist berries, and was more pronounced on immature than on mature berries. The segment isolation and freezing studies showed that infections in grape berry cheeks established by this infection mode were few. Extended incubation periods did not lead to substantially higher rates of surface colonization and skin penetration. Disease symptoms did not develop during the 14-day period on the berries transferred to dry perspex chambers, irrespective of phenological stage, incubation period, or wetness regime. According to these findings, this mode of infection should not contribute to a gradual build-up of secondary inoculum in the vineyard, and to B. cinerea epiphytotics.
Severe late blight epidemics in South Africa in 1995 and 1996 prompted an investigation into the mating type, genotype, and metalaxyl sensitivity of populations of Phytophthora infestans. A country-wide survey was conducted from 1996 to 1998 in which isolates were collected from 101 potato fields (656 isolates) and 16 tomato fields (57 isolates). Six hundred and fifty-seven isolates (600 potato and 57 tomato) were analyzed for mating type, while subsets of isolates were analyzed for genotype at the Glucose-6-phosphate isomerase locus (n = 148), DNA fingerprinting with probe RG-57 (n = 61) and mitochondrial DNA haplotype (n = 20). All isolates tested had the characteristics typical of the pre-1980 population (A1 mating type, 86/100 Gpi genotype, US-1 fingerprint pattern, and mtDNA haplotype I-b) previously found worldwide. Metalaxyl sensitivity testing of 656 potato isolates by the in vitro leaf disk method showed that the frequency of highly resistant isolates (50% effective concentration [EC50] > 200 μg a.i./ml) in potato production regions increased from 35% in 1996 to 51% in 1997. The high frequency of resistant isolates was confined to the southern coastal regions in 1996 and 1997, as well as the western Free State in 1997. Although phenylamides were withdrawn from the southern coastal region in December 1996, screening tests carried out in 1998 indicated that resistance levels remained high (≥83%). Sensitive isolates (EC50 < 40 μg a.i./ml) predominated in the remaining six potato production regions. Screening of 45 isolates collected from tomatoes indicated that no resistant strains were present in the sample tested.
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