Physiological and population genetic analysis of Botrytis field isolates from vineyards in Castilla y Leon, Spain

Grey mould is reported in the vineyards of Castilla y Leon, Spain, every year. However, the natural populations of the pathogen have yet to be properly characterized. Vineyards from six wine-producing areas were surveyed in 2002 and 2007, sampling from bunches of grapes with and without symptoms. A total of 283 Botrytis field isolates were selected for physiological and genetic analyses. Botrytis cinerea isolates predominated in the population, although isolates belonging to Botrytis pseudocinerea and Botrytis prunorum were also identified. These two species are recorded for the first time in Spain in this work. In addition, two isolates closely related to Botrytis californica were identified. Physiologically, the B. cinerea population is very diverse, displaying a normal distribution of aggressiveness values in Vitis vinifera leaves, suggesting a quantitative nature for this trait. Several isolates unable to cause infection were identified, most of them belonging to a mycelial morphotype. Population genetic analysis revealed that genotypic diversity is high and that multiple infections of the same bunch of grapes by different genotypes occur frequently. The high genotypic diversity observed, an even distribution of both mating types and the linkage disequilibrium values detected support a mixed mode of reproduction with low levels of clonality. The wine-producing area in which each isolate was collected imposed a low degree of population differentiation, an effect that does not depend solely on the geographic distances but rather on the management practices used by growers and wine producer associations.