Development of an apparatus and protocol for the efficient isolation of bacteria and yeasts that attach to Botrytis cinerea germlings

Summary An apparatus and protocol for the efficient and consistent isolation of bacteria and yeasts with the ability to attach to germlings of Botrytis cinerea is described. The study focused on minimising microbial contamination by bacteria or yeasts which do not attach to the pathogen B. cinerea but which interact with materials used in the equipment and would otherwise be isolated along with target microbes. After development, the assay reduced the contamination rate to 1–2 cells per 100 added and this was found to be a satisfactory level for the selection of microbial attachers to fungal hyphae. One or more phenotypes of microbes that adhered to B. cinerea germlings were found in 97% of the 70 samples collected from phylloplane washings and processed using this assay.