Botrytis cinerea Isolation Methods.

Efficient and reproducible isolation of Botrytis species, including Botrytis cinerea and Botrytis pseudocinerea, is essential for studies on fungal biology, epidemiology, and host-pathogen interactions. The protocols described in this chapter provide practical methods for obtaining pure cultures from symptomatic and asymptomatic plant tissues, as well as from single spores and hyphal fragments. Each approach is detailed with step-by-step instructions, highlighting critical parameters for successful isolation and culture maintenance. The symptomatic and asymptomatic tissue methods enable rapid recovery of isolates from field material, whereas the single-spore and hyphal-tip protocols ensure clonal purity and reduce contamination risks. Practical notes and troubleshooting tips are also provided to facilitate reproducibility across laboratories. These methods offer a comprehensive toolkit adaptable to different experimental needs, from basic mycological research to applied plant pathology, ensuring reliable access to genetically and phenotypically stable Botrytis isolates for downstream analyses.