Culture and Maintenance Methods for Botrytis cinerea.

Culturing and maintaining Botrytis cinerea in vitro is essential for advancing research on fungal biology, host-pathogen interactions, and plant disease management. Reliable culture methods provide the basis for reproducible experiments, ensuring stable growth and long-term preservation of isolates. This chapter details standardized procedures for establishing fungal colonies on agar and broth media, including Universal Beer Agar, Potato Dextrose Agar, and Czapek-Dox Agar, as well as specialized formulations such as Carrot Agar and Honey Peptone Agar. Protocols for short-term maintenance on agar slants and long-term storage in glycerol at -80°C are described to ensure strain viability over time. Critical variables affecting fungal growth, such as temperature, light, pH, and medium composition, are also discussed, providing guidance for optimizing culture conditions. Together, these methods serve as practical tools for laboratories aiming to maintain pure B. cinerea cultures for physiological, genetic, and pathological studies.