Fluorescence and Ultrastructural Microscopy for Botrytis cinerea

Botrytis cinerea, a notorious pathogenic fungus, leads to postharvest decay in horticultural crops and substantial economic losses, while the pathogenicity of B. cinerea is closely related to its structural characteristics and cellular homeostasis. Among the currently available techniques, fluorescence and ultrastructural observations serve as crucial tools for investigating subcellular structures and cellular events during B. cinerea growth. These techniques enable the monitoring of cell structural integrity, such as variations in the cell membrane and cell wall, as well as intracellular changes, such as reactive oxygen species bursts and cell apoptosis. The results at cytological levels may not only facilitate close-up dissection of B. cinerea cells, but also provide important cues to uncover the inhibitory mechanism of exogenous treatments. This chapter introduces fluorescence staining and observation, as well as the preparation of ultrathin sections of B. cinerea conidia, aiming to provide methodological guidance for cytological examinations of B. cinerea.