Protocol for Collection, Identification, and Artificial Inoculation of Bipolaris sorokiniana for Functional Analyses of Wheat-Spot Blotch Interactions and Germplasm Screening.

Bipolaris sorokiniana, the causal agent of spot blotch in wheat, is a major constraint in warm and humid wheat-growing regions. This chapter outlines a standardized protocol covering diseased sample collection, pure culture isolation, morphological and molecular confirmation, inoculum preparation, and artificial inoculation for reproducible host-pathogen interaction studies, for which complete and standardized procedures are currently limited. Existing reports are often scattered and lack sufficient methodological detail, thereby restricting reproducibility and consistency across studies. This chapter, therefore, consolidates and provides well-tested protocols from a single source. To address these limitations, the present chapter consolidates well-established and validated procedures into a single, coherent workflow. Unlike earlier approaches, this integrated method ensures uniformity in inoculation assays and supports reliable comparative screening of wheat germplasm under controlled conditions. The current protocol facilitates both fundamental research on host-pathogen interactions and applied breeding efforts aimed at improving resistance to spot blotch.