Fecha de publicación: -- Fuente: PubMed "Tomato process" Fungal Biol. 2026 Nov;130(7):101868. doi: 10.1016/j.funbio.2026.101868. Epub 2026 Sep 23.ABSTRACTBotrytis cinerea is a polyphagous fungal pathogen affecting a wide range of ornamental and agricultural crops. Previous studies on natural populations have revealed substantial intraspecific morphological, physiological, and genetic diversity. The field isolate B459 exhibits a distinctive mycelial phenotype, characterized by the lack of sporulation and sclerotia formation, and is unable to infect Vitis vinifera or Phaseolus vulgaris leaves, gerbera petals and tomato fruits. Genetic analysis traced these phenotypes to a mutation in the transcription factor gene Bcin04g03490, introducing a premature stop codon and resulting in a truncated protein. To further investigate the function of Bcin04g03490, CRISPR/Cas9-based strategies were employed to generate B459 transformants either carrying a functional B05.10 allele under the control of the constitutive olicC promoter from Aspergillus nidulans integrated at the niaD locus, or repaired at the endogenous locus through a marker-free approach involving targeted cleavage upstream of the repair template. In both cases, restoration of sporulation and pathogenicity was observed. Notably, complemented and repaired strains also recovered the toxin profile characteristic of the reference aggressive B05.10 isolate, including the production of botrydial (sesquiterpene) and botcinins (polyketides), indicating reactivation of key secondary metabolism pathways. These findings extend the functional scope of Bcin04g03490, demonstrating that it controls developmental processes, pathogenicity, and secondary metabolism pathways critical for virulence. Furthermore, this work provides a proof-of-concept for long-range homology-directed repair in B. cinerea at the Bcin04g03490 locus, offering valuable methodological insights for marker-free genome editing in filamentous fungi.PMID:42875135 | DOI:10.1016/j.funbio.2026.101868