1887

Abstract

We have developed a homologous transformation system for the wheat-pathogenic fungus based on a benomyl-(MBC-) resistant allele of the -tubulin gene. The -tubulin gene was isolated by heterologous hybridization from a cosmid library prepared from an MBC-resistant mutant. Cosmids carrying the gene conferred MBC resistance when introduced into a sensitive strain, demonstrating that resistance to MBC fungicides in may be determined by the -tubulin gene. This MBC resistant allele of the -tubulin gene (A) was subcioned into pUC18 and used as a dominant selectable marker for transformation of wild-type sensitive strains. Transformants arose at frequencies of approximately 5 per μg of DNA, were integrative in nature and were mitotically stable. Some transformants showed a marked reduction in vigour, both in the presence and absence of MBC; this is thought to arise from overproduction of -tubulin. The A gene also conferred MBC resistance on the related species , a pathogen of , following its introduction by cotransformation. Probing digested DNA with A at low stringency revealed only a single -tubulin gene. We anticipate that A will prove a useful tool for the investigation of the pathogenicity of and other fungi.

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1991-09-01
2021-08-01
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