1887

Abstract

Summary: We have developed a homologous transformation system for the wheat-pathogenic fungus based on a benomyl-(MBC-) resistant allele of the β-tubulin gene. The β-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 β-tubulin gene. This MBC resistant allele of the β-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 β-tubulin. The Agene also conferred MBC resistance on the related species , a pathogen of , following its introduction by cotransformation. Probing digested DNA with Aat low stringency revealed only a single β-tubulin gene. We anticipate that Awill prove a useful tool for the investigation of the pathogenicity of and other fungi.

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/content/journal/micro/10.1099/00221287-137-9-2085
1991-09-01
2019-11-17
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http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-137-9-2085
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