4.4 Article

Investigating the role of a Verticillium fungicola β-1,6-glucanase during infection of Agaricus bisporus using targeted gene disruption

Journal

FUNGAL GENETICS AND BIOLOGY
Volume 39, Issue 3, Pages 264-275

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/S1087-1845(03)00061-6

Keywords

Verticillium fungicola; Agaricus bisporus; Agrobacterium; targeted gene disruption; enzyme; glucanase

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Studies on the mycopathogen Verticillium fungicola have shown the up-regulation of beta-1,6-glucanases when grown in the presence of host cell walls and host cell wall components including chitin. These cell-wall-degrading enzymes are hypothesized to contribute to the pathogenic ability of mycopathogens. A beta-1,6-glucanase gene, VfGlu1, showing high similarity to beta-1,6-glucanase genes from Hypocrea virens, Neotyphodium sp., and Trichoderma harzianum, was isolated using degenerate PCR from V. fungicola, a serious mycopathogen of the cultivated mushroom Agaricus bisporus. Agrobacterium-mediated transformation of V fungicola using homologous DNA from VfGlu1 resulted in homologous integration at the VfGlu1 locus in 75% of transformants, generating mutants disrupted in the VfGlu1 gene. VfGlu1 mutants displayed reduced virulence and diminished ability to utilize chitin as a carbon source, implicating VfGlu1 in the disease process. Agrobacterium-mediated transformation affords an efficient technique for the disruption of genes associated with disease symptom development in the complex V. fungicola-A. bisporus interaction. (C) 2003 Elsevier Science (USA). All rights reserved.

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