Gene disruption and characterization of a class V chitin synthase in Botrytis cinerea.
Cell-wall chitin biosynthesis in the phytopathogenic fungus Botrytis cinerea involves 7 classes of chitin synthases. In this study, we disrupted a gene encoding a chitin synthase with a myosin-like motor domain (BcchsV) through Agrobacterium tumefaciens mediated transformation. The resulting mutant was not significantly affected in either growth characteristics or pathogenicity on tomato leaves. Surprisingly, the BcchsV mutant exhibited a 31% (m/m) increase in its chitin content compared with the wild-type strain. In addition, the BcchsV mutant showed increased sensitivity to Calcofluor White and slightly enhanced tolerance to cell-wall disturbing substances and osmosis regulators, including SDS, sorbitol, and NaCl. These results suggest that Bcchs does not play an essential role in the synthesis of cell-wall chitin in B. cinerea. However, disruption of this gene provoked a compensatory mechanism regulating the cellular response to cell-wall damage.
Cui
Zhifeng
Z
College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou 310032, P. R. China.
Ding
Zhenke
Z
Yang
Xiao
X
Wang
Kun
K
Zhu
Tingheng
T
D016960:Agrobacterium tumefaciens / Q000235:genetics; D001557:Benzenesulfonates / Q000494:pharmacology; D020171:Botrytis / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D002473:Cell Wall / Q000737:chemistry; D002686:Chitin / Q000032:analysis / Q000235:genetics; D002687:Chitin Synthase / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D017353:Gene Deletion; D018551:Solanum lycopersicum / Q000382:microbiology; D010802:Phylogeny; D010935:Plant Diseases / Q000382:microbiology
D001557:Benzenesulfonates; D004271:DNA, Fungal; D002686:Chitin; C007061:C.I. Fluorescent Brightening Agent 28; D002687:Chitin Synthase