| Title | Cloning and Expression Analysis of a Chitinase Gene Crchi1 from the Mycoparasitic Fungus Clonostachys rosea (syn. Gliocladium roseum) |
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| Author | Zhongwei Gan, Jinkui Yang, Nan Tao, Zefen Yu, and Ke-Qin Zhang* |
| Address | Laboratory for Conservation and Utilization of Bio-resources, and Key Laboratory for Microbial Resources of the Ministry of Education, Yunnan University, Kunming 650091, P. R. China |
| Bibliography | Journal of Microbiology, 45(5),422-430, 2007, |
| DOI | |
| Key Words | carbon starvation, chitinase, Clonostachys rosea, expression, phylogenetic analysis, real-time PCR |
| Abstract | Clonostachys rosea (syn. Gliocladium roseum) is a well-known biocontrol agent and widely distributed around the world. In this study, an endochitinase gene Crchi1 was isolated from the mycoparasitic fungus C. rosea using the DNA walking strategy. The Crchi1 ORF is 1,746 bp long and interrupted by three introns. The cloned gene Crchi1 encodes 426 amino acid residues and shares a high degree of similarity with other chitinases from entomopathogenic and mycoparasitic fungi. Several putative binding sites for transcriptional regulation of Crchi1 in response to carbon (5''-SYGGRG-3'') and nitrogen (5''-GATA-3'') were identified in the upstream of Crchi1. Expression of Crchi1 gene in different carbon sources was analyzed using real-time PCR (RT-PCR). We found that the Crchi1 expression was suppressed by glucose but strongly stimulated by chitin or solubilized components of the cell wall from Rhizoctonia solani. Phylogenetic analysis of chitinases from entomopathogenic and mycoparasitic fungi suggests that these chitinases have probably evolved from a common ancestor. |
| Download PDF | 45(5)_08.pdf |