Journal
BIOCONTROL SCIENCE AND TECHNOLOGY
Volume 21, Issue 6, Pages 677-686Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/09583157.2011.570430
Keywords
microbial control; cuticle-degrading enzyme; insect fungi; chitinase
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Funding
- National Center for Genetic Engineering and Biotechnology (BIOTEC)
- Thailand Research Fund, under the Royal Golden Jubilee Program
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A gene encoding chitinase was cloned from Ophiocordyceps unilateralis, a Formamidae-specific fungus, collected from Sirindhorn Peat Swamp Forest, Thailand. The O. unilateralis chitinase (OuChi) full-length gene (1311 bp) encodes 436 amino acids with the first 20 amino acids as a putative signal peptide. The gene showed highest identity (78%) to Isaria farinose endochitinase. To investigate if cross-species chitinase expression also enhances fungal toxicity, the mature OuChi gene was subcloned into an Agrobacterium binary vector pPZP-bar and then transformed into Beauveria bassiana strain BCC2659. Chitinase activity was detected using 4-methylumbelliferyl-beta-D-N,N'-diacetylchitobioside. The fungal transformant expressing O. unilateralis chitinase showed higher toxicity against Spodoptera exigua. These results support the hypothesis that chitinolytic enzymes are one of several 'virulence' factors produced by entomopathogenic fungi during host encounter.
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