Journal
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
Volume 163, Issue 8, Pages 937-945Publisher
HUMANA PRESS INC
DOI: 10.1007/s12010-010-9097-7
Keywords
Trichoderma harzianum; Biocontrol; Agrobacterium tumefaciens
Funding
- Chinese High Tech Program (863) [2003AA241140, 2006AA10Z424]
- National Natural Science Foundation of China [81000688]
- China Postdoctoral Science Foundation [20090460911]
- Technology Research Foundation of the Education Department of Heilongjiang Province [11551151]
- Foundation of Key Laboratories of Myocardial Ischemia Mechanism and Treatment (Harbin Medical University), Ministry of Education
- Research Foundation of the Health Department of Heilongjiang Province [2009-217]
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As a soil-borne filamentous fungus, Trichoderma harzianum exhibits biological control properties because it parasitizes a large variety of phytopathogenic fungi. In this study, the vectors pBI121 and pCAMBIA1301 and cloning vector pUC18 were used to successfully construct expression vector pCA-GChiV for filamentous fungi transformation mediated by Agrobacterium tumefaciens.The ChiV gene was successfully transferred into the biocontrol fungus T. harzianum with an efficiency of 90-110 transformants per 10(7) spores using A. tumefaciens-mediated transformation. Putative transformants were analyzed to test the transformation by the southern blot, and the expression of ChiV was detected by reverse transcription PCR. The transformants were co-cultured to assay antifungal activities with Rhizoctonia solani. The inhibition rates of the transformants and no ChiV gene transferred T. harzianum were 98.56% and 82.42%, respectively, on the fourth day.The results showed that the ChiV transformants had significantly higher inhibition activity.
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