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Agrobacterium tumefaciens Mediated Transformation of ChiV Gene to Trichoderma harzianum

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Abstract

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 107 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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Acknowledgements

This work was supported by the Chinese High Tech Program (863) (2003AA241140 and 2006AA10Z424), the National Natural Science Foundation of China (81000688), China Postdoctoral Science Foundation funded project (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 and The Research Foundation of the Health Department of Heilongjiang Province (2009-217).

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Authors and Affiliations

  1. Post-doctoral Research station of Chemical Engineering and Technology, Harbin Institute of Technology, 150001, Harbin, Heilongjiang, China

    Liming Yang & Kening Sun

  2. Department of Pathophysiology, Key Laboratories of Myocardial Ischemia Mechanism and Treatment (Harbin Medical University), Ministry of Education, 150086, Harbin, Heilongjiang, China

    Liming Yang & Ye Tian

  3. Department of Neurobiology, Harbin Medical University, 150086, Harbin, Heilongjiang, China

    Hulun Li

  4. Department of Life Science and Engineering, Harbin Institute of Technology, 150086, Harbin, Heilongjiang, China

    Liming Yang & Qian Yang

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  1. Liming Yang
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  2. Qian Yang
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  3. Kening Sun
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  4. Ye Tian
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Correspondence to Qian Yang.

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Yang, L., Yang, Q., Sun, K. et al. Agrobacterium tumefaciens Mediated Transformation of ChiV Gene to Trichoderma harzianum. Appl Biochem Biotechnol 163, 937–945 (2011). https://doi.org/10.1007/s12010-010-9097-7

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