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