Heterologous expression of chit36 gene from Trichoderma atroviride in canola reduces development of lesion caused by Sclerotinia sclerotiorum

Abstract

An endochitinase gene (chit36) was isolated from the biocontrol fungus Trichoderma atroviride. Chit36 was overexpressed under the CaMV35S constitutive promoter in canola. Transformation of cotyledonary petioles was achieved via Agrobacterium tumefaciens. The insertion of transgene was verified by PCR and DNA dot blotting. RT-PCR analysis indicated that the transgenic canola was able to express chit36 gene. Also, we used a transgenic over-expression approach in order to investigate antifungal activity of expressed chit36 on Sclerotiana sclerotiorum. Antifungal activity was detected in crude protein extracts from transgenic canola using radial diffusion assay. Also, lesion sizes of transgenic canola by S. sclerotiorum were significantly retarded compared to non transgenic canola plant using detached leaf assay.