A comparative study of transgenic canola (Brassica napus L.) harboring either chimeric or native Chit42 genes against phytopathogenic fungi

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• 作者：Mahboobeh Ziaei ; Mostafa Motallebi…
• 关键词：Transgenic canola ; Chit42 ; Chimeric chitinase ; Chitin binding domain ; Antifungal activity
• 刊名：Journal of Plant Biochemistry and Biotechnology
• 出版年：2016
• 出版时间：October 2016
• 年：2016
• 卷：25
• 期：4
• 页码：358-366
• 全文大小：2,165 KB
• 刊物主题：Life Sciences, general; Plant Biochemistry; Protein Science; Receptors; Cell Biology;
• 出版者：Springer India
• ISSN：0974-1275
• 卷排序：25

文摘

Canola (Brassica napus L.), an agro-economically important crop in the world, is sensitive to many fungal pathogens. One strategy to combat fungal diseases is genetic engineering through transferring genes encoding the pathogenesis-related (PR) proteins such as chitinase which cause the chitin degradation of fungal cell wall. Chitinase Chit42 from Trichoderma atroviride (PTCC5220) plays an important role in biocontrol and has high antifungal activity against a wide range of phytopathogenic fungi. This enzyme lacks a chitin binding domain (ChBD) which is involved in binding activity to insoluble chitin. In the present study, we investigated the effect of chitin binding domain fused to Chit42 when compared with native Chit42. These genes were over-expressed under the CaMV35S promoter in B. napus, R line Hyola 308. Transformation of cotyledonary petioles was achieved by pBISM2 and pBIKE1 constructs containing chimeric and native Chit42 genes respectively, via Agrobacterium method. The insertion of transgenes in T₀ generation was verified through polymerase chain reaction (PCR) and Southern blot analysis. Antifungal activity of expressed chitinase in transgenic plants was also investigated by bioassays. The transgenic canola expressing chimeric chitinase showed stronger inhibition against phytopathogenic fungi that indicates the role of chitin binding domain.