Comparative expression analysis of defense-related genes in both wild and transgenic Brassica juncea (var.) Varuna harboring mapk3 gene in response to infection by Albugo candida
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Date
2018-05
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G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand)
Abstract
Mustard and Rapeseed are the third leading source of vegetable oil in the world after soybean and oil palm. According to Department of Agriculture & Farmers welfare 2017-2018 annual production of Rapeseed and mustard is 75.40 Lakh tones. Production in India is still not enough to meet the domestic requirement. The potential yield of Brassica is very high but there is a wide gap between the potential and realized yield of Brassica and it is largely because of biotic & abiotic stresses. White rust is one of the most important disease of Brassica and it is caused by pathogen Albugo candida.Even 60% reduction in yield of Brassica juncea is due to its high susceptibility to white rust. Plants defend themselves against most potent pathogens through various defense mechanisms. Activation of MAPK signalling cascade is one of major defense pathways by which extracellular stimuli are transduced into intracellular responses in all the eukaryotes. MAPK cascade also play key role in regulation of various defense related genes in response to pathogens. Therefore our investigation was to analyse expression level of potential MAPK cascade and defense related genes in wild and transgenic Brassica juncea (var.) Varuna harboring overexpressed MAPK3 gene to access tolerance against Albugo candida. Lower disease index in transgenic Brassica suggests its tolerance to Albugo candida. Higher expression level of MAPK6 than MAPK3 in the transgenic Brassica suggests that MAPK6 mimics a portion of MAPK3 signalling pathway and work in a direct cascade for production of defense related proteins. Overexpression of MAPK3 gene was observed in downregulating the expression of MAPK4 in transgenic Brassica juncea . The expression of WRKY33 and WRKY29 was also analysed in the present study. Expression levels of both WRKY33 and WRKY29 were found to be higher in transgenic plant overexpressing MAPK3. Expression of 3 Defense related genes (OASTL-B, ACD2, and CSD2) were found higher in transgenic plants horboring overexpressed MAPK3 gene & expression level of CYP20-3 was found higher in wild Brassica plant. The defense related genes (OASTL-B, ACD2, CSD2, and CYP20-3) were also subjected to In-silico study. Insilico study showed the Secondary structure, tertiary structure and putative phosphorylation sites of these 4 defense related genes & also derived protein-protein interaction of potential MAPKs cascade and WRKY (29, 33) with OASTL-B, ACD2, CSD2 & CYP20-3.
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