Cloning,expression analysis and subcellular localization of BrADS2 gene in Brassica rapa
[Objective]The present paper aimed to explore the relationship between fatty acid desaturase gene ADS2(△9 fatty acid desatu-rase)and cold resistance in Brassica rapa L.[Method]Based on the previous transcriptome results,the BrADS2 gene was obtained by one-step directional cloning.GFP fusion expression vector was constructed and transiently transformed into tobacco lower epidermal cells by injec-tion method.At the same time,'Zhongshuang 11'was infected by Agrobacterium infection method.The expression pattern of ADS2 gene in the leaves of winter rapeseed varieties with different cold resistance was detected by qRT-PCR,and the morphological changes,antioxidant en-zyme activity and conductivity content of winter rapeseed under low temperature were analyzed.[Result]The BrADS2 gene could encode 307 amino acids,and the protein encoded by this gene had hydrophilic properties.The results of qRT-PCR analysis showed that the expression of BrADS2 gene increased first and then decreased,and the expression level reached the peak at-4 ℃.The results of subcellular localization showed that BrADS2 protein was expressed in cytoplasm and cell membrane.Under low temperature treatment,the relative conductivity and antioxidant enzyme activity of the leaves of the three varieties increased significantly.The antioxidant enzyme activity of the strong cold-resist-ant varieties was higher than that of the weak cold-resistant varieties,but the conductivity was the opposite.The hypocotyls of'Zhongshuang 11'were infected by Agrobacterium tumefaciens,and 15 transgenic positive seedlings were obtained.The expression of BrADS2 gene in the leaves of transgenic rape positive plants was significantly higher than that of wild type.[Conclusion]It is speculated that BrADS2 gene plays an important role in the regulation of membrane lipid unsaturation in winter rapeseed,revealing the relationship between ADS2 gene and crop resistance to low temperature stress.
万方数据
维普
