大丽轮枝菌及激素处理后棉花酵母双杂交文库的构建

大丽轮枝菌是引起棉花、马铃薯等重要作物黄萎病的土传病原真菌,其分泌的胞外蛋白是侵染寄主的重要毒力因子,因此研究胞外蛋白与寄主的相互作用具有重要意义.本研究旨在构建高效的棉花酵母双杂交cDNA文库,并通过已知大丽轮枝菌效应子VCR1互作蛋白的筛选来评价构建文库的质量.分别采用大丽轮枝菌及茉莉酸甲酯、水杨酸和乙烯处理海岛棉...

Construction of a sea-island cotton yeast two-hybrid library under the conditions of infection with Verticillium dahliae and treatment with hormones

Verticillium dahliae is a soil-borne fungal plant pathogen that causes the destructive vascular wilt on many hosts,including economically important cotton and potato crops.Generally,the extracellular proteins secreted by V.dahliae play a crucial role in its virulence on the host.The objective of this study was to construct a yeast two-hybrid library from cotton infected with V.dahliae and treated with hormones,and evaluate the library quality by screening the interaction proteins of a known V.dahliae effector VCR1.To construct the library,the RNA was extracted from the cultivar Gossypium barbadense‘Hai 7124’inoculated with V.dahliae or treated with hormones.The high quality of library was assessed by the high cDNA library titer(1.18×10⁸ cfu/mL),the distribution length of insertion size is 300-2000 bp,and the absolute recombination efficiency.Subsequent screening of this library for the known V.dahliae effector VCR1 yielded 27 candidate interaction proteins,and one of them was proven to interact with VCR1 when validated using the yeast two-hybrid system.In conclusion,a high-quality sea-island cotton yeast two-hybrid library with good efficiency for identifying the interaction proteins of V.dahliae effectors was constructed in this study,which would be helpful for revealing molecular mechanisms underlying the interactions between V.dahliae and cotton.