棉花谷胱甘肽硫转移酶亚基编码基因GhGST3的抗黄萎病功能研究

The Verticillium wilt resistance research of glutathione S-transferase subunit encoding gene GhGST3 in Gossypium hirsutum

棉花黄萎病(Verticillium wilt)是影响产量和品质的最严重病害之一。前期研究表明陆地棉(Gossypium hirsutum)谷胱甘肽硫转移酶亚基编码基因GhGST3(MK179292)参与了棉花抗黄萎病反应。本研究从陆地棉抗病品种‘农大601’(ND601)中克隆GhGST3的开放读码框(Open reading frame,ORF)。对T3代超表达GhGST3拟南芥植株进行表达检测和黄萎病抗性鉴定。通过农杆菌介导的烟草瞬时表达技术进行亚细胞定位。利用酵母双杂交系统,与黄萎病菌诱导下的酵母文库进行杂交,筛选互作蛋白,初步分析其参与的棉花抗病机制。结果表明:超表达GhGST3基因可有效提高植株对黄萎病的抗性。GhGST3定位在细胞核、细胞膜、细胞质。酵母双杂交筛选出11个与GhGST3互作的蛋白。基于这些蛋白的KEGG和GO分析结果,显示GhGST3可能参与植物的柠檬酸盐代谢,氨基糖和核苷酸糖代谢,核糖体代谢,碳代谢以及氧化磷酸化等代谢路径。本研究初步明确了GhGST3的表达及黄萎病抗性机制,为棉花抗病育种提供了基因资源。

Verticillium wilt is one of the most serious diseases to affect yield and fiber quality in cotton(Gossypium hirsutum).In our previous study,GhGST3(MK179292),one glutathione S-transferase subunit encoding gene,participated in the Verticillium wilt resistance in cotton.In this study,the open reading frame of GhGST3 was cloned from resistance variety‘ND601’.T3 generation of overexpressing GhGST3 Arabidopsis plants were tested for expression and resistance to Verticillium wilt.Agrobacterium-mediated tobacco transient expression technology was used for subcellular localization.The yeast two-hybrid system was used to hybridize with the yeast library that induced by Verticillium dahliae to screen the interacting proteins,and to analyze the cotton disease resistance mechanism.The results showed that overexpression of GhGST3 gene could effectively improve the resistance of plants to Verticillium wilt.GhGST3 was located in the nucleus,cell membrane,and cytoplasm.The 11 interaction proteins with GhGST3 were screened by yeast two-hybrid.Based on the KEGG and GO analysis of these proteins,it was shown that GhGST3 may be involved in metabolic pathways of plant citrate metabolism, amino sugar andnucleotide sugar metabolism, ribosome metabolism, carbon metabolism, and oxidative phosphorylation. This studypreliminarily clarified the expression of GhGST3 and the mechanism of Verticillium wilt resistance, which providedgenetic resources for cotton Verticillium wilt resistance breeding.