Screening The Pathogenesis-Related Genes From Verticillium Dahliae And Initial Study On Gene GbEDS1from Cotton

Verticillium dahliae is a fungal disease. It can infect cotton, and great hazard cotton production. Byusing RNA interference technology, V. dahliae pathogenesis-related genes can be detected. This canprovide a new idea using to control of cotton verticillium wilt. Research on disease resistance defensemechanism of signal control systems can dig plant genes of unknown function itself the potential anduse plant own genetic resources to deal with the dangers of Verticillium wilt, this ia also a new idea toimprove the overall resistance of cotton and breeding new varieties resistant to verticillium wilt. In thisstudy, two parts of work ware included: Pathogenesis-related gene screening from Verticillium dahliaeand initial study on GbEDS1gene from cotton. The results are as follows:1. Screening pathogenesis-related gene from Verticillium dahliae:32Pathogenesis-related genefragments from Verticillium dahliae Vd991were cloned, and transformed into N. benthamiana andtransient expressed siRNA in contemporary times by using the virus-induced gene silencing （VIGS）method. We found that ten genes were screened in the course of related genes and plants showed highresistance.2. Cloning GbEDS1gene and its subcellular localization: GbEDS1gene was obtained by RACEtechnology from Gossypium barbadense. GbEDS1is2064bp in length, ORF is1647bp, encode548aa,and there is a phospholipase structure in its N-terminal conserved region. In onion epidermal cellsGbEDS1gene transiently expressed. We found that GbEDS1protein appeared in the cytoplasm and thenucleus. It related with Arabidopsis EDS1-PAD4and EDS1-SAG101protein accumulating in thecytoplasm and the nucleus.3. Induced GbEDS1gene expression and function analysis: We constructed of RNAi vector andeds1gene expression was well interfered in N. benthamiana by using VIGS technology. We found thatplants become more susceptible inoculated V. dahliae, it can be inferred eds1gene which played animportant role resistance to V. dahliae. We also observed GbEDS1gene expression in Gossypiumbarbadense inoculated certain concentration of V. dahliae. The results showed that there is a significantdifference in GbEDS1gene expression, and showed a certain regularity. As time changes, expression ofGbEDS1gene increased and then decreased.