Fuctional Characterization Of Cotton GhCyP、GhDAHPS、 GhCCAOMT Gene Related With Verticillium Wilt Resistance In Cotton

In this study we constructed overexpressing vector about GhCyP、GhDAHPS、GhCCAOMT gene related with verticillium wilt resistance which was obtained from SSH library of resistant cultivar JiMian 20 induced by Verticillium dahlia; GhCyP, GhDAHPS, GhCCAOMT were transformed into Arabidopsis thaliana and cotton; The assay of expressing and regulation for GhCyP、GhDAHPS、GhCCAOMT gene were performed by Real-time PCR; Moreover, the fuction of GhCyP was primarily evaluated. The study will help us to understand the resistence mechanism and the pattern of regulation and expressing of gene related with Verticillium wilt. The main results are as follows:1. The expressing analysis about GhCyP, GhCCAOMT and GhDAHPS by Realtime PCR showed that when stressed with Verticillium dahliae, the susceptible variety Zhong8, the resistant variety Jimian20 and Pima90-53 expressed quite differently for the genes.2. Construction of the over expression vector pBI-GhCyP, pCam-CCoAOMT. pCam-DAHPS were completed.3. The overexpressing vector pBI-GhCyP. pCam-CCoAOMT and pCam-DAHPS were transformed into Arabidopsis thaliana separately through floral dip method, and the T3 plants were obtained after antibiotic screening and the PCR examination.4. The overexpression vector pBI-GhCyP, pCam-GhCCoAOMT, pCam-GhDAHPS were transformed into cotton variety Han 208, Zhong8 and Jimian 11 mediated by the pollen tube method, And transgenic plants were identified by PCR examination.5. After inoculation, the GhCyP transgenic plants improved the resistance to Verticillium wilt.6. After purified by Ni-NTA affinity chromatography, enzyme activity analysis indicated that the recombinatant GhCyP protein has obvious PPIase activity. Inhibit funguals test showed that the recombinatant GhCyP protein obviously suppressed the growth of Verticillium dahliae.