棉花抗黄萎病相关基因GhPP2C的克隆和功能研究-生物化学与分子生物学专业毕业论文.docxVIP

AbstractCotton Abstract Cotton Verticillium wilt is of the primary diseases that affects the production and quality of cotton．so it is importsnt for co,on anti-Verticillium breeding to“=fj渤the resistant related genes and ducidate the mechanism of anti-VerticiUium of cotton．In山is research．the full-length eDNA sequence of Vdr92 from SSH wag cloned．The results of bioinformatics analysis indicated that this gane shares high homologous with protein phosphatase 212(PP2C)that participate in ABA signal Uansduction and was designawA as GhPP2C．The timing expression characmrisfics of GhPP2C were examined in different coUon varieties treated wi血crude extraction toxin from different virulence Verticillium dahliae strains．The Arabidops括thaliana transgenic plants of GhPP2c wb自[-o obtained by tim method of Agrobacterium-mediaed transformation． The 1471bp full—l∞g血eDNA sequence of GhPP2C gene was cloned through 5RACB．The open reading frame(ORF)was 1251bp in length and deduced protein sequence was 416 amino acids．The analysisofDNA sequenceindicatedthatGhPP2Chad 3introns，i．ef 98bp．341bpand96bp．The results of BLAST showed that GhPP2C belonged to the PP2C family and shared 59％similari哆with ABA-Hypersensitive Germination 3似日G3)． The seven-day aged seedlings of three different resistant cotton varieties were used to test the expression characteristics of GhPP2C after treating with different virulence crude extraction toxin from Verticilliilmdah／iae strainsVDKll4 andVDK991 at4timepoints(2h，6h，12h，9Ah)byRT-PCR． The results indicated that when treated wi血high virulence strain VDK991．GhPP2C in resistant variety Island cotton 06 expressed from 12h to 24h，whereas in tolerance variety zhongmian 12 and susceptible variety zhongzhi 86-1 GhPP2C only expressed at 2h．When treated with high virulence strain VDKI 14，the expression of GhPP2C showed more complicated．GhPP2C was expressed at 6h to PAh in resistant variety and 2h to 12h in tolerance variety and only 12h in susceptible variety．Semi RT-PCR re