Cloning Of Two Receptor-Like-Protein Genes And Their Verticillium Wilt Resistance Analysis

Verticillium dahliea is a kind of soil-borne fungal disease, which would seriouslydestroy cotton production.Although lots of researches working on the Verticillium dahlieawere carried out, there were no effective way for disease controlling, because of thecomplexity pathogenicity of Verticillium dahliae and lacking of resistance gene resourcesin cotton. Therefore, cloning resistance genes to Verticillium dahliae has become a hotspotin disease resistance transgenic cotton breeding.Ve1, cloned from Verticillium wilt resistant tomato variety VFN8, is a cell surfacereceptor protein and confers resistance to race1of Verticillium wilt. In this paper，Ve1wascloned from disease resistant tomato variety and inserted into the plant over expressionvector pCAMBIA2301after sequence validation. The recombinant vector was transferredinto Arabidopsis by Agrobacterium-mediated transformation method．5transgenicArabidopsis lines were obtained through kanamycin sulfate screening and PCRmethod．The T1plants of these transgenic lines were subjected to pathogen inoculation toevaluate Verticillium wilt resistance．The results showed that these transgenic lines couldput off symptoms to the defoliating isolates of Verticillium dahliae V991, but nosignificantly enhanced resistance to the non-defoliating Vertucullium wilt Bp2comparedwith the host Arabidopsis plants．In this research, a receptor like protein (RLP) of GhVdr2was cloned from the diseaseresistance cultivar Gossypium hirsutum L.. The full length of GhVdr2is3417bp, whichencodes a protein of1139aa. Although the identity of the amino acids between GhVdr2and Ve1/Ve2is only50%, their structures were similar to the LRR-TM disease resistancegene, which contains leucine-rich repeats (LRRs) and transmembrane domain. There are11LRR domains,2transmembrane domains located in N and C terminus respetively inGhVdr2gene. GhVdr2was upregulated after Verticilliun dahliae V991and Bp2incubation.Overexpressin of GhVdr2was showed to enhance the tolerance of transgenic Arabidopsisto Verticilliun dahliae compared with the wild type. Although some symptoms of leafchlorosis were found in transgenic Arabidopsis, the inflorescence height and seeds wereshown normal. The conclusion can be drawed that the resistance of GhVdr2transgenicplants to V.dahliae was obviously improved.