Screening Of The Promoters Dominantly Expressed In Root And The Functional Analysis Of GhCPK33 In Cotton Response To Verticillium Dahliae

Plant development and resistance are usually antagonistic,the activation of the plant immune system usually affects the growth and development,thus affecting crop yield.So the application of disease resistance genes in disease resistance breeding requires precise time and space expression control elements,on the one hand can lead to crop resistance,on the other hand does not affect the crop yield.Tissue-specific promoters allow the resistance gene to be expressed only in specific tissues,allowing the plants to obtain resistance while being able to grow normally.Calcium-dependent protein kinases(CDPKs)play an important role not only in development,but also in the immune system of plants.Ous previous identify a protein which could interact with Gh OPR3(1,2-Oxygen-plant dienoic acid(OPDA)reductase)in yeast two hybridization.The gene show highly similarity to Gh CPK33 like and named Gh CPK33.In this study,the isolation and identification of the dominant expression promoter in cotton root,and the function of Gh CPK33 in cotton resistance were studied.The main results are as follows:1.Totally,22 genes that highly expressed in the roots through bioinformatic analysis in Gossypium barbadense were identify.Two promoters were obtained by PCR amplification,and the expression vectors were completed and transformed into Arabidopsis thaliana and cotton.2.The expression of GhCPK33 was detected in all tissues of cotton and it did not obviously change after inoculation with Verticillium dahliae.The expression of Gh CPK33 significantly increased after treating 1 h with Me-JA,but for the treatment of exogenous SA,The response of the gene did not obviously change.3.We use virus-induced gene silencing technology to inhibite the expression of Gh CPK33 in Gossypium hirsutum.which enhance the resistance of plant to Verticillium wilt.Meanwhile,the content of isoleucine jasminate that is the activity of endogenous jasmonic acid was increased,and the key gene of JA signal pathway was activated.Plants those expression of Gh CPK33 was increased enchanced the resistant to necrotrophic pathogen-Botrytis cinerea.Therefore,it is speculated that the gene may affect the resistance of cotton to Verticillium wilt though affecting the jasmonic acid signal pathway.The promoters express dominantly in the root,using them to drive the expression of resistance genes,such as Gh CPK33,only in the root,which is a good way to obtain the resistant variety.