Functional Analysis Of CsWRKY30 Gene In Cucumber Under Propamcarb Stress

Cucumber(Cucumis sativus L.) is one of the most popular fruit vegetables in our country, which is mainly cultivated in the greenhouse, there is easily induced downy mildew in cucumber production, because of the special climate conditions in the greenhouse and successive planting. Propamocarb can be taken precautions against downy mildew, but it is volati.and conductive, which is also likely accumulated in cucumber.It causes pesticide residues, food safety, environmental pollution. So, study the problem of propamocarb residues in cucumber will provide safety and pollution-free cucumber products for consumers.The present study on the problem of pesticide residues in vegetables mainly in reducing pesticide inputs and accelerate pesticide metabolism in crops vivo. Whi.the previous low of pesticide(Propamocarb) residues in cucumber varieties D0351 propamocarb stress transcriptome analysis showed that transcription factor CsWRKY30 may be related to cucumber propamocarb threats, the gene expression profiles is up-regulated,these evidence helps us speculate that this gene may be involved in the signal transduction of propamocarb threats, and reduce the residual quantity of cucumber propamocarb. In order to explore the fuctional of CsWRKY30 in low propamocarb, molecular biology technique is mainly used in this study. The main results are as follows:(1) The full-length sequence of CsWRKY30 CDs was amplified by PCR, which is 1014 bp,encoding 337 amino acids contained a WRKY domain structure,belongs to a member of WRKY. And analysised the amino acid sequence of CsWRKY30 encoding, the physicochemical properties of the protein domains,functional domain prediction, homology sequence alignment and phylogenetic analysis;(2) The expression pattern of CsWRKY30 was analysised by qRT-PCR, the results showed that when the cucumber suffered propamocarb stress, expression of CsWRKY30 was up-regulated in D0351(Low propamocarb residue), nearly no difference in D9320(High propamocarb residue); From 0.5 h to 9 h under propamocarb stress, the expression of CsWRKY30 significantly higher than the control, after 24 h, the expression of this gene was no longer notably increased. In additon, CsWRKY30 was mainly expressed in fruit;(3) CsWRKY30 and green fluorescent protein(GFP) fusion, and transferred into Arabidopsis protoplasts, observed by confocal laser scanning microscope, CsWRKY30 protein was localized in the nucleus;(4) Consruct over-expression vector pCAMBIA1303-CsWRKY30, transformed into Arabidopsis. The discovery of propamocarb stress experiments in transgenic Arabidopsis, in untreated conditions, there was no significant difference between CsWRKY30 transgenic and wild type of Arabidopsis thaliana, in 2 mM propamocarb treatment conditions, transgenic Arabidopsis germination rate and root length were significantly higher than that of wild type Arabidopsis;(5) Under other treatments, CsWRKY30 was no response to mational and NaCl, but it is sensitive to propamocarb and Corynespora cassiicola w, thus CsWRKY30 was induced by ABA;(6) pCAMBIA1303-CsWRKY30 was transformed into 649, and obtained F1 seeds.