Interaction Of Spodoptera Exigua And Cabbage Glucosinolates And Preliminary Function Study Of Bra008132 Gene

Glucosinolates are a typical defense metabolite in cruciferous plants.‘hei you tong'cabbage showed high resistanceon the pest feeding.In order to study its insect-resistant mechanism,we tested the content of glucosinolate in ‘hei youtong'cabbage leaves at different growth stages and in different leaf positions,and discussed the distribution of glucosinolatesin physiological and ecological significance in cabbage.The use of interaction between Spodoptera exigua and ‘hei you tong'cabbage,we explored the insect-resistant mechanism of ‘hei you tong'.The gene and promoter analysis of Bra008132 and its related vectors were also carried out based on the dramatic changes in Spodoptera exigua.The main results are as follows:(1)The content of glucosinolate in ‘hei you tong'cabbage leaves changed greatly at different leaf-stage,the content increased from the four-leaf stage to the eight-leaf stage,and decreased at the twelve-leaf stage.Among them,aliphatic glucosinolates showed a gradual downward trend with the growth of leaf stage,the lowest content in the twelve leaf stage.The trend of indole glucosinolates and aromatic glucosinolates was consistent with that of total glucosinolate,with the highest content in the eight-leaf stage and the lower content in the four-leaf stage and twelve-leaf stage.In different leaf positions in period eight,juvenile leaves had higher glucosinolate content than aged leaves.(2)The detection of glucosinolate content of Spodoptera exiguafeeding ‘hei you tong'cabbage at different time points showed that the content of glucosinolate increased with the prolongation of feeding time and the tendency of indole glucosinolates was more obvious than that of aliphatic glucosinolate.Detection of glutathione transferase activity in Spodoptera exigua body showed that its activity increased significantly with the prolongation of feeding time.Quantitative analysis of glucosinolate-related genes,including Br BCAT4-1,BrMAM1-1,BrSUR1-1,BrMYB28-1,BrAOP2-1,BrSOT16-1,BrCYP83A1-1,BrCYP83B1 and BrAPK2-1,found that the feeding of Spodoptera exigua caused the up-regulation of these genes and the variation trend of different genes was inconsistent.(3)The full-length sequence of the Bra008132 gene was cloned from DNA of ‘hei you tong'using PCR technology and it contained a complete open reading frame encoding 339 amino acids.This amino acid has no signal peptide,no transmembrane structure,and its subcellular localization in the cytoplasm.The promoter sequence analysis,founding that it included light response,abscisic acid,defense stress and other cis-acting elements.qRT-PCR results showed that the expression of Bra008132 gene was different at different times.