Cloning Of Potato StDAHPS Gene And Preliminary Study On Its Mediated Resistance To Verticillium Dahliae

Potato(Solanum tuberosum L.)is an important food and economic crop in China,which has great significance to economic development and food security.Verticillium Wilt is one of the most difficult soil-borne diseases to prevent and control.In recent years,Verticillium Wilt has occurred in different degrees in the main potato producing areas,which seriously restricts the development of potato industry in China.In this study,the potato complete sequence of St DAHPS gene was cloned base on the protein omics analysis.After bioinformatics analysis,the group to which it belongs was determined.Using q PCR method,the expression level of St DAHPS gene in different tissues,varieties and inoculated with Verticillium wilt of potato was studied,and its expression pattern was clarified.Construct the VIGS(Virus Induced Gene Silencing)silencing vector to study the effect of St DAHPS gene silencing on potato verticillium wilt resistance.The specific research results were as follows:1.Using online software for bioinformatics analysis of the St DAHPS gene,it was found that the c DNA length of the St DAHPS gene was 1617 bp,encoding a total of 538 amino acids.St DAHPS has a typical Cys-Ser-Ala-Hiss conservative domain and belongs to DAHP＿synth＿2 family members.St DAHPS protein was a hydrophilic protein with54 phosphorylation sites,without transmembrane domain and signal peptide,not belong to transmembrane protein.Phylogenetic tree analysis showed that potato St DAHPS had a relatively genetic distance with wild eggplant and tomato.2.Quantitative Real-time PCR was used to detect the expression of St DAHPS gene in different tissues,which showed that with the highest expression level in the root system and the lowest expression level in the stem.The expression level of St DAHPS gene in different varieties of potatoes was also different,with ‘Xisen 6’ having the highest expression level and ‘Shapoti’ having the lowest expression level.The expression level of other varieties was considered to be between the two.After inoculation with Verticillium dahliae,the expression levels of St DAHPS gene in resistant and susceptible potatoes were both increased,but the expression levels of St DAHPS gene in resistant varieties were significantly higher than those in susceptible varieties.3.The St DAHPS gene in potato was silenced by VIGS technology to investigate the effect of St DAHPS gene on resistance to Verticillium dahlia.The results indicated that the ratio of lesion area of control plants and empty vector PVX-EV plants were 2.5% and 2.6% after being inoculated with Verticillium dahlia.The relative lesion areas of silenced St DAHPS gene plants were 10.6%,14.4%,21.1% and18.1%,which were significantly different from the control plants with and the empty vector PVX-EV plants,with increases of 323%,475%,742%,and 622%.The investigation indicated that St DAHPS gene was silenced to reduce the resistance of potato to Verticillium wilt.