Genome-wide Analisis Of The WRKY Famiy Response To Salt Stress And Histidine Kinase In Beta Vulgaris

Soil salinization and drought are the important factors affecting global crop yield and quality.Therefore,the research on the molecular mechanism of crop salt tolerance has become a hotspot in agricultural science.Recent studies have shown that some genes and transcription factors are involved in the response to salt in plants.WRKY and histidine kinase family genes have been reported to be involved in drought and salt stress response in plants.Sugar beet is an important sugar crop in China.So study the salt tolerance of sugar beet and understand the mechanism of salt toleranc of sugar beet from the molecular level is of great significance for breeding varieties of sugar beet with salt tolerance.In this paper,the expression levels of WRKY and histidine kinase genes in leaves and roots of sugar beet seedlings under salt stress were studied by combining bioinformatics analysis with molecular biology experiments,and the following results were obtained:Bioinformatics method was used to genome-wide screen the WRKY family and the two-component system-related protein and genes in sugar beet.The results showed that there were 45 members of the WRKY family in the sugar beet.There are 10 members of the histidine kinase members in sugar beet.There are 7 members histidine-containing phosphotransfer protein members.There are 24 members response regulator protein members.The bioinformatics analysis of these proteins and genes was carried out by bioinformatics software.Finally,we get the phylogenetic tree of WRKY transcription factor family and two component-related proteins.We also get the chemical properties and subcellular localization of these transcription factors or proteins and the results of their genes' chromosome localization.By analyzing the expression profile data of previous studies of our research group,25 genes related to salt tolerance of WRKY family were found.Root and leaf samples of “O” 68 sugar beet seedlings was treated with 300 m M NaCl solution.These 25 WRKY family genes and all histidine kinase family genes were quantitatively analyzed by qRTPCR.The results showed that 10 genes of WRKY transcription factor in root and 11 genes of WRKY transcription factor in leaf and 4 genes of histidine kinase family were significantly up-regulated.The salt releated WRKY transcription factor(wmfg)and histidine kinase(kucd)'s CDS were cloned and sequenced.The protein sequences' secondary structure and tertiary structure and intereaction and promoters of wmfg and kucd were analyzed.The gene silencing vector of salt releated genes was successfully constructed.The T vector(with the wmfg and kucd CDS fragment)and the pTRV2 vector which was constructed by our laboratory were constructed the gene silencing vector by double enzyme digestion.After the silence vector was verified,it was transformed into agrobacterium.The constructed engineered bacteria can be used in gene functional verification experiments.The salt tolerance control network of sugar beet was established.The network includes different salt signal transduction pathways and transcription factors and two component systems.In this regulatory network,the pathway related to WRKY and beet two-component system was mainly constructed.