Cloning and Expression Analysis of Solanum sisymbriifolium Verticillium Wilt Resistance-Related Genes SsWRKY22 and SsWRKY33
Verticillium wilt is a major disease that seriously affects eggplant yield and quality.WRKY transcription factors play a key role in plant resistance,while the knowledge about the resistance to eggplant verticillium wilt are deficient.In this study,two WRKY genes,named SsWRKY22 and SsWRKY33,were screened from the transcrip-tome database of Solanum sisymbriifolium which is highly resistant to verticillium wilt.The target genes were cloned for bioinformatics analysis.Then the expression pattern of two genes were analyzed by RT-qPCR.The results showed that the full length ORF of SsWRKY22 and SsWRKY33 are 528 and 1 073 bp,encoding 175 and 357 amino acid,respectively.The molecular weights of two proteins are 19.19 and 40.46 kD,respectively,which belong to hydrophilic unstable proteins.The SsWRKY22 protein is acidic protein,while the SsWRKY33 protein is basic protein.Subcellular localization predicted that both proteins are located in the nucleus.In addition,SsWRKY22 and SsWRKY33 with different protein structure belong to class Ⅱ and class Ⅰ WRKY transcription factors,respective-ly.Phylogenetic analysis showed that SsWRKY22 is closely related to WRKY22 in Solanum pennellii,S.lycoper-sicum and S.tuberosum,while the SsWRKY33 is the most closely related to WRKY33 in Capsicum annuum.The RT-qPCR analysis indicated that the expression levels of SsWRKY22 and SsWRKY33 are tissue-specific and the highest in roots.After inoculation by Verticillium dahliae,the expression levels of SsWRKY22 and SsWRKY33 were significantly down-regulated in the root,suggesting that both genes may be negative regulatory factors in re-sponse to verticillium wilt stress.In this study,WRKY transcription factor genes were firstly cloned and their ex-pression patterns were also analyzed in S.sisymbriifolium,which will lay a foundation for the follow-up functional characterization of Ss WRKY22 and Ss WRKY33.
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