Cloning and Expression Analysis of CmWRKY7 Gene in Melon (Cucumis melo)
LI Jia-Qi1, ZHANG Li-Chao2, ZHAO Wang-Long1, CHEN Chun-Lin1, WANG Ji-Qing1, LI Meng1,*, XIAO Huai-Juan1,*
1 College of Horticulture, Henan Agricultural University, Zhengzhou 450046, China; 2 Technology Service Center on Ecological Planting of Chinese Herbal Medicine in Chengde, Chengde 067000, China
Abstract：In plants, WRKY transcription factors play a crucial role in responding to biotic and abiotic stresses. To explore the biological function of WRKY gene in defense against abiotic stress in melon (Cucumis melo), a candidate gene strongly induced by low temperature, CmWRKY7, was screened based on pre transcriptome sequencing. In this study, CmWRKY7 was cloned, and the molecular characteristics of its encoded protein were analyzed using bioinformatics methods. The expression patterns of CmWRKY7 under different stresses were detected by qRT-PCR, including low temperature, drought, salt, and ABA stress. Results showed that the total length of CmWRKY7 was 732 bp, encoding 243 amino acids, with a predicted molecular weight of 27.09 kD and a theoretical isoelectric point of 9.50. It was a hydrophilic protein with an aliphatic amino acid index of 64.20 and high thermal stability. The CmWRKY7 protein sequence contained 2 highly conserved domains: The WRKYGKK and the C2H2 zinc finger structural sequence (C-X4-C-X23-H-X-H). Moreover, the CmWRKY7 protein also contained 26 amino acid residues that might be phosphorylation modification sites, 7 protein kinase C phosphorylation sites, 4 N-myristoylation sites, and 1 tyrosine kinase phosphorylation site. The secondary structure of CmWRKY7 protein was dominated by irregular coils, accounting for 62.14%. Phylogenetic analysis showed that CmWRKY7 had the closest evolutionary relationship with the proteins of cucumber (C. sativus) and horned melon (C. metuliferus), gathering into one branch. Promoter prediction analysis showed that there were multiple hormone and stress-related cis acting elements on the CmWRKY7 promoter, and protein interaction prediction analysis showed that CmWRKY7 mostly interacted with stress-related proteins. Under abiotic stress, the CmWRKY7 gene participated in ABA signal transduction and actively responded to drought, high salt, and low temperature stresses. These results provided theoretical basis for further studying the biological function of CmWRKY7 and its application in melon breeding for abiotic stress resistance.
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