1 College of Agriculture, Guizhou University/Guizhou Branch of National Wheat Improvement Center, Guiyang 550025, China; 2 College of Biological Science and Agriculture, Qiannan Normal University for Nationalities, Duyun 558000, China
Abstract SBP-box (SQUAMOSA PROMOTER BINDING PROTEIN box) gene family encode a class of plant-specific SBP transcription factors, which are widely distributed in plants and play important functional regulation in plant growth, development and reproduction, including embryogenesis, secondary metabolites biosynthesis, stress responses, etc. Therefore, it has application value in crop improvement. This study used bioinformatics and quantitative real-time PCR (qRT-PCR) to identify and analyze the expression of Sorghum bicolor SbSBP15 gene, and the recombinant SbSBP15 protein was obtained by Escherichia coli prokaryotic expression system. The results showed that the full length of SbSBP15 gene (GenBank No. MT634207) was 1 227 bp, encoding 408 amino acids. Bioinformatics analysis showed that SbSBP15 protein contained a typical SBP domain with a relative molecular weight of 42.19 kD and the isoelectric point was 8.87. Phylogenetic results showed the closest family relationship with maize (Zea mays). Subcellular localization was mainly in the nucleus, and it had strong hydrophilicity. Besides, protein interaction analysis demonstrated that SbSBP15 might interact with protein kinase. qRT-PCR analysis demonstrated that SbSBP15 was mainly expressed in the stalk, and drought and salt stresses could induce SbSBP15 expression. Furthermore, the expression level of SbSBP15 was also enhanced after plant hormone indole acetic acid (IAA) treatment, however, salicylic acid (SA) reduced the expression of SbSBP15. The optimal protein expression of SbSBP15 were in E. coli Rosetta (DE3) strain, inducing temperature at 25 ℃ and isopropyl-β-D-thiogalactopyranoside (IPTG) concentration of 0.8 mmol/L. This study provides a candidate gene for SbSBP15-based molecular genetic study of sorghum, and further provides a basis for studying the biological function of plant SBP15 gene.
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