苦荞FtHY5基因克隆、生物信息学及参与花青素合成分析

Cloning and Bioinformatics Analysis of FtHY5 Gene and Its Involvement in Anthocyanin Synthesis in Tartary Buckwheat

HY5 (Elongated hypocotyl 5)转录因子是参与植物光形态建成的重要成员,并调控花青素的生物合成。本研究以苦荞‘晋荞2号’作为材料,利用PCR克隆其HY5基因的完整的CDS序列,并对FtHY5的理化性质、保守功能域、信号肽、亚细胞定位、亲/疏水性、蛋白质结构、系统进化等进行了生物信息学分析,此外还分析了其对苦荞芽菜花青素合成的影响。结果表明,FtHY5基因完整的CDS长度为507 bp,编码的蛋白分子量为18.434 kD,等电点为9.47,空间构象呈拉链形状,属于亲水性蛋白,含有信号肽,亚细胞定位在细胞核里。苦荞FtHY5蛋白的氨基酸序列与藜麦和甜菜的亲缘关系最近。FtHY5基因与花青素生物合成途径上的结构基因在光照条件下培养的芽菜中的表达水平均高于黑暗条件下的,说明Ft HY5可能调控这些花青素合成基因的表达水平,从而参与光诱导花青素合成的过程。综上所述,本研究结果将为进一步研究FtHY5调控苦荞芽菜花青素的生物合成的分子机理提供基础,同时也为利用该基因来改善苦荞芽菜的品质提供理论支撑。

HY5(Elongated hypocotyl 5)transcription factor is the vital member that involves in photomorphogenesis,and regulates anthocyanin biosynthesis.In this study,we cloned the completed CDS sequence of HY5 gene from Tartary buckwheat'Jinqiao2'by PCR method.Then,the physicochemical properties,conversed domains,the signal peptides,hydrophilic/hydrophobic,protein structure and phyletic evolution of FtHY5 were analyzed by bioinformation software.In addition,we also analyzed the effect of FtHY5 on anthocyanin synthesis in Tartary buckwheat sprounts.The results showed that the CDS length of FtHY5 gene was 507 bp,the weight of which coding protein was 18.434 kD and the protien's isoelectric point was 9.47.Its spatial conformation was zip shaped,and it belonged to hydrophilic protein and contained a signal peptide,locating in cell nucleus.Tartary buckwheat FtHY5 protein was most phylogenetically close to Chenopodium quinoa and Beta vulgaris.FtHY5 and structure genes of anthocyanin biosynthesis pathway expressed higher in Tartary buckwheat sprouts under light than that under dark treatment.In conclusion,these results will lay foundation for further study on the molecular mechanism on FtHY5 regulation of anthocyanin biosynthesis in Tartary buckwheat sprouts and could provide theoretical support for the utilization of this gene to improve the quality of Tartary buckwheat sprouts.