红花CtMYB-TF1基因的克隆与表达分析

Cloning and Expression Analysis of CtMYB-TF1 Gene from Carthamus tinctorius

为了探索R2R3-MYB转录因子与植物类黄酮的次生代谢调控之关系,及对植物花青素合成代谢的作用,以红花花瓣为试验材料,采用RT-PCR结合RACE技术,从红花(Carthamus tinctorius)中克隆获得1个全长1260 bp的调控花青素代谢的CtMYB-TF1基因。结果表明,该基因序列可编码249个氨基酸,其蛋白质分子质量为28.08 ku;红花CtMYB-TF1蛋白分子式为C_(1220)H_(1930)N_(358)O_(378)S_(13),是一种不稳定的碱性亲水蛋白;由系统进化树分析可知,红花CtMYB-TF1与葡萄VvMYBPA1亲缘关系较近;通过RT-qPCR技术对表达体系进行基因表达量的检测分析,红花CtMYB-TF1基因在花期的各个时期相对表达量均高于红花CtANS基因,并且随着花期的延长红花CtMYB-TF1基因和CtANS基因的相对表达量也有增高,其中在花期的第5天红花CtMYB-TF1基因的相对表达量出现峰值,说明CtMYB-TF1基因对CtANS基因具明显的上调作用,可为构建高产、稳定的红花基因工程改良细胞株提供参考。

In order to explore the relationship between R2R3-MYB transcription factors and secondary metabolism of plant flavonoids,and the effects on the synthesis and metabolism of plant anthocyanins,then in this paper,we took safflower petals as experimental materials and used the reverse transcription-PCR and rapid amplification of cDNA ends(RACE)techniques,the results showed that the gene sequence could encoded 249 amino acids,and its protein molecular mass was 28.08 ku;the molecular formula of safflower CtMYB-TF1 protein was C_(1220)H_(1930)N_(358)O_(378)S_(13),which was an unstable alkaline hydrophilic protein;the phylogenetic relationship analysis showed that CtMYB-TF1 was closely related to VvMYBPA1 of grape.The expression result reflected that the relative expression of safflower CtMYB-TF1 gene was higher than that of safflower CtANS gene at each stage of flowering,the relative expression of safflower CtMYB-TF1 gene and CtANS gene increased with the flowering time,the relative expression of safflower CtMYB-TF1 gene reached its peak at 5 d of flowering stage.So the overexpression of CtMYB-TF1 gene can enhance CtANS gene expression in Carthamus tinctorius cells,and this can provide reference for high-producing and stable gene motificated Carthamus tinctorius clone.