紫心甘薯转录因子bHLH的基因克隆及功能研究

The Cloning and Functional Analysis of the Transcription Factor bHLH in Purple-Fleshed Sweet Potato

  • 摘要: 通过对紫心甘薯中bHLH类转录因子的基因进行分子克隆,对其结构、表达模式及功能进行研究,明确了其结构特征和生物学功能.通过采用RACE克隆方法,获得了编码bHLH基因且长度分别为2 516 bp和2 304 bp的cDNA全序列.基于DNA序列的分子进化树分析结果表明:两者分别属于植物bHLH1bHLH2基因家族的成员,分别将其命名为IbbHLH1(GenBank登录号:KC708871)和IbbHLH2 (GenBank登录号:JF508437);IbbHLH1和IbbHLH2蛋白均定位于细胞核; 在3个甘薯品种(系)的块根中,IbbHLH2基因与花色素苷合成途径中的酶基因(CHSCHIF3HDFRANS和3GT)的表达量变化趋势相一致,初步推测转录因子IbbHLH2可能参与了紫心甘薯花色素苷合成途径一系列酶基因的表达与调控.

     

    Abstract: The gene of the bHLH transcription factor in purple-fleshed sweet potato was cloned, and its structure, expression pattern and function were studied to reveal its structural characteristics and biological functions. The full cDNA sequences encoding the bHLH gene and having a length of 2 516 bp and 2 304 bp respectively were obtained with the RACE cloning method. The molecular phylogenetic tree analysis based on the DNA sequence showed that the two sequences belonged to the bHLH1 and bHLH2 plant gene families respectively. They were named IbbHLH1 (GenBank accession number: KC708871) and IbbHLH2 (GenBank accession number: JF508437). The IbbHLH1 and IbbHLH2 proteins were localized in the nucleus. The results of semi-quantitative RT-PCR showed that the expression level of IbbHLH2 and that of anthocyanin synthase genes (CHS, CHI, F3H, DFR, ANS and 3GT) were consistent in the roots of three sweet potato cultivars (strains). It is preliminarily speculated that the transcription factor IbbHLH2 is involved in the expression and regulation of a series of enzyme genes in the anthocyanin synthesis pathway of purple-fleshed sweet potato.

     

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