三角梅CHS基因的克隆及表达分析  

Cloning and Expression Analysis of CHS Gene from Bougainvillea spectabilis

作　　者：孙蓉 刘姗[1] 高静雷 刁毅[1] 姜少娟[1] 王胜男 SUN Rong;LIU Shan;GAO Jinglei;DIAO Yi;JIANG Shaojuan;WANG Shengnan(College of Biological and Chemical Engineering,Panzhihua University,Panzhihua Sichuan 617000,China)

机构地区：[1]攀枝花学院生物与化学工程学院,四川攀枝花617000

出　　处：《西北农业学报》2021年第10期1565-1572,共8页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：四川省科技厅科技计划(2019YFH0136);攀枝花学院博士科研启动基金(035200167)。

摘　　要：查尔酮合酶(chalcone synthase,CHS)为花青素苷合成途径的关键酶,是观赏植物花色改良分子育种的研究热点。随着生物技术的发展转录组测序技术成为快速克隆新基因的有效手段,以‘新加坡大白’三角梅苞片转录组数据库为基础,筛选克隆CHS基因,利用生物信息学方法预测分析其蛋白的结构与功能,通过实时荧光定量PCR技术检测该基因在不同花色三角梅中的表达模式。结果表明:获得的CHS基因cDNA全长1176 bp(GenBank ID:MT302539),编码一个43.96 ku的蛋白,定位于细胞质中,含有CHS特征基序及活性位点,三级结构预测显示为二聚体蛋白,系统进化树分析结果显示与同为中央种子目的植物亲缘关系较近,符合系统进化关系。qRT-PCR显示该基因在黄色叶片中的表达量是红色叶片的24倍,表明黄色更适用于花青素途径的研究。研究结果为三角梅花色改良基因转化受体的筛选提供参考。Chalcone synthase is a key enzyme involved in anthocyanin biosynthetic pathway,which is a research focus for the molecular breeding of ornamental plant flower color.With the development of biotechnology,transcriptome sequencing technology has become an effective mean to isolate and clone unknown genes.Based on Bougainville glabra‘Singapore White’transcript tags,gene coding chalcone synthase(CHS)was isolated and identified.Bioinformatic tools were used to predict the structure and function of the mature protein.The quantitative real-time PCR assay was used to examine the gene relative transcription levels in different colors of B.spectabilis.The results showed that the full-length cDNA of BsCHS gene was 1176 bp encoding a 43.96 ku protein.Its GenBank ID was MT302539.Subcellular localization analysis indicated that it was cytoplasmic protein.Comparative analysis showed that it possessed CHS characteristic motifs and active sites.In addition,Swiss-Model online tool was used to predict protein tertiary structure,the results illustrated that BsCHS was dimer protein.Phylogenetic tree analysis indicated that BsCHS had a closer genetic relationship with CHS from Centrospermae,which was in line with the phylogenetic relationship.qRT-PCR analysis showed that the expression of BsCHS in yellow leaves was 24-fold higher than the red leaves,which indicated that yellow varieties were more suitable for anthocyanin pathway research.These findings will provide reference for the B.spectabilis genetic engineering receptors screening.

关 键 词：三角梅 查尔酮合酶基因 生物信息学 荧光定量 

分 类 号：Q781[生物学—分子生物学]