百脉根花青素合酶基因(LcANS)的克隆及表达分析  

Cloning and Expression Analysis of Anthocyanidin Synthase Gene(LcANS)from Lotus corniculatus

作　　者：马兰[1] 

机构地区：[1]生物与环境工程学院、西安文理学院,西安710065

出　　处：《基因组学与应用生物学》2017年第7期2926-2933,共8页Genomics and Applied Biology

基　　金：西安市科技计划项目(CXY1443WL23);陕西省教育厅科研项目(2015DC031)共同资助

摘　　要：花青素合成酶是牧草百脉根中原花青素合成途径的关键酶。在分析百脉根转录组基础上,采用RT-PCR从百脉根中首次克隆到花青素合成酶基因的全长编码c DNA序列,命名为Lc ANS。Lc ANS全长c DNA为1 098 bp,包含一个1 068 bp的开放读码框,编码355个氨基酸。生物信息学分析显示Lc ANS编码蛋白具有植物ANS典型的2OG-FeⅡ_Oxy氧化酶结构域,与豆科的红豆草、苜蓿、大豆等植物中同源ANS具有较高的序列一致性。Lc ANS蛋白定位于细胞质中,没有信号肽和跨膜结构域。实时荧光定量PCR结果表明,Lc ANS基因在百脉根不同组织器官中差异表达,在果荚和花中表达量最高,同时又可以响应ABA和低温的诱导。Anthocyanidin synthase is a key enzyme in the proanthocyanidins biosynthesis in forage Lotus cornicu- latus. Based on the analysisof transcriptome database of L. corniculatus, the full-length coding cDNA sequence of anthocyanidin synthase gene was cloned by RT-PCR for the first time and named as LcANS. The sequence of LcANS was 1 098 bp in full length with an ORF of 1 068 bp, encoding 355 amino acids. Bioinformatics analysis showed that LcANS coding protein contained a 2OG-Fe II -Oxy oxidase domain, a typical domain of plant ANS proteins, which had high identities with homologous proteins from leguminous plants, such as Onobrychis viciifo- lia, Medicago sativa and soybean. Further study demonstrated that LcANS was located in the cytoplasm without signal peptide and trans-membrane domain. Quantitative RT-PCR analysis revealed that LcA NS expressed differ- ently in different organs of L. corniculatus, with the highest expression level in pod and flora. Meanwhile, LcANS also could respond to the inducement of ABA and cold.

关 键 词：百脉根 花青素合酶 基因克隆 生物信息学分析 表达模式 

分 类 号：Q943.2[生物学—植物学] S541.6[农业科学—作物学]