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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 the key enzyme in the synthesis of proanthocyanidins from the roots of forage grasses. Based on the analysis of the genebank of Lotus japonicus, the full-length cDNA encoding the anthocyanin synthase gene was cloned from Lotus japonicus by RT-PCR and named Lc ANS. Lc ANS full-length c DNA is 1 098 bp, contains a 1 068 bp open reading frame, encoding 355 amino acids. Bioinformatics analysis showed that the Lc ANS-encoded protein possesses the 2OG-FeⅡ_Oxy oxidase domain typical of plant ANS and has high sequence identity with homologous ANS in plants of legumes such as sainfoin, alfalfa and soybean. The Lc ANS protein is localized in the cytoplasm with no signal peptide and transmembrane domain. Real-time PCR results showed that Lc ANS gene was differentially expressed in different tissues and organs of Lotus corniculatum, and was the highest in pod and flower, at the same time it could respond to ABA and low temperature induction.