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目的获取金荞麦花青素合成途径关键酶花青素合成酶(anthocyanins synthase,ANS)基因的全长序列,并进行序列分析;对花期金荞麦ANS基因在各组织中表达水平与花青素量的相关性进行分析。方法利用同源克隆技术获得ANS基因cDNA序列;采用半定量RT-PCR对ANS表达量进行分析;采用分光光度法测量花青素量。结果金荞麦ANS基因cDNA包含一个1 077 bp的ORF,编码358个氨基酸,命名为FdANS。生物信息学分析表明,该基因编码蛋白与其他植物ANS蛋白氨基酸序列同源率较高。FdANS在花期金荞麦不同组织中的表达量分析表明,其表达量花>叶>茎>根,花青素量为花>叶>茎>根。结论在金荞麦中首次获得ANS基因的cDNA序列,编码蛋白具有ANS同源蛋白的典型特征。FdANS基因在金荞麦根、茎、叶和花中的表达量与花青素量具有相关性。
OBJECTIVE: To obtain the full-length sequence of the anthocyanins synthase (ANS) gene, which is the key enzyme involved in anthocyanin biosynthesis pathway in Fagopyrum buckwheat, and to analyze the sequence of the ANS gene. The relevance of the analysis. Methods The cDNA sequence of ANS gene was obtained by homologous cloning. The expression of ANS was analyzed by semi-quantitative RT-PCR. The amount of anthocyanin was measured by spectrophotometry. Results The ANS cDNA of F buckle contains a 1 077 bp ORF encoding 358 amino acids, named FdANS. Bioinformatics analysis showed that the gene encoding protein and other plant ANS protein amino acid sequence homology higher. The expression level of FdANS in different tissues of Fagopyrum tataricum showed that the expression level of FdANS was> leaf> stem> root, and the amount of anthocyanins was flower> leaf> stem> root. Conclusion The cDNA sequence of ANS gene was first obtained from Fagopyrum tataricum, and the encoded protein has the typical features of ANS homologous protein. The expression level of FdANS gene in Fagopyrum barbarum, stem, leaf and flower was correlated with anthocyanin content.