金荞麦花青素合酶基因的克隆及其表达与花青素量的相关性研究

Gene cloning of anthocyanin synthase in Fagopyrum dibotrys and correlation between its expression level and anthocyanin content

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 cDNA sequence of anthocyanins synthase （ANS） gene from Fagopyrum dibotrys （FdANS）, and to analyze the expression of FdANS in each tissue and the total anthocyanin content during florescence ofF. dibotrys. Methods The cDNA sequence ofANS gene was cloned by homology cloning from F. dibotrys. The expression of ANS was analyzed by semi-quantitative RT-PCR. The content of total anthocyanin was measured by spectrophotometry. Results Theopenreadingframe（ORF）ofFdANSwas1077 bp and encoded a protein with 358 amino acids named FdANS. Bioinforamtion analysis suggested that the amino acid sequence of FdANS had the higher homology with those in other plants. Both the gene expression FdANS in different tissues during florescence ofF. dibotrys and the total anthocyanin content were in the order of flowers 〉 leaves 〉 stems〉 roots. Conclusion The cDNA sequence of FdANS is firstly obtained from F. dibotrys and its coding protein has the typical characteristic of ANS homologous protein. The gene expression of FdANS shows the correlation to the total anthocyanins content in the flowers, leaves, stems, and roots ofF. dibotrys.