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目的获取金荞麦总黄酮合成途径的关键酶查尔酮异构酶(chalcone isomerase,CHI)基因的全长序列,并进行序列分析;对花期金荞麦CHI基因在各组织中的表达与总黄酮量进行分析。方法利用同源克隆技术获得金荞麦查尔酮异构酶基因(FdCHI)的cDNA序列;采用半定量RT-PCR对CHI表达量进行分析,并采用AlCl3法测量总黄酮量。结果金荞麦FdCHI基因cDNA包含一个750 bp的ORF,编码249个氨基酸,命名为FdCHI。生物信息学分析表明该编码蛋白与其他植物CHI氨基酸序列同源率较高。FdCHI在花期金荞麦不同组织中的表达量分析表明,其表达量花>根>叶>茎,总黄酮量为花>叶>茎>根。结论在金荞麦中首次获得CHI基因的cDNA序列,编码蛋白具有CHI同源蛋白的典型特征。FdCHI基因在金荞麦茎、叶和花中的表达量与总黄酮量具有相关性,但在根中表达量与总黄酮量相关性较小。
OBJECTIVE: To obtain the full-length sequence of the chalcone isomerase (CHI) gene, which is the key pathway for total flavonoid biosynthesis in Fagopyrum tataricum, and to analyze the sequence of CHI gene. Analyze. Methods The cDNA sequence of FdCHI was obtained by homology cloning technique. The expression of CHI was analyzed by semi-quantitative RT-PCR and the total flavonoid content was measured by AlCl3 method. Results F buckle FdCHI gene cDNA contains a 750 bp ORF, encoding 249 amino acids, named FdCHI. Bioinformatics analysis showed that this encoded protein shared high homology with other plant CHI amino acid sequences. The expression levels of FdCHI in different tissues of Fagopyrum tataricum showed that the expression levels of FdCHI were flower> root> leaf> stem and total flavonoids were flower> leaf> stem> root. Conclusion The cDNA sequence of CHI gene was first obtained from Fagopyrum tataricola, and the encoded protein has typical characteristics of CHI homologous protein. The expression of FdCHI gene in stem, leaf and flower of Fagopyrum barbarum had a correlation with the total flavonoids, but the correlation between the expression of FdCHI gene and the total flavonoids in root was small.