金荞麦查尔酮异构酶的基因克隆及其花期表达与黄酮量分析.PDFVIP

中草药 Chinese Traditional and Herbal Drugs 第 44 卷 第 11 期 2013 年 6 月 ·1481· 金荞麦查尔酮异构酶的基因克隆及其花期表达与黄酮量分析 雒晓鹏，白悦辰，高 飞，李成磊，陈 惠，吴 琦* 四川农业大学生命科学与理学院，四川 雅安 625014 摘 要：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。 关键词：金荞麦；查尔酮异构酶；基因克隆；表达水平；总黄酮 中图分类号：R282.12 文献标志码：A 文章编号：0253 - 2670(2013)11 - 1481 - 05 DOI: 10.7501/j.issn.0253-2670.2013.11.023 Gene cloning and expression level of chalcone isomerase during florescence and content of flavonoids in Fagopyrum dibotrys LUO Xiao-peng, BAI Yue-chen, GAO Fei, LI Cheng-lei, CHEN Hui, WU Qi College of Life and Basic Sciences, Sichuan Agricultural University, Ya’an 625014, China Abstract: Objective To clone and analyze the full-length cDNA of chalcone isomerase (CHI) gene from Fagopyrum dibotrys (FdCHI). To analyze the expression of CHI and the total flavonoids content during florescence of F. dibotrys. Methods The cDNA sequence of CHI was cloned by homology cloning from F. dibotrys. The expression of CHI was analyzed in the different tissues during florescence by semi-quantitative RT-PCR. The content of total flavonoids was measured by AlCl3 method. Results The open reading frame (ORF) of FdCHI was 750 bp and encoded a protein with 249 amino acids. Bioinforamtion analysis suggested that the amino acid sequence of FdCHI had the high homology with those in other plants. Gene expression analysis showed that FdCHI was highly expressed in the flowers, followed by the roots and leaves, while lower in the stems. The content of total flavonoids was the highest in the flowers then the leaves and stems, and the lowest was in