天然产物研究与开发 ›› 2018, Vol. 30 ›› Issue (9): 1521-1525.doi: 10.16333/j.1001-6880.2018.9.008

• 研究论文 • 上一篇    下一篇

红花查尔酮异构酶基因的克隆及表达分析

任超翔1,2,唐小慧1,2,何雯1,陈江1,2*,裴瑾1,2,吴清华1,2   

  1. 1成都中医药大学药学院;2中药资源系统研究与开发利用国家重点实验室培育基地,成都 611137
  • 出版日期:2018-10-08 发布日期:2018-10-10
  • 基金资助:

    国家自然科学基金面上项目(81573544);四川省科技厅项目(2014SZ0156);四川省教育厅项目(17ZB0 150);成都中医药大学校基金(ZRYY1610,ZRQN1647)

Cloning and Expression Analysis of Chalcone Isomerase Gene in Safflower

REN Chao-xiang1,2,TANG Xiao-hui1,2,HE Wen1,CHEN Jiang1,2*,PEI Jin1,2,WU Qing-hua1,2   

  1. 1Pharmacy College,Chengdu University of Traditional Chinese Medicine; 2State Key Laboratory Breeding Base of Systematic Research,Development and Utilization of Chinese Medicine Resources,Chengdu 611137,China
  • Online:2018-10-08 Published:2018-10-10

摘要: 实验通过下载已报道查尔酮异构酶基因序列,设计简并引物,利用RACE克隆红花查尔酮异构酶基因全长,克隆了两个红花查尔酮异构酶基因,分别命名为CtCHI1CtCHI2,NCBI登录号分别为MF996507和MF996508。用在线软件对序列进行分析,并用MEGA进行进化树分析,CtCHI1全长967 bp,CtCHI2全长997 bp,属查尔酮异构酶家族基因。定量PCR分析红花查尔酮异构酶基因表达得出,CtCHI1在花中且在时期2表达量较高,而在叶、茎及根中不表达,CtCHI2仅在茎中有少量表达。实验还发现,MeJA显著促进CtCHI1基因的表达,而对CtCHI2无调控作用,推测CtCHI1参与红花花中类黄酮的生物合成。实验成功克隆了两个查尔酮异构酶基因并对其进行表达分析,为红花类黄酮的生物合成及调控机理研究奠定基础。

关键词: 红花, 查尔酮异构酶基因, RACE, 表达分析, 类黄酮

Abstract: The reported chalcone isomerase genes were downloaded from database online,and the degenerate primers were designed.The full length of safflower chalcone isomerase genes was cloned by RACE,named as CtCHI1 and CtCHI2,and NCBI accession numbers were MF996507 and MF996508,respectively.The sequence was analyzed by software online and the evolutionary tree was constructed by MEGA.Sequence analysis showed that CtCHI1 was 967 bp and CtCHI1,2 was 997 bp,and belonged to chalcone isomerase family.The expression pattern of chalcone isomerase gene was analyzed by quantitative RT-PCR.Expression analysis showed that CtCHI1 was mainly expressed in flower with highly expression in the second developmental stages of flower,not expressed in leaves,stems and roots, CtCHI2 was mainly expressed in stem.At the same time,it was found that MeJA could significantly promote the expression of CtCHI1,but had no effect on the expression of CtCHI2.Two chalcone isomerase genes CtCHI1 and CtCHI2 were successfully cloned by RACE,and it was proposed that CtCHI1 was involved in the biosynthesis of flavonoids in safflower.

Key words: safflower, chalcone isomerase gene, RACE, expression analysis, flavonoids

中图分类号: 

R932 R284.3