CYP71AV1和CPR基因在转基因青蒿后代中的遗传分析  

Inheritance Analysis of CYP71AV1 and CPR Genes in Progeny of Transgenic Artemisia annua L.

作　　者：王涛 沈乾[1,2,3] 陆续 江伟民[1,2,3] 唐克轩[1,2,3] 

机构地区：[1]上海交通大学植物生物技术研究中心 [2]复旦-交大-诺丁汉植物生物技术研发中心 [3]上海交通大学农业与生物学院,上海200240

出　　处：《上海交通大学学报（农业科学版）》2012年第4期8-14,30,共8页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：国家"863"高技术研究项目(2011AA100605);上海市科委项目(08391911800);上海市"蔬菜学"重点学科建设项目(B209)

摘　　要：为研究CYP71AV1和CPR基因在转基因青蒿后代中的遗传及表达特性,在获得共转CYP71AV1和CPR基因青蒿(GYR)T0代的基础上,对其T2代用普通PCR和Southern Blot分别检测了目标基因分离比及部分植株中目标基因拷贝数;利用实时定量PCR分析了部分植株中目标基因的转录水平;利用HPLC-ELSD测定了其青蒿素含量;对其关键农艺性状也进行了测定。结果表明虽然目标基因在世代间可以顺利遗传,但其拷贝数则有一些不规则的变化,转基因位点的纯合较难实现;目标基因的表达也受到基因组的修饰限制系统影响。T2代青蒿素含量最高的一组平均含量比对照提高46.9%,植株鲜重和叶片干重则没有显著差异。本研究为进一步获得遗传稳定的转基因青蒿株系打下了基础。In order to investigate the inheritance and expression character of CYP71AV1 and CPR genes in the progenies of previous generated transgenic Artemisia annua（A.annua） plants（GYR） in which CYP71AV1 and CPR were over-expressed.Southern blotting and Quantitative Real-Time PCR methods were used to identify the copy numbers of transgenes and expression stability of the target genes in the T2 generation plants,respectively.Segregation ratio of the introduced genes in T2 generation plants was analyzed by detecting the CYP71AV1 gene by PCR.Artemisinin content and key agronomic trait（biomass） were also analyzed.The results showed that although the CYP71AV1 and CPR genes can be inherited into the progenies and were over-expressed.The segregation ratio of the introduced genes was lower than the expected theoretical ratio.There exists some copy number variation in the transgenic progenies.The highest artemisinin content in transgenic progenies was 46.9% higher than that of the control.No significant difference was observed regarding the plant fresh weight and leaf dry weight between transgenic lines and wild type control lines.This research established a good foundation for developing a stable transgenic A.annua line in application.

关 键 词：青蒿素 CYP71AV1基因 CPR基因 转基因后代 

分 类 号：Q786[生物学—分子生物学]