摘要
为探索白三叶(Trifolium repens)查尔酮合成酶基因对类黄酮含量的影响,根据NCBI上提供的TrCHS基因(Genbank:2247906)序列,采用RT-PCR方法克隆获得白三叶查尔酮合成酶(chalcone synthase,CHS)基因全长cDNA片段,命名为TrCHS;成功构建含有TrCHS的植物表达载体,在烟草(Nicotiana)中进行遗传转化,经抗性筛选与分子检测,获得了转基因烟草株系,并对其类黄酮含量进行了测定。结果表明,与野生型相比,转基因烟草植株TrCHS表达量和类黄酮含量均显著提高(P<0.05),表明过表达TrCHS可提高烟草类黄酮的含量,这为后续解析TrCHS功能以及白三叶新品种选育提供了可靠的理论依据。
To explore the effect of chalcone synthase on the flavonoid content in Trifolium repens,the full-length cDNA fragment named chalcone synthase(TrCHS)gene in Trifolium repens was obtained by RT-PCR cloning,using the sequence of TrCHS(GenBank:No.2247906)provided by NCBI.The expression vector containing TrCHS was successfully constructed in tobacco and the transgenic tobacco lines were obtained and the flavonoid content was determined by resistance screening and molecular detection.The results showed that the expression of TrCHS and flavonoids in the transgenic tobacco were significantly increased compared to the wild type(P<0.05),indicating that over-expression of TrCHS increased the flavonoids content in tobacco.Thus,our results provide a theoretical basis for exploring the function of TrCHS and for breeding new varieties of Trifolium repens.
作者
吴榕
田再民
郑国华
吴华圃
吴艳玲
牛艳芳
WU Rong;TIAN Zaimin;ZHENG Guohua;WU Huapu;WU Yanling;NIU Yanfang(College of Life Science and Technology,Inner Mongolia Normal University,Hohhot 010022,Inner Mongolia,China;College of Agriculture and Forestry Science and Technology,Hebei North University,Zhangjiakou 075131,Hebei,China;Inner Mongolia Hohhot Vegetable Extension Station,Hohhot 010070,Inner Mongolia,China;Saihan District Agriculture and Animal Husbandry Bureau,Hohhot 010020,Inner Mongolia,China)
出处
《草业科学》
CAS
CSCD
2020年第2期305-313,共9页
Pratacultural Science
基金
内蒙古自然科学基金(2017MS0365)
国家自然科学基金(31660157)
内蒙古自治区高等学校研究项目(NJZY17044)
关键词
白三叶
查尔酮合成酶
烟草遗传转化
类黄酮
Trifolium repens
chalcone synthase
transgenetic tobacco
flavonoids
