石柱参人参皂苷合成酶基因的克隆及表达量测定  

The Cloning and Gene-Expression Level Analysis of Ginsenoside Synthase Gene in Shizhu Ginseng

作　　者：王丹丹[1] 冷春玲[1] WANG Dandan;LENG Chunling(College of Agriculture,Eastern Liaoning University,Dandong Liaoning118003,China)

机构地区：[1]辽东学院农学院

出　　处：《西北农业学报》2019年第5期743-752,共10页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：中央财政专项(851800501);丹东市2016科技攻关(2016KJ001);辽东学院青年基金(20175QN050)~~

摘　　要：以人参为对照组,采用HPLC法测定5a生石柱参样品中6种人参皂苷(Rb1、Rb2、Rc、Re、Rg1、Rg2)的质量分数,采用实时荧光定量PCR技术对不同生长期人参皂苷合成酶基因(SS、CYP82D47、CYP716A42)的表达量进行比较,以探究石柱参人参皂苷质量分数较低的原因;并对石柱参CYP82D47基因进行克隆、测序及生物信息学分析,获得石柱参系统发育树。结果表明:石柱参人参皂苷合成酶基因的表达量在根、茎、叶组织中具有高度相似性,却也具有高度组织差异性,比如SS基因在石柱参根、叶中表达活跃,在茎中表达量有限;CYP82D47基因在石柱参中的表达部位主要是茎、叶组织;而CYP82D47基因在在根、茎、叶组织中表达较活跃,尤其是根、叶中;此外,人参皂苷合成酶基因CYP82D47的表达量相对较低是造成石柱参人参皂苷质量分数较低的原因之一;由系统进化树可见,石柱参与竹节参、西洋参、三七及刺五加的亲缘关系较近,结果与物种进化程度相符合。Taking ginseng as the control,the mass fraction of six ginsenosides in 5-year-old Shizhu ginseng(Rb 1,Rb 2,Rc,Re,Rg 1,Rg 2)were determined by HPLC;the expression levels of ginsenoside synthase gene(SS,CYP P82D47,CYP716A42)were determined by real-time quantitative PCR to explore the reasons of low content of ginsenosides in Shizhu ginseng;then the CYP82D47 gene was cloned,sequenced,and bioinformatics analysis was conducted to obtain the phylogenetic tree in Shizhu ginseng.The results showed that the expression of ginsenoside synthase gene in roots,stems and leaves had high similarity,but it also had obvious organizational differences,for example,SS gene was active in roots and leaves,but limited in stem;the main expression of CYP716A42 gene was in stem and leaf tissue;the expression of CYP82D47 gene was active in root,stem and leaf,especially in roots and leaves;the relatively low expression of ginsenoside synthase gene CYP82D47 was one of the reasons why Shizhu ginseng had the lower content of ginsenoside;from the phylogenetic tree,Shizhu ginseng had the close phylogenetic relationship with Panax quinquefolium,Panax notoginseng and Acanthopanax senticosus,its phylogenetic relationship was consistent with the evolutionary degree of species.

关 键 词：石柱参 人参皂苷 克隆 qPCR 基因表达量 

分 类 号：Q663.4[生物学—生物物理学]