摘要
实时荧光定量PCR(qRT-PCR)技术被广泛用于基因表达分析,筛选适宜的内参基因是运用这项技术的前提。本研究采用q RT-PCR筛选适用于越南参变种(Panax vietnamensis var.fuscidiscus)不同组织器官(根、根茎、茎和叶)的内参基因。基于越南参变种转录组数据,筛选出13个候选基因(ACT1、ACT2、bTUB1、bTUB2、aTUB、GADPH、UBQ、elF、EF-1a、F-box、CYP、QCR、TCTP)。利用qRT-PCR技术检测候选内参基因在越南参变种四个组织部位中的表达情况,并用geNorm、NormFinder和BestKeeper综合分析候选基因的表达稳定性,发现ACT1和aTUB在越南参变种不同组织部位中表达稳定性最好。用内参基因ACT1和aTUB分析PvfDS在越南参变种不同组织部位的表达,发现二者表达规律一致。结果表明,越南参变种不同组织部位适宜的内参基因为ACT1和aTUB。
Quantitative real-time PCR(qRT-PCR)is widely used for gene expression analysis.Screening suitable reference genes is the premise of using this technology.This study is aimed at screening suitable reference genes for different tissues(roots,rhizomes,stems and leaves)of Panax vietnamensis var.fuscidiscus.Based on the transcriptomic data of P.vietnamensis var.fuscidiscus,13 candidate reference genes(ACT1,ACT2,bTUB1,bTUB2,aTUB,GADPH,UBQ,elF,EF-1a,F-box,CYP,QCR and TCTP)were selected.The expression trends of these candidate genes in P.vietnamensis var.fuscidiscus were analyzed by qRT-PCR.The geNorm,NormFinder and BestKeeper softwares were used for comprehensive analysis of the expression stability of the candidate genes.It was revealed that ACT1 and aTUB showed the most stable expression in different tissues of P.vietnamensis var.fuscidiscus.The expression of the PvfDS gene was detected by using ACT1 and aTUB as reference genes,respectively.The results show that both ACT1 and aTUB were the suitable internal reference genes for different tissues of P.vietnamensis var.fuscidiscus.
作者
朱灵英
王一博
王宝婕
周青青
曾礼芳
徐福荣
马晓惠
ZHU Lingying;WANG Yibo;WANG Baojie;ZHOU Qingqing;ZENG Lifang;XU Furong;MA Xiaohui(College of Pharmaceutical Science,Yunnan University of Chinese Medicine,Kunming 650500,China)
出处
《植物生理学报》
CAS
CSCD
北大核心
2020年第2期327-335,共9页
Plant Physiology Journal
基金
资助国家自然科学基金(81760690)
云南省应用基础研究计划项目(2018FB144)
云南省科学技术厅-云南中医药大学应用基础研究联合专项[2018FF001(-004)和2019FF002(-059)]
中央本级重大增减支项目-名贵中药资源可持续利用能力建设项目(2060302).
关键词
越南参变种
内参基因
实时荧光定量PCR
Panax vietnamensis var.fuscidiscus
reference gene
quantitative real-time PCR
作者简介
共同通讯作者:徐福荣(xfrong99@163.com);共同通讯作者:马晓惠(maxiaohui1988@126.com)。
