摘要
基于狐臭柴转录组数据,本文筛选出肌动蛋白基因(ACT)、18S核糖体RNA基因(18S rRNA)、α-微管蛋白基因(TUA)、β-微管蛋白基因(TUB)、翻译延伸因子基因(EF-1)和泛素连接酶基因(UBC)等6个候选内参基因,利用实时荧光定量PCR(RT-qPCR)技术检测不同处理(暗培养、乙烯处理、模拟干旱处理)下、狐臭柴不同器官中各候选内参基因的表达情况,并运用Ge Norm、Norm Finder和Best Keeper等软件对候选内参基因的表达稳定性进行综合分析。结果表明:在狐臭柴不同器官中,18S rRNA和ACT的表达最稳定;暗培养条件下可选择UBC、18S rRNA和EF-1作为内参基因,ACT、EF-1、UBC可作为在外源乙烯处理下的内参基因,模拟干旱处理需引入第6个基因进行矫正。此外,ACT和EF-1在所有样品中的表达稳定性均较好,可以用于荧光定量表达分析。本研究结果为狐臭柴相关基因的功能分析及其调控机理研究提供了基础。
Based on the transcriptome data of Premna puberula,six candidate internal reference genes such as ACT,18S rRNA,TUA,TUB,EF-1 and UBC were screened.The expression of each candidate internal reference gene in different organs and treatments(dark culture,ethylene treatment and simulated drought treatment)of P.puberula was detected by RT-qPCR,combined with GeNorm,NormFinder and BestKeeper.The expression stability of candidate internal reference genes were comprehensively analyzed by three softwares.The results showed that the expression of 18S rRNA and ACT was the most stable in different organs of P puberula.UBC,18S rRNA and EF-1 could be selected as internal reference genes under dark culture conditions.ACT,EF-1 and UBC could be used as internal reference genes under exogenous eth-ylene treatment.Under simulated drought treatment,the sixth gene should be required to lead into for cor-rection.In addition,the expression stability of AcT and EF-1 in all samples were good,which could be used for fluorescence quantitative expression analysis.The results of this study provide a basis for the study of gene expression and other related molecular biology in P.puberula.
作者
阳娇
杨宁线
王艳秋
吴洪丽
黄海东
刘淼
张明生
YANG Jiao;YANG Ningxian;WANG Yanqiu;WU Hongli;HUANG Haidong;LIU Miao;ZHANG Mingsheng(School of Life Sciences,Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education),GuizhouUniversity,Guiyang 550025,China)
出处
《植物生理学报》
CAS
CSCD
北大核心
2023年第12期2309-2319,共11页
Plant Physiology Journal
基金
贵州省科技计划重大专项课题(黔科合平台人才[2017]5411-06)
国家喀斯特石漠化防治工程技术研究中心建设项目(2012-FU125X13)
国家重点研发计划(2016YFC0502604)
贵州省中药材现代产业技术体系建设项目(GZCYTX-02)
贵州省生物学一流学科建设项目(GNYL[2017]009FX1KT02)。
关键词
狐臭柴
内参基因
表达稳定性
实时荧光定量PCR
Premna puberula
reference gene
expression stability
quantitative real-time PCR
