香瓜茄多组织部位和病害胁迫条件下qRT-PCR内参基因的选择  

Selection of internal reference genes of qRT-PCR under multi tissue parts and disease stress of Solanum muricatum

作　　者：黄丽萍 李思鸿 钟启文[1,2,3] 王晋民 HUANG Liping;LI Sihong;ZHONG Qiwen;WANG Jinmin(Qinghai University,Xining 810016,China;Academy of Agriculture and Forestry Sciences,Qinghai University,Xining 810016,China;Key Laboratory of Vegetable Genetics and Physiology of Qinghai,Xining 810016,China)

机构地区：[1]青海大学,青海西宁810016 [2]青海大学农林科学院,青海西宁810016 [3]青海省蔬菜遗传与生理重点实验室,青海西宁810016

出　　处：《青海大学学报》2022年第3期33-40,共8页Journal of Qinghai University

基　　金：青海省科学技术厅项目(2020-ZJ-Y02);青海省科学技术协会中青年人才托举工程项目(2019QHSKXRCTJ06);青海省农林科学院创新基金(2021-NKY-02)。

摘　　要：为筛选新兴经济作物香瓜茄的稳定内参基因,本研究以初花期香瓜茄不同组织部位及病害胁迫条件下苗期香瓜茄叶片为样本,选取9个候选内参基因GAPDH1、UBI、UK、β-TUB、GAPDH2、EF1α、ACT、APT、18S rRNA,通过geNorm、Norm Finder和BestKeeper等3个软件对它们在样本中的表达稳定性进行分析和评估。结果表明:通过geNorm软件分析确定最稳定的内参基因是APT;NormFinder软件分析确定稳定性最好的内参基因为APT,其次是18S rRNA和UBI;BestKeeper软件分析确定稳定内参基因为18S rRNA和UBI。综合分析结果,在香瓜茄不同组织部位和病害胁迫条件下APT基因均有良好的表达稳定性,最终确定APT基因是定量检测香瓜茄基因表达的稳定内参基因。Different tissue parts of Solanum muricatum at the initial flowering stage and its leaves at seedling stage under disease stress are taken as samples in order to screen the stable internal reference genes of emerging cash crop Solanum muricatum.There are 9 candidate reference genes selected in the study,including GAPDH1,UBI,UK,β-TUB,GAPDH2,EF1α,ACT,APT and18S rRNA.Their expression stability is analyzed and evaluated by geNorm,Norm Finder and BestKeeper.The results show that the most stable internal reference gene is APT based on the analysis of geNorm;according to the analysis of NormFinder,APT is the most stable internal reference gene,followed by 18S rRNA and UBI;the analysis results of BestKeeper shows that the qualified stable internal reference genes are 18S rRNA and UBI.In general,APT genes in different tissue of Solanum muricatum have good expression stability under disease stress conditions.In conclusion,APT gene is determined to be a stable internal reference gene for quantitative detection of gene expression in Solanum muricatum.

关 键 词：香瓜茄 QRT-PCR 内参基因 GENORM NORMFINDER 

分 类 号：S641[农业科学—蔬菜学]