非生物胁迫下耳叶牛皮消荧光定量PCR内参基因的筛选  

Selection of Reference Genes for qRT-PCR Analysis in Cynanchum auriculatum Under Abiotic Stress

作　　者：朱志鹏 虞健翔 沈敏 刘方方 朱德伟 康贻军 孙淼 ZHU Zhi-peng;YU Jian-xiang;SHEN Min;LIU Fang-fang;ZHU De-wei;KANG Yi-jun;SUN Miao(College of Marine and Biological Engineering,Yancheng Teachers University/Jiangsu Key Laboratory for Bioresources of Saline Soils,Yancheng 224007,China;College of Horticulture,Nanjing Agricultural University/State Key Laboratory of Crop Genetics and Germplasm Enhancement,Nanjing 210095,China)

机构地区：[1]盐城师范学院海洋与生物工程学院/江苏省盐土生物资源研究重点实验室,江苏盐城224007 [2]南京农业大学园艺学院/作物遗传与种质创新国家重点实验室,江苏南京210095

出　　处：《中药材》2023年第10期2430-2436,共7页Journal of Chinese Medicinal Materials

基　　金：江苏省盐土生物资源重点实验室开放项目(JKLBS2016002);国家自然科学基金项目(41977112);江苏省自然科学基金项目(BK20201063);江苏省高等学校自然科学研究面上项目(21KJB210007);江苏省产学研合作项目(BY20221008)。

摘　　要：目的:筛选可用于耳叶牛皮消非生物胁迫条件下的内参基因,掌握耳叶牛皮消中的CaWRKY1基因对不同胁迫影响的表达特性。方法:基于耳叶牛皮消转录组数据,利用荧光定量PCR(qRT-PCR)和4种算法(geNorm、NormFinder、BestKeeper和RefFinder),分析和评价12个常用内参基因在耳叶牛皮消中的表达稳定性,并实测CaWRKY1在不同非生物胁迫下耳叶牛皮消中的表达情况。结果:EF2和SAMDC(高温胁迫)、CYP和TUB-β(低温胁迫)、TUB-α和GAPDH(干旱胁迫)、SAMDC和TUB-α(水涝胁迫)、EF2和ACT7(盐胁迫)的表达最稳定。结论:在不同非生物胁迫条件下,EF2是所有候选基因中表现最稳定的基因,该结果为进一步开展耳叶牛皮消应答非生物胁迫的分子机制研究提供了方法学保障。Objective:To screen the internal reference genes in Cynanchum auriculatum under abiotic stress,to master the expression characteristics of CaWRKY1 gene in Cynanchum auriculatum under different abiotic stresses.Methods:Based on the transcriptome data of Cynanchum auriculatum,the expression stability of 12 commonly used reference genes in Cynanchum auriculatum was analyzed and evaluated by fluorescence quantitative PCR(qRT-PCR)and four algorithms(geNorm,NormFinder,BestKeeper and RefFinder).The expression of CaWRKY1 in Cynanchum auriculatum under different abiotic stresses was measured.Results:The expressions of EF2 and SAMDC(high temperature stress),CYP and TUB-β(low temperature stress),TUB-αand GAPDH(drought stress),SAMDC and TUB-α(waterlogging stress),EF2 and ACT7(salt stress)were the most stable.Conclusion:EF2 is the most stable gene among all candidate genes in Cynanchum auriculatum under different abiotic stress conditions,the result provides methodological guarantee for further research on the molecular mechanism of Cynanchum auriculatum response to abiotic stress.

关 键 词：耳叶牛皮消 内参基因 QRT-PCR 非生物胁迫 基因表达特性 

分 类 号：R282.71[医药卫生—中药学]