中药材2023，Vol.46Issue(10)：2430-2436,7.DOI:10.13863/j.issn1001-4454.2023.10.010
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
摘要
Abstract
Objective:To screen the internal reference genes in Cynanchum auriculatum under abiotic stress,to master the expres-sion 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-α and 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 can-didate 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Key words
Cynanchum auriculatum Royle ex Wight/Reference gene/qRT-PCR/Abiotic stress/Gene expression stability分类
中医学引用本文复制引用
朱志鹏,虞健翔,沈敏,刘方方,朱德伟,康贻军,孙淼..非生物胁迫下耳叶牛皮消荧光定量PCR内参基因的筛选[J].中药材,2023,46(10):2430-2436,7.基金项目
江苏省盐土生物资源重点实验室开放项目(JKLBS2016002) （JKLBS2016002）
国家自然科学基金项目(41977112) （41977112）
江苏省自然科学基金项目(BK20201063) （BK20201063）
江苏省高等学校自然科学研究面上项目(21KJB210007) （21KJB210007）
江苏省产学研合作项目(BY20221008) （BY20221008）
