霜霉病菌胁迫下藜麦内参基因的筛选及其稳定性验证  

Screening of reference genes in Chenopodium quinoa under Peronospora variabilis stress and verification of their stability

作　　者：解宇洁 薛婧 姜晓东[2] 殷辉 赵晓军 冯铸 李新凤[1] XIE Yujie;XUE Jing;JIANG Xiaodong;YIN Hui;ZHAO Xiaojun;FENG Zhu;LI Xinfeng(College of Plant Protection,Shanxi Agricultural University,Taigu,Shanxi 030801,China;College of Agriculture,Shanxi Agricultural University,Taigu,Shanxi 030801,China;Shanxi Agricultural Seed Station,Taiyuan,Shanxi 030006,China)

机构地区：[1]山西农业大学植物保护学院,山西太谷030801 [2]山西农业大学农学院,山西太谷030801 [3]山西省农业种子总站,山西太原030006

出　　处：《福建农林大学学报（自然科学版）》2024年第2期191-198,共8页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：黄土高原特色作物优质高效生产省部共建协同创新中心基金项目(SBGJXTZXKF-11);山西省重点研发计划项目(201803D221012-2);山西农业大学生物育种工程项目(YZGC104)。

摘　　要：【目的】筛选藜麦在霜霉病菌胁迫下稳定表达的内参基因,为深入开展藜麦抗霜霉病菌相关基因表达的研究提供依据。【方法】通过实时荧光定量PCR技术以及geNorm、NormFinder、BestKeeper和RefFinder软件分析和评价8个候选内参基因在藜麦霜霉病菌胁迫下的表达稳定性,并通过对CqSGAT基因表达进行分析,进一步验证候选内参基因的稳定性。【结果】geNorm、NormFinder、BestKeeper和RefFinder软件分析结果均显示,藜麦在霜霉病菌胁迫下稳定性较好的内参基因为CqEF-1a、CqMON1和CqRPS18;geNorm软件分析结果显示,藜麦在霜霉病菌胁迫下最适候选内参基因为CqEF-1a和CqRPS18;以CqSGAT为目标基因对候选内参基因进行验证,发现CqMON1不适合作为藜麦在霜霉病菌胁迫下的内参基因。【结论】藜麦在霜霉病菌胁迫下的最适内参基因为CqEF-1a和CqRPS18。【Objective】The reference genes stably expressed in Chenopodium quinoa under Peronospora variabilis stress was screened so as to further investigate the expression of genes related to resistance against P.variabilis in C.quinoa.【Method】The expression stabilities of 8 candidate genes in C.quinoa were analyzed and evaluated under P.variabilis stress by quantitative real-time PCR and software of geNorm,NormFinder,BestKeeper and RefFinder,and further verified by expression analysis of CqSGAT gene.【Result】The analyses results of geNorm,NormFinder,BestKeeper and RefFinder all showed that CqEF-1a,CqMON1 and CqRPS18 were of good stability as the reference genes in C.quinoa under P.variabilis stress,with CqEF-1a and CqRPS18 being the most suitable reference genes according to the analysis of geNorm.Expression analysis of CqSGAT gene with the candidate reference genes further verified that CqMON1 was not suitable as reference gene.【Conclusion】CqEF-1a and CqRPS18 were the best reference genes in C.quinoa under P.variabilis stress.

关 键 词：藜麦 实时荧光定量PCR 内参基因 生物胁迫 表达稳定性 

分 类 号：S432.4[农业科学—植物病理学]