热胁迫下糙皮侧耳实时荧光定量PCR内参基因的选择  

Selection of Reference Genes for Real-time Quantitative PCR of Pleurotus ostreatus under Heat Stress

作　　者：侯志浩 赵梦然[1] 陈强[1] 张妍 黄晨阳[1] 邬向丽[1] HOU Zhihao;ZHAO Mengran;CHEN Qiang;ZHANG Yan;HUANG Chenyang;WU Xiangli(Institute of Agricultural Resources and Regional Planning,Chinese Academy of Agricultural Sciences/Key Laboratory of Microbial Resources,Ministry of Agriculture and Rural Affairs,Beijing 100081,China)

机构地区：[1]中国农业科学院农业资源与农业区划研究所农业农村部农业微生物资源收集与保藏重点实验室

出　　处：《食用菌学报》2019年第3期11-18,F0003,共9页Acta Edulis Fungi

基　　金：国家自然科学基金资助项目(31601803);国家食用菌产业技术体系(CARS20);农作物种质资源保护与利用项目

摘　　要：在组成型表达基因和热胁迫转录组数据中选择12个基因作为热胁迫下糙皮侧耳(Pleurotus ostreatus)实时荧光定量PCR的候选基因。以40℃热处理0.5、1、3、6、12、24、48 h的糙皮侧耳菌丝体为材料,采用候选基因的特异性引物进行实时荧光定量PCR,运用geNorm、NormFinder和BestKeeper 3种软件对结果进行分析,评估12个候选基因的稳定性。结果显示β-微管蛋白基因、β-肌动蛋白基因和丝/苏氨酸蛋白磷酸酶基因可以作为糙皮侧耳热胁迫下理想的内参基因,而丝氨酸蛋白酶基因、磷酸化酶基因和细胞周期蛋白不适合作为糙皮侧耳热胁迫下的内参基因。From constitutive genes and heat stress responsive transcriptome data,12 genes were selected as candidate genes for quantitative real-time PCR of Pleurotus ostreatus under heat stress.The mycelia of P.ostreatus treated at 40℃ for various time(0.5,1,3,6,12,24,48 h)were used as the templates in quantitative real-time PCR using the sequence-specific primers of the candidate genes.The results were analyzed by geNorm,NormFinder and BestKeeper to evaluate the stability of the 12 candidate genes under different heat stress conditions.The results showed thatβ-tubulin,β-actin and serine/threonine protein phosphatase were suitable reference genes for P.ostreatus under heat stress.On the other hand,serine protease,phosphorylase and cyclin-like-F-box were not recommended as reference genes for P.ostreatus under heat stress.

关 键 词：糙皮侧耳 内参基因 热胁迫 实时荧光定量PCR 

分 类 号：R73[医药卫生—肿瘤]