摘要
根据特定试验材料与条件选择RT-qPCR分析中合适的内参基因,对于准确校正目的基因的表达至关重要。该研究采用RT-qPCR技术对8个内参基因在5个发育时期、3种不同组织器官和4个不同温度处理下的表达情况进行检测,通过ΔCt法、GeNorm和NormFiner对单基因、双基因组合和三基因组合的表达差异情况进行稳定性分析。结果表明:双孢蘑菇不同发育时期、不同组织,以及不同温度处理菌丝中最佳内参基因分别为40 s+actin+Eif 5基因组合、Eif5+ubiquitin+PRL 14基因组合和EF1+ubiquitin+RPL 14基因组合。该研究结果可为双孢蘑菇发育和温度相关基因的定量表达分析提供参考。
According to specific experimental materials and conditions,the selection of appropriate internal reference genes in RT-qPCR analysis was crucial for accurate correction of target genes expression.In present study,eight internal genes were used to analyze expression stability between five development stages,three different tissues or organs,four different temperature by GeNorm,NormFiner softwares andΔCt methed.The results showed that the optimal internal reference genes of Agaricus bisporus at different developmental stages,in different tissues and at different temperatures were 40 s+actin+Eif 5 combination,Eif5+ubiquitin+PRL 14 combination and EF1+ubiquitin+RPL 14 combination,respectively.This study had important practical value for the quantitative expression analysis of key genes related with developmet and temperatural response in Agaricus bisporus.
作者
赵建霞
沈颖越
冯伟林
金群力
宋婷婷
范丽军
蔡为明
ZHAO Jianxia;SHEN Yingyue;FENG Weilin;JIN Qunli;SONG Tingting;FAN Lijun;CAI Weiming(Institute of Horticulture,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;College of Chemistry and Life Science,Zhejiang Normal University,Jinhua 321004,China)
出处
《浙江农业学报》
CSCD
北大核心
2019年第8期1312-1320,共9页
Acta Agriculturae Zhejiangensis
基金
国家现代农业技术体系(CARS-20)
浙江省重大科技专项(2016C02057)
(原)农业部公益性行业(农业)科研专项(201503137)
浙江省农业重大技术协同推广计划(2019XTTGSYJ03)
关键词
RT-QPCR
内参基因
表达稳定性
双孢蘑菇
real time quantitative PCR
internal reference genes
expression stability
Agaricus bisporus
作者简介
赵建霞(1990—),女,河南安阳人,硕士研究生,研究方向为食用菌遗传育种。E-mail:1713205733@qq.com;通信作者:蔡为明,E-mail:caiwm527@126.com
