桉树焦枯病菌内参基因的筛选
Scerening and evaluation of reference genes of Calonectria pseudorte eaudii
陈慧洁 1冯丽贞 1李慧敏 1杨泽慧 1丁奕 1郭朦朦1
作者信息
- 1. 福建农林大学林学院, 福建福州350002;福建农林大学森林保护研究所, 福建福州350002
- 折叠
摘要
为筛选出桉树焦枯病菌在盐胁迫及侵染桉树叶片两种条件下表达稳定的内参基因,采用实时荧光定量聚合酶链式扩增反应(RT-qPCR)技术分析了桉树焦枯病菌中7个常用内参基因(GAPDH、 α-tubulin-1、 α-tubulin-2、18S rRNA、ubqi uitin、 β-actin、 β-tubulin)的mRNA差异表达情况,并采用geNorm软件分析了它们在两种条件下的表达稳定性.结果表明,ubiquitin和α-tubulin-2为桉树焦枯病菌在NaCl胁迫下表达最稳定的基因,且不需要引入第3个基因;18S rRNA和α-tubulin-1为侵染桉树叶片下表达最稳定的基因,但需引入第3个基因GA PDH.因此,ubiquitin和α-tubulin-2为桉树焦枯病菌在NaCl胁迫下较适合的内参基因,而18S rRNA、 α-tubulin-1和GAPDH为侵染桉树叶片较适合的内参基因.
Abstract
In order to screen out the best reference genes for ( real-time quantitative polymerase chain reaction, RT-qPCR ) normalization in Calonectria pseudoreteaudii under salt stress and infection.The mRNA differential expression of 7 traditional reference genes, including GAPDH, α-tbu ulin-1, α-tubulin-2, 18S rRNA, ubiquitin, β-actin andβ-tubulin of C.pseuodr eteaudii, were systematically compared by RT-qPCR.The expression stability of the 7 candidate reference genes were evaluated with geNorm program under different salt stress and injection.The results showed that ubiquitin and α-tubulin-2 were the most stable genes of C.pseudoreteaudii under salt stress while the third gene was not need to introduce;18S rRNA andα-tubulin-1 were the most stable genes under infection when GAPDH was introduced.Therefore, ubiquitin andα-tubulin-2 were the appropriate reference gene as the stability expression under salt stress while 18S rRNA, α-tubulin-1 and GAPDH were the suitable reference gene under infection.
关键词
桉树焦枯病菌/盐胁迫/侵染/实时荧光定量聚合酶链式扩增反应/内参基因Key words
Calonectria pseudoreteaudi/salt stress/infection/real-time quantitative polymerase chain reaction/reference gene引用本文复制引用
基金项目
福建省财政厅项目(Scz10039)
福建省财政厅项目(K8113001A)
福建农林大学科技创新专项(KFA17053A)
出版年
2018
国家科技期刊平台
NSTL
维普
万方数据