摘要
实时荧光定量PCR(qRT-PCR)是研究植物基因定量表达的常用手段之一,选择合适的内参基因是获得可信定量结果的前提。本文以不同干旱胁迫处理下松叶猪毛菜的叶片为材料,应用q RT-PCR技术分析5个常用候选管家基因(18S r RNA、α-tubulin、β-actin、EF1-α和GAPDH)的表达变化,利用ge Norm和Norm Finder软件筛选出在干旱胁迫下表达最稳定的内参基因。结果表明,以不同干旱胁迫处理的所有样品作为总样品池进行内参基因筛选时,候选内参基因表达稳定性顺序依次是β-actin>α-tubulin>GAPDH>EF1-α>18S rRNA。对同一干旱处理的样品进行内参基因筛选时,5个候选内参基因表达稳定性存在明显差异,其中β-actin表达稳定度最高。因此,β-actin是研究松叶猪毛菜干旱胁迫下基因表达最合适的内参基因。
Real-time quantitative PCR（qRT-PCR） is one of the common technologies used for gene expression analysis.Selection of a suitable reference gene is a prerequisite to obtain the reliable results.In this study,five housekeeping genes including ribosomal RNA 18S（18S rRNA）,α-tubulin,β-actin,elongation factor 1 alpha（EFl-a） and glyceraldehyde 3-phosphate dehydrogenase（GAPDH） of the desert plant,Salsola laricifolia,were selected as candidate reference genes in the qRT-PCR,and two softwares,geNorm and NormFinder,were used to evaluate their expression stabilities in the leaves of S.laricifolia under soil drought stress in order to obtain a reliable reference gene.The results showed that there were significant differences among the five candidate reference genes under different soil drought stress treatments.And the order of expression stability was β-actinα-tubulinGAPDHEF1-α18S rRNA.While this order of expression stability was different under the same soil drought stress treatment,but the expression stability of β-actin was highest.In a word,β-actin was the most stable gene for analysis gene expression in S.laricifolia under soil drought stress.
出处
《植物生理学报》
CAS
CSCD
北大核心
2015年第11期2031-2038,共8页
Plant Physiology Journal
基金
国家自然科学基金(31300217)
中国科学院西部之光项目(XBBS201209)
关键词
松叶猪毛菜
实时荧光定量PCR
内参基因
干旱胁迫
藜科
Salsola laricifolia
real-time quantitative PCR
reference gene
soil drought stress
Chenopodiaceae
作者简介
通讯作者（E-mail：zhangml@ibeas．ac．cn；Tel：0991—7823151）。
