摘要
目的通过qRT-PCR分析对比红花开花后不同发育时期的种子中内参基因的表达稳定性,筛选出红花种子成熟过程中最稳定的内参基因。方法应用实时荧光定量PCR技术,分析6个传统内参基因ACT、EF1α、GAPDH、UBI、TUA、TUB在红花种子不同发育时期的mRNA表达差异情况;利用GeNorm和Normfinder软件评价上述内参基因在红花开花后不同发育阶段的种子中的表达稳定性。结果荧光定量及GeNorm软件分析结果显示,6种内参基因的表达性各异,但表达最为稳定的内参基因是EF1α,表达最稳定的内参组合是EF1α和TUB。为了数据的真实可靠又选用了Normfinder软件分析,结果显示UBI基因表达不太稳定,EF1α、ACT、TUB、TUA和GAPDH内参的表达稳定性都小于0.15,相对比较稳定。结论为红花开花后不同发育时期种子的荧光定量PCR分析提供了详尽的内参基因参考。
Objective To select the appropriate reference genes for qRT-PCR by analyzing the stability of housekeeping genes as reference genes in different developmental stages of safflower. Methods Six housekeeping genes such as ACT, EF1α, GAPDH, UBI, TUA, and TUB were selected as candidate reference genes to analyze their expression levels in safflower seeds under different developmental stages by qRT-PCR. The expression stability was evaluated by GeNorm and Norm Finder softwares. Results The expression levels of six reference genes were different, but the best was EF1α and the best combination reference genes were EF1α, TUB, and ACT. Normfinder software was applied to the verification of analysis result by GeNorm software. The Normfinder software results showed that UBI gene was not stable, and other genes were stable. Conclusion This study provides a detailed reference of housekeeping gene for qRT-PCR in different developmental stages of safflower.
出处
《中草药》
CAS
CSCD
北大核心
2017年第9期1845-1850,共6页
Chinese Traditional and Herbal Drugs
基金
吉林省科技计划-优秀青年人才基金项目(20170520035JH)
吉林省教育厅项目(2016179
2016180)
大学生科技创新项目(201707)
关键词
红花
种子
不同发育时期
荧光定量PCR
内参基因
Carthamus tinctorius
seeds
different developmental stages
qRT-PCR
reference gene
