文献类型：期刊文献

中文题名：巨龙竹秆形发育过程实时荧光定量PCR内参基因的筛选

英文题名：Reference Gene Selection for Quantitative Real-Time PCR in Studying Culm Shape Development of Dendrocalamus sinicus

作者：郭晓娟[1] 陈凌娜[1] 杨汉奇[1]

第一作者：郭晓娟

机构：[1]中国林业科学研究院资源昆虫研究所

年份：2018

卷号：31

期号：2

起止页码：120-125

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2017】；CSCD:【CSCD2017_2018】；

基金：中央级公益性科研院所基本科研业务费专项(CAFYBB2017ZX001-8);国家自然科学基金(31270662);云南省科学技术厅(2014HB041)

语种：中文

中文关键词：巨龙竹;秆形发育;内参基因;qRT-PCR

外文关键词：Dendrocalamus sinicus; culm shape development; reference genes; qRT-PCR

分类号：S718

摘要：[目的]利用荧光定量PCR(q RT-PCR)和表达分析软件筛选适用于巨龙竹秆形发育研究的q RT-PCR的稳定内参基因。[方法]应用常规PCR和q RT-PCR,分析EF-1α、GAPDH、Actin、Tubulin、TIP-41、PP2A共6个内参基因在不同秆形巨龙竹("通直型"和"弯曲型")竹笋3个不同发育时期的表达情况。利用Ge Norm、Norm Finder软件对各内参基因的表达稳定性进行评估;以筛选的内参基因对巨龙竹木质素合成中的PTAL基因的表达量进行定量分析,验证所筛选内参基因的有效性。[结果]PCR和q RT-PCR的结果表明,6个候选内参基因PCR目的片段电泳条带单一,熔解曲线具有明显的单一峰。内参基因Actin、EF-1α、GAPDH的稳定性和相关性表现最佳;以它们为内参,对茎秆发育过程中PTAL基因进行定量分析,结果显示它们均具有高效性。[结论]Actin、EF-1α、GAPDH可以作为巨龙竹秆形发育研究中目的基因定量分析的内参基因。为进一步分析巨龙竹秆形发育过程中关键基因表达量的变化提供研究基础。
[Objective] To select suitable quantitative real-time PCR （qRT-PCR） reference genes for studying culm shape development of Dendrocalamus sinicus Chia et J. L. Sun. [Method] PCR and qRT-PCR were used to ana- lyze the mRNA expression stability of six candidate reference genes （EF-1α, GAPDH, Actin, TIP-41, Tubulin and PP2A） in a set of six bamboo shoot samples at three development stages of the straight-culmed and bending-culmed D. sinictts respectively. The software GeNorm and NomFinder were employed to evaluate the data for reference genes. Quantitative analysis of PTAL was used to test the effectiveness of the candidate reference genes. [ Result ] The results showed that six candidate reference genes were of expected size and single peaking melting curve. Of them, Actin, EF-lα and GAPDH had the best stability and correlativity. Meanwhile, these three candidate reference genes had the highest efficiency based on quantitative analysis results of the PTAL gene during the culm develop- ment. [ Conclusion] The Actin, EF-1α and GAPDH were optional inference genes for normalizing purpose genes in studying culm shape development of D. sinicus. The study can provide references for gene expression analysis in the culm development of D. sinicus.