广西植物2018，Vol.38Issue(1)：76-83,8.DOI:10.11931/guihaia.gxzw201702014
黄花大苞姜花药发育qRT⁃PCR内参基因筛选
Selection of reference genes for quantitative real?time PCR during anther development of Caulokaempferia coenobialis(Zingiberaceae)
摘要
Abstract
Quantitative real-time PCR(qRT-PCR)has been widely used in gene expression analysis with quantitative accuracy,high sensitivity and good repeat ability.The stability of the reference genes is very important to accurate analy-sis of the target genes expression.Anther tissues from pollen mother cells stage(PMC),tetrad stage(TET)and mature pollen stage(MP)of Caulokaempferia coenobialis(Zingiberaceae)were used to select reference genes.According to the developing anther transcriptome expression profile data and traditional reference genes reported,Glyceraldehyde 3?phos?phate dehydrogenase(GAPDH),Malate dehydrogenase(MDH),alpha tubulin3(TUA3), beta tubulin7(TUB7)and Actin6(ACT6)were selected as candidate reference genes for qRT-PCR analysis.The expression stability of five candi-date reference genes during the anther development was comprehensive analysis by BestKeeper,geNorm and Normfinder software. The results showed that the expression of MDH and TUB7 were the most stable, and the ACT6 was the worst. The expression patterns of GBE1 during anther development was obtained based on MDH and TUB7 as reference gene respectively. And the expression stability of MDH and TUB7 was further verified by correlation coefficient analysis (Pearson)with the anther transcriptome expression profile data.The results showed that MDH and TUB7 could serve as qRT-PCR reference gene to analyse the gene expression pattern related to anther developing in C.coenobialis.The study would provide research fundamental data for molecular mechanism research of anther development in C.coenobialis,and reference for the selection of reference genes in other Zingiberaceae species during anther development.关键词
黄花大苞姜/花药/内参基因/qRT-PCR/BestKeeper/geNorm/NormfinderKey words
Caulokaempferia coenobialis/anther/reference gene/quantitative realtime PCR(qRT-PCR)/BestKeeper/geNorm/Normfinder分类
农业科技引用本文复制引用
张俊平,王英强..黄花大苞姜花药发育qRT⁃PCR内参基因筛选[J].广西植物,2018,38(1):76-83,8.基金项目
国家自然科学基金委-广东省联合基金重点支持项目(U1301213) （U1301213）
广东省自然科学基金重点项目(7117864) [Supported by the Joint Fund of National Science Foundation of China and Guangdong Provincial Government(U1301213) （7117864）
the Key Pro-gram of Natural Science Foundation of Guangdong, China(7117864)]. （7117864）
