文献类型：期刊文献

英文题名：Validation of Reference Genes Aiming Accurate Normalization of qRT-PCR Data in Dendrocalamus latiflorus Munro

作者：Liu, Mingying[1,2] Jiang, Jing[1,2] Han, Xiaojiao[1,2] Qiao, Guirong[1,2] Zhuo, Renying[1,2]

第一作者：刘明英

通信作者：Zhuo, RY[1]|[a00058eb5603bedf0c2e1]卓仁英;

机构：[1]Chinese Acad Forestry, State Key Lab Tree Genet & Breeding, Beijing, Peoples R China;[2]Chinese Acad Forestry, Res Inst Subtrop Forestry, Fuyang, Peoples R China

年份：2014

卷号：9

期号：2

外文期刊名：PLOS ONE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000330626900067)】；

基金：This work was supported by National Natural Science Foundation of China (No. 31200508), Science and Technology Project of Zhejiang Province (No. 2012C22099 and No. 2012C12908-3). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

语种：英文

摘要：Background: Dendrocalamus latiflorus Munro distributes widely in subtropical areas and plays vital roles as valuable natural resources. The transcriptome sequencing for D. latiflorus Munro has been performed and numerous genes especially those predicted to be unique to D. latiflorus Munro were revealed. qRT-PCR has become a feasible approach to uncover gene expression profiling, and the accuracy and reliability of the results obtained depends upon the proper selection of stable reference genes for accurate normalization. Therefore, a set of suitable internal controls should be validated for D. latiflorus Munro. Results: In this report, twelve candidate reference genes were selected and the assessment of gene expression stability was performed in ten tissue samples and four leaf samples from seedlings and anther-regenerated plants of different ploidy. The PCR amplification efficiency was estimated, and the candidate genes were ranked according to their expression stability using three software packages: geNorm, NormFinder and Bestkeeper. GAPDH and EF1 alpha were characterized to be the most stable genes among different tissues or in all the sample pools, while CYP showed low expression stability. RPL3 had the optimal performance among four leaf samples. The application of verified reference genes was illustrated by analyzing ferritin and laccase expression profiles among different experimental sets. The analysis revealed the biological variation in ferritin and laccase transcript expression among the tissues studied and the individual plants. Conclusions: geNorm, NormFinder, and BestKeeper analyses recommended different suitable reference gene(s) for normalization according to the experimental sets. GAPDH and EF1 alpha had the highest expression stability across different tissues and RPL3 for the other sample set. This study emphasizes the importance of validating superior reference genes for qRT-PCR analysis to accurately normalize gene expression of D. latiflorus Munro.