文献类型：期刊文献

英文题名：Selection and Validation of Reference Genes for Real-Time Quantitative PCR in Hyperaccumulating Ecotype ofSedum alfredii under Different Heavy Metals Stresses

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

第一作者：Sang, Jian

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

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

年份：2013

卷号：8

期号：12

外文期刊名：PLOS ONE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000328707400103)】；

基金：This work was supported by National High-tech Research & Development Program of China (No. 2013AA102701-3), Basic Scientific Research Project of Nonprofit Central Research Institutions (No. CAFYBB2012040 and NO. TGB2013008) and the National Natural Science Foundation of China (NO. 31200465)". 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：Real-time Quantitative PCR (RT-qPCR) has become an effective method for accurate analysis of gene expression in several biological systems as well as under different experimental conditions. Although with high sensitivity, specificity and broad dynamic range, this method requires suitable reference genes for transcript normalization in order to guarantee reproducible and meaningful results. In the present study, we evaluated five traditional housekeeping genes and five novel reference genes in Hyperaccumulating ecotype of Sedum alfredii, a well known hyperaccumulator for heavy metals phytoremediation, under Cd, Pb, Zn and Cu stresses of seven different durations. The expression stability of these ten candidates were determined with three programs - geNorm, NormFinder and BestKeeper. The results showed that all the selected reference genes except for SAND could be used for RT-qPCR normalization. Among them UBC9 and TUB were ranked as the most stable candidates across all samples by three programs together. For the least stable reference genes, however, BestKeeper produced different results compared with geNorm and NormFinder. Meanwhile, the expression profiles of PCS under Cd, Pb, Zn and Cu stresses were assessed using UBC9 and TUB respectively, and similar trends were obtained from the results of the two groups. The distinct expression patterns of PCS indicated that various strategies could be taken by plants in adaption to different heavy metals stresses. This study will provide appropriate reference genes for further gene expression quantification using RT-qPCR in Hyperaccumulator S. alfredii.