文献类型：期刊文献

英文题名：Identification of suitable reference genes in buffalo grass for accurate transcript normalization under various abiotic stress conditions

作者：Li, Wei[1] Qian, Yong-Qiang[1] Han, Lei[1] Liu, Jun-Xiang[1] Sun, Zhen-Yuan[1]

通信作者：Sun, ZY[1]

机构：[1]Chinese Acad Forestry, Res Inst Forestry, State Key Lab ofTree Genet & Breeding, Beijing, Peoples R China; State Forestry Adm, Key Lab Tree Breeding & Cultivat, Beijing, Peoples R China

年份：2014

卷号：547

期号：1

起止页码：55-62

外文期刊名：GENE

收录：;Scopus(收录号：2-s2.0-84903815043);WOS:【SCI-EXPANDED(收录号:WOS:000339537600008)】；

基金：The work presented here was supported by National Non-profit Research Institutions of Chinese Academy of Forestry (CAFYBB2012043), grant for the National Natural Science Foundation of China (31100505) and the Beijing Natural Science Foundation (6122031).

语种：英文

外文关键词：Abiotic stress; Buffalograss; Gene expression; Quantitative real-time PCR; Reference gene

摘要：Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) is a sensitive technique for normalization of the gene expression level of target genes. Buffalograss (Buchloe dactyloides), a warm-season turfgrass with strong abiotic stress resistance, is widely used in North China. Up to now, no work was performed to evaluate the reference genes in buffalograss. In this study, the expression profiles of ten potential reference genes were examined by qRT-PCR in 24 buffalograss samples, which were subjected to a different treatment (salt, osmotic, cold and heat). Three qRT-PCR analysis methods (GeNorm, NormFinder, and Bestkeeper) were used to evaluate the stability of gene expression. The results indicated that DNAJ and beta-ACTIN were the optimal reference genes for salt-treated leaves, and the combination of PP2A and GAPDH was better reference genes for PEG-treated leaves. Under cold stress, DNAJ and beta-ACTIN showed less variety of expression level in leaves. DNAJ and GAPDH exhibited the most stable expression in heat-treated samples. To sum up, glyceral-dehyde-3-phosphate dehydrogenase (GAPDH), beta-ACTIN, DNAJ-like protein (DNAJ) and protein phosphatase 2A (PP2A) were selected as the most stable reference gene among all tested samples. To further validate the suitability of these reference genes, the expression levels of DREB2 (homologs of AtDREB2) were analyzed in parallel. Our results show that the best reference genes differed across different experimental conditions, and these results should enable better normalization and quantification of transcript levels in buffalograss in the future. (C) 2014 Elsevier B.V. All rights reserved.