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Publication Type J
Authors Li, W; Qian, YQ; Han, L; Liu, JX; Sun, ZY
Author Full Name Li, Wei; Qian, Yong-Qiang; Han, Lei; Liu, Jun-Xiang; Sun, Zhen-Yuan
Title Identification of suitable reference genes in buffalo grass for accurate transcript normalization under various abiotic stress conditions
Source GENE
Language English
Document Type Article
Author Keywords Abiotic stress; Buffalograss; Gene expression; Quantitative real-time PCR; Reference gene
Keywords Plus REAL-TIME PCR; QUANTITATIVE RT-PCR; SUPERIOR REFERENCE GENES; INTERNAL CONTROL GENES; HOUSEKEEPING GENES; EXPRESSION NORMALIZATION; VALIDATION; SELECTION; ARABIDOPSIS; QUANTIFICATION
Abstract 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.
Author Address [Li, Wei; Qian, Yong-Qiang; Han, Lei; Liu, Jun-Xiang; Sun, Zhen-Yuan] 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
Reprint Address Sun, ZY (reprint author), Chinese Acad Forestry, Res Inst Forestry, State Key Lab ofTree Genet & Breeding, Beijing, Peoples R China.
E-mail Address sunzy@263.net
Funding Agency and Grant Number National Non-profit Research Institutions of Chinese Academy of Forestry [CAFYBB2012043]; National Natural Science Foundation of China [31100505]; Beijing Natural Science Foundation [6122031]
Funding Text 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).
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Cited Reference Count 55
Times Cited 10
Total Times Cited Count (WoS, BCI, and CSCD) 11
Publisher ELSEVIER SCIENCE BV
Publisher City AMSTERDAM
Publisher Address PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
ISSN 0378-1119
29-Character Source Abbreviation GENE
ISO Source Abbreviation Gene
Publication Date AUG 15
Year Published 2014
Volume 547
Issue 1
Beginning Page 55
Ending Page 62
Digital Object Identifier (DOI) 10.1016/j.gene.2014.06.015
Page Count 8
Web of Science Category Genetics & Heredity
Subject Category Genetics & Heredity
Document Delivery Number AM0LR
Unique Article Identifier WOS:000339537600008
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