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Publication Type J
Authors Zhang, LL; Zhang, QK; Jiang, Y; Li, Y; Zhang, HW; Li, RF
Author Full Name Zhang, Lili; Zhang, Qike; Jiang, Ying; Li, Yang; Zhang, Haiwen; Li, Ruifen
Title Reference genes identification for normalization of qPCR under multiple stresses in Hordeum brevisubulatum
Source PLANT METHODS
Language English
Document Type Article
Author Keywords Hordeum brevisubulatum; Reference gene; Abiotic stress; Quantitative real-time PCR
Keywords Plus REAL-TIME PCR; RT-PCR; EXPRESSION; VALIDATION; SALT
Abstract BackgroundReal-time quantitative PCR has been widely used as the most reliable method to measure gene expression, due to its high accuracy and specificity. Wild barley (Hordeum brevisubulatum (Trin.) Link) is a wild relative species in Triticeae that has strong tolerance to abiotic stresses and extremely wide adaptation. However, suitable references gene have not been documented for standardization of gene expression in wild barley under abiotic stress.ResultsHere we report the first systematic and comprehensive analysis of reference genes for quantitative real-time PCR standardization in wild barley. We selected 11 genes, including ACT (Actin), ADP (ADP-ribosylation factor 1), CYP2 (Cyclophilin 2), EF-1 (Elongation factor 1-alpha), GAPDH (Glyceraldehyde 3-phosphate dehydrogenase), HSP90 (Heat shock protein 90), TUB (Alpha-tubulin), TUB6 (Beta-tubulin 6), UBI (Ubiquitin), 18SrRNA-1 (guanine1575-N7-methyltransferase) and 18SrRNA-3 (adenine1779-N6-dimethyltransferase) from a wild barley transcriptome database and analyzed their expression stabilities in shoots and roots of wild barley seedling under various stress conditions using comparative Ct, BestKeeper, Normfinder and geNorm software. The results demonstrated that ADP was the most suitable reference gene in salt stress while UBI showed peak stability under mannitol and ABA stress; EF-1 was the most appropriate reference gene for PEG, GA(3), ethylene and heat stress; 18SrRNA-3 was the best choice for cold stress; and TUB was the first stable gene across different tissues.ConclusionsOur main contribution was to identify reference genes with suitable and stable expression in wild barley under various stress conditions and in different tissues to provide a useful resource for future studies. The results demonstrate the importance of transcriptome data as a useful resource for the screening of candidate reference genes and highlight the need for specific reference genes for specific conditions. Furthermore, these findings will provide valuable information for wild barley and relative species for future research.
Author Address [Zhang, Lili; Jiang, Ying; Zhang, Haiwen; Li, Ruifen] Beijing Acad Agr & Forestry Sci, Beijing Agrobiotechnol Res Ctr, Beijing Key Lab Agr Genet Resources & Biotechnol, Beijing, Peoples R China; [Zhang, Qike; Li, Yang] Hebei Normal Univ, Coll Life Sci, Shijiazhuang, Hebei, Peoples R China
Reprint Address Li, RF (reprint author), Beijing Acad Agr & Forestry Sci, Beijing Agrobiotechnol Res Ctr, Beijing Key Lab Agr Genet Resources & Biotechnol, Beijing, Peoples R China.
E-mail Address liruifen@aliyun.com
Funding Agency and Grant Number China Postdoctoral Science Foundation [2017M620 676]; Beijing Postdoctoral Research Foundation [2018-ZZ-065]; Special Program for Innovation of Beijing Academy of Agriculture and Forestry Sciences [KJCX20180201]; Beijing Academy of Agriculture and Forestry Sciences Postdoctoral Research Fund; National Natural Science Foundation of China [3177100015]
Funding Text This work was funded by the China Postdoctoral Science Foundation (Grant No. 2017M620 676), Beijing Postdoctoral Research Foundation (Agreement No. 2018-ZZ-065), Special Program for Innovation of Beijing Academy of Agriculture and Forestry Sciences (Grant No. KJCX20180201), Beijing Academy of Agriculture and Forestry Sciences Postdoctoral Research Fund, and the National Natural Science Foundation of China (Grant No. 3177100015).
Cited Reference Count 51
Publisher BMC
Publisher City LONDON
Publisher Address CAMPUS, 4 CRINAN ST, LONDON N1 9XW, ENGLAND
ISSN 1746-4811
29-Character Source Abbreviation PLANT METHODS
ISO Source Abbreviation Plant Methods
Publication Date DEC 18
Year Published 2018
Volume 14
Article Number 110
Digital Object Identifier (DOI) 10.1186/s13007-018-0379-3
Page Count 14
Web of Science Category Biochemical Research Methods; Plant Sciences
Subject Category Biochemistry & Molecular Biology; Plant Sciences
Document Delivery Number HF0BT
Unique Article Identifier WOS:000453822400002
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