Evaluation and validation of housekeeping genes in two contrast species of thyme plant to drought stress using real-time PCR

Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Bibliographische Detailangaben
Veröffentlicht in:Plant physiology and biochemistry : PPB. - 1991. - 132(2018) vom: 24. Nov., Seite 54-60
1. Verfasser: Ashrafi, Mohsen (VerfasserIn)
Weitere Verfasser: Azimi Moqadam, Mohammad Reza, Moradi, Parviz, Mohsenifard, Ehsan, Shekari, Farid
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2018
Zugriff auf das übergeordnete Werk:Plant physiology and biochemistry : PPB
Schlagworte:Journal Article Abiotic stress Gene expression analysis Reference genes Thymus kotschyanus Thymus vulgaris
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100 1 |a Ashrafi, Mohsen  |e verfasserin  |4 aut 
245 1 0 |a Evaluation and validation of housekeeping genes in two contrast species of thyme plant to drought stress using real-time PCR 
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500 |a Date Completed 26.11.2018 
500 |a Date Revised 30.09.2020 
500 |a published: Print-Electronic 
500 |a Citation Status MEDLINE 
520 |a Copyright © 2018 Elsevier Masson SAS. All rights reserved. 
520 |a To decrease errors and increase accuracy and reliability of quantitative real-time PCR (qRT-PCR) results, the use of a reference gene is inevitable. Despite the industrial importance of genus Thymus, not any validated reference gene has not been reported for T. kotschyanus and T. vulgaris which could limit such investigations. In this study, the expression stability of seven housekeeping genes including Actin, Cyclophilin-18, elongation factor-1A, glyceraldehyde-3-phosphate dehydrogenase, 18S ribosomal RNA, Cullin, and Polypyrimidine tract-binding protein were evaluated in T. kotschyanus and T. vulgaris which grown at four levels of drought stress using geNorm, NormFinder, and BestKeeper algorithms. Histone deacetylase-6 (HDA-6) gene was also used for validation of evaluated reference genes. In T. vulgaris, all of the algorithms similarly ranked elongation factor-1A and glyceraldehyde-3-phosphate dehydrogenase as the two most stably expressed genes. In T. kotschyanus, only NormFinder and BestKeeper had a similar ranking and identified Actin and glyceraldehyde-3-phosphate dehydrogenase as the two most stably expressed genes, but geNorm algorithm ranked elongation factor-1A and glyceraldehyde-3-phosphate dehydrogenase as the best two reference genes. On the other hand, all algorithms ranked 18S rRNA and Cyclophilin-18 as the least stable genes in T. kotschyanus and T. vulgaris, respectively. Validation results indicated that there was a significant change (0.53-3.19 fold change) in relative expression of HDA-6 normalized by the best stable gene compare to the least ranked gene. Our study presented the first systematic validation of reference gene(s) selection in T. kotschyanus and T. vulgaris and provided useful information to obtain more accurate qRT-PCR results in these species 
650 4 |a Journal Article 
650 4 |a Abiotic stress 
650 4 |a Gene expression analysis 
650 4 |a Reference genes 
650 4 |a Thymus kotschyanus 
650 4 |a Thymus vulgaris 
700 1 |a Azimi Moqadam, Mohammad Reza  |e verfasserin  |4 aut 
700 1 |a Moradi, Parviz  |e verfasserin  |4 aut 
700 1 |a Mohsenifard, Ehsan  |e verfasserin  |4 aut 
700 1 |a Shekari, Farid  |e verfasserin  |4 aut 
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773 1 8 |g volume:132  |g year:2018  |g day:24  |g month:11  |g pages:54-60 
856 4 0 |u http://dx.doi.org/10.1016/j.plaphy.2018.08.007  |3 Volltext 
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