Identification of reference genes for quantitative RT-PCR analysis of microRNAs and mRNAs in castor bean (Ricinus communis L.) under drought stress

Copyright © 2016. Published by Elsevier Masson SAS.

Bibliographische Detailangaben
Veröffentlicht in:Plant physiology and biochemistry : PPB. - 1991. - 106(2016) vom: 01. Sept., Seite 101-7
1. Verfasser: Cassol, Daniela (VerfasserIn)
Weitere Verfasser: Cruz, Fernanda P, Espindola, Kauê, Mangeon, Amanda, Müller, Caroline, Loureiro, Marcelo Ehlers, Corrêa, Régis L, Sachetto-Martins, Gilberto
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2016
Zugriff auf das übergeordnete Werk:Plant physiology and biochemistry : PPB
Schlagworte:Journal Article Drought Internal control genes RT-qPCR Ricinus communis L. Transcript normalization MicroRNAs RNA, Messenger
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245 1 0 |a Identification of reference genes for quantitative RT-PCR analysis of microRNAs and mRNAs in castor bean (Ricinus communis L.) under drought stress 
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520 |a Quantitative real-time PCR (RT-qPCR) is one of the most powerful and sensitive techniques to the study of gene expression. Several factors influence RT-qPCR performance though, including the stability of the reference genes used for data normalization. While the selection of appropriate reference genes is crucial for accurate and reliable gene expression analysis, no suitable reference genes have been previously identified in castor bean under drought stress. In this study, the expression stability of eleven mRNAs, thirteen microRNAs (miRNAs) and one small nuclear RNA were analyzed in roots and leaves across different levels of water deficit. Three different algorithms were employed to analyze the RT-qPCR data, and the resulting outputs were merged using a non-weighted unsupervised rank aggregation method. Our analysis indicated that the Elongation factor 1-beta (EF1B), Protein phosphatase 2A (PP2A) and ADP-ribosylation factor (ADP) ranked as the best candidates across diverse samples submitted to different levels of drought conditions. EF1B and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and EF1B and SKP1/ASK-interacting protein 16 (SKIP16) were found as the most suitable reference genes for expression analysis in roots and leaves, respectively. In addition, miRNAs miR168, miR160 and miR397 were selected as optimal reference genes across all tissues and treatments. miR168 and miR156 were recommended as reference for roots, while miR168 and miR160 were recommended for leaves. Together, our results constitute the first attempt to identify and validate the most suitable reference genes for accurate normalization of gene expression in castor bean under drought stress 
650 4 |a Journal Article 
650 4 |a Drought 
650 4 |a Internal control genes 
650 4 |a RT-qPCR 
650 4 |a Ricinus communis L. 
650 4 |a Transcript normalization 
650 7 |a MicroRNAs  |2 NLM 
650 7 |a RNA, Messenger  |2 NLM 
700 1 |a Cruz, Fernanda P  |e verfasserin  |4 aut 
700 1 |a Espindola, Kauê  |e verfasserin  |4 aut 
700 1 |a Mangeon, Amanda  |e verfasserin  |4 aut 
700 1 |a Müller, Caroline  |e verfasserin  |4 aut 
700 1 |a Loureiro, Marcelo Ehlers  |e verfasserin  |4 aut 
700 1 |a Corrêa, Régis L  |e verfasserin  |4 aut 
700 1 |a Sachetto-Martins, Gilberto  |e verfasserin  |4 aut 
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773 1 8 |g volume:106  |g year:2016  |g day:01  |g month:09  |g pages:101-7 
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