Validation of novel reference genes for reverse transcription quantitative real-time PCR in drought-stressed sugarcane

One of the most challenging aspects of RT-qPCR data analysis is the identification of reliable reference genes. Ideally, they should be neither induced nor repressed under different experimental conditions. To date, few 
One of the most challenging aspects of RT-qPCR data analysis is the identification of reliable reference genes. Ideally, they should be neither induced nor repressed under different experimental conditions. To date, few reference genes have been adequately studied for sugarcane (Saccharum spp.) using statistical approaches. In this work, six candidate genes (αTUB, GAPDH, H1, SAMDC, UBQ, and 25S rRNA) were tested for gene expression normalization of sugarcane root tissues from drought-tolerant and -sensitive accessions after continuous dehydration (24 h). By undergoing different approaches (GeNorm, NormFinder, and BestKeeper), it was shown that most of them could be used in combinations for normalization purposes, with the exception of SAMDC. Nevertheless three of them (H1, αTUB, and GAPDH) were considered the most reliable reference genes. Their suitability as reference genes validated the expression profiles of two targets (AS and PFPα1), related to SuperSAGE unitags, in agreement with results revealed by previous in silico analysis. The other two sugarcane unitags (ACC oxidase and PIP1-1), after salt stress (100 mM NaCl), presented their expressions validated in the same way. In conclusion, these reference genes will be useful for dissecting gene expression in sugarcane roots under abiotic stress, especially in transcriptomic studies using SuperSAGE or RNAseq approaches.
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Author:Roberta Lane de Oliveira Silva, Manassés Daniel Silva, José Ribamar Costa Ferreira Neto, Claudia Huerta de Nardi, Sabrina Moutinho Chabregas, William Lee Burnquist, Günter Kahl, Ana Maria Benko-Iseppon, Ederson Akio Kido
URN:urn:nbn:de:hebis:30:3-371790
DOI:http://dx.doi.org/10.1155/2014/357052
ISSN:1537-744X
Pubmed Id:http://www.ncbi.nlm.nih.gov/pubmed?term=24987730
Parent Title (English):The scientific world journal
Publisher:Hindawi Publishing Corp.
Document Type:Article
Language:English
Date of Publication (online):2014/06/02
Date of first Publication:2014/06/02
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2015/03/22
Volume:2014
Issue:Article ID 357052
Pagenumber:13
First Page:1
Last Page:12
Note:
Copyright © 2014 Roberta Lane de Oliveira Silva et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
HeBIS PPN:368924238
Institutes:Biowissenschaften
Dewey Decimal Classification:570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
Sondersammelgebiets-Volltexte
Licence (German):License LogoCreative Commons - Namensnennung 4.0

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