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Reference genes for quantitative real-time PCR analysis in the model plant foxtail millet (Setaria italica L.) subjected to abiotic stress conditions

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Title Reference genes for quantitative real-time PCR analysis in the model plant foxtail millet (Setaria italica L.) subjected to abiotic stress conditions
 
Creator Kumar, Karunesh
Muthamilarasan, Mehanathan
Prasad, Manoj
 
Subject Abiotic stress
Foxtail millet
Reference genes
Quantitative real time PCR (qRT-PCR)
GeNorm
NormFinder
Setaria italica
 
Description Accepted date: 30 April 2013
Reference genes are standards for quantifying gene expression through quantitative real-time PCR (qRT-PCR); however, the variation observed in their expression levels is the major hindrance towards realising their effective use. Hence, a systematic validation of reference genes is required to ensure proper normalization. However, no such study has been conducted in foxtail millet [Setaria italica (L.)], which has recently emerged as a model crop for genetic and genomic studies. In the present study, 8 commonly used reference genes were evaluated, including 18S ribosomal RNA, elongation factor-1α, Actin2, alpha tubulin, beta tubulin, translation factor, RNA polymerase II and adenine phosphoribosyl transferase. Expression stability of candidate internal control genes was investigated under salinity and dehydration treatments. The results obtained suggested a wide range of Ct values and variable expression of all reference genes. geNorm and NormFinder analysis had revealed that Act2 and RNA POL II are suitable reference genes for salinity stress-related studies and EF-1α and RNA POL II are appropriate internal controls for dehydration stress-related expression analyses. These qualified reference genes has also been validated for relative quantification of 14-3-3 expression analysis which demonstrated their applicability. Thus, this is the first report on selection and validation of superior reference genes for qRT-PCR in foxtail millet under different abiotic stress conditions.
Grateful thanks are due to the Director,
National Institute of Plant Genome Research (NIPGR), New Delhi,
India for providing facilities. The authors work in this area was
supported by the core Grant of NIPGR. Mr. Karunesh Kumar and Mr.
Mehanathan Muthamilarasan acknowledge the award of Senior
Research Fellowship and Junior Research Fellowship from Council of
Scientific and Industrial Research and University Grants Commission,
New Delhi, India, respectively.
 
Date 2015-11-05T09:06:34Z
2015-11-05T09:06:34Z
2013
 
Type Article
 
Identifier Plant Cell Tiss. Organ Cult., 115(1): 13-22
1573-5044
http://172.16.0.77:8080/jspui/handle/123456789/329
http://link.springer.com/article/10.1007%2Fs11240-013-0335-x
10.1007/s11240-013-0335-x
 
Language en_US
 
Publisher Springer