Selection and validation of reference genes for quantitative gene expression studies by real-time PCR in eggplant (Solanum melongena L)

BMC Research Notes

Uma Rao³  Rohini Sreevathsa¹  Divya Kamaraju³  Prasoon Kumar Thakur³  Pradeep Kumar Papolu²  Nagavara Prasad Gantasala² 

[1] National Research Centre for Plant Biotechnology, IARI, Pusa Campus, New Delhi, India;Department of Genetic Engineering, SRM University, Chennai, India;Division of Nematology, Indian Agricultural Research Institute, New Delhi, 110012, India

关键词: Gene expression;    Normalization;    Quantitative real-time PCR (qRT-PCR);    Solanum melongena L;    Eggplant (or) brinjal;    Housekeeping genes;    Reference genes;   

Others  :  1141948
DOI  :  10.1186/1756-0500-6-312

 received in 2013-01-18, accepted in 2013-07-29,  发布年份 2013

【 摘 要 】

Background

Analysis of gene expression patterns leads to functional understanding of biological processes. Quantitative real-time PCR has become the most commonly used technique for in-depth studies of gene expression. To quantify variation in specific gene expression, accurate and reliable normalization across different samples and tissues is necessary. This can be achieved by selecting one or more suitable reference genes to compare the target mRNA transcript levels. In the present work, we illustrate the first evaluation of potential internal control or reference genes across different developmental stages of eggplant for reliable quantification of transcripts by real-time PCR.

Results

We have evaluated the stability in expression of six candidate reference genes (18s rRNA, APRT, GAPDH, Cyclophilin, Actin, and RuBP) in a set of tissues representing six developmental stages of eggplant. The candidate genes were cloned from cDNA and analysed by real-time PCR. The expression data analyzed by three statistical methods (geNorm, NormFinder and BestKeeper) identified 18s rRNA, Cyclophilin and APRT as the most stable and suitable reference genes in eggplant. This was further confirmed in four different varieties, two representative lines of transgenic eggplant as well as in nematode infected eggplant.

Conclusion

18s rRNA, Cyclophilin and APRT have been found to be appropriate for the normalization of real-time PCR data for gene expression studies in eggplant.

【 授权许可】

   
2013 Gantasala et al.; licensee BioMed Central Ltd.

【 图 表 】

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