期刊
MOLECULAR BREEDING
卷 29, 期 1, 页码 261-274出版社
SPRINGER
DOI: 10.1007/s11032-010-9544-8
关键词
Ascochyta blight; Chickpea; Normalisation; Quantitative PCR; Reference genes
资金
- Spanish Ministerio de Ciencia e Innovacion
- [RTA2007-00009]
Normalisation to a reference gene is the most common method of internally controlling for error in quantitative PCR (qPCR) experiments. Studies based on qPCR in chickpea have been carried out using potential reference genes exclusively. Inappropriate normalisation may result in the acquisition of biologically irrelevant data. We have tested the expression of 12 candidate internal control genes in 36 samples representing different organs/developmental stages, genotypes and stress conditions. The most stably expressed genes were PUBQ, GAPDH, UBQ and bHLH, whereas 18S rRNA and EF-1a showed considerable regulation. The most suitable combination of reference genes for the particular experimental sets tested is provided. To illustrate the use of chickpea reference genes, we checked the expression of a putative defence gene in two different genotypes infected with Ascochyta rabiei (Pass.) Lab. The set of reference genes presented here will enable the more accurate and reliable normalisation of qPCR results for gene expression studies in this important legume crop. Our findings can be used as a starting point for reference gene selection in experimental conditions different from those tested here.
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