Selection of reference genes in Hedysarum coronarium under various stresses and stages of development

Córdoba, Eva María; Die, José V.; González-Verdejo, Clara Isabel; Nadal, Salvador; Román, Belén

doi:10.1016/j.ab.2010.10.031

Cited by 28 publications

(25 citation statements)

References 29 publications

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“…This implies that there is no reference gene universally suited for every species and tissues under all types of treatments. It is really noteworthy that another best reference gene TUA has also got its suitability confirmed in Hedysarum coronarium under two conditions of abiotic stress and at various stages of development (Cordoba et al 2011), in cucumber sample pools, including those from different tissues and from the plants treated with hormones and abiotic stresses (Wan et al 2010), and in poplar infected tissues (Hacquard et al 2011). These results strongly support that TUA might be a proper reference gene in plants treated with different stresses.…”

Section: Discussionmentioning

confidence: 98%

Selection and validation of reference genes for studying stress-related agarwood formation of Aquilaria sinensis

et al. 2012

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Section: Discussionmentioning

confidence: 98%

Selection and validation of reference genes for studying stress-related agarwood formation of Aquilaria sinensis

et al. 2012

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“…Gene (Cordoba et al, 2011). Therefore, for accurate normalization, qRT-PCR data must be normalized with one or more suitable and stable reference genes.…”

Section: Discussionmentioning

confidence: 99%

“…Most frequently used reference genes in plant gene expression studies include ACT, GAPDH,TUB,UBQ and 18s,25s rRNA (Czechowski et al, 2005;Jian et al, 2008, Cordoba et al, 2011. If expression of the reference gene is altered by the experimental conditions, the results obtained may be incorrect and leads to misinterpretation (Dheda et al, 2005;Gutierrez et al, 2008).…”

Section: Quantification Of Gene Expression Levels Has Been Determinedmentioning

confidence: 97%

Cloning and validation of reference genes for normalization of gene expression studies in pearl millet [Pennisetum glaucum (L.) R. Br.] by quantitative real-time PCR

Reddy

Sharma

et al. 2015

Plant Gene

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Cloning and validation of reference genes for normalization of gene expression studies in pearl millet [Pennisetum glaucum (L.) R. Br.] by quantitative real-time PCR Cloning and validation of reference genes for normalization of gene expression studies in pearl millet [Pennisetum glaucum (L.) R. Br.] by quantitative real-time PCR, (2015), AbstractTo facilitate gene expression studies in pearl millet (Pennisetum glaucum (L.) R. Br.), the key reference genes including ACP, ACT, TUB, CYP, EIF4A, GAPDH, MDH, PP2C, UBC and S24 were selected based on the available literature, and their expression stabilities were studied to determine their suitability for normalizing gene expression in pearl millet. Sequence information of the reference genes were obtained from the closely related species and cloned from pearl millet using homology based cloning strategy. Further, expression stabilities were validated for their accurate expression in different tissues, genotypes and abiotic stress treatments using three statistical algorithms including geNorm, NormFinder and RefFinder.Analysis showed that while the expression of EF-1α and EIF4A were most stable in different plant tissues, MDH and EIF4A were stable under different abiotic stress conditions. Amongst the different genotypes of pearl millet tested, while UBC and MDH genes exhibited most stable expression, MDH and ACP showed greater stability in all samples set. Interestingly, the widely used reference genes S24 and TUB were found to be least stable across all the tested samples.Pair-wise analysis showed that two reference genes were sufficient for proper normalization, except when analyzing the gene expression studies in all samples set. Results of this study can help in the selection of reference genes for quantitative real time PCR (qRT-PCR) normalization in pearl millet that will contribute towards more accurate and reliable quantification of transcripts in this important crop of the drylands.

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“…However, several studies have reported variable expression of these types of genes under different conditions and tissues (Thellin et al 1999;Cordoba et al 2011;Czechowski et al 2005). With this, researchers have agreed on the need to perform basic procedures, such as the selection of genes characterized by low variation in expression levels, or typical housekeeping genes to allow for the determination of the best set of internal controls for certain treatments (Andersen et al 2004;Tunbridge et al 2011).…”

Section: Introductionmentioning

confidence: 99%

Validation of reference genes for RT-qPCR normalization in common bean during biotic and abiotic stresses

2011

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Selection of reference genes is an essential consideration to increase the precision and quality of relative expression analysis by the quantitative RT-PCR method. The stability of eight expressed sequence tags was evaluated to define potential reference genes to study the differential expression of common bean target genes under biotic (incompatible interaction between common bean and fungus Colletotrichum lindemuthianum) and abiotic (drought; salinity; cold temperature) stresses. The efficiency of amplification curves and quantification cycle (C (q)) were determined using LinRegPCR software. The stability of the candidate reference genes was obtained using geNorm and NormFinder software, whereas the normalization of differential expression of target genes [beta-1,3-glucanase 1 (BG1) gene for biotic stress and dehydration responsive element binding (DREB) gene for abiotic stress] was defined by REST software. High stability was obtained for insulin degrading enzyme (IDE), actin-11 (Act11), unknown 1 (Ukn1) and unknown 2 (Ukn2) genes during biotic stress, and for SKP1/ASK-interacting protein 16 (Skip16), Act11, Tubulin beta-8 (β-Tub8) and Unk1 genes under abiotic stresses. However, IDE and Act11 were indicated as the best combination of reference genes for biotic stress analysis, whereas the Skip16 and Act11 genes were the best combination to study abiotic stress. These genes should be useful in the normalization of gene expression by RT-PCR analysis in common bean, the most important edible legume.

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Selection of reference genes in Hedysarum coronarium under various stresses and stages of development

Cited by 28 publications

References 29 publications

Selection and validation of reference genes for studying stress-related agarwood formation of Aquilaria sinensis

Selection and validation of reference genes for studying stress-related agarwood formation of Aquilaria sinensis

Cloning and validation of reference genes for normalization of gene expression studies in pearl millet [Pennisetum glaucum (L.) R. Br.] by quantitative real-time PCR

Validation of reference genes for RT-qPCR normalization in common bean during biotic and abiotic stresses

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