ArticleFrontiers in plant science2015
Selection of appropriate reference genes for quantitative real-time PCR in Oxytropis ochrocephala Bunge using transcriptome datasets under abiotic stress treatments.
Article in Frontiers in plant science, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 42 papers.
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42 citing papers in PubMed, 77 citations in OpenAlex.
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- Screening of stable internal reference gene of Quinoa under hormone treatment and abiotic stress.Physiology and molecular biology of plants : an international journal of functional plant biology · 2021Article
- Reference gene selection for real-time quantitative PCR assays in different tissues ofPlant direct · 2021Article
- An optimized protocol for stepwise optimization of real-time RT-PCR analysis.Horticulture research · 2021Article
- Using RNA-Seq to Identify Reference Genes of the Transition from Brown to White Adipose Tissue in Goats.Animals : an open access journal from MDPI · 2020Article
- Selection and Validation of Appropriate Reference Genes for Quantitative RT-PCR Analysis inBioMed research international · 2020Article
- Selection of reliable reference genes for gene expression analysis in seeds at different developmental stages and across various tissues in Paeonia ostii.Molecular biology reports · 2019Article
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- Validation of Reference Genes for Gene Expression Normalization in RAW264.7 Cells under Different Conditions.BioMed research international · 2019Article
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- Reference genes selection for Calotropis procera under different salt stress conditions.PloS one · 2019Article
- Identification of suitable internal control genes for transcriptional studies inPhysiology and molecular biology of plants : an international journal of functional plant biology · 2018Article
- Transcriptome-wide identification of optimal reference genes for expression analysis of Pyropia yezoensis responses to abiotic stress.BMC genomics · 2018Article
- Selection and evaluation of reference genes for qRT-PCR analysis inPlant methods · 2018Article
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5 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
backgroundOxytropis ochrocephala Bunge, an indigenous locoweed species in China, poses great threats to livestock on grasslands. There is a need for further genetic study in the plants per se, for understanding the basis of its acclimation mechanism in various unfavorable environmental conditions and to implement effective control measures. Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is the most commonly used method for gene expression analysis. To facilitate gene expression studies and obtain more accurate qRT-PCR data, normalization relative to stable reference genes is required. The aim of this study was to select the most stable reference genes for transcriptional analysis in O. ochrocephala.
resultsWe selected 12 candidate reference genes, 18S ribosomal RNA (18S RNA), actin2/7 (ACT7), β-actin (ACTB), actin101 (ACT101), actin11 (ACT11), β-tubulin (TUB), α-tubulin (TUA), glyceraldehyde-3-phosphate dehydrogenase-1 (GAPDH1), GAPDH2, metallothionein-like protein (MET), fructose-bisphosphate aldolase (FBA) and histone H3 (HIS), from the transcriptome datasets of O. ochrocephala and determined the suitability by analyzing their expression levels when exposed to a range of abiotic stress conditions. By employing software packages including geNorm, NormFinder and BestKeeper, HIS, ACT7, and ACT101 were assessed as the most suitable set for normalization in all samples. When normalized with the most stable reference genes, the expression patterns of the three target genes were in accordance with those in the transcriptome data, indicating that the reference genes selected in this study are suitable.
conclusionsThe study provided appropriate reference genes for accurate normalization in qRT-PCR analysis in O. ochrocephala and emphasized the importance of validating reference genes for gene expression analysis under specific experimental condition. The usage of inappropriate reference gene would cause misinterpretation.
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