ArticlePloS one2015
Selection of reference genes for qPCR- and ddPCR-based analyses of gene expression in Senescing Barley leaves.
Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed.
- Analyzing qPCR data: Better practices to facilitate rigor and reproducibility.Biochemistry and biophysics reports · 2025Article
- Mechanistic Insights into Cytokinin-Regulated Leaf Senescence in Barley: Genotype-Specific Responses in Physiology and Protein Stability.International journal of molecular sciences · 2025Article
- Cathepsin B- and L-like Protease Activities Are Induced During Developmental Barley Leaf Senescence.Plants (Basel, Switzerland) · 2024Article
- Article
- Quantitative evaluation of endogenous reference genes for ddPCR under salt stress using a moderate halophile.Extremophiles : life under extreme conditions · 2023Article
- Regulation of Phenolic Compound Production by Light Varying in Spectral Quality and Total Irradiance.International journal of molecular sciences · 2022Article
- Evaluation of FMR4, FMR5 and FMR6 Expression Levels as Non-Invasive Biomarkers for the Diagnosis of Fragile X-Associated Primary Ovarian Insufficiency (FXPOI).Journal of clinical medicine · 2022Article
- A matter of origin - identification of SEMA3A, BGLAP, SPP1 and PHEX as distinctive molecular features between bone site-specific human osteoblasts on transcription level.Frontiers in bioengineering and biotechnology · 2022Article
- Quantification of Actaea racemosa L. (black cohosh) from some of its potential adulterants using qPCR and dPCR methods.Scientific reports · 2021Article
- Genes regulating development and behavior exhibited altered expression in Drosophila melanogaster exposed to bisphenol A: use of real-time quantitative PCR (qRT-PCR) and droplet digital PCR (ddPCR) in genotoxicity study.Environmental science and pollution research international · 2021Article
- Dark-Induced Barley Leaf Senescence - A Crop System for Studying Senescence and Autophagy Mechanisms.Frontiers in plant science · 2021Review
- Digital PCR: What Relevance to Plant Studies?Biology · 2020Review
- Relationships between UBE3A and SNORD116 expression and features of autism in chromosome 15 imprinting disorders.Translational psychiatry · 2020Article
- Molecular Cloning and Functional Characterization ofPlants (Basel, Switzerland) · 2020Article
- Identification and validation of superior housekeeping gene(s) for qRT-PCR data normalization inPhysiology and molecular biology of plants : an international journal of functional plant biology · 2020Article
- Quantitative evaluation of the molecular marker using droplet digital PCR.Genomics & informatics · 2020Article
- Digital PCR as an Emerging Tool for Monitoring of Microbial Biodegradation.Molecules (Basel, Switzerland) · 2020Review
- Contribution of epigenetic variation to adaptation in Arabidopsis.Nature communications · 2018Article
- Physio-Genetic Dissection of Dark-Induced Leaf Senescence and Timing Its Reversal in Barley.Plant physiology · 2018Article
- Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Leaf senescence is a tightly regulated developmental or stress-induced process. It is accompanied by dramatic changes in cell metabolism and structure, eventually leading to the disintegration of chloroplasts, the breakdown of leaf proteins, internucleosomal fragmentation of nuclear DNA and ultimately cell death. In light of the global and intense reorganization of the senescing leaf transcriptome, measuring time-course gene expression patterns in this model is challenging due to the evident problems associated with selecting stable reference genes. We have used oligonucleotide microarray data to identify 181 genes with stable expression in the course of dark-induced senescence of barley leaf. From those genes, we selected 5 candidates and confirmed their invariant expression by both reverse transcription quantitative PCR and droplet digital PCR (ddPCR). We used the selected reference genes to normalize the level of the expression of the following senescence-responsive genes in ddPCR assays: SAG12, ICL, AGXT, CS and RbcS. We were thereby able to achieve a substantial reduction in the data variability. Although the use of reference genes is not considered mandatory in ddPCR assays, our results show that it is advisable in special cases, specifically those that involve the following conditions: i) a low number of repeats, ii) the detection of low-fold changes in gene expression or iii) series data comparisons (such as time-course experiments) in which large sample variation greatly affects the overall gene expression profile and biological interpretation of the data.
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