ArticleMolecular genetics and genomics : MGG2010
Stable internal reference genes for normalization of real-time RT-PCR in tobacco (Nicotiana tabacum) during development and abiotic stress.
Article in Molecular genetics and genomics : MGG, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 243 papers.
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243 citing papers in PubMed, 551 citations in OpenAlex.
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- A mutant allele of the long-day floral activator NtFT5 is associated with the short-day-specific flowering of tobacco cultivar Maryland Mammoth.Communications biology · 2026Article
- Pollen-expressed methionine synthases in Nicotiana tabacum regulate pollen development and germination.Plant physiology · 2026Article
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- High-frequency biparental inheritance of plant mitochondria upon chilling stress and loss of a genome-degrading nuclease.Nature plants · 2026Article
- Heterologous Production of Forskolin in Tobacco (Nicotiana tabacum) via Glandular Trichome Specific Engineering and Metabolic Flux Redirection.Plant biotechnology journal · 2026Article
- Screening and Stability Validation of RT-qPCR Reference Genes inInternational journal of molecular sciences · 2026Article
- Genome-wide analysis of BAHD gene family and in vivo characterization of HQT-like genes involved in chlorogenic acid synthesis in tobacco (Nicotiana tabacum L.).BMC plant biology · 2026Article
- Integrative transcriptomic and metabolomic analyses provide insights into the effects of overexpression and knockout ofFrontiers in plant science · 2026Article
- Time-course transcriptomic information unravels the adaptation strategies ofFrontiers in plant science · 2026Article
- Transcriptomic analysis reveals light quality response and systemic nutrient allocation under monochromatic light environments in Nicotiana tabacum.BMC plant biology · 2025Article
- qPCR-Based Reference Gene Validation inCurrent issues in molecular biology · 2025Article
- T-DNA orientation, distance between two T-DNAs, and the transformation target cells significantly impact vector backbone integration and efficiency of generating marker-free transgenic plants in a co-transformation system.The Plant journal : for cell and molecular biology · 2025Article
- Unraveling Paracetamol Metabolism and Its Circadian Regulation: Insights from Tobacco Hairy Roots as a Model System.Plants (Basel, Switzerland) · 2025Article
- Stabilizing the Baseline: Reference Gene Evaluation in Three InvasiveInternational journal of molecular sciences · 2025Article
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183 more citing papers are in PubMed but not listed here.
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Authors and funding
2 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Real-time RT-PCR is a powerful technique for the measurement of gene expression, but its accuracy depends on the stability of the internal reference gene(s) used for data normalization. Tobacco (Nicotiana tabacum) is an important model in studies of plant gene expression, but stable reference genes have not been well-studied in the tobacco system. We address this problem by analysing the expression stability of eight potential tobacco reference genes. Primers targeting each gene (18S rRNA, EF-1alpha, Ntubc2, alpha- and beta-tubulin, PP2A, L25 and actin) were developed and optimized. The expression of each gene was then measured by real-time PCR in a diverse set of 22 tobacco cDNA samples derived from developmentally distinct tissues and from plants exposed to several abiotic stresses. L25 and EF-1alpha demonstrated the highest expression stability, followed by Ntubc2. Measurement of L25 and EF-1alpha was sufficient for accurate normalization in either the developmental or stress-treated samples, but Ntubc2 was also required when considering the entire sample set. Analysis of a tobacco circadian gene (NTCP-23) verified these reference genes in an additional context, and all techniques were optimized to enable a high-throughput approach. These results provide a foundation for the more accurate and widespread use of real-time RT-PCR in tobacco.
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