ArticlePloS one2014
Selection and evaluation of reference genes for expression studies with quantitative PCR in the model fungus Neurospora crassa under different environmental conditions in continuous culture.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 23 citations in OpenAlex.
- Total fungal RNA extraction from complex solid- state fermentation products.Applied microbiology and biotechnology · 2026Article
- Article
- Identification and evaluation of suitable reference genes for RT-qPCR analyses in Trichoderma atroviride under varying light conditions.Fungal biology and biotechnology · 2023Article
- Calmodulin, Calcium/Calmodulin-Dependent Kinases-1 and 2 Regulate Expression of the Heat Shock Proteins for Heat Shock Tolerance and Pheromone Signaling Genes for Sexual Development inIndian journal of microbiology · 2023Article
- Adaptation to glucose starvation is associated with molecular reorganization of the circadian clock ineLife · 2023Article
- Shannon entropy as a metric for conditional gene expression in Neurospora crassa.G3 (Bethesda, Md.) · 2021Article
- No Evidence That Homologs of Key Circadian Clock Genes Direct Circadian Programs of Development or mRNA Abundance inFrontiers in microbiology · 2020Article
- Selection and Evaluation of Appropriate Reference Genes for RT-qPCR Normalization ofBioMed research international · 2018Article
- Identification of suitable reference genes during the formation of chlamydospores in Clonostachys rosea 67-1.MicrobiologyOpen · 2017Article
- Establishment of Neurospora crassa as a host for heterologous protein production using a human antibody fragment as a model product.Microbial cell factories · 2017Article
- TheFrontiers in microbiology · 2017Article
- Identification of novel and robust internal control genes from Volvariella volvacea that are suitable for RT-qPCR in filamentous fungi.Scientific reports · 2016Article
- Identification and Evaluation of Reliable Reference Genes in the Medicinal Fungus Shiraia bambusicola.Current microbiology · 2016Article
- Suppressing the Neurospora crassa circadian clock while maintaining light responsiveness in continuous stirred tank reactors.Scientific reports · 2015Article
- Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Neurospora crassa has served as a model organism for studying circadian pathways and more recently has gained attention in the biofuel industry due to its enhanced capacity for cellulase production. However, in order to optimize N. crassa for biotechnological applications, metabolic pathways during growth under different environmental conditions must be addressed. Reverse-transcription quantitative PCR (RT-qPCR) is a technique that provides a high-throughput platform from which to measure the expression of a large set of genes over time. The selection of a suitable reference gene is critical for gene expression studies using relative quantification, as this strategy is based on normalization of target gene expression to a reference gene whose expression is stable under the experimental conditions. This study evaluated twelve candidate reference genes for use with N. crassa when grown in continuous culture bioreactors under different light and temperature conditions. Based on combined stability values from NormFinder and Best Keeper software packages, the following are the most appropriate reference genes under conditions of: (1) light/dark cycling: btl, asl, and vma1; (2) all-dark growth: btl, tbp, vma1, and vma2; (3) temperature flux: btl, vma1, act, and asl; (4) all conditions combined: vma1, vma2, tbp, and btl. Since N. crassa exists as different cell types (uni- or multi-nucleated), expression changes in a subset of the candidate genes was further assessed using absolute quantification. A strong negative correlation was found to exist between ratio and threshold cycle (CT) values, demonstrating that CT changes serve as a reliable reflection of transcript, and not gene copy number, fluctuations. The results of this study identified genes that are appropriate for use as reference genes in RT-qPCR studies with N. crassa and demonstrated that even with the presence of different cell types, relative quantification is an acceptable method for measuring gene expression changes during growth in bioreactors.
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