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Cited by 46Molecular Biology Techniques and ApplicationsPlant Molecular Biology ResearchPlant Stress Responses and Tolerance

Reference genes selection and normalization of oxidative stress responsive genes upon different temperature stress conditions in Hypericum perforatum L.

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Velada, Isabel, Ragonezi, Carla, Arnholdt-Schmitt, Birgit, Cardoso, Hélia·2014·PLoS ONE
DOI 10.1371/journal.pone.0115206PMC PMC4263753
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Abstract

Reverse transcription-quantitative real-time PCR (RT-qPCR) is a widely used technique for gene expression analysis. The reliability of this method depends largely on the suitable selection of stable reference genes for accurate data normalization. Hypericum perforatum L. (St. John's wort) is a field growing plant that is frequently exposed to a variety of adverse environmental stresses that can negatively affect its productivity. This widely known medicinal plant with broad pharmacological properties (anti-depressant, anti-tumor, anti-inflammatory, antiviral, antioxidant, anti-cancer, and antibacterial) has been overlooked with respect to the identification of reference genes suitable for RT-qPCR data normalization. In this study, 11 candidate reference genes were analyzed in H. perforatum plants subjected to cold and heat stresses. The expression stability of these genes was assessed using GeNorm, NormFinder and BestKeeper algorithms. The results revealed that the ranking of stability among the three algorithms showed only minor differences within each treatment. The best-ranked reference genes differed between cold- and heat-treated samples; nevertheless, TUB was the most stable gene in both experimental conditions. GSA and GAPDH were found to be reliable reference genes in cold-treated samples, while GAPDH showed low expression stability in heat-treated samples. 26SrRNA and H2A had the highest stabilities in the heat assay, whereas H2A was less stable in the cold assay. Finally, AOX1, AOX2, CAT1 and CHS genes, associated with plant stress responses and oxidative stress, were used as target genes to validate the reliability of identified reference genes. These target genes showed differential expression profiles over time in treated samples. This study not only is the first systematic analysis for the selection of suitable reference genes for RT-qPCR studies in H. perforatum subjected to temperature stress conditions, but may also provide valuable information about the roles of genes associated with temperature stress responses.

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  • The AOX pathway is a means of relieving stress from temperature and drought.

    “This conclusion agrees with the literature consensus that the AOX pathway is a means of relieving stress from temperature and drought (Wang and Vanlerberghe, 2013; Velada et al., 2014; Dahal and Vanlerberghe, 2017; Campos et al., 2021).”

    From Exploring the Applicability of Calorespirometry to Assess Seed Metabolic Stability Upon Temperature Stress Conditions—Pisum sativum L. Used as a Case Study (2022) · cites this paper as Velada et al., 2014

    The Introduction (page unavailable) says AOX limits reactive oxygen species arising from environmental stress. The Discussion (page unavailable) reports evidence that AOX improves cold tolerance and finds AOX1 responsive to cold and heat. The paper does not specifically address drought, so it supports only part of the attributed claim. (LLM, conf=0.86)

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Authors
Velada, I., Ragonezi, C., Arnholdt-Schmitt, B., & Cardoso, H.
Affiliations
University of Évora (PT)
Corresponding author
Cardoso, Hélia
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2014-12-11
Volume
9
Issue
12
Pages
e115206
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0115206
PMC
PMC4263753
OpenAlex
W2151069820
Cited by
46
References
1
Citations by year
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FWCI
1.78
Type
journal article
Language
en
Update notices
correction (2015-03-26)
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • Portuguese National Funding Agency for Science, Research and Technology (RCAAP Project by FCT) (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Portuguese National Funding Agency for Science, Research and Technology (RCAAP Project by FCT) (submittedVersion, other-oa)
  • Figshare (submittedVersion, cc-by)
Funders
  • University of Sussex
  • Directorate for Biological Sciences
  • Fundação para a Ciência e a Tecnologia (COMPETE, PEst-C/AGR/UI0115/2011, UI0115, PEst-OE/AGR/UI0115/2014)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Molecular Biology Research
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Stress Responses and Tolerance
Keywords
Hypericum perforatum, reference gene selection, qRT-PCR normalization, temperature stress, cold stress, heat stress, oxidative stress, gene expression stability, AOX1, AOX2, catalase, chalcone synthase, glyceraldehyde-3-phosphate dehydrogenase, 26S rRNA
MeSH terms
Cold Temperature, Hot Temperature, Plant Proteins, Stress, Physiological, Oxidative Stress, Gene Expression Regulation, Plant, Hypericum

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