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Identification and evaluation of reference genes for quantitative analysis of brazilian pine (Araucaria angustifolia Bertol. Kuntze) gene expression

PLoS ONE, ISSN: 1932-6203, Vol: 10, Issue: 8, Page: e0136714
2015
  • 11
    Citations
  • 0
    Usage
  • 26
    Captures
  • 0
    Mentions
  • 47
    Social Media
Metric Options:   Counts1 Year3 Year

Metrics Details

  • Citations
    11
  • Captures
    26
  • Social Media
    47
    • Shares, Likes & Comments
      47
      • Facebook
        47

Article Description

Quantitative analysis of gene expression is a fundamental experimental approach in many fields of plant biology, but it requires the use of internal controls representing constitutively expressed genes for reliable transcript quantification. In this study, we identified fifteen putative reference genes from an A. angustifolia transcriptome database. Variation in transcript levels was first evaluated in silico by comparing read counts and then by quantitative realtime PCR (qRT-PCR), resulting in the identification of six candidate genes. The consistency of transcript abundance was also calculated applying geNorm and NormFinder software packages followed by a validation approach using four target genes. The results presented here indicate that a diverse set of samples should ideally be used in order to identify constitutively expressed genes, and that the use of any two reference genes in combination, of the six tested genes, is sufficient for effective expression normalization. Finally, in agreement with the in silico prediction, a comprehensive analysis of the qRT-PCR data combined with validation analysis revealed that AaEIF4B-L and AaPP2A are the most suitable reference genes for comparative studies of A. angustifolia gene expression.

Bibliographic Details

http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84942866858&origin=inward; http://dx.doi.org/10.1371/journal.pone.0136714; http://www.ncbi.nlm.nih.gov/pubmed/26313945; https://dx.plos.org/10.1371/journal.pone.0136714.g003; http://dx.doi.org/10.1371/journal.pone.0136714.g003; https://dx.plos.org/10.1371/journal.pone.0136714.g004; http://dx.doi.org/10.1371/journal.pone.0136714.g004; https://dx.plos.org/10.1371/journal.pone.0136714.t004; http://dx.doi.org/10.1371/journal.pone.0136714.t004; https://dx.plos.org/10.1371/journal.pone.0136714.g001; http://dx.doi.org/10.1371/journal.pone.0136714.g001; https://dx.plos.org/10.1371/journal.pone.0136714.t003; http://dx.doi.org/10.1371/journal.pone.0136714.t003; https://dx.plos.org/10.1371/journal.pone.0136714; https://dx.plos.org/10.1371/journal.pone.0136714.t001; http://dx.doi.org/10.1371/journal.pone.0136714.t001; https://dx.plos.org/10.1371/journal.pone.0136714.g002; http://dx.doi.org/10.1371/journal.pone.0136714.g002; https://dx.plos.org/10.1371/journal.pone.0136714.t002; http://dx.doi.org/10.1371/journal.pone.0136714.t002; https://dx.plos.org/10.1371/journal.pone.0136714.t005; http://dx.doi.org/10.1371/journal.pone.0136714.t005; https://dx.doi.org/10.1371/journal.pone.0136714.t005; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0136714.t005; https://dx.doi.org/10.1371/journal.pone.0136714.t003; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0136714.t003; https://dx.doi.org/10.1371/journal.pone.0136714; https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0136714; https://dx.doi.org/10.1371/journal.pone.0136714.g003; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0136714.g003; https://dx.doi.org/10.1371/journal.pone.0136714.t004; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0136714.t004; https://dx.doi.org/10.1371/journal.pone.0136714.t002; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0136714.t002; https://dx.doi.org/10.1371/journal.pone.0136714.g002; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0136714.g002; https://dx.doi.org/10.1371/journal.pone.0136714.t001; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0136714.t001; https://dx.doi.org/10.1371/journal.pone.0136714.g004; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0136714.g004; https://dx.doi.org/10.1371/journal.pone.0136714.g001; https://journals.plos.org/plosone/article/figure?id=10.1371/journal.pone.0136714.g001; http://journals.plos.org/plosone/article/metrics?id=10.1371/journal.pone.0136714; https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0136714&type=printable; http://dx.plos.org/10.1371/journal.pone.0136714.t002; http://dx.plos.org/10.1371/journal.pone.0136714.g002; http://dx.plos.org/10.1371/journal.pone.0136714.g001; http://dx.plos.org/10.1371/journal.pone.0136714.t004; http://dx.plos.org/10.1371/journal.pone.0136714.g003; http://dx.plos.org/10.1371/journal.pone.0136714.t003; http://dx.plos.org/10.1371/journal.pone.0136714.t001; http://dx.plos.org/10.1371/journal.pone.0136714.g004; http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0136714; http://dx.plos.org/10.1371/journal.pone.0136714; http://dx.plos.org/10.1371/journal.pone.0136714.t005; http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0136714; http://www.plosone.org/article/metrics/info:doi/10.1371/journal.pone.0136714; http://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0136714&type=printable

Paula Elbl; Bruno V. Navarro; Leandro F. de Oliveira; Juliana Almeida; Amanda C. Mosini; André L. W. dos Santos; Magdalena Rossi; Eny I. S. Floh; Debasis Chakrabarty

Public Library of Science (PLoS)

Multidisciplinary

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