Selection and validation of reference genes for qRT-PCR analysis during biological invasions: The thermal adaptability of Bemisia tabaci MED

PLoS One. 2017 Mar 21;12(3):e0173821. doi: 10.1371/journal.pone.0173821. eCollection 2017.

Abstract

The Bemisia tabaci Mediterranean (MED) cryptic species has been rapidly invading to most parts of the world owing to its strong ecological adaptability, which is considered as a model insect for stress tolerance studies under rapidly changing environments. Selection of a suitable reference gene for quantitative stress-responsive gene expression analysis based on qRT-PCR is critical for elaborating the molecular mechanisms of thermotolerance. To obtain accurate and reliable normalization data in MED, eight candidate reference genes (β-act, GAPDH, β-tub, EF1-α, GST, 18S, RPL13A and α-tub) were examined under various thermal stresses for varied time periods by using geNorm, NormFinder and BestKeeper algorithms, respectively. Our results revealed that β-tub and EF1-α were the best reference genes across all sample sets. On the other hand, 18S and GADPH showed the least stability for all the samples studied. β-act was proved to be highly stable only in case of short-term thermal stresses. To our knowledge this was the first comprehensive report on validation of reference genes under varying temperature stresses in MED. The study could expedite particular discovery of thermotolerance genes in MED. Further, the present results can form the basis of further research on suitable reference genes in this invasive insect and will facilitate transcript profiling in other invasive insects.

Publication types

  • Validation Study

MeSH terms

  • Acclimatization / genetics
  • Acclimatization / physiology
  • Algorithms
  • Animals
  • Gene Expression Profiling / methods*
  • Gene Expression Profiling / standards
  • Gene Expression Profiling / statistics & numerical data
  • Genes, Insect*
  • Genomic Instability
  • Global Warming
  • Hemiptera / genetics*
  • Hemiptera / physiology*
  • Insect Proteins / genetics
  • Introduced Species*
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stress, Physiological
  • Temperature
  • Transient Receptor Potential Channels / genetics

Substances

  • Insect Proteins
  • Transient Receptor Potential Channels

Grants and funding

This work was funded by the National Key Research and Development Program (2016YFC1200600), the Ministry of Science and Technology, China, and the National Natural Science Foundation of China (31672088), and the Special Fund for Scientific Research in Environmental Protection Public Interest (201409061), and the Common Wealth Special Fund for the Agricultural Industry (No. 201303019), and the International Science & Technology Cooperation Program of China (2015DFG32300).