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Measuring 'normalcy' in plant gene expression after herbivore attack.

Erin MacNeal Rehrig1, Heidi M Appel, Jack C Schultz

  • 1Division of Plant Sciences and Christopher Bond Life Sciences Center, The University of Missouri, 105 Bond Life Sciences Center, Columbia, MO 65211, USA. erin@rehrig.net

Molecular Ecology Resources
|March 25, 2011
PubMed
Summary

Traditional reference genes are unreliable for measuring plant gene expression after insect attack. A new method using RNA quantification and external mRNA spikes is proposed for accurate normalization in plant stress studies.

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Area of Science:

  • Plant biology
  • Molecular biology
  • Genomics

Background:

  • Plants alter gene expression to survive environmental stresses like herbivory.
  • Accurate gene expression measurement requires stable reference genes for normalization.
  • Traditional reference genes often fail in plant-herbivore interaction studies.

Purpose of the Study:

  • To evaluate the stability of commonly used reference genes in plants under insect attack.
  • To identify the limitations of traditional reference genes in plant stress research.
  • To propose a novel normalization method for gene expression analysis in herbivory studies.

Main Methods:

  • Tested the expression stability of 12 traditional reference genes in plants post-insect attack.
  • Analyzed the impact of insect attack on whole-cell metabolism and gene expression.
  • Developed and validated a normalization method using RNA quantification and an external mRNA spike.

Main Results:

  • None of the 12 traditional reference genes exhibited stable expression after insect attack.
  • Insect herbivory significantly affects plant metabolism, impacting reference gene stability.
  • The proposed method using RNA quantification and external mRNA spikes provides reliable normalization.

Conclusions:

  • Traditional reference genes are inadequate for normalizing gene expression in plants subjected to insect attack.
  • A robust normalization strategy is crucial for accurate gene expression studies in plant stress research.
  • The novel RNA quantification and external spike method offers a reliable alternative for normalization in challenging plant studies.