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Plant-Insect Ecosystems

10-Minute Paper

P-IE: Ecology On-Demand Presentations

Reference genes for normalizing gene expression in selected tissues of an ambrosia beetle, Xylosandrus germanus (Coleoptera: Curculionidae)

On-Demand
On-Demand

    Presenting Author(s)

  • Nisha Patwa

    Post-Doctoral Research Associate
    USDA-ARS, Horticultural Insects Research Lab
    Wooster, Ohio

    • Co-Author(s)

  • Christopher M. Ranger

    Research Entomologist
    USDA-Agricultural Research Service
    Wooster, Ohio

  • MR

    Michael E. Reding

    Research Entomologist
    USDA-ARS, Horticultural Insects Research Lab
    Wooster, Ohio

  • ML

    Maximilian Lehenberger

    3Max Planck Institute for Chemical Ecology, Department of Biochemistry
    Jena, Thuringen, Germany

  • PB

    Peter Biedermann

    University of Wurzburg
    Würzburg, Baden-Wurttemberg, Germany

The wood-boring ambrosia beetle Xylosandrus germanus transports and cultivates a fungal symbiont that provides the sole source of nourishment for larvae and adults. This study sought to identify reference genes necessary for future studies aimed at characterizing gene expression within selected tissues of X. germanus, particularly the specialized structure (i.e., mycetangia) used for transporting their fungal symbiont. Total RNA was extracted from X. germanus adults using whole body, head, abdomen, early developed mycetangia with scutellum from lab-reared adults, and fully developed mycetangia with and without scutellum from lab-reared and field-collected specimens. Real time-quantitative polymerase chain reaction and RefFinder tool were used to measure the expression and ranked the stability of the following selected reference genes: 28S rRNA, AK, EF1α, CO1, and CAD. The CO1 reference gene was the most stable gene expressed in whole body and head tissues of lab-reared and field-collected X. germanus. The AK and EF1α reference genes were the most stable expressed in the abdomen of lab-reared and field-collected specimens, respectively. AK was the most stable reference gene identified in early and fully developed mycetangia with scutellum in lab-reared specimens, showed highest expression stability of reference gene. CAD was the most stable gene expressed in the mycetangia with and without the scutellum from field-collected specimens but 28S rRNA was the most stable reference gene in mycetangia without scutellum from lab-reared specimens. These results will facilitate future efforts to characterize gene expression patterns in mycetangia of X. germanus for transcriptomic and functional genomic analyses.