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Defensive Spiroketals from Asceles glaber (Phasmatodea): Absolute Configuration and Effects on Ants and Mosquitoes

Abstract

Insects are the largest and most diverse group of organisms on earth, with over 1,000,000 species identified to date. Stick insects (“walkingsticks” or “phasmids”, Order Phasmatodea) are known for and name-derived from their camouflage that acts as a primary line of defense from predation. However, many species also possess a potent chemical defense spray. Recently we discovered that the spray of Asceles glaber contains spiroketals [a confirmed major component: (2S,6R)-(−)(E)-2-methyl-1,7-dioxaspiro[5.5]undecane, and a tentatively identified minor component: 2-ethyl-1,6-dioxaspiro[4.5]decane] and glucose. In this paper, we: 1) illustrate the identification of spiroketals and glucose in the defense spray of A. glaber by using Nuclear Magnetic Resonance (NMR), Gas Chromatography/Mass Spectrometry (GC/MS), and comparison with a synthetic reference sample; 2) provide the elucidation of the absolute configuration of the major spiroketal in that defense spray; and 3) demonstrate the effect of this compound and its enantiomer on both fire ants (Solenopsis invicta) and mosquitoes (Aedes aegypti).

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Acknowledgement

We thank James R. Rocca at the Advanced Magnetic Resonance Imaging and Spectroscopy, University of Florida, for technical assistance on NMR experiments. We also thank David Milne and Michele Hosack for conducting fire ant bioassays. A.T.D. thanks Dr. Spencer S. Walse (USDA-ARS) for lending the enantiomer selective GC column. NMR data were collected in the AMRIS facility in the McKnight Brain Institute of the University of Florida. Funding was provided by the NSF-supported National High Magnetic Field Laboratory (University of Florida) and NIH Grant GM026782 (WRR, Scripps Florida).

Supplemental Material Available online: S1) GC/MS Chromatograms of all natural samples of Asceles glaber defense spray analyzed; S2) Mass spectra of A. glaber defense spray samples and synthetic spiroketal 1; S3) TIC for A. glaber chemical defense spray; S4) The EI mass spectra for A. glaber chemical defense spray; S5) TIC and mass spectrum of synthetic spiroketal 1; S6) The NIST EI Mass Spectral Library search identified the minor peak in A. glaber defense spray; S7) The NIST EI Mass Spectral Library search for the major peak for A. glaber chemical defense spray; S8) 1D 1H NMR spectral overlays of natural A. glaber defense spray and synthetic spiroketal 1 In benzene-d6; S9) NMR spectra of A. glaber defense spray extracted with benzene-d6; S10) NMR spectra of synthetic spiroketal 1; S11) EI mass spectra from enantiomer selective GC/MS analysis of A. glaber chemical defense spray and synthetic spiroketal 1; S12) External calibration curves from the GC/MS of synthetic spiroketal 1 for quantification of that compound in A. glaber chemical defense spray; S13) Extended table of concentrations of spiroketal 1 in A. glaber chemical defense spray; S14) 1D 1H NMR spectral stack plots of natural A. glaber chemical defense spray and authentic D-Glucose dissolved in D2O, S15) NMR spectra of A. glaber defense spray dissolved in D2O; S16) NMR spectra of synthetic spiroketal 1 dissolved in D2O; and S17) NMR spectra of authentic D-Glucose dissolved in D2O.

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Dossey, A.T., Whitaker, J.M., Dancel, M.C.A. et al. Defensive Spiroketals from Asceles glaber (Phasmatodea): Absolute Configuration and Effects on Ants and Mosquitoes. J Chem Ecol 38, 1105–1115 (2012). https://doi.org/10.1007/s10886-012-0183-x

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Keywords

  • Spiroketal
  • Phasmatodea
  • Asceles glaber
  • Solenopsis invicta
  • Aedes aegypti
  • Defense
  • Phasmatodea.