Higher Activities of Defense-Associated Enzymes may Contribute to Greater Resistance of Manchurian Ash to Emerald Ash Borer Than A closely Related and Susceptible Congener

Abstract

Emerald ash borer (EAB) is an invasive beetle native to Asia that infests and kills ash (Fraxinus spp.) in North America. Previous experiments indicated that larvae feeding on co-evolved, resistant Manchurian ash (F. mandshurica) have increased antioxidant and quinone-protective enzyme activities compared to larvae feeding on susceptible North American species. Here, we examined mechanisms of host-generated oxidative and quinone-based stress and other putative defenses in Manchurian ash and the closely related and chemically similar, but susceptible, black ash (F. nigra), with and without exogenous application of methyl jasmonate (MeJA) to induce resistance mechanisms. Peroxidase activities were 4.6–13.3 times higher in Manchurian than black ash, although both species appeared to express the same three peroxidase isozymes. Additionally, peroxidase-mediated protein cross-linking activity was stronger in Manchurian ash. Polyphenol oxidase, β-glucosidase, chitinase, and lipoxygenase activities also were greater in Manchurian ash, but only lipoxygenase activity increased with MeJA application. Phloem H2O2 levels were similar and were increased by MeJA application in both species. Lastly, trypsin inhibitor activity was detected in methanol and water extracts that were not allowed to oxidize, indicating the presence of phenolic-based trypsin inhibitors. However, no proteinaceous trypsin inhibitor activity was detected in either species. In response to MeJA application, Manchurian ash had higher trypsin inhibitor activity than black ash using the unoxidized water extracts, but no treatment effects were detected using methanol extracts. Based on these results we hypothesize that peroxidases, lignin polymerization, and quinone generation contribute to the greater resistance to EAB displayed by Manchurian ash.

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Acknowledgments

We thank Brittini Hill, Elizabeth Sancomb, Lauren Shewhart, and Jennifer Jessie (Wright State University) for assistance in the field and in the laboratory, Dr. Michael Leffak and Dr. Joanna Barthelemy (Wright State University) for use of their gel imager, and Diane Hartlzer and Paul Snyder (The Ohio State University) for maintenance of the ash common garden. This project was funded by the USDA-APHIS Accelerated Emerald Ash Borer Research Program, and by state and federal funds appropriated to the Ohio Agricultural Research and Development Center, The Ohio State University, and Wright State University.

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Correspondence to Chad M. Rigsby.

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Rigsby, C.M., Herms, D.A., Bonello, P. et al. Higher Activities of Defense-Associated Enzymes may Contribute to Greater Resistance of Manchurian Ash to Emerald Ash Borer Than A closely Related and Susceptible Congener. J Chem Ecol 42, 782–792 (2016). https://doi.org/10.1007/s10886-016-0736-5

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Keywords

  • Host resistance
  • Phloem defenses
  • Phenolic oxidation
  • Oxidative stress
  • Monolignol polymerization
  • Peroxidase
  • Coleoptera
  • Buprestidae