Skip to main content
SpringerLink
Log in

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

  • Published:
Journal of Chemical Ecology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Appel HM (1993) Phenolics in ecological interactions: The importance of oxidation. J Chem Ecol 19:1521–1552

    Article  CAS  PubMed  Google Scholar 

  • Bai X, Rivera-Vega L, Mamidala P, Bonello P, Herms DA, Mittapalli O (2011) Transcriptomic signatures of ash (Fraxinus spp.) phloem. PLoS One 6:e16368

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bhonwong A, Stout MJ, Attajarusit J, Tantasawat P (2009) Defensive role of tomato polyphenol oxidases against cotton bollworm (Helicoverpa armigera) and beet armyworm (Spodoptera exigua). J Chem Ecol 35:28–38

    Article  CAS  PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principal of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Broadway RM (1993) Purification and partial characterization of trypsin/chymotrypsin inhibitors from cabbage foliage. Phytochemistry 33:21–27

    Article  CAS  Google Scholar 

  • Camillo LR, Filadelfo CR, Monzani PS, Corrêa RX, Gramacho K, Micheli F, Pirovani CP (2013) Tc-cAPX, a cytosolic ascorbate peroxidase of Theobroma cacao L. engaged in the interaction with Moniliophthora perniciosa, the causing agent of witches' broom disease. Plant Physiol Biochem 73:254–265

    Article  CAS  PubMed  Google Scholar 

  • Carvalho RH, Lemos F, Lemos MANDA, Vojinović V, Fonseca LP, Cabral JMS (2006) Kinetic modeling of phenol co-oxidation using horseradish peroxidase. Bioprocess Biosyst Eng 29:99–108

    Article  CAS  PubMed  Google Scholar 

  • Chakraborty S, Whitehill JGA, Hill AL, Opiyo O, Cipollini D, Herms DA, Bonello P (2014) Effects of water availability on emerald ash borer larval performance and phloem phenolics of Manchurian and black ash. Plant Cell Environ 37:1009–1021

    Article  CAS  PubMed  Google Scholar 

  • Cheeseman JM (2007) Hydrogen peroxide and plant stress: A challenging relationship. Plant Stress 1:4–15

    Google Scholar 

  • Cipollini D, Bergelson J (2000) Environmental and developmental regulation of trypsin activity in Brassica napus. J Chem Ecol 26:1411–1422

    Article  CAS  Google Scholar 

  • Cipollini D, Wang Q, Whitehill JGA, Powell JR, Bonello P, Herms DA (2011) Distinguishing defensive characteristics in the phloem of ash species resistant and susceptible to emerald ash borer. J Chem Ecol 37:450–459

    Article  CAS  PubMed  Google Scholar 

  • Corrado G, Alagna F, Rocco M, Renzone G, Varricchio P, Coppola V, Garonna A, Baldoni L, Scaloni A, Rao R (2012) Molecular interactions between the olive and the fruit fly Bactrocera oleae. BMC Plant Biol 12:86

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dowd PF (1994) Enhanced maize (Zea mays L.) pericarp browning: Associations with insect resistance and involvement of oxidizing enzymes. J Chem Ecol 20:2777–2803

    Article  CAS  PubMed  Google Scholar 

  • Erbilgin N, Krokene P, Christiansen E, Zeneli G, Gershenzon J (2006) Exogenous application of methyl jasmonate elicits defenses in Norway spruce (Picea abies) and reduces host colonization by the bark beetle Ips typographus. Oecologia 148:426–436

    Article  PubMed  Google Scholar 

  • Eyles A, Jones W, Riedl K, Cipollini D, Schwartz S, Chan K, Herms DA, Bonello P (2007) Comparative phloem chemistry of Manchurian (Fraxinus mandshurica) and two North American ash species (Fraxinus americana and Fraxinus pennsylvanica). J Chem Ecol 33:1430–1448

    Article  CAS  PubMed  Google Scholar 

  • Felton GW, Summers CB, Mueller AJ (1994) Oxidative responses in soybean foliage to herbivory by bean leaf beetle and three-cornered alfalfa hopper. J Chem Ecol 20:639–650

    Article  CAS  PubMed  Google Scholar 

  • Feussner I, Waternack C (2002) The lipoxygenase pathway. Annu Rev Plant Biol 53:275–297

    Article  CAS  PubMed  Google Scholar 

  • Galati G, Sabzevari O, Wilson JX, O’Brien PJ (2002) Prooxidant activity and cellular effects of the phenoxyl radicals of dietary flavonoids and other polyphenolics. Toxicology 177:91–104

    Article  CAS  PubMed  Google Scholar 

  • Ginzberg I (2008) Wound-periderm formation. In: Schaller A (ed) Induced plant resistance to herbivory. Springer, Netherlands, pp. 131–146

    Chapter  Google Scholar 

  • Goldwasser Y, Hershenhorn J, Plakhine D, Kleifeld Y, Rubin B (1999) Biochemical factors involved in vetch resistance to Orobanche aegyptiaca. Physiol Mol Plant Pathol 54:87–96

    Article  CAS  Google Scholar 

  • Guo H, Sun Y, Ren Q, Zhu-Salzman K, Kang C, Li C, Ge F (2012) Elevated CO2 reduces the resistance and tolerance of tomato plants to Helicoverpa armigera by suppressing the JA signaling pathway. PLoS One 7:e41426

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hebard FV, Griffin GJ, Elkins JR (1984) Developmental histopathology of cankers incited by hypovirulent and virulent isolates of Endothia parasitica on susceptible and resistant chestnut trees. Phytopathology 74:140–149

    Article  Google Scholar 

  • Herms DA (2015) Host range and host resistance. In: Van Driesche RG, Reardon R (eds) Biology and control of emerald ash borer, Technical Bulletin FHTET 2014–09. USDA Forest Service, Morgantown, West Virginia, USA, pp. 153–163

    Google Scholar 

  • Herms DA, McCullough DG (2014) Emerald ash borer invasion of North America: history, biology, ecology, impacts, and management. Annu Rev Entomol 59:13–30

    Article  CAS  PubMed  Google Scholar 

  • Hildebrand DF, Rodriguez JGm Brown GC, Luu KT, Volden CS (1986) Peroxidative responses of leaves in two soybean genotypes injured by twospotted spider mites (Acari: Tetranychidae). J Econ Entomol 79:1459–1465

    Article  Google Scholar 

  • Junglee S, Urban L, Sallanon H, Lopez-Lauri F (2014) Optimized assay for hydrogen peroxide determination in plant tissue using potassium iodide. Am J Anal Chem 5:730–736

    Article  CAS  Google Scholar 

  • Komsta L (2011) Outliers: Tests for outliers. R package version 0.14. http://CRAN.R-project.org/package=outliers

  • Konno K, Hirayama C, Yasui H, Nakamura M (1999) Enzymatic activation of oleuropein: a protein crosslinker used as a chemical defense in the privet tree. Proc Natl Acad Sci U S A 96:9159–91640

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kwon KS, Lee J, Kang HG, Hah YC (1994) Detection of β-glucosidase activity in polyacrylamide gels with esculin as substrate. Appl Environ Microbiol 60:4584–4586

    CAS  PubMed  PubMed Central  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  • Lawrence SD, Novak NG, CJT J, Cooke JEK (2008) Potato, Solanum tuberosum, defense against Colorado potato beetle, Leptinotarsa decemlineata (say): microarray gene expression profiling of potato by Colorado potato beetle regurgitant treatment of wounded leaves. J Chem Ecol 34:1013–1025

    Article  CAS  PubMed  Google Scholar 

  • Leatham GF, King V, Stahmann MA (1980) In vitro protein polymerization by quinones or free radicals generated by plant or fungal oxidative enzymes. Phytopathology 70:1134–1140

    Article  CAS  Google Scholar 

  • Lee BR, Kim KY, Jung WJ, Avice JC, Ourry A, Kim TH (2007) Peroxidases and lignification in relation to the intensity in water-deficit stress in white clover (Trifolium repens L. J Exp Bot 58:1271–1279

    Article  CAS  PubMed  Google Scholar 

  • Levine A, Tenhaken R, Dixon R, Lamb C (1994) H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79:583–593

    Article  CAS  PubMed  Google Scholar 

  • Makkar HPS, Blummel M, Becker K (1995) Formation of complexes between polyvinylpyrrolidones or polyethylene glycols with tannins, and their implications in gas production and true digestibility in in vitro techniques. Br J Nutr 73:897–913

    Article  CAS  PubMed  Google Scholar 

  • Marri C, Frazzoli A, Hochkoeppler A, Poggi V (2003) Purification of a polyphenol oxidase isoform from potato (Solanum tuberosum) tubers. Phytochemistry 63:745–752

    Article  CAS  PubMed  Google Scholar 

  • Mittapalli O, Bai X, Mamidala P, Rajarapu SP, Bonello P, Herms DA (2010) Tissue-specific transcriptomics of the exotic invasive insect pest emerald ash borer (Agrilus planipennis). PLoS One 5:e13708

    Article  PubMed  PubMed Central  Google Scholar 

  • Muilenburg VL, Herms DA (2012) A review of bronze birch borer (Agrilus anxius, Coleoptera: Buprestidae) life history, ecology, and management. Environ Entomol 41:1372–1385

    Article  PubMed  Google Scholar 

  • Paulillo LCMS, Sebbenn AM, de Carvalho Derbyshite MTV, Góes-Neto A, de Paula Brotto MA, Figueira A (2012) Evaluation of in vitro and in vivo effects of semipurified proteinase inhibitors from Theobroma seeds on midgut protease activity of lepidopteran pest insects. Arch. Insect Biochem 81:34–52

    Article  CAS  Google Scholar 

  • Pedraza-Reyes M, Lopez-Romero E (1991) Chitinase activity in germinating cells of Mucor rouxii. Anton Leeuw 59:183–189

    CAS  Google Scholar 

  • Peiffer M, Tooker JF, Luthe DS, Felton GW (2009) Plants on early alert: glandular trichomes as sensors for insect herbivores. New Phytol 184:644–656

    Article  CAS  PubMed  Google Scholar 

  • R Core Team (2015) A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. ISBN 3–900051–07-0, URL http://www.R-project.org/

  • Rebek EJ, Herms DA, Smitley DR (2008) Interspecific variation in resistance to emerald ash borer (Coleoptera: Buprestidae) among north American and Asian ash (Fraxinus spp. Environ Entomol 37:242–246

    Article  PubMed  Google Scholar 

  • Rigsby CM, Showalter DN, Herms DA, Koch JL, Bonello P, Cipollini D (2015) Physiological responses of emerald ash borer larvae to feeding on different ash species reveal putative resistance mechanisms and insect counter-adaptations. J Insect Physiol 78:47–54

    Article  CAS  PubMed  Google Scholar 

  • Shahwar D, Raza MA, Rehman SU, Abbasi MA, Rahman AU (2012) An investigation of phenolic compounds from plant sources as trypsin inhibitors. Nat Prod Res 26:1087–1093

    Article  CAS  PubMed  Google Scholar 

  • Siemens DH, Mitchell-Olds T (1998) Evolution of pest defenses in Brassica plants: tests of theory. Ecology 79:632–646

    Article  Google Scholar 

  • Sollai F, Zucca P, Sanjust E, Steri D, Rescigno A (2008) Umbelliferone and Esculetin: inhibitors or substrates for polyphenol oxidases?. Biol Pharm Bull 31:2187–2193

  • Spadafora A, Mazzuca S, Chiappetta FF, Parise A, Perri E, Innocenti AM (2008) Oleuropein-specific-β-glucosidase activity marks the early response of olive fruits (Olea europaea) tomimed insect attack. Agric Sci China 7:703–712

  • Summers CB, Felton GW (1994) Prooxidant effects of phenolic acids on the generalist herbivore Helicoverpa zea (Lepidoptera: Noctuidae): potential mode of action for phenolic compounds in plant anti-herbivore chemistry. Insect Biochem Mol Biol 24:943–953

    Article  CAS  Google Scholar 

  • Vance CP, Kirk TK, Sherwood RT (1980) Lignification as a mechanism of disease resistance. Annu Rev Phytopathol 18:259–288

    Article  CAS  Google Scholar 

  • Villari C, Herms DA, Whitehill JGA, Cipollini D, Bonello P (2016) Progress and gaps in understanding mechanisms of ash tree resistance to emerald ash borer, a model for wood boring insects that kill angiosperm trees. New Phytol 209:63–79

    Article  CAS  PubMed  Google Scholar 

  • Wainhouse D, Cross D, Howell R (1990) The role of lignin as a defence against the spruce bark beetle Dendroctonus micans: effect on larvae and adults. Oecologia 85:257–265

    Article  Google Scholar 

  • Wallander E (2008) Systematics of Fraxinus (Oleaceae) and evolution of dioecy. Plant Syst Evol 273:25–49

    Article  Google Scholar 

  • Wang XY, Yang ZQ, Gould JR, Zhang YN, Liu GJ, Liu ES (2010) The biology and ecology of the emerald ash borer, Agrilus planipennis, in China. J Insect Sci 10:128

  • Wang Y, Chantreau M, Sibout R, Hawkins S (2013) Plant cell wall lignification and monolignol metabolism. Front Plant Sci 4:1–14

    Google Scholar 

  • War AR, Paulraj MG, Ahmad T, Buhroo AA, Hussain B, Ignacimuthu S, Sharma HC (2012) Mechanisms of plant defense against insect herbivores. Plant Signal Behav 7:1306–1320

    Article  PubMed  PubMed Central  Google Scholar 

  • Whitehill JGA, Opiyo SO, Koch JL, Herms DA, Cipollini DF, Bonello P (2012) Interspecific comparison of constitutive ash phloem phenolic chemistry reveals compounds unique to Manchurian ash, a species resistant to emerald ash borer. J Chem Ecol 38:499–511

    Article  CAS  PubMed  Google Scholar 

  • Whitehill JGA, Rigsby C, Cipollini D, Herms DA, Bonello P (2014) Decreased emergence of emerald ash borer from ash (Fraxinus spp.) treated with methyl jasmonate is associated with induction of general defense traits and the toxic phenolic compound verbascoside. Oecologia 176:1047–1059

    Article  PubMed  Google Scholar 

  • Whitehill JGA, Henderson H, Schuetz M, Skyba O, Yuen MMS, King J, Samuels AL, Mansfield SD, Bohlmann J (2015) Histology and cell wall biochemistry of stone cells in the physical defence of conifers against insects. Plant Cell Environ 39:1646–1661

    Article  PubMed  Google Scholar 

  • Zhu-Salzman K, Salzman RA, Ahn JE, Koiwa H (2004) Transcriptional regulation of sorghum defense determinants against a phloem-feeding aphid. Plant Physiol 134:420–431

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

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.

Author information

Author notes

  1. Chad M. Rigsby

    Present address: Department of Biological Sciences, The University of Rhode Island, Kingston, RI, 02881, USA

Authors and Affiliations

  1. Department of Biological Sciences and Environmental Sciences PhD Program, Wright State University, Dayton, OH, 45435, USA

    Chad M. Rigsby & Don Cipollini

  2. Department of Entomology, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster, OH, 44691, USA

    Daniel A. Herms

  3. Department of Plant Pathology, The Ohio State University, Columbus, OH, 43210, USA

    Pierluigi Bonello

Authors
  1. Chad M. Rigsby
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daniel A. Herms
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pierluigi Bonello
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Don Cipollini
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chad M. Rigsby.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10886-016-0736-5

Keywords

Search

Navigation