Journal of Chemical Ecology

, Volume 45, Issue 5–6, pp 515–524 | Cite as

Pathogen-Mediated Tritrophic Interactions: Baculovirus-Challenged Caterpillars Induce Higher Plant Defenses than Healthy Caterpillars

  • Qinjian PanEmail author
  • Ikkei Shikano
  • Kelli Hoover
  • Tong-Xian LiuEmail author
  • Gary W. Felton


Although the tritrophic interactions of plants, insect herbivores and their natural enemies have been intensely studied for several decades, the roles of entomopathogens in their indirect modulation of plant-insect relationships is still unclear. Here, we employed a sublethal dose of a baculovirus with a relatively broad host range (AcMNPV) to explore if feeding by baculovirus-challenged Helicoverpa zea caterpillars induces direct defenses in the tomato plant. We examined induction of plant defenses following feeding by H. zea, including tomato plants fed on by healthy caterpillars, AcMNPV-challenged caterpillars, or undamaged controls, and subsequently compared the transcript levels of defense related proteins (i.e., trypsin proteinase inhibitors, peroxidase and polyphenol oxidase) and other defense genes (i.e., proteinase inhibitor II and cysteine proteinase inhibitor) from these plants, in addition to comparing caterpillar relative growth rates. As a result, AcMNPV-challenged caterpillars induced the highest plant anti-herbivore defenses. We examined several elicitors and effectors in the secretions of these caterpillars (i.e., glucose oxidase, phospholipase C, and ATPase hydrolysis), which surprisingly did not differ between treatments. Hence, we suggest that the greater induction of plant defenses by the virus-challenged caterpillars may be due to differences in the amount of these secretions deposited during feeding or to some other unknown factor(s).


Plant defense Induce defense Immune responses Saliva Oral secretions Ventral eversible gland Herbivore perception 



This research was financially supported by the U.S. National Science Foundation (IOS-1645548) awarded to GWF, IS, and KH. QJP acknowledges financial support from China Scholarship Council (Grant 201506300111). IS acknowledges financial support from Natural Sciences and Engineering Research Council of Canada Postdoctoral Fellowship (NSERC PDF-488105-2016). The financial support from Northwest A&F University’ Special Talent Fund to TXL is greatly appreciated. We appreciate all technical assistance and suggestions from M. Peiffer and F. McCullough. We thank Dr. D. Luthe (Department of Plant Science, Pennsylvania State University) for sharing her laboratory equipment.

Supplementary material

10886_2019_1077_Fig9_ESM.png (13 kb)
Fig. S1

Dose response of virus (AcMNPV)-challenged fifth instar Trichoplusia ni and fourth instar Helicoverpa zea caterpillars. (PNG 12 kb)

10886_2019_1077_MOESM1_ESM.eps (730 kb)
High resolution image (EPS 729 kb)
10886_2019_1077_MOESM2_ESM.docx (13 kb)
Table S1 Tomato primers used for RT-PCR in this study. (DOCX 13 kb)


  1. Acevedo FE, Rivera-Vega LJ, Chung SH, Ray S, Felton GW (2015) Cues from chewing insects-the intersection of DAMPs, HAMPs, MAMPs and effectors. Curr Opin Plant Biol 26:80–86. CrossRefGoogle Scholar
  2. Acevedo FE, Peiffer M, Tan CW, Stanley BA, Stanley A, Wang J, Jones AG, Hoover K, Rosa C, Luthe D, Felton G (2017) Fall armyworm-associated gut bacteria modulate plant defense responses. Mol Plant-Microbe Interact 30:127–137. CrossRefGoogle Scholar
  3. Ali MI, Felton GW, Meade T, Young SY (1998) Influence of interspecific and intraspecific host plant variation on the susceptibility of Heliothines to a baculovirus. Biol Control 12:42–49. CrossRefGoogle Scholar
  4. Ali MI, Bi JL, Young SY, Felton GW (1999) Do foliar phenolics provide protection to Heliothis virescens from a baculovirus? J Chem Ecol 25:2193–2204. CrossRefGoogle Scholar
  5. Appel HM, Cocroft RB (2014) Plants respond to leaf vibrations caused by insect herbivore chewing. Oecologia 175:1257–1266. CrossRefGoogle Scholar
  6. Bostock RM (2005) Signal crosstalk and induced resistance: straddling the line between cost and benefit. Annu Rev Phytopathol 43:545–580. CrossRefGoogle Scholar
  7. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. CrossRefGoogle Scholar
  8. Chaudhary R, Atamian HS, Shen Z, Briggs SP, Kaloshian I (2014) GroEL from the endosymbiont Buchnera aphidicola betrays the aphid by triggering plant defense. PNAS 111:8919–8924. CrossRefGoogle Scholar
  9. Chung SH, Felton GW (2011) Specificity of induced resistance in tomato against specialist lepidopteran and coleopteran species. J Chem Ecol 37:378–386. CrossRefGoogle Scholar
  10. Chung SH, Rosa C, Scully ED, Peiffer M, Tooker JF, Hoover K, Luthe DS, Felton GW (2013) Herbivore exploits orally secreted bacteria to suppress plant defenses. PNAS. 110:15728–15733. CrossRefGoogle Scholar
  11. Chung SH, Scully ED, Peiffer M, Geib SM, Rosa C, Hoover K, Felton GW (2017) Host plant species determines symbiotic bacterial community mediating suppression of plant defenses. Sci Rep 7:39690. CrossRefGoogle Scholar
  12. Clark EL, Karley AJ, Hubbard SF (2010) Insect endosymbionts: manipulators of insect herbivore trophic interactions? Protoplasma 244:25–51. CrossRefGoogle Scholar
  13. Cory JS, Hoover K (2006) Plant-mediated effects in insect-pathogen interactions. Trends Ecol Evol 21:278–286. CrossRefGoogle Scholar
  14. Cory JS, Myers JH (2003) The ecology and evolution of insect baculoviruses. Annu Rev Ecol Evol Syst 34:239–272. CrossRefGoogle Scholar
  15. Cusumano A, Zhu F, Volkoff AN, Verbaarschot P, Bloem J, Vogel H, Dicke M, Poelman EH (2018) Parasitic wasp-associated symbiont affects plant-mediated species interactions between herbivores. Ecol Lett 21:957–967. CrossRefGoogle Scholar
  16. Dillon RJ, Dillon VM (2004) The gut bacteria of insects: nonpathogenic interactions. Annu Rev Entomol 49:71–92. CrossRefGoogle Scholar
  17. Eichenseer H, Mathews MC, Bi JL, Murphy JB, Felton GW (1999) Salivary glucose oxidase: multifunctional roles for Helicoverpa zea? Arch Insect Biochem Physiol 42:99–109.<99::AID-ARCH10>3.0.CO;2-B CrossRefGoogle Scholar
  18. Elderd BD, Rehill BJ, Haynes KJ, Dwyer G (2013) Induced plant defenses, host-pathogen interactions, and forest insect outbreaks. PNAS. 110:14978–14983. CrossRefGoogle Scholar
  19. Felton GW (2005) Indigestion is a plant's best defense. PNAS 102:18771–18772. CrossRefGoogle Scholar
  20. Felton GW (2008) Caterpillar secretions and induced plant responses. In: Schaller A (ed) Induced plant resistance to herbivory. Springer, New York, pp 369–389CrossRefGoogle Scholar
  21. Felton GW, Duffey SS (1991) Protective action of midgut catalase in lepidopteran larvae against oxidative plant defenses. J Chem Ecol 17:1715–1732. CrossRefGoogle Scholar
  22. Felton GW, Duffey SS, Vail PV, Kaya HK, Manning J (1987) Interaction of nuclear polyhedrosis virus with catechols: potential incompatibility for host-plant resistance against noctuid larvae. J Chem Ecol 13:947–957. CrossRefGoogle Scholar
  23. Heil M (2009) Damaged-self recognition in plant herbivore defence. Trends Plant Sci 14:356–363. CrossRefGoogle Scholar
  24. Helms AM, De Moraes CM, Tooker JF, Mescher MC (2013) Exposure of Solidago altissima plants to volatile emissions of an insect antagonist (Eurosta solidaginis) deters subsequent herbivory. PNAS 110:199–204. CrossRefGoogle Scholar
  25. Hilker M, Fatouros NE (2015) Plant responses to insect egg deposition. Annu Rev Entomol 60:493–515. CrossRefGoogle Scholar
  26. Hoover K, Stout MJ, Alaniz SA, Hammock BD, Duffey SS (1998) Influence of induced plant defenses in cotton and tomato on the efficacy of baculoviruses on noctuid larvae. J Chem Ecol 24:253–271. CrossRefGoogle Scholar
  27. Hoover K, Washburn JO, Volkman LE (2000) Midgut-based resistance of Heliothis virescens to baculovirus infection mediated by phytochemicals in cotton. J Insect Physiol 46:999–1007. CrossRefGoogle Scholar
  28. Ikeda M, Yamada H, Hamajima R, Kobayashi M (2013) Baculovirus genes modulating intracellular innate antiviral immunity of lepidopteran insect cells. Virology 435:1–13. CrossRefGoogle Scholar
  29. Keating ST, McCarthy WJ, Yendol WG (1989) Gypsy moth (Lymantria dispar) larval susceptibility to a baculovirus affected by selected nutrients, hydrogen lons (pH), and plant allelochemicals in artificial diets. J Invertebr Pathol 54:165–174. CrossRefGoogle Scholar
  30. Le Chevalier F, Cascioferro A, Frigui W et al (2015) Revisiting the role of phospholipases C in virulence and the lifecycle of Mycobacterium tuberculosis. Sci Rep 5:16918. CrossRefGoogle Scholar
  31. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔC T method. Methods 25:402–408. CrossRefGoogle Scholar
  32. Pan Q, Shikano I, Hoover K, Liu T-X, Felton GW (2019a) Enterobacter ludwigii, isolated from the gut microbiota of Helicoverpa zea, promotes tomato plant growth and yield without compromising anti-herbivore defenses. Arthropod Plant Interact 13:271–278. CrossRefGoogle Scholar
  33. Pan Q, Shikano I, Hoover K, Liu T-X, Felton GW (2019b) Host permissiveness to baculovirus influences time-dependent immune responses and fitness costs. Manuscript submitted for publication.Google Scholar
  34. Peiffer M, Felton GW (2005) The host plant as a factor in the synthesis and secretion of salivary glucose oxidase in larval Helicoverpa zea. Arch Insect Biochem Physiol 58:106–113. CrossRefGoogle Scholar
  35. Peiffer M, Felton GW (2009) Do caterpillars secrete “oral secretions”? J Chem Ecol 35:326–335. CrossRefGoogle Scholar
  36. 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. CrossRefGoogle Scholar
  37. Price PW, Ce B, Gross P, McPheron BA, Thompson JN, Weis AE (1980) Interactions among three trophic levels: influence of plants on interactions between insect herbivores and natural enemies. Annu Rev Ecol Syst 11:41–65CrossRefGoogle Scholar
  38. Rotenberg D, Thompson TS, German TL, Willis DK (2006) Methods for effective real-time RT-PCR analysis of virus-induced gene silencing. J Virol Methods 138:49–59. CrossRefGoogle Scholar
  39. Schmelz EA (2015) Impacts of insect oral secretions on defoliation-induced plant defense. Curr Opin Insect Sci 9:7–15. CrossRefGoogle Scholar
  40. Shikano I (2017) Evolutionary ecology of multitrophic interactions between plants, insect herbivores and entomopathogens. J Chem Ecol 43:586–598. CrossRefGoogle Scholar
  41. Shikano I, Rosa C, Tan CW, Felton GW (2017a) Tritrophic interactions: microbe-mediated plant effects on insect herbivores. Annu Rev Phytopathol 55:313–331. CrossRefGoogle Scholar
  42. Shikano I, Shumaker KL, Peiffer M, Felton GW, Hoover K (2017c) Plant-mediated effects on an insect-pathogen interaction vary with intraspecific genetic variation in plant defences. Oecologia 183:1121–1134. CrossRefGoogle Scholar
  43. Shikano I, McCarthy EM, Elderd BD, Hoover K (2017d) Plant genotype and induced defenses affect the productivity of an insect-killing obligate viral pathogen. J Invertebr Pathol 148:34–42CrossRefGoogle Scholar
  44. Shikano I, McCarthy EM, Hayes-Plazolles N, Slavicek JM, Hoover K (2018) Jasmonic acid-induced plant defenses delay caterpillar developmental resistance to a baculovirus: slow-growth, high-mortality hypothesis in plant-insect-pathogen interactions. J Invertebr Pathol 158:16–23CrossRefGoogle Scholar
  45. Tan CW, Peiffer M, Hoover K, Rosa C, Acevedo FE, Felton GW (2018) Symbiotic polydnavirus of a parasite manipulates caterpillar and plant immunity. PNAS 115:5199–5204. CrossRefGoogle Scholar
  46. Trudeau D, Washburn JO, Volkman LE (2001) Central role of Hemocytes in Autographa californica M Nucleopolyhedrovirus pathogenesis in Heliothis virescens and Helicoverpa zea. J Virol 75:996–1003. CrossRefGoogle Scholar
  47. Vilaplana L, Wilson K, Redman EM, Cory JS (2010) Pathogen persistence in migratory insects: high levels of vertically-transmitted virus infection in field populations of the African armyworm. Evol Ecol 24:147–160. CrossRefGoogle Scholar
  48. Waldbauer GP (1968) The consumption and utilization of food by insects. Adv Insect Physiol 5:229–288. CrossRefGoogle Scholar
  49. Wang J, Peiffer M, Hoover K, Rosa C, Zeng R, Felton GW (2017) Helicoverpa zea gut-associated bacteria indirectly induce defenses in tomato by triggering a salivary elicitor(s). New Phytol 214:1294–1306. CrossRefGoogle Scholar
  50. Wang J, Yang M, Song Y, Acevedo FE, Hoover K, Zeng R, Felton GW (2018) Gut-associated bacteria of Helicoverpa zea indirectly trigger plant defenses in maize. J Chem Ecol 44:690–699. CrossRefGoogle Scholar
  51. Warnecke F, Luginbühl P, Ivanova N, Ghassemian M, Richardson TH, Stege JT, Cayouette M, McHardy AC, Djordjevic G, Aboushadi N, Sorek R, Tringe SG, Podar M, Martin HG, Kunin V, Dalevi D, Madejska J, Kirton E, Platt D, Szeto E, Salamov A, Barry K, Mikhailova N, Kyrpides NC, Matson EG, Ottesen EA, Zhang X, Hernández M, Murillo C, Acosta LG, Rigoutsos I, Tamayo G, Green BD, Chang C, Rubin EM, Mathur EJ, Robertson DE, Hugenholtz P, Leadbetter JR (2007) Metagenomic and functional analysis of hindgut microbiota of a wood-feeding higher termite. Nature. 450:560–565CrossRefGoogle Scholar
  52. Zavala JA, Patankar AG, Gase K, Baldwin IT (2004) Constitutive and inducible trypsin proteinase inhibitor production incurs large fitness costs in Nicotiana attenuata. PNAS 101:1607–1612. CrossRefGoogle Scholar
  53. Zebelo SA, Maffei ME (2012) The ventral eversible gland (VEG) of Spodoptera littoralis triggers early responses to herbivory in Arabidopsis thaliana. Arthropod Plant Interact 6:543–551. CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Crop Stress Biology for Arid Areas, and Key Laboratory of Integrated Pest Management on the Loess Plateau of Ministry of AgricultureNorthwest A&F UniversityYanglingChina
  2. 2.Department of Entomology and Center for Chemical EcologyPennsylvania State UniversityUniversity ParkUSA

Personalised recommendations