Endosymbionts Differentially Alter Exploratory Probing Behavior of a Nonpersistent Plant Virus Vector


Insect endosymbionts (hereafter, symbionts) can modify plant virus epidemiology by changing the physiology or behavior of vectors, but their role in nonpersistent virus pathosystems remains uninvestigated. Unlike propagative and circulative viruses, nonpersistent plant virus transmission occurs via transient contamination of mouthparts, making direct interaction between symbiont and virus unlikely. Nonpersistent virus transmission occurs during exploratory intracellular punctures with styletiform mouthparts when vectors assess potential host-plant quality prior to phloem feeding. Therefore, we used an electrical penetration graph (EPG) to evaluate plant probing of the cowpea aphid, Aphis craccivora Koch, an important vector of cucurbit viruses, in the presence and absence of two facultative, intracellular symbionts. We tested four isolines of A. craccivora: two isolines were from a clone from black locust (Robinia pseudoacacia L.), one infected with Arsenophonus sp. and one cured, and two derived from a clone from alfalfa (Medicago sativa L.), one infected with Hamiltonella defensa and one cured. We quantified exploratory intracellular punctures, indicated by a waveform potential drop recorded by the EPG, initiation speed and frequency within the initial 15 min on healthy and watermelon mosaic virus-infected pumpkins. Symbiont associations differentially modified exploratory intracellular puncture frequency by aphids, with H. defensa-infected aphids exhibiting depressed probing, and Arsenophonus-infected aphids an increased frequency of probing. Further, there was greater overall aphid probing on virus-infected plants, suggesting that viruses manipulate their vectors to enhance acquisition-transmission rates, independent of symbiont infection. These results suggest facultative symbionts differentially affect plant-host exploration behaviors and potentially nonpersistent virus transmission by vectors.

This is a preview of subscription content, access via your institution.

We’re sorry, something doesn't seem to be working properly.

Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Fig. 1


  1. 1.

    Oliver KM, Degnan PH, Burke GR, Moran NA (2010) Facultative symbionts in aphids and the horizontal transfer of ecologically important traits. Annu Rev Entomol 55:247–266

    Article  PubMed  CAS  Google Scholar 

  2. 2.

    Ezenwa VO, Gerardo NM, Inouye DW, Medina M, Xavier JB (2012) Animal behavior and the microbiome. Science 338:198–199

    Article  PubMed  CAS  Google Scholar 

  3. 3.

    Gottlieb Y, Zchori-Fein E, Mozes-Daube N, Kontsedalov S, Skaljac M, Brumin M, Sobol I, Czosnek H, Vavre F, Fleury F (2010) The transmission efficiency of tomato yellow leaf curl virus by the whitefly Bemisia tabaci is correlated with the presence of a specific symbiotic bacterium species. J Virol 84:9310–9317

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. 4.

    Rana VS, Singh ST, Priya NG, Kumar J, Rajagopal R (2012) Arsenophonus GroEL interacts with CLCuV and is localized in midgut and salivary gland of whitefly B. tabaci. PLoS One 7:e42168

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  5. 5.

    Su Q, Oliver KM, Pan H, Jiao X, Liu B, Xie W, Wang S, Wu Q, Xu B, White JA, Zhou X, Zhang Y (2013) Facultative symbiont Hamiltonella confers benefits to Bemisia tabaci (Hemiptera: Aleyrodidae), an invasive agricultural pest worldwide. Environ Entomol 42:1265–1271

    Article  PubMed  Google Scholar 

  6. 6.

    Kliot A, Cilia M, Czosnek H, Ghanim M (2014) Implication of the bacterial endosymbiont Rickettsia spp. in interactions of the whitefly Bemisia tabaci with Tomato yellow leaf curl virus. J Virol 88:5652–5660

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  7. 7.

    Pinheiro PV, Kliot A, Ghanim M, Cilia M (2015) Is there a role for symbiotic bacteria in plant virus transmission by insects? Curr Opin Insect Sci 8:69–78

    Article  Google Scholar 

  8. 8.

    Fereres A, Collar J (2001) In: Harris KF, Smith OP, Duffus JE (eds) Analysis of noncirculative transmission by electrical penetration graphs. Virus-insect-plant interactions. Academic Press, San Diego

    Google Scholar 

  9. 9.

    Powell G (1991) Cell membrane punctures during epidermal penetrations by aphids: consequences for the transmission of two potyviruses. Ann Appl Biol 119:313–321

    Article  Google Scholar 

  10. 10.

    Wensler RJD, Filshie BK (1969) Gustatory sense organs in the food canal of aphids. J Morphol 129:473–492

    Article  Google Scholar 

  11. 11.

    Tjallingii W (1995) Regulation of phloem sap feeding by aphids. Regulatory mechanisms in insect feeding. Springer, New York

    Google Scholar 

  12. 12.

    Martin B, Collar JL, Tjallingii WF, Fereres A (1997) Intracellular ingestion and salivation by aphids may cause the acquisition and inoculation of non-persistently transmitted plant viruses. J Gen Virol 78:2701–2705

    Article  PubMed  CAS  Google Scholar 

  13. 13.

    Tjallingii WF, Hogen Esch T (1993) Fine structure of aphid stylet routes in plant tissues in correlation with EPG signals. Physiol Entomol 18:317–328

    Article  Google Scholar 

  14. 14.

    Powell G, Pirone T, Hardie J (1995) Aphid stylet activities during potyvirus acquisition from plants and an in vitro system that correlate with subsequent transmission. Eur J Plant Pathol 101:411–420

    Article  Google Scholar 

  15. 15.

    Fereres A, Moreno A (2009) Behavioural aspects influencing plant virus transmission by homopteran insects. Virus Res 141:158–168

    Article  PubMed  CAS  Google Scholar 

  16. 16.

    Angelella G, Egel D, Holland J, Nemacheck J, Williams C, Kaplan I (2015) Differential life history trait associations of aphids with nonpersistent viruses in cucurbits. Environ Entomol 44:562–573

    Article  PubMed  CAS  Google Scholar 

  17. 17.

    Angelella GM, Holland JD, Kaplan I (2016) Landscape composition is more important than local management for crop virus–insect vector interactions. Agric Ecosyst Environ 233:253–261

    Article  Google Scholar 

  18. 18.

    Brunt AA, Crabtree K, Dallwitz MJ, Gibbs AJ, Watson L, Zurcher EJE (1996 onwards) Plant viruses online: descriptions and lists from the VIDE database. vol. Version: 16th January 1997

  19. 19.

    Dallwitz MJ (1980) A general system for coding taxonomic descriptions. Taxon 29:41–46

    Article  Google Scholar 

  20. 20.

    Dallwitz MJ, Paine TA, Zurcher EJ (1993) User’s guide to the DELTA system: a general system for processing taxonomic descriptions. CSIRO Division of Entomology, Canberra

    Google Scholar 

  21. 21.

    Paulsrud B (2005) Workshop summary. Midwest pest management strategic plan for processing & jack-o-lantern pumpkins: Illinois, Indiana, Iowa and Missouri. University of Illinois Extension, Urbana, Illinois, pp. 109

  22. 22.

    Brady C, Asplen M, Desneux N, Heimpel G, Hopper K, Linnen C, Oliver K, Wulff J, White J (2014) Worldwide populations of the aphid Aphis craccivora are infected with diverse facultative bacterial symbionts. Microb Ecol 38:1–10

    Google Scholar 

  23. 23.

    Brady CM, White JA (2013) Cowpea aphid (Aphis craccivora) associated with different host plants has different facultative endosymbionts. Ecol Entomol 38:433–437

    Article  Google Scholar 

  24. 24.

    Wagner SM, Martinez AJ, Ruan YM, Kim KL, Lenhart PA, Dehnel AC, Oliver KM, White JA (2015) Facultative endosymbionts mediate dietary breadth in a polyphagous herbivore. Funct Ecol 29:1402–1410

    Article  Google Scholar 

  25. 25.

    White JA, McCord JS, Jackson KA, Dehnel AC, Lenhart PA (2017) Differential aphid toxicity to ladybeetles is not a function of host plant or facultative bacterial symbionts. Funct Ecol 31:334–339

    Article  Google Scholar 

  26. 26.

    Jousselin E, Cœur d’Acier A, Vanlerberghe-Masutti F, Duron O (2013) Evolution and diversity of Arsenophonus endosymbionts in aphids. Mol Ecol 22:260–270

    Article  PubMed  Google Scholar 

  27. 27.

    Dykstra HR, Weldon SR, Martinez AJ, White JA, Hopper KR, Heimpel GE, Asplen MK, Oliver KM (2014) Factors limiting the spread of the protective symbiont Hamiltonella defensa in the aphid Aphis craccivora. Appl Environ Microbiol 80:5818–5827

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. 28.

    Eigenbrode SD, Ding H, Shiel P, Berger PH (2002) Volatiles from potato plants infected with potato leafroll virus attract and arrest the virus vector, Myzus persicae (Homoptera: Aphididae). Proceedings of the Royal Society of London Series B. Biol Sci 269:455–460

    Article  CAS  Google Scholar 

  29. 29.

    Prado E, Tjallingii WF (1994) Aphid activities during sieve element punctures. Entomol Exp Appl 72:157–165

    Article  Google Scholar 

  30. 30.

    R Core Team (2016) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna

    Google Scholar 

  31. 31.

    Ingwell LL, Eigenbrode SD, Bosque-Perez NA (2012) Plant viruses alter insect behavior to enhance their spread. Sci Rep 578:1–6

    Google Scholar 

  32. 32.

    Mauck KE, Bosque-Perez NA, Eigenbrode SD, De Moraes CM, Mescher MC (2012) Transmission mechanisms shape pathogen effects on host-vector interactions: evidence from plant viruses. Funct Ecol 26:1162–1175

    Article  Google Scholar 

  33. 33.

    Mauck KE, De Moraes CM, Mescher MC (2010) Deceptive chemical signals induced by a plant virus attract insect vectors to inferior hosts. Proc Natl Acad Sci 107:3600–3605

    Article  PubMed  Google Scholar 

  34. 34.

    Visôtto L, Oliveira M, Guedes R, Ribon A, Good-God P (2009) Contribution of gut bacteria to digestion and development of the velvetbean caterpillar, Anticarsia gemmatalis. J Insect Physiol 55:185–191

    Article  PubMed  CAS  Google Scholar 

  35. 35.

    Barr KL, Hearne LB, Briesacher S, Clark TL, Davis GE (2010) Microbial symbionts in insects influence down-regulation of defense genes in maize. PLoS One 5:e11339

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  36. 36.

    Boone CK, Keefover-Ring K, Mapes AC, Adams AS, Bohlmann J, Raffa KF (2013) Bacteria associated with a tree-killing insect reduce concentrations of plant defense compounds. J Chem Ecol 39:1003–1006

    Article  PubMed  CAS  Google Scholar 

  37. 37.

    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. Proc Natl Acad Sci 110:15728–15733

    Article  PubMed  Google Scholar 

  38. 38.

    Zhu F, Poelman EH, Dicke M (2014) Insect herbivore-associated organisms affect plant responses to herbivory. New Phytol 204:315–321

    Article  Google Scholar 

  39. 39.

    Su Q, Xie W, Wang S, Wu Q, Liu B, Fang Y, Xu B, Zhang Y (2014) The endosymbiont Hamiltonella increases the growth rate of its host Bemisia tabaci during periods of nutritional stress. PLoS One 9:e89002

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  40. 40.

    Staudacher H, Schimmel BC, Lamers MM, Wybouw N, Groot AT, Kant MR (2017) Independent effects of a herbivore’s bacterial symbionts on its performance and induced plant defences. Int J Mol Sci 18:182

    Article  PubMed Central  CAS  Google Scholar 

  41. 41.

    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

    Article  PubMed  CAS  Google Scholar 

  42. 42.

    Hansen AK, Moran NA (2014) The impact of microbial symbionts on host plant utilization by herbivorous insects. Mol Ecol 23:1473–1496

    Article  PubMed  Google Scholar 

  43. 43.

    Moran PJ, Thompson GA (2001) Molecular responses to aphid feeding in Arabidopsis in relation to plant defense pathways. Plant Physiol 125:1074–1085

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  44. 44.

    Morkunas I, Gabryś B (2011) Phytohormonal signaling in plant responses to aphid feeding. Acta Physiol Plant 33:2057–2073

    Article  CAS  Google Scholar 

  45. 45.

    Argandona V, Chaman M, Cardemil L, Munoz O, Zuniga G, Corcuera L (2001) Ethylene production and peroxidase activity in aphid-infested barley. J Chem Ecol 27:53–68

    Article  PubMed  CAS  Google Scholar 

  46. 46.

    Katis NI, Tsitsipis JA, Stevens M, Powell G (2007) Transmission of plant viruses. In: van Emden HF, Harrington R (eds) Aphids as crop pests. CABI, Cambridge

    Google Scholar 

Download references


We would like to thank Allison Dehnel for shipping Aphis craccivora from J. White’s lab at the University of Kentucky so often to initiate colonies at Purdue for use in this study. We would also like to thank an anonymous reviewer for helpful comments on an earlier version of this article. Funding provided by United States Department of Agriculture-National Institute of Food and Agriculture Predoctoral Fellowship grant 107483. Aphid colonies at the University of Kentucky were maintained on funds from United States Department of Agriculture-National Institute of Food and Agriculture grant 2014-67013-21576.

Author information



Corresponding author

Correspondence to G. Angelella.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Electronic supplementary material


(PDF 89 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Angelella, G., Nalam, V., Nachappa, P. et al. Endosymbionts Differentially Alter Exploratory Probing Behavior of a Nonpersistent Plant Virus Vector. Microb Ecol 76, 453–458 (2018). https://doi.org/10.1007/s00248-017-1133-5

Download citation


  • Nonpersistent virus transmission
  • Aphis craccivora
  • Arsenophonus
  • Hamiltonella defensa
  • Watermelon mosaic virus
  • Endosymbionts