Journal of Pest Science

, Volume 87, Issue 2, pp 371–378 | Cite as

Identification of the most common predatory hoverflies of Mediterranean vegetable crops and their parasitism using multiplex PCR

  • Priscila Gomez-Polo
  • Michael Traugott
  • Oscar Alomar
  • Cristina Castañé
  • Santos Rojo
  • Nuria AgustíEmail author
Original Paper


The larvae of many hoverflies (Diptera: Syrphidae) are important polyphagous predators used in integrated pest management programs. Because the accurate identification of preimaginal stages by morphological characters is difficult, we have developed a multiplex PCR to identify the immature and/or adult stages of the most common syrphid species in Mediterranean vegetable crops: Episyrphus balteatus, Scaeva pyrastri, Eupeodes corollae, Meliscaeva auricollis, Sphaerophoria scripta, and Sphaerophoria rueppellii. The latter two species were amplified by the same primer pair due to the high similarity of their cytochrome oxidase subunit I sequences. Additionally, the assay included a primer pair targeting Diplazon laetatorius, a common koinobiont ichneumonid endoparasitoid of predatory syrphid larvae. The multiplex PCR assay proved to be highly specific and sensitive, and it was used to study the assemblage of hoverfly species in larval stage in two Mediterranean lettuce crops in two consecutive years. The molecular analysis revealed that Eu. corollae, Ep. balteatus, and Sph. scripta/Sph. rueppellii were the species present in the investigated fields. Species composition differed depending on sampling date and whether the larvae were collected on the plants or on the ground. The parasitoid D. laetatorius was not detected in any of the analyzed hoverfly larvae, suggesting low-parasitism pressure in the studied syrphid populations. The wide distribution of most of these syrphid species makes this multiplex PCR assay an ideal tool to deepen our knowledge on the ecology of these polyphagous hoverfly species in preimaginal stages and to improve the use of hoverflies to control insect pests.


Syrphidae COI Diplazon laetatorius Molecular species identification Lettuce crops Diagnostic PCR 



The authors thank Thaïs Aznar and Vanesa Vega for their technical support. The authors also thank Drs. R. Meyhöfer and P. Hondelmann (Leibniz University, Hannover, Germany) for providing syrphid samples, as well as Dr. A. Fereres (Institute of Agricultural Sciences-Spanish National Research Council (ICA-CSIC), Madrid, Spain) for providing specimens of several aphid species. This study has been funded by the Spanish Ministry of Economy and Competitiveness (MINECO) (Projects AGL2008-00546 and AGL2011-24349). P. Gomez-Polo was supported by a FPI Grant funded by the MINECO.

Supplementary material

10340_2013_550_MOESM1_ESM.docx (24 kb)
Supplementary material 1 (DOCX 23 kb)


  1. Amorós-Jiménez R, Pineda A, Fereres A, Marcos-García MA (2012) Prey availability and abiotic requirements of immature stages of the aphid predator Sphaerophoria rueppellii. Biol Control 63:17–24CrossRefGoogle Scholar
  2. Bastian O (1986) Schwebfliegen (Syrphidae). Die Neue Brehm-Bücherei, Bd. 576. 168 S, 352 Abb (z T farb)—A Ziemsen Verlag, WittenbergGoogle Scholar
  3. Benefer CM, van Herk WG, Ellis JS, Blackshaw RP, Vernon RS, Knight ME (2013) The molecular identification and genetic diversity of economically important wireworm species (Coleoptera: Elateridae) in Canada. J Pest Sci 86:19–27CrossRefGoogle Scholar
  4. Bordera S, Agulló P, Rojo S (2000) Nuevos Diplazontinae (Hymenoptera, Ichneumonidae) para la entomofauna iberbalear y potenciales sírfidos hospedadores (Diptera, Syrphidae). Bol Asoc Esp Ent 24(1–2):131–139Google Scholar
  5. Bordera S, Agulló P, Rojo S (2001) Catálogo de los Diplazontinae iberobaleares (Hymenoptera, Ichneumonidae) y potenciales sírfidos hospedadores (Diptera, Syrphidae). Bol Asoc Esp Ent 25(1–2):153–174Google Scholar
  6. Branquart E, Hemptinne JL (2000) Selectivity in the exploitation of floral resources by hoverflies (Diptera: Syrphinae). Ecography 23:732–742CrossRefGoogle Scholar
  7. Dusek J, Laska P (1961) Beitragzur Kenntnis der Schwebfliegen-Larven III (Syrphidae, Diptera). Prirod cas slezsky 22:513–541Google Scholar
  8. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol 3:294–299PubMedGoogle Scholar
  9. Ghahari H, Hayat R, Tabari M, Ostovan H (2008) Hoverflies (Diptera: Syrphidae) from rice fields and around grasslands of norther Iran. Munis Entomol Zool 3:275–284Google Scholar
  10. Gilbert FS (1993) Hoverflies. Richmond Publishing Co. Ltd., SloughGoogle Scholar
  11. Greco CF (1997) Specificity and instar preference of Diplazon laetatorius (Hym.: Ichneumonidae) parasitizing aphidophagous syrphids (Dipt: Syrphidae). Entomophaga 42:315–318CrossRefGoogle Scholar
  12. Haenke S, Scheid B, Schaefer M, Tscharntke T, Thies C (2009) Increasing syrphid fly diversity and density in sown flower strips within simple vs. complex landscapes. J Appl Ecol 46:1106–1114CrossRefGoogle Scholar
  13. Hagen KS, Mills NJ, Gordh G, McMurtry JA (1999) Terrestrial arthropod predators of insect and mite pests. In: Bellows TS, Fisher TW (eds) Handbook of biological control. Academic Press, San Diego, pp 383–503CrossRefGoogle Scholar
  14. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98Google Scholar
  15. Haslett JR (1983) A photographic account of pollen digestion by adult hoverflies. Physiol Entomol 8:167–171CrossRefGoogle Scholar
  16. Hazell SP, Wenlock C, Bachel S, Fellowes MDE (2005) The costs and consequences of parasitoid attack for the predatory hoverfly, Episyrphus balteatus. Evol Ecol Res 7:669–679Google Scholar
  17. Hebert PDN, Cywinska A, Ball SL, DeWaard JR (2003) Biological identifications through DNA barcodes. Proc R Soc A 270:313–321CrossRefGoogle Scholar
  18. Hopper JV, Nelson EH, Daane KM, Mills NJ (2011) Growth, development and consumption by four syrphid species associated with the lettuce aphid, Nasonovia ribisnigri, in California. Biol Control 58:271–276CrossRefGoogle Scholar
  19. Jankowska B (2004) Parasitoids of aphidophagous Syrphidae occurring in cabbage aphid (Brevicoryne brassicae L.) colonies on cabbage vegetables. J Plant Prot Res 44:299–305Google Scholar
  20. Jansen JP (2000) A three-year field study on the short-term effects of insecticides used to control cereal aphids on plant-dwelling aphid predators in winter wheat. Pest Manag Sci 56:533–539CrossRefGoogle Scholar
  21. Jauker F, Bondarenko B, Becker HC, Steffan-Dewenter I (2012) Pollination efficiency of wild bees and hoverflies provided to oilseed rape. Agric Forest Entomol 14:81–87CrossRefGoogle Scholar
  22. Jinbo U, Kato T, Ito M (2011) Current progress in DNA barcoding and future implications for entomology. Entomol Sci 14:107–124CrossRefGoogle Scholar
  23. Kehlmaier C, Martínez de Murguía L (2004) Syrphidae recorded in a heterogenous forest ecosystem at the “Finca de Artikutza” (Navarre, Northern Spain) (Diptera). Fragm Entomol 36:253–274Google Scholar
  24. King RA, Moreno-Ripoll R, Agustí N, Shayler SP, Bell JR, Bohan DA, Symondson WOC (2011) Multiplex reactions for the molecular detection of predation on pest and nonpest invertebrates in agroecosystems. Mol Ecol Resour 11:370–373PubMedCrossRefGoogle Scholar
  25. Krawczyk A, Hurej M, Jackowski J (2011) Syrphids and their parasitoids from maize crop. J Plant Prot Res 51:93–97CrossRefGoogle Scholar
  26. Laska P, Perez-Bañón C, Mazanek L, Rojo S, Ståhls G, Marcos-García MA, Bicik V, Dusek J (2006) Taxonomy of the genera Scaeva, Simosyrphus and Ischiodon (Diptera: Syrphidae): Descriptions of immature stages and status of taxa. Eur J Entomol 103:637–655CrossRefGoogle Scholar
  27. Lee W, Lee S (2012) Unexpected problem in aphid DNA barcoding by universal primers. Entomol Sci 15:121–126CrossRefGoogle Scholar
  28. Marcos-García MA, Isidro PM, Rojo S, Pérez-Bañón C (1998) Catálogo y distribución geográfica de las especies de Syrphidae (Diptera) en la Península Ibérica. I.—Syrphinae y Microdontinae. Bol Asoc Esp Entomol 22:37–61Google Scholar
  29. Marcos-García MA, Rojo S, Pérez-Bañón C (2002) Family Syrphidae. In: Catálogo de los dípteros de España, Portugal y Andorra (Insecta), vol 8. Monografías SEA, Zaragoza, pp 132–136Google Scholar
  30. Marshall EJP, West TM (2007) Impacts of field margins, landscape and crop on the distributions of Syrphidae on an arable farm. Asp Appl Biol 81:91–100Google Scholar
  31. Mayadunnage S, Wijayagunasekara HNP, Hemachandra KS, Nugaliyadde L (2009) Occurrence of aphidophagous syrphids in apphid colonies on cabbage (Brassica oleracea) and their parasitoids. J Trop Agri Res 21:99–109Google Scholar
  32. Mengual X, Stahls G, Rojo S (2008) First phylogeny of predatory flower flies (Diptera, Syrphidae, Syrphinae) using mitochondrial COI and nuclear 28S rRNA genes: conflict and congruence with the current tribal classification. Cladistics 24:543–562CrossRefGoogle Scholar
  33. Morales I, Díaz BM, Nebreda M, López-Lastra C, Goldarazena A, Sánchez JA, Pineda A, Marcos-García MA, Fereres A (2007) Principales agentes de biocontrol en cultivos de lechuga en la zona centro de España. Revista Horticultura 49:46–49Google Scholar
  34. Moreno-Ripoll R, Agustí N, Berruezo R, Gabarra R (2012) Conspecific and heterospecific interactions between two omnivorous predators on tomato. Biol Control 62:189–196CrossRefGoogle Scholar
  35. Pascual-Villalobos MJ, La casa A, González A, Varo P, García MJ (2006) Effect of flowering plant strips on aphid and syrphid populations in lettuce. Eur J Agron 24:182–185CrossRefGoogle Scholar
  36. Pineda A, Marcos-García MA (2008) Seasonal abundance of aphidophagous hoverflies (Diptera : Syrphidae) and their population levels in and outside Mediterranean sweet pepper greenhouses. Ann Entomol Soc Am 101:384–391CrossRefGoogle Scholar
  37. Rojo S (1995) Biología de los sírfidos afidófagos (Diptera, Syrphidae), presentes en cultivos hortofrutícolas mediterráneos. Implicaciones en el control biológico de pulgones (Homoptera, Aphididae). PhD dissertation, University of Alicante, Alicante, SpainGoogle Scholar
  38. Rojo S, Marcos-García MA (1998) Catálogo de los sírfidos (Diptera, Syrphidae) afidófagos (Homoptera, Aphididae) presentes en cultivos y plantas herbáceas de España y Portugal. Boll Zool Agrar Bachic 30:39–54Google Scholar
  39. Rojo S, Gilbert FS, Marcos-García MA, Nieto JM, Mier MP (2003) A world review of predatory hoverflies (Diptera, Syrphidae, Syrphinae) and their prey. CIBIO (Centro Iberoamericano de la Biodiversidad) ediciones, AlicanteGoogle Scholar
  40. Rotheray GE (1993) Colour guide to hoverfly larvae (Diptera, Syrphidae) in Britain and Europe. Dipterists Digest 9:156Google Scholar
  41. Rotheray GE, Gilbert F (2011) The natural history of hoverflies. Forrest, TresaithGoogle Scholar
  42. Sajjad A, Saeed S, Masood A (2008) Pollinator community of onion (Allium cepa L.) and its role in crop reproductive success. Pak J Zool 40:451–456Google Scholar
  43. Scott EI (1939) An account of the developmental stages of some aphidophagous Syrphidae (Dipt) and their parasites (Hymenopt). Ann Appl Biol 26:509–532CrossRefGoogle Scholar
  44. Smith HA, Chaney WE (2007) A survey of syrphid predators of Nasonovia ribisnigri in organic lettuce on the central coast of California. J Econ Entomol 100:39–48PubMedCrossRefGoogle Scholar
  45. Speight MCD (2011) Species accounts of European Syrphidae (Diptera). Syrph the Net, the database of European Syrphidae, vol 695. Syrph the Net publications, DublinGoogle Scholar
  46. Stahls G, Vujic A, Pérez-Bañón C, Radenkovic S, Rojo S, Petanidou T (2009) COI barcodes for identification of Merodon hoverflies (Diptera, Syrphidae) of Lesvos Island, Greece. Mol Ecol Resour 9:1431–1438PubMedCrossRefGoogle Scholar
  47. Staudacher K, Pitterl P, Furlan L, Cate PC, Traugott M (2011) PCR-based species identification of Agriotes larvae. Bull Entomol Res 101:201–210PubMedCrossRefGoogle Scholar
  48. Stubbs AE, Falk SJ (1983) British hoverflies: an illustrated identification guide. British Entomology and Natural History Society, LondonGoogle Scholar
  49. Traugott M, Symondson WOC (2008) Molecular analysis of predation on parasitized hosts. Bull Entomol Res 98:223–231PubMedGoogle Scholar
  50. Traugott M, Zangerl P, Juen A, Schallhart N, Pfiffner L (2006) Detecting key parasitoids of lepidopteran pests by multiplex PCR. Biol Control 39:39–46CrossRefGoogle Scholar
  51. Traugott M, Kamenova S, Ruess L, Seeber J, Plantegenest M (2013) Empirically characterising trophic networks: what emerging DNA-based methods, stable isotope and fatty acid analyses can offer. Adv Ecol Res 49:177–224CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Priscila Gomez-Polo
    • 1
  • Michael Traugott
    • 2
  • Oscar Alomar
    • 1
  • Cristina Castañé
    • 1
  • Santos Rojo
    • 3
  • Nuria Agustí
    • 1
    Email author
  1. 1.EntomologyInstitute for Food and Agricultural Research and Technology (IRTA)CabrilsSpain
  2. 2.Mountain Agriculture Research Unit, Institute of EcologyUniversity of InnsbruckInnsbruckAustria
  3. 3.University Research Institute CIBIO/Department of Environmental SciencesUniversity of AlicanteAlicanteSpain

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