Molecular analysis of predation on parasitized hosts

Abstract Predation on parasitized hosts can significantly affect natural enemy communities, and such intraguild predation may indirectly affect control of herbivore populations. However, the methodological challenges for studying these often complex trophic interactions are formidable. Here, we evaluate a DNA-based approach to track parasitism and predation on parasitized hosts in model herbivore-parasitoid-predator systems. Using singleplex polymerase chain reaction (SP-PCR) to target mtDNA of the parasitoid only, and multiplex PCR (MP-PCR) to additionally target host DNA as an internal amplification control, we found that detection of DNA from the parasitoid, Lysiphlebus testaceipes, in its aphid host, Aphis fabae, was possible as early as 5 min. post parasitism. Up to 24 h post parasitism SP-PCR proved to be more sensitive than MP-PCR in amplifying parasitoid DNA. In the carabid beetles Demetrias atricapillus and Erigone sp. spiders, fed with aphids containing five-day-old parasitoids, parasitoid and aphid DNA were equally detectable in both predator groups. However, when hosts containing two-day-old parasitoids were fed to the predators, detection of parasitoid prey was possible only at 0 h (immediately after consumption) and up to 8 h post consumption in carabids and spiders, respectively. Over longer periods of time, post-feeding prey detection success was significantly higher in spiders than in carabid beetles. MP-PCR, in which parasitoid and aphid DNA were simultaneously amplified, proved to be less sensitive at amplifying prey DNA than SP-PCR. In conclusion, our study demonstrates that PCR-based parasitoid and prey detection offers an exciting approach to further our understanding of host-parasitoid-predator interactions.

[1]  S. Scheu,et al.  The effects of temperature on detection of prey DNA in two species of carabid beetle , 2008, Bulletin of Entomological Research.

[2]  M. Traugott,et al.  INVITED REVIEW: Molecular analysis of predation: a review of best practice for DNA‐based approaches , 2008, Molecular ecology.

[3]  U. Kuhlmann,et al.  Parasitoids, predators and PCR: the use of diagnostic molecular markers in biological control of Arthropods , 2007 .

[4]  M. Payton,et al.  Feeding mode and prey detectability half-lives in molecular gut-content analysis: an example with two predators of the Colorado potato beetle , 2007, Bulletin of Entomological Research.

[5]  M. Traugott,et al.  Revealing species‐specific trophic links in soil food webs: molecular identification of scarab predators , 2007, Molecular ecology.

[6]  X. Pons,et al.  Seasonal parasitism of cereal aphids in a Mediterranean arable crop system , 2007, Journal of Pest Science.

[7]  M. Traugott,et al.  Detecting key parasitoids of lepidopteran pests by multiplex PCR , 2006 .

[8]  B. Deagle,et al.  Quantification of damage in DNA recovered from highly degraded samples – a case study on DNA in faeces , 2006, Frontiers in Zoology.

[9]  Michael Traugott,et al.  Amplification facilitators and multiplex PCR : Tools to overcome PCR-inhibition in DNA-gut-content analysis of soil-living invertebrates , 2006 .

[10]  M. Traugott,et al.  Earthworm primers for DNA-based gut content analysis and their cross-reactivity in a multi-species system , 2006 .

[11]  W. Symondson,et al.  Molecular detection of predation by soil micro‐arthropods on nematodes , 2006, Molecular ecology.

[12]  M. Greenstone Molecular methods for assessing insect parasitism , 2006, Bulletin of Entomological Research.

[13]  J. Bell,et al.  Detection of secondary predation by PCR analyses of the gut contents of invertebrate generalist predators , 2005, Molecular ecology.

[14]  S. Sheppard,et al.  The significance of facultative scavenging in generalist predator nutrition: detecting decayed prey in the guts of predators using PCR , 2005, Molecular ecology.

[15]  T. Haye,et al.  A single-step multiplex PCR assay for the detection of European Peristenus spp., parasitoids of Lygus spp. , 2005 .

[16]  M. Bruford,et al.  Rapid screening of invertebrate predators for multiple prey DNA targets , 2005, Molecular ecology.

[17]  M. Jervis Insects as natural enemies : a practical perspective , 2005 .

[18]  K. Giles,et al.  Estimation of hymenopteran parasitism in cereal aphids by using molecular markers. , 2005, Journal of economic entomology.

[19]  J. Harwood,et al.  Prey selection by linyphiid spiders: molecular tracking of the effects of alternative prey on rates of aphid consumption in the field , 2004, Molecular Ecology.

[20]  C. G. Jackson,et al.  Potential of Detection and Identification of Nymphal Parasitoids (Hymenoptera: Braconidae) of Lygus Bugs (Heteroptera: Miridae) by Using Polymerase Chain Reaction and ITS2 Sequence Analysis Techniques , 2004 .

[21]  P. Dang,et al.  Detection and Differentiation of Parasitoids (Hymenoptera: Aphidiidae and Aphelinidae) of the Brown Citrus Aphid (Homoptera: Aphididae): Species-Specific Polymerase Chain Reaction Amplification of 18S rDNA , 2004 .

[22]  M. Hoy,et al.  HIGH-FIDELITY PCR ASSAY DISCRIMINATES BETWEEN IMMATURE LIPOLEXIS OREGMAE AND LYSIPHLEBUS TESTACEIPES (HYMENOPTERA: APHIDIIDAE) WITHIN THEIR APHID HOSTS , 2004 .

[23]  M. Traugott,et al.  Detecting predation and scavenging by DNA gut-content analysis: a case study using a soil insect predator-prey system , 2004, Oecologia.

[24]  K. Sunderland,et al.  Composition, abundance and pest control potential of spider communities in agroecosystems: a comparison of European and US studies , 2003 .

[25]  E. Friedman Life cycle. , 2003, Health Forum journal.

[26]  W. Symondson Molecular identification of prey in predator diets , 2002, Molecular ecology.

[27]  Rainer Meyhöfer,et al.  Intraguild predation by aphidophagous predators on parasitised aphids: the use of multiple video cameras , 2001 .

[28]  A. Ives,et al.  GENERALIST PREDATORS DISRUPT BIOLOGICAL CONTROL BY A SPECIALIST PARASITOID , 2001 .

[29]  J. Rosenheim,et al.  Intraguild interactions in aphid parasitoids , 2000 .

[30]  K. Pike,et al.  Aphid parasitoids (Hymenoptera: Braconidae: Aphidiinae) of Northwest USA. , 2000 .

[31]  T. A. Hall,et al.  BIOEDIT: A USER-FRIENDLY BIOLOGICAL SEQUENCE ALIGNMENT EDITOR AND ANALYSIS PROGRAM FOR WINDOWS 95/98/ NT , 1999 .

[32]  P. Brenchley,et al.  Populations and communities , 1998 .

[33]  J. Rosenheim Higher-order predators and the regulation of insect herbivore populations. , 1998, Annual review of entomology.

[34]  J. Axelsen,et al.  Pest control by a community of natural enemies. , 1997 .

[35]  M. Walton,et al.  Populations and Communities , 1996 .

[36]  R. Vrijenhoek,et al.  DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. , 1994, Molecular marine biology and biotechnology.

[37]  M. Walton,et al.  Electrophoretic «keys» for the identification of parasitoids (Hymenoptera: Braconidae: Aphelinidae) attacking Sitobion avenae (F.) (Hemiptera: Aphididae) , 1990 .

[38]  M. Walton,et al.  Electrophoresis as a tool for estimating levels of hymenopterous parasitism in field populations of the cereal aphid, Sitobion avenue , 1990 .

[39]  G. Snodgrass,et al.  Emergence and survival of the parasitoid Lysiphlebus testaceipes from Aphis gossypii exposed to aphicides. , 1990 .

[40]  P. Starý,et al.  Biocontrol of aphids by the introduced Lysiphlebus testaceipes (Cress.) (Hym., Aphidiidae) in Mediterranean France , 1988 .

[41]  A. F. Bennett,et al.  Foraging Strategy and Metabolic Rate in Spiders , 1980 .

[42]  K. Sunderland,et al.  Aphid feeding by some polyphagous predators in relation to aphid density in cereal fields. , 1980 .

[43]  J. Anderson,et al.  Metabolic rates of spiders. , 2015, Comparative biochemistry and physiology.