title: Un-nesting DNA Russian dolls - the potential for constructing food webs using residual DNA in empty aphid mummies.
abstract: Constructing food-web assemblages comprising parasitoid wasps involves large field collections of hosts followed by labour-intensive rearing of the insects to evaluate the rates of parasitism along with morphological or molecular identification of the parasitoid species. This article presents research towards a new molecular method for the practical and accurate construction of aphid-based food webs. We hypothesize that parasitoid and hyperparasitoid DNA left inside aphid mummies after emergence of these third and fourth trophic-level guilds can be simultaneously detected using universal polymerase chain reaction (PCR) primers for nonspecific DNA amplification in combination with single-stranded conformation polymorphism (SSCP) analysis. Such a protocol theoretically allows food-web construction to be performed with no a priori knowledge of the species present. Moreover, the use of empty mummies circumvents rearing and minimizes labour and time in the field and laboratory. To test our hypothesis, we conducted DNA analyses on laboratory-produced parasitized aphids (mummies) from Myzus persicae and Brevicoryne brassicae (two important aphid pest species) after exposure to the parasitoid Diaeretiella rapae and the hyperparasitoid Asaphes vulgaris. DNA is amplifiable in empty aphid mummies for as long as 3 weeks after parasitoid emergence. However, the simultaneous identification of several species in a single mummy sample was rare, which hinders the accurate inference of trophic links. DNA quality and relative quantity, together with preferential amplification, are potential explanations of current results. Technical refinements are needed to ensure full reliability and detection of complex trophic links. The use of PCR-SSCP for food-web construction is novel, and its potential to include an important number of different species is yet to be fully explored.
authors.affiliation: Bio-Protection Research Centre, Lincoln University, PO Box 85084, Lincoln, 7647, New Zealand.
authors.forename: Y-D
authors.initials: YD
authors.lastname: Varennes
authors.forename: S
authors.initials: S
authors.lastname: Boyer
authors.forename: S D
authors.initials: SD
authors.lastname: Wratten
mesh_terms: D000818:Animals; D001042:Aphids / Q000469:parasitology*; D004247:DNA / Q000032:analysis; D020387:Food Chain; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D017422:Sequence Analysis, DNA / Q000379:methods*; D014863:Wasps / Q000235:genetics / Q000502:physiology*
chemical_list: D004247:DNA
keywords: Brevicoryne brassicae; Myzus persicae; biological control; environmental DNA