Evaluation of temperature gradient gel electrophoresis for the analysis of prey DNA within the guts of invertebrate predators.
The utility of temperature gradient gel electrophoresis (TGGE) as a means of analysing the gut contents of predators was evaluated. Generalist predators consume multiple prey species and a species-specific primer approach may not always be a practical means of analysing predator responses to prey diversity in complex and biodiverse ecosystems. General invertebrate primers were used to amplify the gut contents of predators, generating banding patterns that identified component prey remains. There was no evidence of dominance of the polymerase chain reaction (PCR) by predator DNA. When applied to field samples of the carabid predator Pterostichus melanarius (Illiger) nine banding patterns were detected, including one for aphids. To further distinguish between species, group-specific primers were designed to separate species of earthworm and aphid. TGGE of the earthworm PCR products generated banding patterns that varied with haplotype in some species. Aphid and earthworm DNA could be detected in the guts of carabids for up to 24 h using TGGE. In P. melanarius, with low numbers of prey per insect gut (mean<3), interpretation of banding patterns proved to be tractable. Potential problems of interpretation of TGGE gels caused by multiple prey bands, cryptic bands, haplotype variation, taxonomic uncertainties (especially with regard to earthworms), secondary predation, scavenging and presence of parasites and parasitoids in the prey or the predators, are discussed. The results suggest that PCR, using combinations of general invertebrate and group-specific primers followed by TGGE, provides a potentially useful approach to the analysis of multiple uncharacterized prey in predators.
Harper
G L
GL
Cardiff School of Biosciences, Cardiff University, PO Box 915, Cardiff, CF10 3TL, UK.
Sheppard
S K
SK
Harwood
J D
JD
Read
D S
DS
Glen
D M
DM
Bruford
M W
MW
Symondson
W O C
WO
D000818:Animals; D001042:Aphids / Q000235:genetics; D001517:Coleoptera / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D004063:Digestion / Q000502:physiology; D003576:Electron Transport Complex IV / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000662:veterinary*; D005260:Female; D007413:Intestinal Mucosa / Q000378:metabolism; D007448:Invertebrates / Q000145:classification* / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D009835:Oligochaeta / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012335:RNA, Ribosomal / Q000235:genetics; D013997:Time Factors
D017931:DNA Primers; D012335:RNA, Ribosomal; C075526:RNA, ribosomal, 12S; D003576:Electron Transport Complex IV