PCR-DHPLC assay for the identification of predator-prey interactions

• Denaturing high-performance liquid chromatography (DHPLC) is a relatively new method for separating amplicons in a mixture, and was recently developed for parasite detection in the blue crab Callinectes sapidus. That assay used a peptide nucleic acid (PNA) PCR hybridization blocking probe (PNAPCRDHPLC) to decrease the generic PCR bias of dominant templates (the host) in the mixture prior to separation on the DHPLC column, thus enhancing the less abundant parasite DNA. The same assay and rational can be used to investigate predatorprey interactions. However, in ecosystem studies with many predatorprey relationships, development of specific PNA-blocking probes for each predator would be too laborious. Here, we have developed a PCRDHPLC assay excluding the dominant predator amplicons in a first DHPLC run, followed by re-amplification of the non-predator retention volumes and further separation and characterization in a second DHPLC run. This assay generated data on the specific trophic interactions between the calanoid copepod Limnocalanus macrurus and its prey from a seasonal sampling programme. The assay provides an efficient way for an unbiased screening of predatorprey relationships, and although developed for L. macrurus in this study, the approach has wide applicability for any predatorprey interaction.

Ämnesord

NATURVETENSKAP  -- Geovetenskap och miljövetenskap -- Oceanografi, hydrologi och vattenresurser (hsv//swe)

NATURAL SCIENCES  -- Earth and Related Environmental Sciences -- Oceanography, Hydrology and Water Resources (hsv//eng)

NATURVETENSKAP  -- Biologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences (hsv//eng)

Nyckelord

trophic interaction

biodiversity

feeding

zooplankton

18S rDNA

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)