IMIS

Publications | Institutes | Persons | Datasets | Projects | Maps
[ report an error in this record ]basket (0): add | show Print this page

Quantification of copepod gut content by differential length amplification quantitative PCR (dla-qPCR)
Troedsson, C.; Simonelli, P.; Nägele, V.; Nejstgaard, J.C.; Frischer, M.E. (2009). Quantification of copepod gut content by differential length amplification quantitative PCR (dla-qPCR). Mar. Biol. (Berl.) 156(3): 253-259. http://dx.doi.org/10.1007/s00227-008-1079-8
In: Marine Biology: International Journal on Life in Oceans and Coastal Waters. Springer: Heidelberg; Berlin. ISSN 0025-3162; e-ISSN 1432-1793, more
Peer reviewed article  

Available in  Authors 

Keyword
    Marine/Coastal

Authors  Top 
  • Troedsson, C.
  • Simonelli, P.
  • Nägele, V.
  • Nejstgaard, J.C.
  • Frischer, M.E.

Abstract
    Quantification of feeding rates and selectivity of zooplankton is vital for understanding the mechanisms structuring marine ecosystems. However, methodological limitations have made many of these studies difficult. Recently, molecular based methods have demonstrated that DNA from prey species can be used to identify zooplankton gut contents, and further, quantitative gut content estimates by quantitative PCR (qPCR) assays targeted to the 18S rRNA gene have been used to estimate feeding rates in appendicularians and copepods. However, while standard single primer based qPCR assays were quantitative for the filter feeding appendicularian Oikopleura dioica, feeding rates were consistently underestimated in the copepod Calanus finmarchicus. In this study, we test the hypothesis that prey DNA is rapidly digested after ingestion by copepods and describe a qPCR-based assay, differential length amplification qPCR (dla-qPCR), to account for DNA digestion. The assay utilizes multiple primer sets that amplify different sized fragments of the prey 18S rRNA gene and, based on the differential amplification of these fragments, the degree of digestion is estimated and corrected for. Application of this approach to C. finmarchicus fed Rhodomonas marina significantly improved quantitative feeding estimates compared to standard qPCR. The development of dla-qPCR represents a significant advancement towards a quantitative method for assessing in situ copepod feeding rates without involving cultivation-based manipulation.

All data in the Integrated Marine Information System (IMIS) is subject to the VLIZ privacy policy Top | Authors