A quantitative real-time polymerase chain reaction assay for the seagrass pathogen Labyrinthula zosterae.

Bergmann, Nina, Fricke, Birgit, Schmidt, Martina, Tams, Verena, Beining, Katrin, Schwitte, Hildegard, Boettcher, Anne A., Martin, Daniel L., Bockelmann, Anna-Christina, Reusch, Thorsten B.H. and Rauch, Gisep (2011) A quantitative real-time polymerase chain reaction assay for the seagrass pathogen Labyrinthula zosterae. Molecular Ecology Resources, 11 (6). pp. 1076-1081. DOI 10.1111/j.1755-0998.2011.03051.x.

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Abstract

The protist Labyrinthula zosterae (Phylum Bigyra, sensu Tsui et al. 2009) has been identified as a causative agent of wasting disease in eelgrass (Zostera marina), of which the most intense outbreak led to the destruction of 90% of eelgrass beds in eastern North America and western Europe in the 1930s. Outbreaks still occur today, albeit at a smaller scale. Traditionally, L. zosterae has been quantified by measuring the necrotic area of Z. marina leaf tissue. This indirect method can however only lead to a very rough estimate of pathogen load. Here, we present a quantitative real-time polymerase chain reaction (qPCR) approach to directly detect and quantify L. zosterae in eelgrass tissue. Based on the internal transcribed spacer (ITS) sequences of rRNA genes, species-specific primers were designed. Using our qPCR, we were able to quantify accurately and specifically L. zosterae load both from culture and eelgrass leaves using material from Europe and North America. Our detection limit was less than one L. zosterae cell. Our results demonstrate the potential of this qPCR assay to provide rapid, accurate and sensitive molecular identification and quantification of L. zosterae. In view of declining seagrass populations worldwide, this method will provide a valuable tool for seagrass ecologists and conservation projects.

Document Type: Article
Keywords: Food Webs; Evolutionary Ecology of Marine Fishes; Labyrinthula; pathogen quantification; qPCR; seagrass
Research affiliation: OceanRep > GEOMAR > FB3 Marine Ecology > FB3-EOE-N Experimental Ecology - Food Webs
OceanRep > GEOMAR > FB3 Marine Ecology > FB3-EV Marine Evolutionary Ecology
OceanRep > The Future Ocean - Cluster of Excellence
Refereed: Yes
Open Access Journal?: No
DOI etc.: 10.1111/j.1755-0998.2011.03051.x
ISSN: 1755-098X
Projects: Future Ocean
Date Deposited: 30 Nov 2011 14:40
Last Modified: 27 Feb 2017 12:57
URI: http://oceanrep.geomar.de/id/eprint/12646

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