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

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.

journal_name

Mol Ecol Resour

authors

Bergmann N,Fricke B,Schmidt MC,Tams V,Beining K,Schwitte H,Boettcher AA,Martin DL,Bockelmann AC,Reusch TB,Rauch G

doi

10.1111/j.1755-0998.2011.03051.x

subject

Has Abstract

pub_date

2011-11-01 00:00:00

pages

1076-81

issue

6

eissn

1755-098X

issn

1755-0998

journal_volume

11

pub_type

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