Development of a real-time PCR assay for detection of Mytilus species specific alleles: Application to a sampling survey in Scotland

P. J. Dias, L. Sollelis, E. J. Cook, S. B. Piertney, I. M. Davies, M. Snow

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Shellfish aquaculture is a growing industry in Scotland, dominated by the production of the mussel Mytilus edulis, the native species. Recently the discovery of Mytilus galloprovincialis and Mytilus trossulus together with M. edulis and all 3 hybrids in cultivation in some Scottish sea lochs led to questions regarding the distribution of mussel species in Scotland. The establishment of an extensive sampling survey, involving the collection of mussels at 34 intertidal sites and 10 marinas around Scotland, motivated the development of a high-throughput method for identification of Mytilus alleles from samples. Three Taqman (R)-MGB probes and one set of primers were designed, based on the previously described Me 15/16 primers targeting the adhesive protein gene sequence, and samples were screened for the presence of M. edulis, M. galloprovincialis and M. trossulus alleles using real-time PCR. Mytilus edulis alleles were identified in samples from all 44 sites. Mytilus galloprovincialis alleles were found together with M. edulis alleles extensively in northern parts of the west and east coasts. Mytilus trossulus alleles were identified in samples from 6 sites in the west and southwest of Scotland. Because M. trossulus is generally undesirable in cultivation and therefore preventing the geographical spread of this species across Scotland is considered beneficial by the shellfish aquaculture industry, these 6 samples were further analysed for genotype frequencies using conventional PCR. Although distribution of the non-native species M. galloprovincialis and M. trossulus have proven to be more widespread than previously thought, there is no evidence from our study of either M. trossulus or M. galloprovincialis acting as an invasive species in Scotland. The real-time PCR method developed in this study has proven to be a rapid and effective tool for the identification of M. edulis, M. galloprovincialis and M. trossulus alleles from samples and should prove useful in future surveys, ecological or aquaculture management related studies in both unispecific and mixed species areas of these species. Crown Copyright (C) 2008 Published by Elsevier B.V. All rights reserved.

Original languageEnglish
Pages (from-to)253-258
Number of pages6
JournalJournal of Experimental Marine Biology and Ecology
Volume367
Issue number2
Early online date11 Nov 2008
DOIs
Publication statusPublished - 15 Dec 2008

Keywords

  • mytilus
  • real-time PCR
  • Scotland
  • shellfish aquaculture
  • indigenous perna-perna
  • cod gadus-morhua
  • mussel mytilus
  • genetic-structure
  • simultaneous identification
  • plankton samples
  • edulis complex
  • blue mussels
  • hybrid zone
  • galloprovincialis

Cite this

Development of a real-time PCR assay for detection of Mytilus species specific alleles : Application to a sampling survey in Scotland. / Dias, P. J.; Sollelis, L.; Cook, E. J.; Piertney, S. B.; Davies, I. M.; Snow, M.

In: Journal of Experimental Marine Biology and Ecology, Vol. 367, No. 2, 15.12.2008, p. 253-258.

Research output: Contribution to journalArticle

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AU - Davies, I. M.

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AB - Shellfish aquaculture is a growing industry in Scotland, dominated by the production of the mussel Mytilus edulis, the native species. Recently the discovery of Mytilus galloprovincialis and Mytilus trossulus together with M. edulis and all 3 hybrids in cultivation in some Scottish sea lochs led to questions regarding the distribution of mussel species in Scotland. The establishment of an extensive sampling survey, involving the collection of mussels at 34 intertidal sites and 10 marinas around Scotland, motivated the development of a high-throughput method for identification of Mytilus alleles from samples. Three Taqman (R)-MGB probes and one set of primers were designed, based on the previously described Me 15/16 primers targeting the adhesive protein gene sequence, and samples were screened for the presence of M. edulis, M. galloprovincialis and M. trossulus alleles using real-time PCR. Mytilus edulis alleles were identified in samples from all 44 sites. Mytilus galloprovincialis alleles were found together with M. edulis alleles extensively in northern parts of the west and east coasts. Mytilus trossulus alleles were identified in samples from 6 sites in the west and southwest of Scotland. Because M. trossulus is generally undesirable in cultivation and therefore preventing the geographical spread of this species across Scotland is considered beneficial by the shellfish aquaculture industry, these 6 samples were further analysed for genotype frequencies using conventional PCR. Although distribution of the non-native species M. galloprovincialis and M. trossulus have proven to be more widespread than previously thought, there is no evidence from our study of either M. trossulus or M. galloprovincialis acting as an invasive species in Scotland. The real-time PCR method developed in this study has proven to be a rapid and effective tool for the identification of M. edulis, M. galloprovincialis and M. trossulus alleles from samples and should prove useful in future surveys, ecological or aquaculture management related studies in both unispecific and mixed species areas of these species. Crown Copyright (C) 2008 Published by Elsevier B.V. All rights reserved.

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KW - indigenous perna-perna

KW - cod gadus-morhua

KW - mussel mytilus

KW - genetic-structure

KW - simultaneous identification

KW - plankton samples

KW - edulis complex

KW - blue mussels

KW - hybrid zone

KW - galloprovincialis

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JF - Journal of Experimental Marine Biology and Ecology

SN - 0022-0981

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