Development and Multiplexed Amplification of SSR Markers for Thuja occidentalis (Cupressaceae) Using Shotgun Pyrosequencing

Premise of the study: Sixteen novel, polymorphic, multiplexed microsatellite loci were developed for eastern white cedar (Thuja occidentalis) using simple sequence repeat (SSR)–enriched shotgun pyrosequencing. Methods and Results: Sixteen loci were tested on a panel of 24 individuals from different...

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Published inApplications in plant sciences Vol. 1; no. 5; pp. 1200427 - n/a
Main Authors Xu, Huaitong, Tremblay, Francine, Bergeron, Yves
Format Journal Article
LanguageEnglish
Published United States Botanical Society of America 01.05.2013
John Wiley & Sons, Inc
Wiley
Subjects
454 GS-FLX Titanium
Alleles
Canada
Cupressaceae
Deoxyribonucleic acid
DNA
Endangered & extinct species
forests
Genetic diversity
genetic variation
Heterozygosity
islands
loci
microsatellite marker
microsatellite repeats
next-generation sequencing
Plant sciences
polymerase chain reaction
population
Population genetics
PRIMER NOTE
sequence analysis
shotgun pyrosequencing
Thuja occidentalis
Canada
James Bay Canada
United States > US
Tennessee
Quebec Canada
Montreal Quebec Canada
California
North America
Germany
Thuja occidentalis
population genetics
microsatellite marker
shotgun pyrosequencing
next-generation sequencing
454 GS-FLX Titanium
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Summary:Premise of the study: Sixteen novel, polymorphic, multiplexed microsatellite loci were developed for eastern white cedar (Thuja occidentalis) using simple sequence repeat (SSR)–enriched shotgun pyrosequencing. Methods and Results: Sixteen loci were tested on a panel of 24 individuals from different populations. The number of observed alleles ranged from four to 22. Four sets of multiplex PCR for the 16 loci were then carried out on 60 individuals of two populations from islands of FERLD Duparquet Forest, Canada. Mean number of alleles, observed heterozygosity, and expected heterozygosity were respectively 5.75, 0.594, and 0.574 for Island 58, and 5.50, 0.704, and 0.624 for Island 134. Conclusions: Four sets of multiplex microsatellite loci can be used for future genetic studies, which includes investigating genetic diversity and structure, and fragmentation and regeneration studies.
Bibliography:http://dx.doi.org/10.3732%2Fapps.1200427
The authors thank V. Paul, A. Leduc, G. Trudeau, R. Libert, and S. Chauchard for help in the field and Dr. W. F. J. Parsons for corrections to the English. This research was funded by the Natural Sciences and Engineering Research Council of Canada (NSERC; STPGP 336871) to F.T., and also supported by an Industrial Innovation Doctoral Scholarship from NSERC, a BMP innovation doctoral scholarship from the Fonds de recherche du Québec–Nature et technologies (FRQNT) and Tembec, and a scholarship from the Forêt d'enseignement et de recherche du lac Duparquet (FERLD) to H.X.
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The authors thank V. Paul, A. Leduc, G. Trudeau, R. Libert, and S. Chauchard for help in the field and Dr. W. F. J. Parsons for corrections to the English. This research was funded by the Natural Sciences and Engineering Research Council of Canada (NSERC; STPGP 336871) to F.T., and also supported by an Industrial Innovation Doctoral Scholarship from NSERC, a BMP innovation doctoral scholarship from the Fonds de recherche du Québec–Nature et technologies (FRQNT) and Tembec, and a scholarship from the Forêt d’enseignement et de recherche du lac Duparquet (FERLD) to H.X.
ISSN:2168-0450
2168-0450
DOI:10.3732/apps.1200427