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Int. J. Mol. Sci. 2015, 16(9), 21330-21341; doi:10.3390/ijms160921330

Development of Seven Microsatellite Markers Using Next Generation Sequencing for the Conservation on the Korean Population of Dorcus hopei (E. Saunders, 1854) (Coleoptera, Lucanidae)

1,†,* , 2,†,* and 3,†,*
1
Plant Quarantine Technology Center, Animal and Plant Quarantine Agency, 234-3, Mangpo-dong, Yeongtong-gu, Suwon, Gyeonggi-do 443-400, Korea
2
Department of Life Science, College of Natural Science, Kyonggi University, 154-42, Gwanggyeongsan-ro, Yeongtong-gu, Suwon, Gyeonggi-do 443-780, Korea
3
Biological Resources Research Department, National Institute of Biological Resources, 42, Hwangyeong-ro, Seo-gu, Incheon 404-170, Korea
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Academic Editor: Ritva Tikkanen
Received: 2 June 2015 / Revised: 27 August 2015 / Accepted: 28 August 2015 / Published: 7 September 2015
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
View Full-Text   |   Download PDF [1668 KB, uploaded 7 September 2015]   |  

Abstract

We developed microsatellite markers for genetic structural analyses of Dorcus hopei, a stag beetle species, using next generation sequencing and polymerase chain reaction (PCR)-based genotyping for regional populations. A total of 407,070,351 base pairs of genomic DNA containing >4000 microsatellite loci except AT repeats were sequenced. From 76 loci selected for primer design, 27 were polymorphic. Of these 27 markers, 10 were tested on three regional populations: two Chinese (Shichuan and Guangxi) and one Korean (Wanju). Three markers were excluded due to inconsistent amplification, genotyping errors, and Hardy-Weinberg equilibrium (HWE). By multi-locus genotyping, the allele number, observed heterozygosity and polymorphism information content of seven microsatellite loci were ranged 2‒10, 0.1333‒1.0000, and 0.1228‒0.8509, respectively. In an analysis on the genetic differentiation among regional populations including one Japanese population and one cross-breeding population, the individual colored bar-plots showed that both Chinese populations were closer to each other than to the Far East Asian populations. In Far East Asian populations, Wanju and Nirasaki populations could not be distinguished from each other because the frequency of genetic contents was very similar in some individuals of two populations. Moreover, the cross-breeding population contained all patterns of genetic contents shown in Chinese, Korean, and Japanese populations, compared with the genetic content frequency of each regional population. As a result, we examined whether the cross-breeding population might be a hybrid population, and might contain a possibility of interbreeding with Chinese populations in parental generations. Therefore, these markers will be useful for analyses of genetic diversity in populations, genetic relationships between regional populations, genetic structure analyses, and origin tests. View Full-Text
Keywords: Coleoptera; Dorcus hopei; hybrid population; Lucanidae; microsatellite marker; MiSeq; next generation sequencing; pet insect Coleoptera; Dorcus hopei; hybrid population; Lucanidae; microsatellite marker; MiSeq; next generation sequencing; pet insect
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Kang, T.H.; Han, S.H.; Park, S.J. Development of Seven Microsatellite Markers Using Next Generation Sequencing for the Conservation on the Korean Population of Dorcus hopei (E. Saunders, 1854) (Coleoptera, Lucanidae). Int. J. Mol. Sci. 2015, 16, 21330-21341.

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