Development of microsatellite markers for blue duiker ( Cephalophus monticola ) using next-generation sequencing and cro
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TECHNICAL NOTE
Development of microsatellite markers for blue duiker (Cephalophus monticola) using next-generation sequencing and cross-amplification in other duikers Etienne Franc¸ois Akomo-Okoue • Eiji Inoue Christopher Adenyo • Azusa Hayano • Miho Inoue-Murayama
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Received: 5 March 2013 / Accepted: 8 March 2013 / Published online: 14 March 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract We developed nine microsatellite markers for duikers by next-generation sequencing using a muscle tissue sample of blue duiker (Cephalophus monticola). For blue duikers, the number of alleles ranged from 4 to 14 (mean 9.4), and the observed and expected heterozygosities were 0.333–0.833 (mean 0.716) and 0.625–0.878 (mean 0.817), respectively. Cumulative probability of identity was low (1.5 9 10-12), indicating that these markers are enough for individual discrimination. All nine markers were successfully amplified in three other duikers (C. dorsalis, C. ogilbyi and C. sylvicultor), and almost all loci were polymorphic. These markers will be useful for clarifying genetic structure of duikers. Keywords Duiker Next-generation sequencing Microsatellite markers Genetic analysis
Duikers (genus Cephalophus) are small ungulates widely distributed in African rainforests and are important sources of protein and income for local people (Newing 2001; Schenck et al. 2006). Several species of duikers, belonging to the same genus, normally live in the same habitat
E. F. Akomo-Okoue E. Inoue (&) Department of Zoology, Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan e-mail: [email protected] E. F. Akomo-Okoue Institut de Recherche en Ecologie Tropicale (IRET/ CENAREST), BP 13354, Libreville, Gabon C. Adenyo A. Hayano M. Inoue-Murayama Wildlife Research Center of Kyoto University, Kyoto 606-8203, Japan
in Central Africa areas (Kingdon 1997). Blue duiker (C. monticola) is the smallest current duiker inhabiting all forest types (Dubost, 1980). Across Western and Central African forests, blue duiker is one of the major mammal species for bushmeat (Wilkie and Carpenter 1999; Newing 2001). It is important to monitor genetic diversity for their conservation, although current hunting pressure on this species was estimated to remain within a sustainable level (Fa et al. 1995; van Vliet and Nasi 2008). Microsatellite markers are useful for describing genetic diversity and genetic structure of wild animals. Some microsatellite markers which were developed in other bovine species were amplified and were polymorphic in duikers (Ntie et al. 2010a). We tried those markers using DNA extracted from fecal samples of duikers, but the amplification of several markers failed probably due to low quality and quantity of DNA. We therefore found it necessary, to develop original microsatellite markers for duikers. We collected 11 muscle tissue samples of blue duiker and two muscle tissue samples each of bay duiker (C. dorsalis) and Ogilby’s duiker (C. ogilbyi) in Gabonese meat markets. The samples were pre
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