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Genetic Characteristics of the Japanese Serow Capricornis

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Genetic Characteristics of the Japanese Serow Capricornis Genetic Characteristics of the Japanese Serow Capricornis crispus in the Kii Mountain Range, Central Japan Authors: Arisa Iwahori, Jyun-ichi Kitamura, and Kouichi Kawamura Source: Zoological Science, 36(4) : 306-315 Published By: Zoological Society of Japan URL: https://doi.org/10.2108/zs180187 BioOne Complete (complete.BioOne.org) is a full-text database of 200 subscribed and open-access titles in the biological, ecological, and environmental sciences published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Complete website, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/terms-of-use. Usage of BioOne Complete content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder. BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research. Downloaded From: https://bioone.org/journals/Zoological-Science on 07 Aug 2019 Terms of Use: https://bioone.org/terms-of-use Access provided by Zoological Society of Japan ZOOLOGICAL306 SCIENCE 36: 306–315 (2019) A. Iwahori et al. © 2019 Zoological Society of Japan Genetic Characteristics of the Japanese Serow Capricornis crispus in the Kii Mountain Range, Central Japan Arisa Iwahori1, Jyun-ichi Kitamura2, and Kouichi Kawamura1* 1Faculty of Bioresources, Mie University, Kurimamachiya 1577, Tsu, Mie 514-8507, Japan 2Mie Prefectural Museum, 3060 Isshinden-kouzubeta, Tsu, Mie 514-0061, Japan The Japanese serow, Capricornis crispus, is an indigenous bovid species exclusively inhabiting mountain regions in the main Japanese islands, excepting Hokkaido. It had decreased in abun- dance to its lowest level due to overhunting and deforestation, with its distribution severely frag- mented from the middle of the 20th century, many populations of C. crispus currently facing the risk of extinction. The Kii Mountain Range (KM) on Honshu is one such location that has seen a drastic population decline of C. crispus. In this study, we examined genetic characteristics of C. crispus in KM and neighboring regions of the Chubu district, using mtDNA and microsatellite mark- ers, in order to devise strategies for its conservation. Results for mtDNA were characterized by low nucleotide diversity with five endemic and two dominant haplotypes shared by individuals in neighboring regions. A Bayesian skyline plot indicated a gradual increase after the last glacial maximum. For microsatellites, the genetic diversity of C. crispus in KM was comparable to Shi- zuoka and higher than Shikoku. Recent genetic bottlenecks were strongly suggested in C. crispus in KM. Bayesian clustering showed a genetic cline between KM and neighboring regions, where multivariate analysis suggested three local populations. A Mantel test indicated male-biased dis- persal. These results indicate that C. crispus in KM and neighboring regions constitute multiple local populations, connected through restricted gene flow. For the conservation of C. crispus, it is important to define small-scale conservation units, among which genetic connectivity should be facilitated to prevent further loss of genetic diversity. Key words: Capricornis crispus, conservation unit, dispersal, genetic bottleneck, genetic cline decrease of C. crispus to the lowest levels (Ochiai, 2016). In INTRODUCTION 1955 the species was designated as a “Special National The Japanese serow, Capricornis crispus, is an indige- Monument of Japan” by the Japanese Government. Owing nous bovid species in Japan. Capricornis species are widely to the subsequent strict protection, some populations of C. distributed in East and Middle Asia, ranging from the tropical crispus on Honshu showed signs of recovery. However, fol- to the subarctic zone (Ochiai, 2016). The genus was formerly lowing increases problems associated with damage to agri- considered to be primitive in the subfamily Caprinae, belong- cultural and forestry sites emerged in some prefectures, ing to the tribe Rupicaprini (Simpson, 1945). However, recent such as Gifu and Aichi in central Honshu (Ochiai, 2016). molecular studies suggest that Capricornis is not a member Consequently, in 1979 the Japanese government desig- of the Rupicaprini, but genetically related to species of the nated legally protected areas for C. crispus while allowing tribe Ovibovini (Ropiquet and Hassanin, 2009). The genus culling for population control in other regions under special Capricornis comprises three species, C. sumatraensis, C. license (MEGJ, 2010). Populations in Shikoku and Kyushu swinhoei and C. crispus (Novak, 1999). Although C. swinhoei have remained small and fragmented, and continue to face in Taiwan had long been considered to be closely related to the risk of extinction (Okumura, 2003). C. crispus, recent studies using mtDNA have shown it to be The Kii Mountain Range (KM) in central Honshu is one a sister species of C. sumatraensis (Min et al., 2004). of the principal habitats of C. crispus. In this region, compre- Formerly, C. crispus was widely distributed in mountain- hensive research on C. crispus has been periodically con- ous regions of the main Japanese islands, excepting ducted since 1986 by the collaboration of Mie, Nara and Hokkaido (Ochiai, 2016). However, hunting and deforesta- Wakayama Prefectures to elucidate the ecology, distribution tion resulted in its decline, resulting not only the fragmenta- and population dynamics of the species (PBEMNW, 2010). tion of populations but also their disappearance from Based on the results of the first tranche of research in 1986– Chugoku, Fukuoka and Izu districts. Indiscriminate hunting 1987, KM was registered as a protected area for C. crispus in the 19th and 20th centuries in particular drove the in 1989. However, the fourth phase of research on C. crispus in the KM protected area (FSRCPK, conducted in 2008– * Corresponding author. E-mail: [email protected] 2009) reported a decrease in numbers and the fragmenta- doi:10.2108/zs180187 tion of habitat, despite an increase in the distribution of C. Downloaded From: https://bioone.org/journals/Zoological-Science on 07 Aug 2019 Terms of Use: https://bioone.org/terms-of-use Access provided by Zoological Society of Japan Conservation genetics of Japanese serow 307 crispus. Results also suggested that KM supported the low- prior to DNA extraction. DNA was extracted using DNeasy Blood est population density of C. crispus among the protected and Tissue Kit (Qiagen, USA). areas of C. crispus in Japan, along with the Southern Ou- Sex of collected individuals was primarily judged from external Mountain Range (PBEMNW, 2010). genital features. For sexually ambiguous individuals, PCR-based sex identification was employed as described below. Most collected In an analysis of the cytochrome b region of the mtDNA individuals were adult, and only three individuals (two females and of C. crispus (sequence data not published), PBEMNW one male) in KM were young (less than three years), judging from (2010) reported that the dominant haplotype of C. crispus in horn size (Kishimoto, 1988). KM was found in individuals from Gifu, Shizuoka and Yamagata Prefectures, although some haplotypes were PCR-based Sex identification endemic to KM. These findings suggested that C. crispus in For sexually ambiguous individuals (n = 30) PCR-based sex KM and that of neighboring regions were not genetically identification was used. In mammals, the amelogenin gene is separated, but were connected through a small number of widely used for sex identification, because it has a locus on both occasional migrants. Yamashiro and Yamashiro (2012) com- sex chromosomes (X and Y) and respective base sequences (Pfei- pared the genetic diversity of C. crispus in Shikoku, KM ffer and Brenig, 2005). This gene was found to be effective for the sex identification of C. crispus (Nishimura et al., 2010). We ampli- (including the Shiga population) and Shizuoka, using five fied the amelogenin gene via PCR, using the primer pairs: SE47 microsatellite markers (genotype data not published). They and SE38 and PCR protocols described by Nishimura et al. (2010). reported that the Shikoku population expressed the lowest The PCR products were electrophoresed on 2% agarose gel with genetic diversity, while the other two populations showed TAE buffer. The gel was stained by GelStar nucleic acid stain (FMC approximately the same level of genetic diversity. In the pre- Fig. 1 IwahoriBioproducts, et al. USA) and visualized under UV illumination to identify ceding studies of C. crispus (e.g., PBEMNW, 2010; Nishimura et al., 2011; Yamashiro and Yamashiro, 2012), genetic infor- mation was exclusively restricted to one kind of genetic N marker, which is insufficient for the elucidation of the genetic 36°N characteristics of the species (Gompert et al., 2006). Hokkaido HM In conservation management, it is important to clarify Japan the genetic characteristics of the targeted species, because it enables inference of their genetic structure, genetic rela- Yamagata Gifu tionships and demographic history. Such information is especially indispensable for the identification of conserva- Honshu tion units, which are a requisite for the design of a rational Shizuoka
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