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II: Chromosome Analyses of Stripe Tree Skink (Lipinia Vittigera)

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II: Chromosome Analyses of Stripe Tree Skink (Lipinia Vittigera) © 2017 The Japan Mendel Society Cytologia 82(1) Special Issue: 83–90 Cytogenetic of Skink (Reptilia, Scincidae) from Thailand: II: Chromosome Analyses of Stripe Tree Skink (Lipinia vittigera) Isara Patawang1, Alongklod Tanomtong2*, Sarun Jumrusthanasan3, Hathaipat Khongcharoensuk2, Sarawut Kaewsri3 and Krit Pinthong4 1 Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, Muang 50200, Thailand 2 Applied Taxonomic Research Center (ATRC), Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen, Muang 40002, Thailand 3 Program in Biology, Department of Science, Faculty of Science, Buriram Rajabhat University, Buriram, Muang 31000, Thailand 4 Program in Biology, Department of Fundamental Science, Faculty of Science and Technology, Surindra Rajabhat University, Surin, Muang 32000, Thailand Received February 19, 2015; accepted June 4, 2015 Summary Chromosomal analyses of the stripe tree skink (Lipinia vittigera Boulenger, 1894) from mitotic and meiotic cell divisions were studied. Bone marrow and testis samples were taken from five male and five female skinks. Skink chromosome preparations were conducted by squash technique from bone marrow and testis. The chromosomes were stained by conventional staining and Ag-NOR banding techniques. The results showed that the diploid chromosome number of L. vittigera was 2n=28, and the fundamental number (NF) was 44 in both males and females. The types of chromosomes were present as 8 large metacentric, 2 medium metacentric, 6 small metacentric macrochromosomes, and 12 microchromosomes. There was no observation of strange size chromosomes related to sex. Nucleolar organizer regions (NORs) appear to the telomere on the long arm of the largest metacentric chromosome pair 1 and medium metacentric chromosome pair 5. We found that during meta- phase I on meiosis, the homologous chromosomes showed synapsis, which can be defined as the 14 bivalents and 14 haploid chromosomes at metaphase II as diploid species. The karyotype formula of L. vittigera is as follows: m mm 2n (diploid) 28=L8 +M 26 +S +12 microchromosomes Key words Stripe tree skink, Lipinia vittigera, Karyotype, Chromosome. The stripe tree skink (Lipinia vittigera) belongs to the size; lower eyelid with a clear window; auricular lobules class Reptilia, order Squamata, Suborder Lacertilia, in- absent; body scales smooth; longitudinal scale rows fraorder Scincomorpha, family Scincidae, and subfam- at midbody ≤28; basal subdigital lamellae expanded; ily Lygosominae (Fig. 1). The subfamily Lygosominae vomers fused; pterygoid teeth absent; dorsal colour pat- contains over 600 species (Matsui 1992, Zug 1993). tern typically comprising a pale middorsal stripe at least Within this subfamily, three evolutionary lineages (i.e., anteriorly; visceral fat bodies absent and brood size (Das Eugongylus, Mabuya and Sphenomorphus groups) are and Greer 2002). recognized on morphological, karyological and immu- Chromosome number in the family Scincidae var- nogenetic grounds (Donnellan 1991a, Ota et al. 1996, ies from 2n=24 to 2n=36 (Table 1). In addition, the Honda et al. 1999). diploid number tends to be approximately genus spe- The scincid genus Lipinia Gray, 1845, which was re- cific, for example 2n=24 in Lygosoma (Bhatnagar 1959), vived from the synonymy of Lygosoma Hardwicke and 2n=26 in Plestiodon (Caputo et al. 1994), 2n=28 in Gray, 1827 by Mittleman (1952), contains 26 nominal Cryptoblepharus (Makino and Momma 1949), 2n=30 in species ranging from the Andaman and Nicobar Island Eulamprus (King 1973b) and 2n=32 in Cyclodomorphus of India, east to Thailand and Cambodia, the Malay (Donnellan 1991a). General karyotypes of the scincid Peninsula, Sumatra, Borneo, the Philippines, to New lizards are composed of 2n=26, 28, 30, or 32 chromo- Guinea and the Republic of Belau (Austin 1995, Das and somes that form three distinct size-groups (large and Greer 2002). Members of the genus Lipinia are charac- small biarmed macrochromosomes and a series of mi- terized by the following suite of characters: small body crochromosomes). Most scincid karyotypes so far stud- ied can be assigned to one of the two main groups: the * Corresponding author, e-mail: [email protected] first group, represented by most species in the subfamily DOI: 10.1508/cytologia.82.83 Lygosominae and a number of species of the subfamily 84 I. Patawang et al. Cytologia 82(1) Special Issue Fig. 1. General characteristic of the stripe tree skink, Lipinia vittigera (Squamata, Scincidae) from northeast Thailand (scale bar indicates 1 cm). Scincinae (Greer 1970), is distinguished by four pairs of Results and discussion distinctly large biarmed chromosomes, while the second group, accommodating some other scincine species, is Chromosome number, fundamental number and karyo- characterized by no more than two pairs of distinctly type of L. vittigera large metacentric chromosomes (Giovannotti et al. 2009, The Scincidae are a large family of lizards and until Castiglia et al. 2013). now, no study investigated the karyotype of L. vittigera, This present study is the first report of the chro- and this is the first report on cytogenetic characteriza- mosomal characteristics of L. vittigera accomplished tion by conventional staining and Ag-NOR banding with the conventional staining and Ag-NOR banding techniques in this species. For L. vittigera, we found techniques. The results obtained can provide increasing a diploid chromosome number of 2n=28 in all studied cytogenetic information for future studies on taxonomy samples, of which 16 pairs were macrochromosomes and and evolutionary relationships. In addition, it provides 12 pairs were microchromosomes. The chromosomes useful basic information for the conservation and breed- of some reptile groups are highly variable in terms of ing practices as well as analyses of chromosome evolu- size and morphology, and are characterized by bimodal tion of this species. or asymmetric karyotypes composed of macrochromo- somes and microchromosomes (Pinthong et al. 2013, Materials and methods Phimphan et al. 2013, Siripiyasing et al. 2013). The types of chromosomes of L. vittigera were 16 The L. vittigera samples were obtained from Surin metacentric macrochromosomes and 12 microchromo- Province, northeast Thailand. The lizards, five males somes (Table 2). This is in agreement with previous and five females, were transferred to a laboratory and reports of cytogenetic information of lizards in subfam- were kept under standard conditions for seven days prior ily Lygosominae which show 14–16 macrocrhomosomes to experimentation. Preparation of their chromosome and 12–16 microchromosomes (Makino and Momma was conducted by squash technique from bone marrow 1949, Bhatnagar 1959, Becak et al. 1972, King 1973a, and testis (Kasiroek et al. 2017). The chromosomes were b, Hardy 1979, De Smet 1981, Kupriyanova 1973, 1974, stained with 10% Giemsa’s for 30 min (Chooseangjaew 1984, Adegoke 1985, Colus and Ferrari 1988, Adegoke et al. 2017) and identified for NORs by Ag-NOR stain- and Ejere 1991, Donnellan 1991a, Eremchenko et al. ing (Howell and Black 1980, Sangpakdee et al. 2017). 1992, Ota and Lue 1994, Ota et al. 1991, 1996, Norris The length of short arm (Ls) and long arm (Ll) chromo- 2003). some were measured and calculated for the length of No sex chromosome heteromorphism was observed. total arm chromosome (LT, LT=Ls+Ll). Relative length It is a consistent characteristic from previous reports of (RL) and centromeric index (CI) were also calculated. the cytogenetics of lizards in family Scincidae (Makino CI was estimated to classify the types of chromosomes and Momma 1949, Bhatnagar 1959, Dallai and Tal- according to Chaiyasut (1989) and Chooseangjaew et al. luri 1969, Deweese and Wright 1970, Becak et al. 1972, (2017). All parameters were used in karyotyping and King 1973a, b, Wright 1973, Hardy 1979, De Smet 1981, idiograming. Kupriyanova 1973, 1974, 1984, 1986, Branch 1980, Adegoke 1985, Cano et al. 1985, Capriglione 1987, Co- lus and Ferrari 1988, Guo and Dong 1988, Caputo and 2017 Cytogenetic of Skink (Reptilia, Scincidae) from Thailand: II: Chromosome Analyses of Stripe Tree Skink (Lipinia vittigera) 85 Table 1. Review of cytogenetic reports of lizard in family Scincidae (Reptilia, Squamata). Genera TSN SSN 2n mac mi NORs (p) Reference Lygosominae Ablepharus 9 2 28–30 14–16bi 14 4 Kupriyanova (1973), Eremchenko et al. (1992) Asymblepharus 8 1 30 12bi 18 4 Kupriyanova (1984), Eremchenko et al. (1992) Bassiana 3 1 30 16bi 14 ̶ King (1973b) Brasiliscincus 3 1 32 18bi 14 2 Colus and Ferrari (1988) Corucia 1 1 32 16bi+2t 14 2 Donnellan (1991a) Cryptoblepharus 53 2 28 10bi 18 ̶ Makino and Momma (1949) Ctenotus 101 2 30 16bi 14 ̶ King (1973b) Cyclodina 11 1 30 14bi+XY 16 ̶ Hardy (1979) Cyclodomorphus 8 2 32 16bi+2t 14 2 Donnellan (1991a) Dasia 9 1 30 ̶ ̶ ̶ Ota et al. (1996) Egernia 17 17 32 16bi+2t 14 2 (mi), 6, 8 King (1973b), Donnellan (1991a) Eulamprus 15 1 30 16bi 14 ̶ King (1973b) Eutropis 31 7 26–32 16–28 4–14 2, 4 De Smet (1981), Donnellan (1991a), Eremchenko (10–24bi, 0–6t) et al. (1992), Ota et al. (1996) Hemispheridion 1 1 32 16bi+2t 14 2 (mi) Donnellan (1991a) Lampropholis 11 9 28–30 12–16bi+2–6t 10–12 ̶ Donnellan (1991b) Leiolopisma 5 1 30 14bi 16 ̶ Hardy (1979) Lipinia 27 1 28 16bi 12 1, 5 Present study Lygosoma 31 1 24 12 12 ̶ Bhatnagar (1959) Mabuya 8 1 30 16bi 14 ̶ Becak et al. (1972) Mochlus 13 2 30 18 12 ̶ De Smet (1981), Adegoke (1985) Oligosoma 40 2 30 12 18 ̶ Norris (2003) Pseudoemoia 6 4 30 14–16bi+0–2t 14 ̶ King (1973b), Hardy (1979) Psychosaura 2 1 32 18bi 14 2 Colus and Ferrari (1988) Saproscincus 11 4 30 14bi+4t 12 ̶ Donnellan (1991b) Sphenomorphus 120 2 28–30 10–14bi 16–18 2 Makino and Momma (1949), Ota and Lue (1994) Tiliqua 7 6 32 18–26 6–14 2 (mi) King (1973a, b), De Smet (1981), Donnellan (16–24bi, 0–2t) (1991a) Trachylepis 78 10 28–34 16–26 0–22 2, 4 Kupriyanova (1974), De Smet (1981), Colus and (10–28bi, 0–6t) Ferrari (1988), Adegoke and Ejere (1991), Erem- chenko et al.
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