Genetic and Morphological Variability in South American Rodent Oecomys (Sigmodontinae, Rodentia): Evidence for a Complex of Species
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c Indian Academy of Sciences RESEARCH ARTICLE Genetic and morphological variability in South American rodent Oecomys (Sigmodontinae, Rodentia): evidence for a complex of species C. C. ROSA1, T. FLORES2, J. C. PIECZARKA1, R.V.ROSSI3, M.I.C.SAMPAIO2, J. D. RISSINO1, P. J. S. AMARAL1 and C. Y. NAGAMACHI1∗ 1Laboratório de Citogenética, Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém 66.075-900, Brazil 2Laboratório de Genética e Biologia Molecular, Universidade Federal do Pará, Campus Universitário de Bragança, Pará 68.370-000, Brazil 3Departamento de Biologia e Zoologia, Instituto de Biociências, Universidade Federal do Mato Grosso 79.070-900, Brazil Abstract The rodent genus Oecomys (Sigmodontinae) comprises ∼16 species that inhabit tropical and subtropical forests in Central America and South America. In this study specimens of Oecomys paricola Thomas, 1904 from Belém and Marajó island, northern Brazil, were investigated using cytogenetic, molecular and morphological analyses. Three karyotypes were found, two from Belém (2n = 68, fundamental number (FN) = 72 and 2n = 70, FN = 76) and a third from Marajó island (2n = 70, FN = 72). No molecular or morphological differences were found between the individuals with differing cytotypes from Belém, but differences were evident between the individuals from Belém and Marajó island. Specimens from Belém city region may represent two cryptic species because two different karyotypes are present in the absence of significant differences in morphology and molecular characteristics. The Marajó island and Belém populations may represent distinct species that have been separated for some time, and are in the process of morphological and molecular differentiation as a consequence of reproductive isolation at the geographic and chromosomal levels. Thus, the results suggest that O. paricola may be a complex of species. [Rosa C. C., Flores T., Pieczarka J. C., Rossi R. V., Sampaio M. I. C., Rissino J. D., Amaral P. J. S. and Nagamachi C. Y. 2012 Genetic and morphological variability in South American rodent Oecomys (Sigmodontinae, Rodentia): evidence for a complex of species. J. Genet. 91, 265–277] Introduction first described as a subgenus of Oryzomys (Thomas 1906), and later accepted as a full genus (Thomas 1909). In the Rodents are important members of most faunal communi- only taxonomic review of Oecomys, Hershkovitz (1960)fol- ties, as they are cosmopolitan and native to most terrestrial lowed Thomas (1906) in considering the group as a sub- areas. Wide variation in their morphology, and diversity of genus of Oryzomys, but others disagreed with this taxonomic their habitats, climatic tolerance and food sources make them arrangement. Only after the study of Musser and Carleton the most numerous and evolutionarily successful among the (1993) was the generic status of the genus Oecomys largely orders of mammals (Emmons and Feer 1997). accepted. Recent phylogenetic studies including species of The subfamily Sigmodontinae, which includes most of Oecomys have confirmed the monophyly of this genus the species of South American rodents, occurs only in (Patton and Da Silva 1995; Smith and Patton 1993; Patton Americas. It includes 386 species in 81 genera and nine et al. 2000; Andrade and Bonvicino 2003; Weksler 2003, tribes: Abrotrichini, Akodontini, Ichthyomyini, Oryzomyini, 2006), which now includes 16 valid species (Musser and Phyllotini, Reithrodontini, Sigmodontini, Thomasomyini Carleton 2005; Carleton et al. 2009). and Wiedomyini (Reig 1980; Smith and Patton 1999; Musser Genus Oecomys occurs in tropical and subtropical rain- and Carleton 2005; Weksler et al. 2006; D’Elía et al. forests in Central America and South America. In the 2007). The tribe Oryzomyini comprises 28 genera (Weksler Brazilian Amazon region there are nine species, of which et al. 2006) and includes the genus Oecomys,whichwas only five have had their karyotype described (Andrade and Bonvicino 2003; Langguth et al. 2005). Cytogenetic stud- ∗ For correspondence. E-mail: [email protected]. ies of Oecomys show that the diploid number ranges from Keywords. cytogenetics; chromosome; speciation; cryptic species; rodent; Oecomys paricola. Journal of Genetics, Vol. 91, No. 3, December 2012 265 C. C. Rosa et al. Table 1. Chromosomal characterization of the genus Oecomys. 58 to 86 (table 1), suggesting a high level of chromosomal rearrangement in this species (Gardner and Patton 1976; Species 2n FN Reference Patton et al. 2000). This karyotypic variability demonstrates the importance of chromosomal studies in taxonomic identi- Oecomys sp. 86 98 Gardner and Patton (1976) O. bicolor 80 134–136 Gardner and Patton (1976) fication of the species (Langguth et al. 2005). O. bicolor 80 140 Patton et al. (2000) Oecomys paricola Thomas, 1904 is an Amazonic species O. roberti 80 114 Patton et al. (2000) with an incompletely defined distribution south of the O. roberti 82 106 Langguth et al. (2005) Amazon river and east of the Xingu river (Voss et al. 2001; O. superans 80 108 Gardner and Patton (1976) Musser and Carleton 2005; Rocha et al. 2011). No karyotype Oecomys sp. 72 90 Andrade and Bonvicino (2003) O. paricola 70 76 Present work information has been reported for this species. O. paricola O. paricola 68 72 Present work has predominantly grey-based ventral fur, except for chin and O. paricola 70 72 Present work throat, which are usually selfcoloured (all pale), sometimes O. concolor 60 62 Patton et al. (2000) with a ventral self-coloured midline extending caudally to O. bahiensis 60 62 Langguth et al. (2005) the groin (Voss et al. 2001). The tail is uniformly dark, with O. trinitatis 58 96 Patton et al. (2000) 2n, diploid number; FN, fundamental number. Table 2. Specimens analysed, with localities, specimen preparation type, collection and GenBank number. Material Locality Skin Skull Fluid Cyt b Kar. Collection number GenBank number Barcarena (01◦30S48◦40W) X X BAR006 X X X BAR013 JF759675 X X X BAR017 JF759676 X X X BAR023 JF759677 X X X BAR029 JF759678 Belém, Parque Ambiental de Belém X X MPEG2477 (Utinga) (01◦27S48◦29W) X X MPEG2584 X X MPEG2593 X X MPEG2605 X MPEG2610 X X MPEG2614 X X MPEG2615 X MPEG2632 X MPEG2635 X X MPEG2647 X X X MPEG38659 JF759679 X X X MPEG38664 JF759680 X X X X MPEG39699 JF759681 X X X X MPEG39701 JF759682 X X X MPEG39703 X X X X MPEG39705 JF759683 X X X MPEG39708 JF759684 X X MPEG39709 X MPEG40732 BR-010, km 87–97 (01◦40S47◦47W) X X MG8382 X X MG8438 Marajó island, Chaves, X X X MPEG40842 JF759668 Fazenda Tauarí (00◦39S50◦11W) X X X MPEG40843 JF759669 X X X MPEG40844 JF759671 X X X MPEG40845 JF759670 X X X MPEG40846 X X X MPEG40848 JF759672 X X X MPEG40849 JF759673 X X MPEG40850 X X X X MPEG40851 JF759674 Kar., karyotype; MPEG, Museu Paraense Emilio Goeldi; BAR, specimens from Universidade Federal do Pará (Barcarena project) X, material available. 266 Journal of Genetics, Vol. 91, No. 3, December 2012 Genetic and morphological variability in Oecomys a terminal tuft of hairs 6–10 mm long. Cranially, O. pari- region and Marajó island, in Pará, Brazil. This is the first cola has a primitive pattern of carotid arterial supply, sep- description of the karyotype of the species, and provides new arated accessory oval and buccinator–masticatory foramina; information on molecular and morphologic variation in the the subsquamosal fenestra are absent. species. The identification of rodent species using morphological traits is very complex (Patton and Gardner 1972; Gardner and Emmons 1984), leading to uncertainties in the taxonomy of many genera of this group. Studies combining morpho- Material and methods logical, molecular and karyotype analysis have the potential Karyotype analysis to demonstrate occurrence of a greater diversity of Oecomys species, resulting in identification of new species, and reval- We karyotyped six specimens of O. paricola (table 2), of idation of previously described ones (Patton and Da Silva which four were collected in the Environmental Park in 1995; Smith and Patton 1999; Andrade and Bonvicino 2003; Belém (01◦27S, 48◦29W) and two (males) were collected at Weksler 2006). Molecular studies using the cytochrome b Marajó island (01◦00S, 49◦30W), in the region of the Ama- gene as a marker in phylogenetic studies of Sigmodonti- zon river mouth (figure 1). The specimens were collected nae rodents are particularly relevant (Smith and Patton 1991, using pitfall traps (buried 60 L buckets) connected by plas- 1993, 1999; Patton et al. 2000; Andrade and Bonvicino tic tapes, and with conventional traps including the Sherman 2003; D’Elía 2003; Miranda et al. 2007; D’Elía et al. 2008; trap and cages on the ground containing baits of peanuts, Catzeflis and Tilak 2009). This gene has been reported to flour and canned fish. Samples for metaphase chromosome have arrangements consistent with the species boundaries analysis were taken in the field using the bone marrow tech- based on classic taxonomic studies (Johns and Avise 1998; nique (Ford and Hamerton 1956). We also made laboratory Avise and Walker 1999), making it highly appropriate for culture of fibroblasts. biodiversity studies (Bradley and Baker 2001). The chromosome analysis involved conventional stain- In this study we undertook chromosomal, molecular and ing, G-banding and C-banding (Seabright 1971; Sumner morphological analysis of O. paricola from the Belém city 1972), and Ag-NOR staining (Howell and Black 1980). The Figure 1. Map with the collect locations. Journal of Genetics, Vol. 91, No. 3, December 2012 267 C. C. Rosa et al. karyotypes were organized by size and morphology of the bisection-reconnection (TBR) algorithm. Clade support was chromosome pairs. performed for both analysis using bootstrap with 1000 repli- cates. Estimates of evolutionary divergence of sequence pairs between and within Oecomys species and populations were Molecular analysis conducted using the Kimura-2 parameter method in MEGA4 (Kimura 1980;Tamuraet al. 2007).