Comparative chromosomal studies in Rhinolophus formosae and R. luctus from China and Vietnam: elevation of R. l. lanosus to species rank Author Volleth, Marianne Department of Human Genetics, Otto von Guericke University, Leipziger Strasse 44, 39120 Magdeburg, Germany Author Son, Nguyen Truong Department of Vertebrate Zoology, Institute of Ecology and Biological Resources and Graduate University of Sciences and Technology, Vietnam Academy of Sciences and Technology, 18 Hoang Quoc Viet, Cau Giay, Hanoi, Vietnam Author Wu, Yi College of Life Sciences, Guangzhou University, 230 Wei Huan Xi Road, Guangzhou, 510006, China Author Li, Yuchun Marine College, Shandong University, Weihai, 264209, China Author Yu, Wenhua College of Life Sciences, Guangzhou University, 230 Wei Huan Xi Road, Guangzhou, 510006, China Author Lin, Liang-Kong Laboratory of Wildlife Ecology, Department of Biology, Tunghai University, No. 1727, Sec. 4. Taiwan Boulevard, Xitun District, Taichung 40704, Taiwan Author Arai, Satoru Infectious Disease Surveillance Center, National Institute of Infectious Diseases, Toyama 1 - 23 - 1, Shinjuku, Tokyo 162 - 8640, Japan Author Trifonov, Vladimir Institute of Human Genetics, Jena University Hospital, Friedrich-Schiller University, Kollegiengasse 10, 07743 Jena, Germany Author Liehr, Thomas Institute of Molecular and Cellular Biology, Siberian Branch of RAS, 8 Ac. Lavrentieva Avenue, Novosibirsk 630090, Russia Author Harada, Masashi Laboratory Animal Center, Graduate School of Medicine, Osaka City University, Abeno, Osaka 545 - 8585, Japan Corresponding author: E-mail: Marianne. Volleth @ med. ovgu. de olleth@med.ovgu.de text Acta Chiropterologica 2017 2017-06-01 19 1 41 50 journal article 21475 10.3161/15081109ACC2017.19.1.003 4e1384ef-ef30-4f65-95df-aa5eb3f09571 1733-5329 3944816 Rhinolophus formosae Two males and one female from Taiwan were studied cytogenetically. All specimens showed a karyotype with 2 n = 52 and a FNa = 60 ( Fig. 6 ). There were five bi-armed and 19 acrocentric autosomal pairs. In addition, the smallest, dot-like pair no. 25 is likely also bi-armed. Due to the minuteness of this element, the metacentric condition was visible only in a small percentage of metaphase spreads. Therefore, this chromosome was counted as one arm only for the FNa. The medium-sized X chromosome FIG. 2. Metaphase spreads of R . cf. luctoides after CBG-staining (A) and silver-staining (B), respectively. The arrows in B indicate the active NORs on pair 8 FIG. 3. FISH examples showing the hybridization patterns of A. stoliczkanus (AST) chromosome-specific painting probes onto metaphase spreads of R. cf. luctoides (A, D), R. lanosus (B, E) and R. formosae (C, F). A–C — hybridization signals of AST1 (red) and AST8 (green) meet in chromosome 5 in R . cf. luctoides (A) and R. lanosus (B) but not in R. formosae (C). In B and C AST X (yellow) was applied in addition. D, E — hybridization signals of AST2 (green) and AST5 (red) meet in chromosome 3 of R . cf. luctoides (D), but not in R. lanosus (E), where AST2 (green) is found in combination with AST1 (red) on chromosome 4 instead. F — chromosome 1 of R. formosae showed homology to AST1 (green) and AST12 (red) is submetacentric and the dot-like Y chromosome is probably bi-armed. Concerning the amount of centromeric heterochromatin, the acrocentric pairs differed from the bi-armed ones. In contrast to the faint staining of centromeric regions of bi-armed pairs 1 to 5 and 25, the acrocentric chromosomes showed dark stained pericentromeric regions after CBG-banding, which were also GTG-, QFQ- and DAPI-positive ( Fig. 7 A–B). The amount of centromeric heterochromatin of the X chromosome was similar to that of other Rhinolophus species and not enlarged as in R. cf. luctoides and R. lanosus . The Y chromosome of R. formosae was hardly distinguishable from the autosomal pair 25 after CBG-banding and showed no clear centromeric heterochromatin ( Fig. 7A ). The secondary constriction of chromosomal pair 18 (homologous to MMY21) was shown to bear active NORs by silver-staining ( Fig. 7C ). Analysis of 20 metaphase spreads of one male specimen revealed a frequency of 2.0 NORs per cell. The complete set of AST painting probes and some selected MMY probes (MMY8, 14, 23) were applied on R. formosae . The results are given on the G-banded karyogram ( Fig. 6 ) and examples of FISH experiments are shown in Fig. 3C and 3F . Of the chromosomal pairs 1 to 5, only two, i.e., 4 and 5, show the same combination of chromosomal arms as found in R. luctoides , R. lanosus and R. morio . Pairs 1 to 3 show a unique combination hitherto found in no other rhinolophid species.