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66. Karyoty Pes O F Two Species O F Frogs from Taiwan, Rana Sauteri

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66. Karyoty Pes O F Two Species O F Frogs from Taiwan, Rana Sauteri No. 9] Proc. Japan Acad., 58, Ser. B (1982) 279 66. Karyoty pes o f Two Species of Frogs from Taiwan, Rana sauteri Boulenger and Rana latouchii Boulenger By Takeshi SET0,*) Yasuaki UTSUNOMIYA,** and Taeko UTSUNOMIYA** (Communicatedby Sajiro MAKINO,M. J. A., Nov. 12, 1982) The amphibian fauna of the Ryukyu Islands and Taiwan (For- mosa) is quite interesting to ecological, systematic and cytogenetic studies, because of the geographical feature which forms a chain of archipelago fringing the eastern border of the Asiatic continent. Thus the unique distribution of anuran and urodelan species has been known in this region. Chromosome studies of anurans indigenous to the Ryukyu Islands and Taiwan have been carried out by Kuramoto (1972, 1980) , Kura- moto and Teshima (1970), Kuramoto et al. (1973), and Lin and Huang (1979). The chromosomes of the ranids from Taiwan have, however, remained unknown in some species. The present paper deals with a chromosome study of two species of the genus Rana, R. sauteri and R. latouchii, endemic to Taiwan, since the chromosome study has not well been done. Materials and methods. A male of Rana sauteri, endemic to Taiwan and chromosomally untouched, and a female of R. latouchii, distributed in both Taiwan and the People's Republic of China, provided material for this study. The two individuals under study were procured in Chi-Tou, southern district of Nan Tow Hsien, Taiwan. The fully grown frogs were intraperitoneally injected with 0.1 ml of colchicine solution (2 mg/ml; SIGMA) per gram of body weight. Following 48 hr of the treatment, the animals were sacrificed and entire digestive tracts were taken out. Somatic chromosomes were prepared from the gut epithelium by the method of Kezer and Sessions (1979). Chromosomes were stained according to the con- ventional Giemsa technique. The C-band technique (Sumner 1972) was applied in order to identify the individual chromosomes in R. tat ouchii. Observations and remarks. The diploid number of R. sauteri and R. latouchii was determined as 26, common to that of the ma- Department of Biology, Faculty of Education, Shimane University, Matsue, Japan 690. **' Faculty of Applied Biological Science, Hiroshima University, Fukuyama, Japan 720. 280 T. SETO, Y. UTSUNOMIYA, and T. UTSUNOMIYA [Vol. 58(B), Figs. 1-3. Karyotypes of Rana sauteri (1), and Rana latouchii (2); each showing 26 chromosomes. 3 ; C-banding karyotype of Rana latouchii 2. Bar indicates 10 jim. j ority of Rana species (Morescalchi 1973). Karyotype analysis was made with 6 well-spread metaphasic cells in each species. Karyotype of Rana sauteri (Fig. 1) : The karyotype consisted of 13 pairs of somatic chromosomes containing 5 pairs of large-sized elements and 8 pairs of small-sized homologues. The chromosomes of the large group were characterized by metacentric (nos. 1, 4 and No. 9] Karyotypes of Ranids from Taiwan 281 5) and submetacentric structure (nos. 2 and 3) , as demonstrated in other Rana species so far examined. The small chromosome group includes 3 metacentric (nos. 8, 11 and 12) , 3 submetacentric (nos. 7, 9 and 10) , and 2 subtelocentric pairs (nos. 6 and 13) . No. 6 is easily identified by a secondary constriction on the long arm. The morphology of this chromosome group of R. sauteri, especially of nos. 6 and 13, was dissimilar from that of the other Taiwan and Ryukyu ranids reported by Kuramoto (1972) , Kuramoto et al. (1973) , and Lin and Huang (1979). Boulenger (1909) described on the basis of a single reserved speci- men that R. sauteri showed striking resemblance in the external char- acters to R. japonica. However, our observation in a living specimen at the habitat is incompatible with Boulenger's view (1909) . In addi- tion, the present chromosomal findings supported our view on this respect. Karyotypes of R. japonica (Seto 1965) and R. longicrus (Kuramoto et al. 1973) , which are known to be closely related to R. japonica, were dissimilar to that of R. sauteri. With the larval characters such as bearing a sucking disk on the ventral side and abundance of the dental formula (3 :4 + 4/7 :1 + 1), R. sauteri is thought to be an intermediate type of the brown-frog group and the genus Amalops. Further karyological investigations have been needed to prove the above feature. Karyotype of Rana latouchii (Fig. 2) : A brief study on the chromosomes of this species was given by Kuramoto (1980), though more precise knowledge on the chromosomal details has been needed. The chromosome morphology of the large-sized group (nos. 1-5) was quite similar to that of R. sauteri and other common members of the genus Rana. In contrast, the small-sized group (nos. 6-13) was specific in the chromosome morphology ; this group comprised 2 metacentric pairs (nos. 7 and 12) and 6 pairs of submetacentric elements (nos. 6, 8, 9,10,11 and 13). There are no subtelocentric and telocentric element in the karyotype of this form. Nos. 10 and 11 were frequently charac- terized by a secondary constriction on their long arms. The present observation in less-condensed metaphasic chromosomes revealed a considerable dissimilarity to the karyotype presented by Kuramoto (1980) for this species, particularly for the small chromosome group (nos. 6-13). According to Pope (1931) and Utsunomiya (un- published), R. latouchii appeared to be a particular species in the genus Rana by having several external features in both larval and adult individuals. A comparison of karyotypes was made between typical brown frogs occurring in Japan such as R. chensinensis, R. ornativentris (formerly R. t. temporaria and R. t. ornativentris, re- spectively), R. japonica, R. okinavana (Kobayashi 1962: Seto 1965; 282 T. SETO,Y. UTSUNOMIYA,and T. UTSUNOMIYA [Vol. 58(B), Kuramoto 1972) and R. latouchii, and we noted the existence of an obvious karyological dissimilarity in the latter species. C-stained chromosomes were analysed in R. latouchii (Fig. 3). The C-banding patterns of this species revealed the presence of centromeric heterochromatin in all chromosomal pairs. In addition, the long arm of no. 2 and the short arms of nos. 11 and 13 had telomeric heterochromatin. The telomeric and interstitial constitu- tive heterochromatin appeared heterogeneously in each species of Ranidae aside from the main centric heterochromatin (Schmid 1978). The present observation, however, was unable to provide any signifi- cant evidence to discuss the interspecific heterogeneity of the telomeric C-band pattern. Summary. By means of the squash technique of the intestinal epithelium, the karyotypes of Rana sauteri and Rana latouchii from Taiwan are investigated. These two species, like many other mem- bers of the genus Rana, show 26 chromosomes which consist of 5 large-sized pairs and 8 small-sized pairs. The chromosome mor- phology of the large pairs is similar to that of the other common Rana species, leaving the small pairs which show a characteristic feature in respective species. The morphology of 2 small pairs of R. sauteri is different from that of any other Taiwan and Japanese ranids. Based on genotypic and phenotypic evidence we suppose that R, sauteri could be an intermediate type of the brown-frog group and the genus Amalops. The chromosomes of Rana latouchii are also dissimilar to those of typical brown frogs in Japan. Evidence presented here corresponds to the results of the external morphology in both larval and adult individuals of this species. Acknowledgement. The authors are deeply indebted to Pro- fessor Emeritus Dr. Saj iro Makino, M. J. A., for improvement of the manuscript with invaluable advice. References Boulenger, G. A. (1909) : Ann. Mag. Nat. Hist., ser. 8, 4, 493. Kezer, J., and Sessions, S. K. (1979) : Chromosoma (Berl.), 71, 65-80. Kobayashi, M. (1962) : J. Sci. Hiroshima Univ., Ser. B, Div. 1, 20, 157-179. Kuramoto, M. (1972) : Cytologia, 25, 547-559. (1980) : Experientia, 36, 826-827. Kuramoto, M., and Teshima, R. (1970) : Bull. Fukuoka Univ. Educ., III, 20, 99-104. Kuramato, M., Furuya, E., Takegami, M., and Yano, K. (1973) : ibid., 23, 67-78. Lin, F.-Y., and Huang, C.-C. (1979) : Proc. Natl. Sci. Counc. ROC, 3, 429-436. Morescalchi, A. (1973) : Cytotaxonomy and Vertebrate Evolution (ed. Chiarelli, A. B., and Capanna, E.). Academic Press, N. Y., pp. 233-348. Pope, C. H. (1931) : Bull. Am. Mus. Nat. Hist., 61, Art. VIII, 531. Schmid, M. (1978) : Chromosoma (Berl.), 68, 131-148. Seto, T. (1965) : Cytologia, 30, 437-446. Sumner, A. T. (1972) : Exp. Cell Res., 75, 304-306..
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