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(2) Journal of Hokkaido University of Education (Section HB) Vol. 33, No. 1 September, 1982. »?a^ir^NSS (lt2^B) ^33^ ^1-?- Bgffi57^9J1. Chromosomes of New Parthenogenetic Races of Callirhopalus. bifasciatus Roelofs and Callirkopalus obesus Roelofs. (Curculionidae : Coleoptera). Yasushi TAKENOUCHI Biological Laboratory, Sapporo College, Hokkaido University of Education,. Sapporo 064. ^ ^: x^'ij y^A^^^^y^^^o^rL^-^-^^^a^^^-fa^. wMCT^^Lt^m^w^ Abstract The chromosomes of the curculionid weevil species Callirhopalus (= Psetidocneorhimis} bifasciatus Roelofs and Callirhopaliis obesus Roelofs were studied in early cleavage nuclei using squash preparations. This revealed that the former has three parthenogenetic races, 3x, 4x, and a rare 5x, and that the latter has four parthenogenetic races, 2x, 3x, 4x, and 6x, each with. distinct geographical distributions. Callirhopalus (= Pseudocneorkinus) bifasciatus Roelofs and Callirhopalus obesus Roelofs are weevil species belonging to the tribe Callirhopalini, subfamily Eremninae. I have previously reported that in several localities of Hokkaido and Aomori City and Setagaya, Tokyo, JapanHondo, C. bifasciatits had two parthenogenetic races, the one a tetraploid carrying 44 chromo-. somes, and the other a pentaploid with 55 chromosomes (Takenouchi, 1957, 1959, 1981a.c). No males have so far been recorded in Japan. On the other hand, in August of 1974, I visited the British Museum (Natural Science and History), London, and with the kind help of Dr. R. Thomson we found a couple of male and female specimens among the many specimens of the. species which were kept in the Kritish Museum. The male and the female specimens had been collected together in Chou-Shan Island, China. No cytological studies have been carried out but, of course, they are no doubt members of a diploid bisexual race when compared to the other bisexual weevil species that have hitherto been studied cytologically (Takenouchi, 1972,. 1981b). Callirhopalus obesus Roelofs is a more rare weevil species than C. bifasciatns and I have. (D.
(3) 2 Yasushi TAKENOUCHI. reported that in Hokkaido the species has two parthenogenetic races, 4x and 6x (Takenouchi, 1972, 1976, 1981c). This time I have had a chance to study the chromosomes of different parthenogenetic races of both species. This paper reports the details.. Materials and Methods One live Callirhopalus (= Pseudocneorhinus} bifasciatus female and a single Callirhopaltts obesus female were collected by Dr. H. Sasaji from Mt. Taishiyama, Katsuyama City, Fukui Prefecture, in mid-May, 1978, and were kindly sent to me for cytological study. Four C.. bifasciatus females collected by Miss K. Takada and Miss M. M^urayama from Gunji (A), Saigoshi, Shingo-mura, Sannohe-gun, Aomori Prefecture, in late May, 1978, and two C. obesus. females collected by Miss K. Takada from the same address, (Gunji B), in late-July, comprised the material for the study. The distance between Gunji (A) and Gunji (B) is approximately 800 m. Every specimen was reared in a petri-dish and given a piece of a food plant as usual. (Takenouchi, 1959). The eggs were then squashed at various hourly intervals after oviposition. Squash slides were made according to Smith's method (Smith, 1943) and stained by basic fuchsin and methyl-green solution as usual (Smith and Takenouchi, 1959). Photomicrographs were taken according to Professor Shimakura's movifocal method using Olympus M.6.. Observations Callirhopalus bifasciatus Roelofs Tetraploid Parthenogenetic race A single female from Mt. Taishiyama, Fukui Prefecture, showed without exception 44 chromosomes in early cleavage mitotic metaphases (Fig. 1). Most of the elements were of meta- or submetacentric nature and none similar to sex chromosomes were observed. A single. female collected from Gunji (A), Aomori Prefecture, also shows 44 elements in the early cleavage mitotic nucleus at metaphase (Fig. 2). Triploid parthenogenetic race Among the four females from Gunji (B) one showed exactly 33 chromosomes in the early cleavage mitotic prophase (Fig. 3). Callirhopalus obesus Roelofs Diploid parthenogenetic race A single female from Mt. Taishiyama, Fukui Prefecture, showed 22 chromosomes of. univalent nature in both early cleavage mitotic metaphase and anaphase (Figs. 4 and 5). These were meta- or submetacentric chromosomes. No sex chromosomes were detectable. Further, no sperms were observable in her seminal recetacle.. Triploid parthenogenetic race. (2).
(4) New Parthenogenetic Races of Two Japanese Weevils. Figs. 1—3. Chromosomes of Callirhopahis bifasciatus. Fig. 1. Metaphase plate of early cleavage in a Mt. Taishiyama female (4x = 44). Fig. 2. Metaphase plate of early cleavage in a Gunji (A) female (4x == 44). Fig. 3. Late prophase of early cleavage in a Gunji (B) female (3x = 33). Figs. 4—6. Chromosomes of Callirhopahts obesus. Fig. 4. Metaphase plate of early cleavage in a Mt. Taishiyama female (2n = 2x = 22). Fig. 5. Anaphase plate of early cleavage in the same specimen. Fig. 6. Metaphase plate of early cleavage in a Gunji (B) female (3x = 33). X 3,600.. (3).
(5) 4 Yasushi TAKENOUCHI. Two Gunji (B) females showed without exception exactly 33 chromosomes in early cleavage mitotic metaphase (Fig. 6). Most of the elements were of meta- or submetacentri'c nature.. Discussion. To date a comparatively large number of curculionid weevil species have been studied cytologically. Among them 61 species or races are parthenogenetic and are members of 6 closely allied subfamilies, Brachydermae, Otiorrhynchinae, Eremninae, Leptopinae, Cylindror-. rhninae, and Curculioninae. The cytological studies on the parthenogenetic weevils have proved that the phenomenon of parthenogenesis generally occurs in connection with polyploidy (Suomalainen, 1958, 1969 ; Suomalinen, Saura, and Lokki, 1976 ; Takenouchi, 1972, 1976, 1978, 1981a). Excluding the European Polydrosus mollis and Japanese Scepticus insnlans and Catapionus gracilicornis with 22 chromosomes (2n = 2x = 22), 34 triploids have 33 chromosomes (3x), 16 tetraploids have 44 chromosomes (4x), 5 pentaploids have 55 chromosomes (5x), and 2 hexaploids (6x) found in Japan have 65, 66, and 68 chromosomes. These biotypes are characterized by the basic number of 11. 3 exceptional triploid species, Liophloeus tessulatm, Listroderes costirostris and Eusomits ovulum, have 30 chromosomes suggesting a basic number. of 10 (Takenouchi, 1969 ; Mikulska, 1953 ; Sanderson, 1973 ; Suomalainen, 1955). All the parthenogenetic weevils so far studied cytologically are of an apomictic or thelytokous type and no arrhenotokous type is found in the family. In these apomictic weevils the eggs undergo only one maturation division and the chromosomes divide equationally ; consequently, no. reduction takes place. Callirhopalus bifasciatus, studied here, had chromosomally different races with 33 chromosomes in addition to a common tetraploid parthenogenetic race (44 chromosomes) and a rare pentaploid race (55 chromosomes). The chromosome number 33. clearly means that the race is a triploid parthenogenetic one. Callirhopahis obesus of Gunji (B), Aomori Prefecture has 33 chromosomes. The race is also a triploid parthenogenetic race.. Further, the female in Mt. Taishiyama, Fukui Prefecture, with 22 chromosomes is diploid parthenogenetic one (2n =- 2x = 22). The specimens of both species are all females and they have no sperm in their seminal receptacles. Further, so far, no males of either species have been recorded in Japan.. Acknowledgements I wish to express my appreciation to Dr. H. Sasaji, Fukui University, Miss K. Takada, and M:iss M. Yamamura, for their co-operation in collecting the material used in the study. My thanks are also due to Dr. K. Morimoto, Department of Entomology, Faculty of Agriculture,. Kyfishu University, Fukuoka, for identification of the species.. 4).
(6) New Parthenogenetic Races of Two Japanese Weevils. References. Mikulska, I. 1953. The chromosomes of the parthenogenetic and thelytokian weevil Eusomi(s ovnlum germ. (Curculionidae, Coleoptera). Bull. Acad. pol. Sci. Ser. B : (II) 1951 : 293-307. Sanderson, A. R. 1973. The cytology of the parthenogenetic Australian weevil Listroderes costirostris Schonh. Trans. Roy. Soc. Edin. 69 : 71—89. Smith, S. G. 1943. Techniques for the study of insect chromosomes. Can. Entomol. 75 : 21—34. Smith, S. G. and Takenouchi, Y. 1969. Chromosomal polymorphism in Pissodes weevils : Further on incompatibility in Pissodes terminalis. Canad. J. Genet. Cytol. 11 : 761—782. Suomalainen, E. 1955. A further instance of geographical parthenogenesis and polyploidy in the weevils, Curculionidae. Arch Soc. Zool. Bot. Fenn, 'Vanamo' (Suppl.). 9 : 350—354. Suomalainen, E. 1958. On polyploidy in animals. Proc. Finn. Acad. Sci. Lett. 1958 : 1—15. Suomalainen, E. 1969. Evolution in parthenogenetic Curculionidae. In Evolutionary Bi.ology, Vol. 3. Edited by T. Dobzhansky, M. K. Hecht and W. C. Steere. Appleton-Century-Crofts, New York. pp. 261-296. Suomalainen, E., Saura, A. & Lokki, J. 1976. Evolution of parthenogenetic insects. Evolutionary Biology, 9 (Hecht. Steere, and Wallace, eds.). pp. 209-257. Plenum Press. Takenouchi, Y. 1957. Polyploidy in some parthenogenetic weevils (A preliminary note). Annot. Zool. Japan. 30 : 38-41. Takenouchi, Y. 1959. Some oecological observations of three species of curculionid weevils, with special reference to parthenogenetic reproduction. J. Hokkaido Gakugei Univ., Section B, 10 : 297—340. Takenouchi, Y. 1969. A further study on the chromosomes of the parthenogenetic weevil, Listroderes costirostris Schonherr, from Japan. Cytologia 34 : 360—368. Takenouchi, Y. 1972. Chromosome numbers of Japanese weevils of Curculionoidea (Coleoptera). Kontyfl 40 : 123-132. Takenouchi, Y. 1976. On the chromosomes of parthenogenetic curculionid weevils in Japan. Proc. Jap. Acad. 52 : 126-129. Takenouchi, Y. 1978. A chromosome study of the parthenogenetic rice water weevil, Lissorhoptms o-iyzo-. philns Kuschel (Coleoptera : Curculionidae), in Japan. Experientia 34 : 444—445. Takenouchi, Y. 1981a. A further chromosome survey on a parthenogenetic weevil, Callirhopalus bifasciatns. Roelofs (Coleoptera : Curculionidae). Genetica 55 : 141—145. Takenouchi, Y. 1981b. A diploid parthenogenetic race of the weevil species Catapionns gracilicornis from Japan (Coleoptera : Curculionidae). Entomol. Genera. 6 : 367—369. Takenouchi, Y. 1981c. Chromosome numbers of Japanese weevils of Curculionoidea (Coleoptera) II. Seibutsu-kyozai 16 : 155—170.. (5).
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