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Studies on the Spermatogenesis of Some Indian Homoptera Part II 1955 225 Studies on the Spermatogenesis of Some Indian Homoptera Part II. Chromosomes of the Sugarcane Leaf Hopper Pyrilla perpusilla Wlk (Fulgoridae) S. R. Venkatasubba Rao Department of Zoology, Birla College of Science , Pilani, India Received January 6, 1955 Introduction Our knowledge of the cytology of Fulgoridae is very fragmentary. The only work in this field is that by Boring ('07) on the spermatogenesis of three species of Poeciloptera, viz., P. septentrionalis; P. pruinosa; P. bivittata and of Amphiscapa bivittata. She recorded in all these insects an XO (_??_) type of sex chromosome mechanism. In these species the X-chromosome occupies a position removed from the autosomal bivalents on the metaphase plate of primary spermatocyte division and it lags behind the autosomes during anaphase I. She also described in P. septentrionalis and P. pruinosa the occurence of in-chromosomes. The interesting XY mechanism in two species of this family Fulgoridae, belonging to the genus Eurybrachis has already been described in a previous paper (Rao, S. R. V. 1955). In view of the interesting cytology of this family a study of Indian Fulgorid bugs was undertaken. The present paper refers to the chromosomes of Pyrilla perpusilla Wlk, the Indian sugar-cane leaf hopper. Material and methods Specimens of Pyrilla perpusilla were collected from three different localities in India viz., Mandya (South India), Delhi (North India), and Pilani (Rajasthan, North India). Collections were made during several months of the year. The testes were fixed in Bouin's (Allen's modification) and Carnoy's fluids. After the usual method of dehydration through a graded series of alcohols and embedding in paraffin, sections were cut at a thickness of 10ƒÊ. Squash preparations were also made. Sections of material fixed in Bouin's were stained with Heidenhain's iron hematoxylin. Sections of Carnoy material were stained with the Feulgen technique. Observations The testes are orange red in colour. They occur between the 4th and 7th abdominal segments. The follicles of the testis are elongated and range from 6 to 8 in number. The spermatogonia are present at the apices of the follicles and are closely 226 S. R. V. Rao Cytologia 20 packed together. The diploid chromosome number (2n) is 27 and can be easily counted on the metaphase plates of spermatogonial divisions. Of these, four chromosomes (A1, A2, A3, A4, fig. 1) are distinct ly larger than the rest of the autosomes. The sex chromosomes, however, does not exhibit any stain ing peculiarity and cannot be identified at this stage. But it becomes ap parent, being posi tively heteropycno tic, at the beginn ing of meiosis (fig. 2). As meiosis pro ceeds, there is a considerable in crease in the vol ume of the nucleus and of the cell (compare figs. 2 and 3). During the "diffuse" stage Figs. 1-10. 1. Polar view of a spermatogonial metaphase show when the nucleus ing 27 chromosomes out of which there are 4 comparatively large has reached the chromosomes (A, to A,). The X-chromosome cannot be identified. maximum size, the 2. Beginning of meiosis. The X-chromosome is condensed over sex chromosome the rest of the autosomes. 3. The 'diffuse' nucleus. The X chromosome stands out clearly as a darkly staining body. The appears as a darkly nucleolus is lightly staining and is larger than that of X. 4. The staining body while diakinetic nucleus showing the dumb-bell shaped autosomal biva the autosomes can lents. In some the chiasma still persists. The X-chromosome apears not be distinguish as a round body. 5. Metaphase I (Polar view) showing the X, distributed away from the autosomal bivalent cluster. Two large ed (fig. 3). A autosomal bivalents are recognisable (A1, A2). 6. Same (side lightly staining view). The presence of sex-chromosome is marked out by its shape nucleolus is present which is different from the characteristic dumb-bell shaped auto at this stage, but somal bivalents. The X-chromosome is attached to only one pole (compare the spindle fibre on either side of the X-chromosome). disappears at the 7. Anaphase I (side view). The X-chromosome is lagging on the onset of diskinesis. spindle. 8. Telophase I. (side view). The X-chromosome is still on its poleward movement. 9 and 10. Metaphase II (polar views) During diskinesis showing 13 and 14 chromosomes. the autosomes are 1955 Studies on the Spermatogenesis of Some Indian Homoptera 227 dumb-bell shaped while the sex-chromosome is rounded body . The terminal isation of the chiasmata is not complete in all the bivalents (fig . 4). How ever, by the time metaphase I is reached, all the chiasmata become com pletely terminalised. On the metaphase plate whereas the autosomal bivalents form a cluster, the sex-chromosome occupies a position outside the autosome bivalents (fig. 5). Careful observation of the spindle fibres reveals that the sex-chromosome is attached to one pole only. This is further substantiated by the fact that while continuous fibres of a uniform thickness run from one pole to the other, in the case of the sex-chromosome only on one side there appears a thick fibre, while on the opposite side the fibre is very thin (fig. 6). The fine fibre is obviously a continuous one while the thicker fibre is the sex chromosomal fibre superimposed on the continuous fibre. In anaphase I (fig. 7), the autosomes are seen to be very near the pole while the sex-chromosome still occupies a position near the equator. In early telophase (fig. 8), the sex-chromosome has not yet reached the pole while the autosomes have clustered at the pole. The sex-chromosome occurs as a dis tinct darkly staining body outside the assemblage of the autosomes. An examination of the metaphase plates of second division shows that there are two types of secondary spermatocytes. In some there are 13 dyads while in other there are 13 dyads plus the sex-chromosome (figs. 9 and 10). Conclusions The present study reveals a close resemblance between the spermato genesis of P. perpusilla and that in the species described by Boring ('07). The metaphases of spermatogonial divisions in all the species described by Boring ('07) and also in Pyrilla perpusilla show the presence of a few chromosomes larger than the others. Secondly, the species described by Boring and the one described in this paper show the peculiar behaviour of the sex-chromosome occupying a position outside the group of autosomal bivalents in metaphase I, and its special quality of lagging behind the rest of the autosomes. But unlike, Boring's species there is no rn-chromosome in Pyrilla perpusilla. The above study throws interesting light on the taxonomy of Indian species of Pyrilla. Three species of Pyrilla have been described from India, P. perpusilla Wlk, P. pusana Dist and P. aberrans Kirby and were con sidered morphologically distinct by Distant ('06, '16), LeFroy ('09) and Quadri and Aziz ('50). Recent breeding experiments by Mukerji and Prased ('54) have shown that these three should be considered as varieties of a single species. My collection of animals coming from three different localities would lead me to the same conclusion. There were no cytological differences among them and it is perhaps correct to regard them as belonging to a single species. 228 S. R. V. Rao Cytologia 20 Summary 1. The diploid number in the male species is 27 (26 autosomes plus X-chromosome). 2. The X-chromosome is always outside the group of autosomes. A distinct tendency for lagging in anaphase I is noticed in the X-chromosome. 3. The presence of large autosomes (macrochromosomes) is recorded. 4. A brief discussion on the systamatics of Pyrilla is given. Acknowledgement The work was done under the guidence and encouragement of Prof. M. A. Moghe, Professor and Head of the Department of Zoology, Birla College of Science, Pilani, to whom I am indebted. I am also indebted to Prof. B. R. Seshachar, Professor and Head of the Department of Zoology, Central College, Bangalore, for help at different stages of my work. My thanks are also due to Dr. M. Puttarudraiah (Government Entomologist, Department of Agriculture, Bangalore) and Mr. B. R. Subba Rao (Division of Entomology, Indian Agricultural Research Institute, New Delhi), for their kind help in the collection of material. References Boring, A. M. 1907. A study of spermatogenesis in the Membracidae, Jassidae, Cercopidae, Fulgoridae with special reference to the behaviour of the odd chromosome. Jour. Exp. Zool. 4: 469-512. Distant, W. L. 1906. Fauna of British India, Rhynchota, 3. - 1916. Fauna of British India, Rhynchota, 6. LeFroy, H. M. 1909. Indian Insect life. Thacker, Spink, Co., London. Makino, S. 1951. An atlas of chromosome number in animals. Iowa Coll. Press. Mukerji, S and Prased, V. G. 1954. The present systematic position of the sugarcane leaf hopper-Pyrilla perpu-silla Wlk. Curr. Sci. 23: 193-194. Quadri, M. A. H, and Aziz, A. 1950. Biology, life-history and external and internal anatomy of Pyrilla perpu.silla Walker. Monographs. On Indian Insect types. (Aligaran, Muslim University Publication-Zoological series). Rao, S. R. V. 1955. Studies on the spermatogenesis of Indian Homoptera. I. Meiosis in two species of Eurybraehis (Fulgoridae). Chromosoma, 7: 170-180..
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