Cytogenetic report on Cordulegaster brevistigma and Watanabeopetalia atkinsoni1st June 2019101
Cytogenetic report onCordulegaster brevistigma and Watanabeopetalia atkinsoni (Odonata: Cordulegastridae, Chlorogomphidae)
Gurinder Kaur Walia1 & Sarabjit Singh Chahal
Department of Zoology and Environmental Sciences, Punjabi University, Patiala 147002, Punjab, India; 1 corresponding author, [email protected]
Received 26th October 2018; revised and accepted 8th April 2019
Abstract. Live adult male specimens of C. brevistigma and W. atkinsoni have been collected from Shimla, Himachal Pradesh (India). Male germ cell chromosomes of the species are de- scribed on the basis of conventional staining, C-banding, silver nitrate staining and sequence specific staining. Both the species possesses 2n = 25m, as a diploid chromosome number and XO (♂)/XX (♀) type sex determination. In both the species, dark terminal C-bands are present on all the autosomal bivalents and X chromosome is C-positive throughout the length. Terminal light/dark NORs (Nucleolar Organizer Regions) are present on all auto- somal bivalents, while X chromosome also possesses terminal NORs. During sequence spe- cific staining, all the autosomal bivalents show prominent terminal DAPI (4',6-diamidino-
2-phenylindole) and CMA3 (Chromomycin A3) bright regions and X chromosome also pos- sesses both DAPI and CMA3 signals. In addition, a brief review of the size of X element in the allied families Cordulegastridae, Corduliidae and Macromiidae is given. Further key words. Dragonfly, Anisoptera, chromosomes, conventional staining, C-band- ing, silver nitrate staining, sequence specific staining, India
Introduction Cytogenetic data on eight species of the Cordulegastridae is available world- wide (Oguma 1930; Oksala 1939a, b; Kichijo 1942b; Kiauta 1968a, b, 1969; Cruden 1968, 1969; Kiauta & Kiauta 1976; Zhu & Wu 1986; Sandhu & Malhotra 1994; Perepelov et al. 2001), whereas no report is available for the Chlorogomphidae. In the present study, Cordulegaster brevistigma Selys, 1854, (Cordulegastridae) and Watanabeopetalia atkinsoni (Selys, 1878), (Chlorogomphidae) have been studied cytogentically. Struc- ture and behaviour of chromosomes during meiosis, detection of constitu- tive heterochromatin, localization of NORs (Nucleolar Organizer Regions)
Odonatologica 48(1/2) 2019: 101-113Odonatologica – DOI:10.5281/zenodo.2677693 48(1/2) 2019: 101-113 102 G. Kaur Walia & S. Singh Chahal and distribution of AT- and GC-rich regions were investigated and com- pared in both species.
Material and methods Live adult male specimens of Cordulegaster brevistigma and Watanabeopeta lia atkinsoni were collected from Summer Hills, Shimla, Himachal Pradesh, India, in June 2015. Specimens were dissected in 0.67 % saline solution in the field and testes were taken out. Subsequently, testes were put in sodium citrate (0.9 %) for 45 minutes and then fixed in freshly prepared Carnoy’s fixative (3:1, absolute alcohol: glacial acetic acid). Testes were teased on grease free slides and slides were air dried. Slides were prepared for con- ventional staining (Carr & Walker 1961), C-banding (Sumner 1972), sil- ver nitrate staining (Howell & Black 1980) and sequence specific staining (Rebagaliati et al. 2003). Relevant meiotic and mitotic stages were micro- photographed.
Results Conventional staining In both studied species, 25 elements were present during spermatogonial metaphase; out of these, 24 were autosomes, also including a pair of m chro- mosomes and one X chromosome. X chromosome was the 2nd smallest ele- ment in Cordulegaster brevistigma (Fig. 1a), while it was the largest element in the whole complement in Watanabeopetalia atkinsoni (Fig. 1e). Dur- ing the diplotene stage of W. atkinsoni autosomal bivalents showed cross- shaped structures due to the presence of single chiasma per bivalent, while a large bipartite X chromosome and small m bivalent were clearly visible (Fig. 1f). In both species, during diakinesis and metaphase I, 13 elements were visible; among these, 12 were autosomal bivalents and one was an X chromosome (Figs 1b, c, g, h). During metaphase II in C. brevistigma the chromosomes were half the size of metaphase I chromosomes, while X and m chromosomes were clearly distinguishable (Fig. 1d).
Banding patterns In C. brevistigma, during diplotene and diakinesis, all the autosomal biva- lents including m bivalent showed terminal C-bands and the X chromosome
Odonatologica 48(1/2) 2019: 101-113 Cytogenetic report on Cordulegaster brevistigma and Watanabeopetalia atkinsoni 103 was C-positive and showed bipartite behaviour in the diplotene (Figs 2a, b). In W. atkinsoni, in metaphase I, all the autosomal bivalents showed termi- nal C-bands, while m bivalent and X chromosome were C-positive because of condensation of elements (Figs 2c, d). In C. brevistigma, during diplo- tene, terminal NORs were present on all the autosomal bivalents and the X chromosome showed light NOR bands on both the chromatids (Fig. 2e). During diakinesis, all the autosomal bivalents showed terminal NORs and the X chromosome was rich in NORs (Fig. 2f). In W. atkinsoni, during dia- kinesis, terminal NORs were present on autosomal bivalents and the large X chromosome showed dark NOR bands (Figs 2g, h). In both the species, the interphase cell showed DAPI and CMA3 bright regions (Figs 3a, b, e, f) and all the autosomal bivalents showed terminal DAPI (4',6-diamidino-2- phenylindole) and CMA3 (Chromomycin A3) bright regions and the X chro- mosome was also DAPI and CMA3 bright in diakinesis (Figs 3c, d, g, h). Discussion Cytogenetically, Odonata are characterized by holokinetic chromosomes, post-reductional meiosis for sex chromosome, XX/XO (Female/Male) sex determining mechanism, with m chromosomes present and a single chi- asma per bivalent. So far, only eight species of Cordulegastridae have been cytogenetically studied world-wide (Table 1). The majority of the species possesses the chromosome number 2n(♂) = 25 (Oguma 1930; Oksala 1939a, b; Kichijo 1942b; Kiauta 1968a, b, 1969; Cruden 1968, 1969; Ki- auta & Kiauta 1976; Zhu & Wu 1986; Perepelov et al. 2001). The only exception is Anotogaster basalis with a chromosome number 2n = 23, which originates by the fusion of m chromosomes with another pair of autosomes (Sandhu & Malhotra 1994). Among these, four species of Cordulegaster possess 2n = 25m, which is the type number of the family. In the present study, C. brevistigma also possesses 2n (♂) = 25m, with X0(♂)/XX(♀) type sex determination, which is in accordance with reports of the chromosome number of other species in the genus. X is the 2nd smallest element in the complement after m chromosomes. No cytogenetic report was hitherto available on any species of the Chlorogomphidae. This study shows Watan abeopetalia atkinsoni possesses 2n (♂) = 25m, with X0(♂)/XX(♀) sex deter- mination and X is the largest element in the complement.
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Figure 1. a−d, Conventional staining in Cordulegaster brevistigma; e−h, conven- tional staining in Watanabeopetalia atkinsoni. a − Spermatogonial metaphase; b, c – diakinesis; d − metaphase I; e − spermatogonial metaphase; f – diplotene; g, h − diakinesis. X and m marked with arrows. Bar: 0.01 mm
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Figure 2. a−b, C-banding in Cordulegaster brevistigma; c−d, C-banding in Watana beopetalia atkinsoni; e−f, silver nitrate staining in Cordulegaster brevistigma; g−h, silver nitrate staining in Watanabeopetalia atkinsoni. a – Diplotene; b – diakinesis; c, d − metaphase I; e – diplotene; f, g, h − diakinesis. X and m marked with arrows. Bar: 0.01 mm
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Figure 3. a−d, Sequence specific staining in Cordulegaster brevistigma; e−h, se- quence specific staining in Watanabeopetalia atkinsoni. a, c, e, g − DAPI staining; b, d, f, h − CMA3 staining; a, b, e, f − interphase cell; c, d, g, h − diakinesis. X and m marked with arrows. Bar: 0.01 mm
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Size of the X chromosome in Cordulegastridae, Corduliidae and Macro miidae In the Cordulegastridae, the size of X chromosome has been estimated in six out of eight cytogentically studied species. It is smallest when m chro- mosomes are absent (Kiauta & Kiauta 1976; Zhu & Wu 1986; Sandhu & Malhotra 1994) and 2nd smallest when m chromosomes are present (Oguma 1930; Cruden 1968, 1969). The only exception is Anotogaster sie boldii, in which X chromosome shows variation in size, as it is 2nd smallest (Oguma 1930) and medium sized (Perepelov et al. 2001). In the Cordulii- dae, the size of the X chromosome has been reported only in 15 out of 23 cy- togenetically studied species. It is smallest or 2nd smallest in majority of spe- cies. The only exception is Somatochlora borisi with chromosome comple- ment (n(♂) = 11/neo-XY) which originated from two fusions, one between two small pairs of autosomes and another between a pair of autosomes and the X element (Grozeva & Marinov 2007). In family Macromiidae, among the four cytogenetically studied species, the X chromosome is the 2nd small- est in three species (Dasgupta 1957; Cruden 1968; Kiauta 1977; Walia & Chahal 2018). The size of the X chromosome has been reviewed in the three allied fami- lies Cordulegastridae, Corduliidae and Macromiidae (Table 1). The size of X chromosome in majority of the species of all the allied families is 2nd small- est or smallest. In the presently studied species, Cordulegaster brevistigma has a X chromosome that is 2nd smallest as reported in earlier studied spe- cies of the family. However, in Watanabeopetalia atkinsoni it is largest in size, which might be considered as the cytotaxonomic feature of the family as no other species of this family has hitherto been studied. Perepelov et al. (2001) studied the distribution of constitutive hetero- chromatin in Anotogaster sieboldii (2n = 25m) (Cordulegastridae) for the first time. They observed terminal C bands on all the autosomes, including the m chromosomes and X chromosome also possesses C-blocks at both ends. Similarly, during the present study, C bands are present at terminal ends of all the autosomal bivalents including m bivalent and X chromosome is C positive in C. brevistigma and W. atkinsoni. In the Anisoptera, few reports are available on silver nitrate staining and sequence specific staining. Nokkala et al. (2002) performed silver nitrate
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Table 1. Cytogenetic data of Chlorogomphidae, Cordulegastridae, Corduliidae and Macromiidae with chromosome complement, locality and size of X chromosome. Nomenclature largely follows the ‘World Odonata List’ by Schorr & Paulson (2018). Names in square parentheses refer to the taxon used in references cited. CC – chromosome complement.
Taxon CC Locality Size of X References chromosome Chlorogomphidae Watanabeopetalia n=13(m) India largest Present study atkinsoni (Selys, 1878) element
Cordulegastridae Anotogaster kuchenbeiseri n=13 China smallest Zhu & Wu (1986) (Förster, 1899) element Anotogaster sieboldii n=13(m) Japan 2nd smallest Oguma (1930) (Selys, 1854) element – Kichijo (1942b); Kiauta (1969) n=13(m) Russia medium in Perepelov et al. (2001) size Anotogaster basalis n=12 India smallest Sandhu & Malhotra Selys, 1854 element (1994) Cordulegaster boltonii n=13(m) Finland – Oksala (1939a, b) (Donovan, 1807) n=13 Austria – Kichijo (1942b) Sweden – Kiauta (1968a, b, 1969) Cordulegaster brevistigma n=13(m) India 2nd smallest Present study Selys, 1854 element Cordulegaster diastatops n=13(m) U.S.A. – Cruden (1968) (Selys, 1854) Cordulegaster dorsalis n=13(m) U.S.A. 2nd smallest Cruden (1968, 1969) Hagen in Selys, 1858 element [Cordulegaster deserticola Cruden, 1969] Cordulegaster maculata n=13(m) U.S.A. 2nd smallest Cruden (1968) Selys, 1854 element Neallogaster hermionae n=13 Nepal smallest Kiauta & Kiauta (1976) (Fraser, 1927) element
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Taxon CC Locality Size of X References chromosome Corduliidae Cordulia aenea n=13 Finland – Oksala (1939a) (Linnaeus, 1758) U.S.S.R. – Makalowskaja (1940) Nether- – Kiauta (1968c, 1969) lands n=13 Russia smallest Perepelov et al. (1998) element n=13 Bulgaria – Grozeva & Marinov (2007) n=13 Russia smallest Kuznetsova et al. element (2017) Cordulia shurtleffii n=13 U.S.A. smallest Cruden (1968) Scudder, 1866 element Dorocordulia libera n=6+7; U.S.A. – Cruden (1968); (Selys, 1871) n=7 Kiauta (1969) Epicordulia princeps n=13(m) U.S.A. – Hung (1971) (Hagen, 1861) Epitheca bimaculata n=13(m) Russia 2nd smallest Kuznetsova et al. (Charpentier, 1825) element (2017) Epitheca canis n=13(m) U.S.A. 2nd smallest Cruden (1968) McLachlan, 1886 element Epitheca cynosura n=10+11 U.S.A. – Cruden (1968); (Say, 1840) n=11 Kiauta (1969) Epitheca semiaquea n=13 U.S.A. – Cruden (1968) (Burmeister, 1839) Epitheca spinigera n=13(m) U.S.A. – Cruden (1968) (Selys, 1871) Procordulia grayi n=13(m) New 2nd smallest Jensen (1980) (Selys, 1871) Zealand element Procordulia smithii n=13(m) New 2nd smallest Jensen (1980) (White, 1846) Zealand element Rialla villosa n=13 Argen- – De Gennaro (2004) (Rambur, 1842) tina Somatochlora alpestris n=13 Switzer- smallest Kiauta & Kiauta (1980) (Selys, 1840) land element n=14(m) 2nd smallest element
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Taxon CC Locality Size of X References chromosome Somatochlora borisi n=11/ Bulgaria X fused with Grozeva & Marinov Marinov, 2001 neo-XY one auto- (2007) some and large in size Somatochlora flavomacu n=13 U.S.S.R. – Makalowskaja (1940) lata (Vander Linden, 1825) Somatochlora graeseri n=13m Russia 2nd smallest Perepelov et al. 2001 Selys, 1887 element Somatochlora meridionalis n=13 Slovenia smallest Kiauta & Kiauta (1995) Nielsen, 1935 element n=13 Bulgaria – Grozeva & Marinov (2007) Somatochlora metallica n(♀)=13 Finland – Oksala (1945) (Vander Linden, 1825) n=13(m) Switzer- 2nd smallest Kiauta & Kiauta (1995) land element n=13 Finland smallest Nokkala et al. (2002) element n=13(m) Bulgaria – Grozeva & Marinov (2007) Somatochlora petechialis n=11 U.S.A. – Cumming (1964) (Muttkowski, 1911) [Tetragoneuria petechialis Muttkowski, 1911] Somatochlora n=13 U.S.A. smallest Cruden (1968) semicircularis (Selys, 1871) element Somatochlora uchidai n=13(m) Japan 2nd smallest Oguma (1915, 1930); Förster, 1909 element Kichijo (1942a) Somatochlora viridiaenea n=13 Japan smallest Oguma (1915, 1930); (Uhler, 1858) element Kichijo (1942a) Tetragoneuria spinigera n=14 U.S.A. – Hung (1971) (Selys, 1871) Macromiidae Didymops transversa n=13(m) U.S.A. 2nd smallest Cruden (1968) (Say, 1839) element Epophthalmia frontalis n=13(m) India 2nd smallest Dasgupta (1957) frontalis Selys, 1871 element Macromia magnifica n=13(m) U.S.A. – Cruden (1968) (McLachlan, 1874) n=13 Macromia moorei n=13(m) Nepal 2nd smallest Kiauta (1977); Selys, 1874 India element Walia & Chahal (2018)
Odonatologica 48(1/2) 2019: 101-113 Cytogenetic report on Cordulegaster brevistigma and Watanabeopetalia atkinsoni 111 staining for the first time in the order Odonata on Aeshna grandis (Lin- naeus, 1758). Walia & Chahal (2014, 2018), Walia et al. (2016, 2018) and Kuznetsova et al. (2017) studied the distribution of NORs in the species of Gomphidae, Aeshnidae and Macromiidae and observed terminal NORs. In the present study, in both species terminal NORs are present on all the autosomal bivalents and the X chromosome is NOR rich. De Gennaro et al. (2008) performed sequence specific staining in the aeshnid Coryph aeschna perrensi (McLachlan, 1887), and observed more CMA3 bright re- gions. Walia et al. (2018) and Walia & Chahal (2018) studied sequence specific staining in Aeshnidae and Macromiidae and reported DAPI and
CMA3 bright regions on all the autosomal bivalents and X chromosome. During the present study, in sequence specific staining, all the autosomal bivalents show prominent terminal DAPI and CMA3 bright regions and X chromosome also possesses both DAPI and CMA3 signals. These results in- dicate that complement of both species possess interspersed AT and GC rich regions, which correspond to the results of C banding and silver nitrate staining.
References Carr D.H. & Walker J.E. 1961. Carbol- ings of the Zoological Society, Calcutta, 10: fuchsin as a stain for human chromosomes. 1-66 Stain Technology 30: 233-236 De Gennaro D. 2004. Análisis meiótic o y Cruden R.W. 1968. Chromosome numbers caracterización de la heterocromatina en of some North American dragonflies (Odo- especies argentinas de Anisoptera (Odo- nata). Canadian Journal of Genetics and Cy nata). Tesis de Licenciatura, Facultad de tology 10: 200-214 Ciencias Exactas y Naturales, U.B.A., Bue- nos Aires Cruden R.W. 1969. A new species of Cor dulegaster from the Great Basin Region of De Gennaro D., Rebagliati P.J. & Mola L.M. the United States (Odonata: Cordulegastri- 2008. Fluorescent banding and meiotic be- dae). Pan-Pacific Entomologist 45: 126-132 haviour in Erythrodiplax nigricans (Libelluli- dae) and Coryphaeschna perrensi (Aeschni- Cumming R.B. 1964. Cytogenetic studies in dae) (Anisoptera, Odonata). Caryologia 61: the order Odonata. PhD thesis, University 60-67 of Texas, Austin Grozeva S.M. & Marinov M.G. 2007. Cy- Dasgupta J. 1957. Cytological studies on togenetic study of Somatochlora borisi the Indian dragonflies. II. A study of the Marinov, 2001 (Odonata: Corduliidae), and chromosomes during meiosis in thirty spe- three relative species. Acta zoologica bul cies of Indian Odonata (Insecta). Proceed garica 59: 53-58
Odonatologica 48(1/2) 2019: 101-113 112 G. Kaur Walia & S. Singh Chahal
Howell M. & Black D.A. 1980. Controlled Kiauta B. & Kiauta M.A.J.E. 1980. The silver staining of nucleolar organizer re- karyotypes of Aeshna subarctica elisa gions with protective colloidal developer: bethae Djak. and Somatochlora alpestris I step method. Experientia 36: 1014-1015 (Sel.) from Switzerland (Anisoptera, Aeshni- Notulae odonatologicae Hung A.C.F. 1971. Cytological studies of dae, Corduliidae). five dragonflies (Odonata: Anisoptera). En 1 (6): 104-105 tomological News 82: 103-106 Kiauta B. & Kiauta M. 1995. The karyo- types of Somatochlora meridionalis Nielsen Jensen A.L. 1980. The karyotypes of five from Slovenia and S. metallica (Vander L.) species of Odonata endemic to New Zea- from Switzerland, with a tentative note on land. Odonatologica 9: 29-33 the origin of Central European S. metallica Kiauta B. 1968a. Variation in size of the (Odonata: Corduliidae). Opuscula zoologica dragonfly m-chromosome, with considera- fluminensia 137: 1-5 tions on its significance for the chorogeo Kichijo H. 1942a. Konchu no Senshokutai. graphy and taxonomy of the order Odona- III. Tombo-Moku 1 [Insect chromosomes. ta, and notes on the validity of the rule of III. Order dragonflies, pt. 1]. Nagasaki Iga Reinig. Genetica 39: 64-74 kukai Zasshi [Nagasaki medical Journal] 20: Kiauta B. 1968b. The chromosomes of the 1084-1092 [in Japanese] male dragonfly, Cordulegaster boltoni (Do- Kichijo H. 1942b. Konchu no Senshokutai. novan, 1807) (Odonata: Cordulegastridae). IV. Tombo-Moku 2 [Insect chromosomes. Biološki Vestnik, Ljubljana, 16: 87-94 IV. Order dragonflies, pt. 2]. Nagasaki Iga Kiauta B. 1968c. Morphology and kinetic kukai Zasshi [Nagasaki medical Journal] 20: behaviour of the odonate sex chromo- 1639-1648 [in Japanese] somes, with a review of the distribution of Kuznetsova V.G., Maryańska-Nadachows sex determining mechanisms in the order. ka A., Shapoval N.A., Anokhin B.A. & Sha Genen en Phaenen 12: 21-24 poval A.P. 2017. Cytogenetic characteriza- Kiauta B. 1969. Sex chromosomes and sex tion of eight Odonata species originating determining mechanism in Odonata, with a from the Curonian Spit (the Baltic Sea, review of the cytological conditions in the Russia) using C-Banding and FISH with 18S family Gomphidae and references to the rDNA and telomeric (TTAGG)n Probes. Cy karyotypic evolution of the order. Genetica togenetics and Genome Research 153: 147- 40: 127-157 157 Kiauta B. 1977. Notes on new or little Makalowskaja W.N. 1940. Comparative known dragonfly karyotypes. V. The male karyological studies of dragonflies (Odona germ cell chromosomes of Macromia ta). Archives russes d’Anatomie, d’Histolo moorei Selys from Nepal (Anisoptera: Cor gie et d’Embryologie 25: 24-39 duliidae, Epophthalmiinae). Genen en Nokkala S., Laukkanen A. & Nokkala C. Phaenen 19: 49-51 2002. Mitotic and meiotic chromosomes Kiauta B. & Kiauta M.A.J.E. 1976. The chro- in Somatochlora metallica (Cordulidae, mosomes of some dragonflies from the Odonata). The absence of localized centro- Langtang valley, Central Nepal. Odonato meres and inverted meiosis. Hereditas 136: logica 5: 347-354 7-12
Odonatologica 48(1/2) 2019: 101-113 Cytogenetic report on Cordulegaster brevistigma and Watanabeopetalia atkinsoni 113
Oguma K. 1915. [A study of the chromo- biodiversity-resources/dragonflies/world- somes of dragonflies].Zoological Magazine odonata-list2/ 27: 241-250 [in Japanese] Sumner A.T. 1972. A simple technique for Oguma K. 1930. A comparative study of the demonstrating centromeric heterochroma- spermatocyte chromosome in allied spe- tin.Experimental Cell Research 75: 304-306 cies of the dragonfly. Journal of Faculty of Walia G.K. & Chahal S.S. 2014. Distribution Sciences, Hokkaido University VI: 1-32 of constitutive heterochromatin and nu- Oksala T. 1939a. Über Tetraploidie der cleolar organizer regions in two species of Binde-und Fettgewebe bei den Odonaten. family Gomphidae (Odonata: Anisoptera). Hereditas 25: 132-144 The Nucleus 57: 223-227 Oksala T. 1939b. Über die somatische Poly- Walia G.K. & Chahal S.S. 2018. Cytogenetic ploidie bei Insekten. Annales entomologici characterization ofMacromia moorei Selys, fennici 5: 208-218 1874 of family Macromidae (Odonata: An isoptera) from India by C-banding, Silver ni- Oksala T. 1945. Zytologische Studien an trate staining and Sequence specific stain- Odonaten. III. Die Ovogenese. Annales Aca ing. International Journal of Life Sciences demiae Scientiarum fennicae (A IV, Biologi- Research 6: 64-68 ca) 9: 1-132 Walia G.K., Chahal S.S. & Babu R. 2016. Perepelov E.A., Bugrov A.G. & Warcha Cytogenetic report on Gynacanthaeschna łowska-Śliwa E. 1998. C-banded karyo- sikkima from India (Odonata: Aeshnidae). types of some dragonfly species from Rus- Odonatologica 45: 87-94 sia. Folia biologica, Kraków, 46: 137-142 Walia G.K., Chahal S.S. & Somal D.S. Perepelov E.A., Bugrov A.G. & Warcha 2018. Chromosome observations based łowska-Śliwa E. 2001. C-banded karyo- on C-banding, Ag-NOR and Sequence spe- types of some dragonfly species from Rus- cific staining in twoAnax species from India sia. II. The families Cordulegastridae, Cor- (Odonata: Aeshnidae). Odonatologica 47: duliidae and Gomphidae. Folia biologica, 145-160 Kraków, 49: 175-178 Zhu H.-q. & Wu J.-i. 1986. Notes on the Rebagliati P.J., Papeschi A.G. & Mola L.M. male germ cell karyotypes of some Odona- 2003. Meiosis and fluorescent banding in ta from the Shanxi Province, China. Notulae Edessa meditabunda and E. rufomarginata odonatologicae 2: 118-120 (Heteroptera: Pentatomidae: Edessinae). European Journal of Entomology 100: 11-18 Sandhu R. & Malhotra I. 1994. New chro- mosome count in male dragonfly, Anato gaster s. basalis. Bionature 14: 69-70 Schorr M. & Paulson D. 2018. World Odonata list, last revision: 24 March 2019. Online on the internet, URL [02-iv-2019]: https://www.pugetsound.edu/academ- ics/academic-resources/slater-museum/
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