NORTH-WESTERN JOURNAL OF ZOOLOGY 10 (2): 355-358 ©NwjZ, Oradea, Romania, 2014 Article No.: 141301 http://biozoojournals.ro/nwjz/index.html
The first record of karyotypes in Leiurus abdullahbayrami and Compsobuthus matthiesseni (Scorpiones: Buthidae) from Turkey
Frantisek STAHLAVSKY1,*, Halil KOÇ2 and Ersen Aydın YAĞMUR3
1. Charles University in Prague, Faculty of Science, Department of Zoology, Viničná 7, CZ-12844 Prague, Czech Republic. 2. Sinop University, Faculty of Arts and Science, Department of Biology, TR-57000 Sinop, Turkey. 3. Alaşehir Vocational School, Celal Bayar University, Alaşehir, Manisa, Turkey. *Corresponding author, F. Stahlavsky, E-mail: [email protected]
Received: 22. January 2014 / Accepted: 24. March 2014 / Available online: 22. July 2014 / Printed: December 2014
Abstract. The karyotypes of Leiurus abdullahbayrami Yağmur, Koç & Kunt, 2009 and Compsobuthus matthiesseni (Birula, 1905) from Turkey are presented for the first time. Both analyzed species have achiasmatic meiosis and the same number of holocentric chromosomes (2n=22) gradually decreasing in size. During the first meiotic division we observed tetravalent in one male of L. abdullahbayrami and octovalent in one male of C. matthiesseni. These types of multivalents may increase genetic variability and indicate reciprocal translocations within these species.
Key words: Arachnid, cytogenetics, holocentric chromosomes, multivalents.
Introduction Uroplectes have more than three analyzed species (Newlands & Martindale 1980, Moustafa et al. The Scorpiones is the fifth largest order of the class 2005, Mattos et al. 2013). That was a reason why Arachnida with approx. 2091 described species we focused on karyotype analysis of buthid scor- divided into 197 genera (Rein 2012) and 15-19 pions Leiurus abdullahbayrami and Compsobuthus families (Soleglad & Fet 2003, Prendini & Wheeler matthiesseni from Turkey using standard Giemsa 2005). Despite their worldwide distribution and staining. From the cytogenetic point of view just L. high abundance in subtropical and tropical areas quinquestriatus has been analyzed (Qumsiyeh et al. the cytogenetic data has been available only in 80 2013) whereas there is no information about the species and more than half of this number belongs chromosomes in genus Compsobuthus. Both genera to the family Buthidae (Schneider et al. 2013). Spe- have a wide range of distribution all over North cies of the family Buthidae have holocentric chro- Africa and from Near East to India (Yağmur et al. mosomes and relatively low diploid chromosome 2008, Rein 2012). Compsobuthus is a diverse genus numbers, ranging from 5 in Tityus bahiensis (Perty, with 43 described species (Kovařík & Ojanguren- 1833) to 56 in Buthus occitanus (Amoreux, 1789) Affilastro 2013). In the genus Leiurus there are five (Schneider et al. 2013). In addition, high intras- species described but high cryptic diversity is sus- pecific variability was observed in some species, pected, especially in L. quinquestriatus (Ehrenberg, especially in T. bahiensis (2n=5-19) (Schneider et al. 1828) (Kovařík, pers. comm. 2013). This high di- 2009) and Hottentotta tamulus (Fabricius, 1798) versity and wide distribution enable using these (2n=20-28) (Venkatanarasimhiah & Rajaseka- genera as a model group for subsequent analysis rasetty 1964). This variability may reflect the holo- of dynamics of chromosomal changes in buthid centric nature of their chromosomes that tolerate scorpions. frequent fragmentation or fission/fusion and that is also responsible for odd diploid chromosome numbers in some individuals (Schneider et al. Materials and Methods 2009). Despite known intraspecific variability, the majority of the karyotyped scorpions display spe- For the analysis we used three males and three females of Leiurus abdullahbayrami and three males and one female of cific diploid numbers. The potential application of Compsobuthus matthiesseni from Turkey. The living scorpi- this information in taxonomy and reconstruction ons were collected during the night with UV lamp. One of karyotype evolution in this family is reduced male of L. abdullahbayrami was collected at: Gaziantep due to the number of cytogenetic analyses that fo- Province, Cerityeniyapan Village, 37°10'55.2"N, cused only on a few geographic regions (mainly 37°08'42.0"E, 1052 m, June 2013, Lgt., Det. et Coll. E.A. Brazil) (see Schneider et al. 2013). Moreover there Yağmur. All other material was collected at: Gaziantep is a limited number of analyzed species within Province, Şehitkamil District, Çaybaşı Village, 36°59'57.5"N 37°31'09.8"E, 772 m, June 2013, Lgt., Det. et particular genera. Only Androctonus, Tityus and 356 F. Stahlavsky et al.
Coll. E.A. Yağmur. regions that confirm holocentric constitution of For the karyotype analysis we used chromosome chromosomes. We found only the standard biva- preparations made by spreading technique described by lents in two males of L. abdullahbayrami (Fig. 1a) Traut (1976). For the hypotonisation of the gonads we and in two males of C. matthiesseni (Fig. 1d). used 0.075 M KCl for 20 mins. and then the tissue was fixed in methanol: glacial acetic acid (3:1) for at least 20 Moreover we also detected one tetravalent (Fig. mins. After this time the tissue was dissociated in a drop 1b) in all observed nuclei of one another male L. of 60% acetic acid on a microscope slide. This drop was abdullahbayrami from Çaybaşı Village and one oc- moved to a warm histological plate for evaporation tovalent (Fig. 1e) in all observed nuclei of one (45oC). The chromosomes were stained by 5% Giemsa so- other male C. matthiesseni from Cerityeniyapan lution in Sörensen phosphate buffer (pH = 6.8) for 20 Village during the first meiotic division. The size mins. The chromosome preparations were observed with of the chromosomes in males with only bivalents Olympus AX70 Provis and photographed with Olympus DP72. The measurements were taken from photographs gradually decreases from 6.09% (SD ±0.17) to using the software ImageJ 1.45r (http://rsbweb.nih. 3.24% (SD ±0.21) DSL in L. abdullahbayrami and gov/ij) with the plugin Levan (Sakamoto & Zacaro 2009). from 6.28% (SD ±0.60) to 3.12% (SD ±0.11) DSL in The relative diploid set length (DSL) was calculated for C. matthiesseni, respectively. The size of the chro- each chromosome as a percentage of the diploid set based mosomes in males with the multivalents also on ten postpachytene or metaphase I. We also analyzed gradually decreases from 5.99% (SD ±0.31) to meiosis that may reveal unusual pairing and separation 3.18% (SD ±0.21) DSL in L. abdullahbayrami and of chromosomes. from 6.68% (SD ±0.34) to 3.15% (SD ±0.18) DSL in C. matthiesseni, respectively. The reciprocal trans- Results locations form chromosomes of different sizes in tetravalent (5.72%, 5.20%, 4.78%, 4.42% of DSL) as The diploid number of chromosomes is 22 in all well as in octovalent (6.68%, 6.24%, 5.90%, 5.75%, analyzed specimens of Leiurus abdullahbayrami 5.43%, 4.91%, 3.95%, 3.29% of DSL). According to (Fig. 1a-c) and Compsobuthus matthiesseni (Fig. 1d- the gradual decreasing of the chromosome size it f). The number of the chromosomes and their size is not possible to detect the chromosomes from are similar in males (Fig. 1a,b,d,e) and females multivalents precisely in mitotic metaphases (Fig. (Fig. 1c,f) in both species. In observed chromo- 1f). During the stages of first meiotic division somes we did not detect prominent centromeric (pachytene, postpachytene, metaphase I) we did
Figure 1. Chromosomes of Leiurus abdullahbayrami, 2n=22 (a-c) and Compsobuthus matthiesseni, 2n=22 (d-f). (a) postpachytene of male; (b) metaphase I of male, arrowhead shows tetravalent; (c) mitotic metaphase of fe- male; (d) postpachytene of male; (e) metaphase I of male, arrows show chromosomes of octovalent; (f) mitotic metaphase of the male with octovalent configuration in meiosis. Bar = 5 μm.
Karyotypes of scorpions from Turkey 357
not observe any evidence of the crossing over be- the family Buthidae. This type of rearrangement is tween homologous chromosomes. That is a reason documented within Buthidae in the genera An- why we considered this type of meiosis as achias- anteris (Mattos et al. 2013), Androctonus (Moustafa matic in males. et al. 2005), Gint (Kovařík et al. 2013), Hottentotta (Sharma et al. 1959), Isometrus, Isometroides (Shanahan, 1989), Lychas (Shanahan & Hayman Discussion 1990), Odontobuthus (Sharma et al. 1959), Rhopalu- rus (Mattos et al. 2013) and Tityus (e.g. Schneider In accord with the previous analyses (e.g. et al. 2009, Mattos et al. 2013). The specific impact Shanahan 1989, Schneider et al. 2009, Mattos et al. of this rearrangement should be observed by mo- 2013) we found holocentric chromosomes and lecular analysis in the future. achiasmatic meiosis in males of two other buthid species. This special trait seems to define this evo- lutionary branch of scorpions (Schneider et al. 2009). Our data in Leiurus abdullahbayrami and Acknowledgements. We wish to thank Mehmet Özkörük Compsobuthus matthiesseni (in both species 2n=22) and Erman Tezcan for their help in the field trip. We are also supports another characteristic of buthid also grateful to Ivana Hynkova, Sergio Gustavo Rodríguez Gil and Marielle Cristina Schneider for karyotypes – low number of chromosomes. The valuable comments on the manuscript. diploid number in this family ranges from 2n=5 to
2n=56 but the average is 20 (see Schneider et al.
2013). Within buthid scorpions a different type of karyotype variability can be found. Some buthid References genera posses interspecific variability as was documented in the genera Hottentotta (2n=16 and Goyffon, M., Chovet, G., Deloince, R., Vachon, M. (1971): Étude du caryotype de quelques scorpions Buthides. Arachnologorum 24) (Newlands & Martindale 1980, Qumsiyeh et al. Congressus Internationalis V. Brno. 23-25. 2013), Parabuthus (2n=18, 20 and 36) and Uroplectes Kovařík, F., Ojanguren-Affilastro, A.A. (2013): Illustrated catalog of (2n=20, 22, 24 and 26) (Newlands & Martindale scorpions, part II. Prague, Czech Republic: Clarion Production. Kovařík, F., Lowe, G., Plíšková, J., Šťáhlavský, F. (2013): A new 1980). Moreover, high intraspecific variability was scorpion genus, Gint gen.n., from the Horn of Africa documented in some species, especially in Tityus (Scorpiones: Buthidae). Euscorpius 173: 1-19. (e.g. T. bahiensis 2n=5-19) (Schneider et al. 2009) Mattos, V.F., Cella, D.M., Carvalho, L.S., Candido, D.M., Schneider, M.C. (2013): High chromosome variability and the presence of and Hottentotta tamulus (2n=20-28) (Venkatanara- multivalent associations in buthid scorpions. Chromosome simhiah & Rajasekarasetty 1964). On the other Resereach 21: 121-136. hand in five species of genus Androctonus were Moustafa, M.A., Alaa, A.M., Sarhan, M.H., Yaseen, A.E. (2005): Chromosomal studies on four Egyptian scorpion species of found similar karyotypes 2n=24 (Goyffon et al. genus Androctonus (Family: Buthidae). Cytologia 70: 161-165. 1971, Moustafa et al. 2005). Comparing our results Newlands, G., Martindale, C.B. (1980): The buthid scorpion fauna with L. quinquestriatus (2n=22) (Qumsiyeh et al. of Zimbabwe-Rhodesia with checklists and keys to the genera and species, distributions and medical importance (Arachnida: 2013) it seems that the interspecific karyotype Scorpiones). South African Institute for Medical Research 67: 51- variability is low in genus Leiurus. But higher 77. number of analyzed species would be recom- Prendini, L., Wheeler, W.C. (2005): Scorpion higher phylogeny and classification, taxonomic anarchy, and standards for peer review mended to test this conclusion. in online publishing. Cladistics 21: 446-494. The holocentric nature of buthid chromo- Qumsiyeh, M.B., Salman, I.N.A., Salsaa, M., Amr, Z.S. (2013): somes leads to the hypothesis that the variability Records of scorpions from the Palestinian Territories, with the first chromosomal data (Arachnida: Scorpiones). Zoology in the of chromosomes is generated mainly by fis- Middle East 59: 70-76. sion/fusion of chromosomes. These mechanisms Rein, J.O. (2012): The Scorpion Files. Trondheim: Norwegian lead to the high intraspecific variability in diploid University of Science and Technology. Available in
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