Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics 73(2): 121-126, 2020 Firenze University Press www.fupress.com/caryologiaCaryologia International Journal of Cytology, Cytosystematics and Cytogenetics ISSN 0008-7114 (print) | ISSN 2165-5391 (online) | DOI: 10.13128/caryologia-610 Citation: R. Monteiro do Nascimento, A. Freire Carvalho, W. C. Santana, A. Barth, M. A. Costa (2020) Karyotype diversity of stingless bees of the genus Frieseomelitta (Hymenoptera, Apidae, Meliponini). Caryologia 73(2): 121-126. doi: 10.13128/caryologia-610 Received: August 26, 2019 Accepted: April 13, 2020 Published: July 31, 2020 Copyright: © 2020 R. Monteiro do Nascimento, A. Freire Carvalho, W. C. Santana, A. Barth, M. A. Costa. This is an open access, peer-reviewed article published by Firenze University Press (http://www.fupress.com/caryologia) and distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distri- bution, and reproduction in any medi- um, provided the original author and source are credited. Data Availability Statement: All rel- evant data are within the paper and its Supporting Information files. Competing Interests: The Author(s) declare(s) no conflict of interest. Karyotype diversity of stingless bees of the genus Frieseomelitta (Hymenoptera, Apidae, Meliponini) Renan Monteiro do Nascimento1, Antonio Freire Carvalho1, Weyder Cristiano Santana2, Adriane Barth3, Marco Antonio Costa1,* 1 Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brazil 2 Departamento de Entomologia, Universidade Federal de Viçosa, Viçosa, MG, Brazil 3 Instituto Federal de Educação Ciência e Tecnologia de Mato Grosso, Campus Rondonó- polis, Brazil *Corresponding author. E-mail: costama@uesc.br Abstract. Frieseomelitta (Ihering, 1912) is a genus of stingless bees, distributed in the Nearctic and Neotropical regions. Specimens can be found in forests, cerrado, caat- inga and mountainous regions. This genus has 16 species, of which 13 are recorded in Brazil. Cytogenetics has contributed to evolutionary studies of some Hymenoptera groups and although many Frieseomelitta species have been described, few species have been studied cytogenetically. The present study aims to contribute to the knowledge of the karyotype diversity of this genus, seeking to understand the possible evolutionary mechanisms that occurred in the diversification of the karyotype of this genus. Frie- seomelitta portoi and Frieseomelitta trichocerata and Frieseomelitta doederleini showed diploid karyotypes with 2n = 30 chromosomes, similarly to all the species previously analyzed in the genus. Unprecedentedly, Frieseomelitta longipes showed 2n = 34. These results confirm that the frequent diploid number of 30 chromosomes is typical of this genus. The finding of 2n = 34 chromosomes in F. longipes comprises the first record of a diploid chromosome number different from 2n=30 in this group, which suggests that it can be the result of a recent chromosome change event. An interspecific comparative analysis was developed involving present and previous studies, as well as a discussion on the mechanisms involved in the karyotypic evolution in the genus. Keywords: heterochromatin, karyotype evolution, centric fission, chromosome, varia- tion. INTRODUCTION Frieseomelitta (Ihering, 1912) is a Neartic and Neotropical genus of stingless bees, and occurs from Mexico to southeast Brazil (Camargo and Pedro, 2013). These bees present small to medium size, with body meas- uring about 10 mm (Silveira et al., 2002). According to Camargo and Pedro (2013), 16 species are known to be valid in the genus Frieseomelit- 122 Renan Monteiro do Nascimento et al. ta, of which 13 are found in Brazilian territory: Frie- seomelitta dispar (Moure, 1950), Frieseomelitta doed- erleini (Friese, 1900), Frieseomelitta flavicornis (Fab- ricius, 1798), Frieseomelitta francoi (Moure, 1946), Frieseomelitta freiremaiai (Moure, 1963), Frieseomelitta languida (Moure, 1989), Frieseomelitta longipes (Smith, 1854), Frieseomelitta meadewaldoi  (Cockerell, 1915), Frieseomelitta paranigra  (Schwarz, 1940), Frieseomelit- ta portoi (Friese, 1900), Frieseomelitta silvestrii (Friese, 1902), Frieseomelitta trichocerata (Moure, 1988) and Frieseomelitta varia (Lepeletier, 1836). Cy togenetic analysis has been a widely used tool in evolutionary and taxonomic studies in some groups of Hymenop- tera, however, for the genus Frieseomelitta, the chro- mosomal analysis are hitherto restricted the species F. doederleini (Kerr and Silveira, 1972; Tarelho, 1973; Rocha et al., 2003; Santos et al., 2018), F. languida (Rocha et al., 2003), F. varia (Kerr, 1969; Kerr and Sil- veira, 1972; Tarelho, 1973; Rocha et al., 2003; Santos et al., 2018), F. dispar (Carvalho and Costa, 2011; Santos et al., 2018), F. francoi (Carvalho and Costa, 2011; San- tos et al., 2018), Frieseomelitta meadewaldoi and Frie- seomelitta sp. n. (Santos et al., 2018). During the karyotype analyzes of several species of bees, Kerr (1972a, b) recorded the haploid chromosome number n = 15 chromosomes in the species F. varia, F. doederleini and F. ghilianii. Rocha et al. (2003) reported the same chromosome number and described the karyo- type of the species F. doederleini, F. languida and F. var- ia as part of an analysis of different genera of stingless bees. Carvalho & Costa (2011) also described the karyo- types of F. dispar and F. francoi, and Santos et al. (2018) developed a comparative analysis of the hybridization patterns of microsatellite DNA probes in karyotypes of five species, F. dispar, F. doederleini e F. francoi, F. mead- ewaldoi, Frieseomelitta sp. n. and F. varia. In addition, other karyotypic features were reported. In all these cases the chromosome number was consistently n = 15 or 2n = 30. These differ from the chromosome numbers determined for other less closely related genera, such as n = 9 for Melipona, and n = 17, common to several of the other genera of stingless bees. However, groups closely related to Frieseomelitta such as genus Duckeola also have shown n = 15 chromosomes (Kerr 1972a, b). The taxonomy and phylogeny of Friesomelitta is still not well resolved, and there are some species with broad geographic distribution. In this context, the present study aimed to contribute to the knowledge of the kar- yotype diversity of this genus, including characterizing samples of new points in the distribution of species and new species. Furthermost, we search for data that may aid in the taxonomic resolution of the group and in the understanding of the possible evolutionary mechanisms that occurred in the diversification of the karyotype of the group. MATERIAL AND METHODS We analyzed samples of four species from different localities of Brazil, Frieseomelitta doederleini, from the municipality of Canavieiras, state of Bahia, (15º 61 ‘S, 39º42’ W), Frieseomelitta longipes, from the municipality of Belém, state of Pará, 1:30 ‘S, 48 ° 73’ W); Frieseomelit- ta portoi, from the municipality of Rio Branco, state of Acre (9º98 ‘S, 67º90’ W); and Frieseomelitta aff. tricho- cerata, from the municipality of Juína, state of Mato Grosso (11º52 ‘S, 60º50’ W). Taxonomist of bees identi- fied the collected specimens and adult specimens of each of the species were mounted on entomological pins and deposited in the entomological collection of the Univer- sidade Estadual de Santa Cruz, Ilheus, BA. The slide preparations were made from cells of the cerebral ganglia of specimens in the prepupa stage, according to the protocol described by Imai et al. (1988). The prepared slides were stained conventionally with 3% Giemsa/Sorensens’s Buffer and the selected metaphases were photographed on an Olympus CX-41 microscope with attached Olympus C7070 digital camera. Staining with the base-specif ic f luorochromes 4,6-diamidino-2-phenylindole (DAPI) and chromomy- cin A3 (CMA3) to evidence the chromosomal regions rich in AT (DAPI) and CG (CMA3), respectively, were performed according to Schweizer (1980), with modi- fications proposed by Guerra and Souza (2002). Cover- slips were mounted on slides with antifading Vectash- ield (Vector Laboratories, Burlingame, USA). The images were captured on a Leica DMRA2 epifluores- cence microscope using the Leica IM50 software (Leica Microsystems Imaging Solutions Ltda, Cambridge, UK). To allow comparison with previous studies, we fol- lowed chromosomal nomenclature proposed by Imai (1991). (M) Metacentric chromosome: the arms of approximately equal sizes and euchromatic, the het- erochromatin restricted to the centromeric region; (A) Acrocentric chromosome: centromeric region and short heterochromatic arms; (AM) Pseudoacrocentric chromo- some: centromeric region, middle or long heterochro- matic arms and short eucrotic arm. The Karyograms were organized with the use of Adobe Photoshop® CS6 13.0x 64 software. From the kar- yotypes, the chromosome pairs, diploid (2n) and haploid (n) numbers and karyotype formulas (2k) were defined. 123Karyotype diversity of stingless bees of the genus Frieseomelitta (Hymenoptera, Apidae, Meliponini) RESULTS AND DISCUSSION The chromosome number found for the species F. doederleini, F. portoi and F. aff. trichocerata was 2n = 30 for females. In F. longipes, females showed 2n = 34 chro- mosomes (Table 1, Fig. 1). Kerr and Silveira, 1972 regis- tered 2n = 30 chromosomes for F. doederleini and F. var- ia and Rocha et al. (2003) found 2n = 30 chromosomes for F. doederleini, F. languida, F. varia. The same number was found by Carvalho and Costa (2011) for F. dispar and F. portoi. Together these results indicate that the fre- quent diploid number of 30 chromosomes is characteris- tic of the genus. The finding of 2n = 34 chromosomes in F. longipes comprises the first record of a diploid chro- mosome number different from 30 in this genus. In the species analyzed here, the following karyotypic formu- las were found: F. doederleini, 2K = 4M + 4A + 22AM, F. portoi, 2K = 4M + 26A, F. aff. Trichocerata, 2K = 6M + 20A + 4AM, and F. longipes, 2K = 8M + 12A + 14AM (Tab. 1). The karyotypic formula observed in F. doederleini is similar to that cited by Rocha et al., (2003) for another population of the same species, evidencing intraspecific karyotypic stability. The predominance of acrocentric and pseudoac- rocentric (AM - which contains a long heterochromatic arm) chromosomes in karyotypes was consistent with that observed in previous studies for other species of the genus, such as F. dispar. F. francoi, F. languida, and F. varia (Rocha et al., 2003; Carvalho and Costa, 2011). However, in F. portoi and F. aff. trichocerata was observed a reduced number of AM chromosomes. The classical cytogenetics using conventional Giem- sa staining and C-banding allowed observing hetero- chromatin in studies of several meliponine genera such as Friesella (Mampumbu, 2002), Leurotrigona (Pompolo and Campos, 1995), Melipona (Hoshiba, 1988; Rocha & Pompolo, 1998; Rocha et al., 2002); Nannotrigona (Hoshiba & Imai, 1993), Partamona (Costa et al., 1992; Martins et al., 2012), Plebeia (Caixeiro & Pompolo, 1999), Tetragonisca (Barth et al., ), Trigona (Hoshiba and Imai, 1993; Costa, 2004; Domingues et al., 2005), among others. The distribution of heterochromatin has still been the focus of comparative studies in stingless bees (e.g. Travenzoli et al., 2019), and for being variable Table 1. Available karyotype data of Frieseomelitta species. 2n = diploid number, 2k = diploid karyotype formula, M = metacentric, A = acrocentric, AM = pseudoacrocênctrico chromosomes. Species Sampling location 2n 2k References F. dispar Ilhéus/BA 30 4M + 2M + 4A + 20AM  Carvalho & Costa (2011) F. doederleini - Santana do Seridó/RN Canavieiras/BA 30 30 30 - 4M + 4A + 22AM  4M + 4A + 22AM Kerr & Silveira, (1972) Rocha et al., (2003) Present study F. francoi Cairú/BA 30 4M + 2M + 4A + 20AM  Carvalho & Costa (2011) F. languida Divinópolis/MG 30 4M + 4A + 22AM Rocha et al., (2003) F. longipes Belém/PA 34 8M + 12A + 14AM Present study F. portoi Rio Branco/AC 30 4M + 26A Present study F. aff. trichocerata Juína/MT 30 6M + 20A + 4AM  Present study F. varia - Divinópolis/MG 30 30 - 4M + 4A + 22AM  Kerr & Silveira, (1972) ROCHA et al., (2003) Figure 1. Karyotypes of workers of Frieseomelitta species stained with Giemsa: (A) F. doederleini (B) F. portoi; (C) F. aff. trichocerata; (D) F. longipes. Bar = 10um. 124 Renan Monteiro do Nascimento et al. among genera and species is a useful cytological infor- mation for the characterization of species or populations of these bees. Imai et al. (1988) have suggested that the tandem addition of terminal heterochromatin in chromosomes is a relatively rapid way of restoring telomeric stability after centric fission events, leading to the formation of AM chromosomes. Elimination of any excess heterochroma- tin addition may follow this addition by deletion mecha- nisms, as a tendency to reduce non-specific heterochro- matin interactions of the long pseudoacrocentric chro- mosomes. The high content of heterochromatin found, however, contrasts with the numerical or even structural stability observed in Frieseomelitta, especially considering species such as F. doederlaini, analyzed in different studies and localities of its geographic distribution. The CMA3/DAPI fluorochrome staining in F. doed- erleini, showed that the centromeric region of the meta- centric, acrocentric and the heterochromatic arm of the pseudoacrocentric chromosomes are rich in AT base pairs (DAPI+). This same region was also weakly stained with CMA3 (Fig. 2A). These results were simi- lar to the findings of Rocha et al. (2003) for F. varia and this marking pattern may be related to the presence of a nucleolus organizing region. These regions are often labeled by CMA3 in karyotypes of bees. NOR banding or in situ hybridization of ribosomal probes are likely to confirm this location. In the F. portoi, CMA3+/DAPI- bands were identified in short heterochromatic arms in chromosomal pairs 1, 2, 3, 4, 5, 6, 8, 9, 11, 12 and 14. Pairs 7, 10 and 13 were totally CMA3-/DAPI+ in this species (Fig. 2B). Frieseomelitta aff. trichocerata showed CMA3+/DAPI- bands in chromosomal pairs 1, 2, 3, 5, 7, 8, 10 and 11, and pairs 4, 6, 9, 12, 13, 14 and 15 were homogeneously CMA3-/DAPI+ (Fig. 2C). We observed in the four species analyzed here that the first pair showed the CMA3+/DAPI- label on the short arm. Although in F. longipes only one of the hom- ologues was labeled by CMA3, suggesting the presence of a heteromorphism, this labeling, which may be asso- ciated with the presence of NOR, seems to be common in the genus. This can be confirmed in further analyzes that include new species. Diverging from the other spe- cies, F. longipes had the long arms of pairs 6 and 11 and pair 14 strongly labeled CMA3+/DAPI- (Fig. 2D). The results obtained in the present analysis togeth- er with the previous karyotypic descriptions show that the karyotype differentiation in Frieseomelitta mostly involved minor structural alterations such as hetero- chromatin gain and loss. However, numerical change occurred in the differentiation of F. longipes, possibly due to centric fission. Since this is the only record of a chromosome number other than 2n = 30 in this genus, this is possibly a derived characteristic in this group. The lack of a better taxonomic definition and a more resolved and complete phylogeny for Frieseomelitta leaves this question open. If a derived position F. lon- gipes is found, the recent origin of this numerical differ- ence will be confirmed. From the observation of the high frequency of the chromosome number 2n = 30 (n = 15) in the Frie- seomelitta, it is possible to suggest that this chromo- some number is basal for this genus. The n = 15 chro- mosomes was also found in the species Duckeola ghilia- nii Kerr, 1972a, b; Kerr and Silveira, 1972). According to the Meliponini phylogeny proposed by Rasmussen and Cameron (2010) Duckeola is closely related to the genus Frieseomelitta. However, without a more detailed phy- logenetic assessment, the hypothesis that the 2n = 34 found in F. longipes, alternatively, represents the basal condition for this group can not be discarded, since this chromosome number is found in most genera of neo- tropical Meliponini (Rocha et al., 2003). More complete phylogenetic analyzes, including a representative sam- pling of Friesemelitta diversity, however, would be neces- sary to better clarify these questions. ACKNOWLEDGMENTS This study was supported by CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico – Process number 310178/2015-0). RMN and AFC were Figure 2. Karyotypes of workers of Frieseomelitta species, stained with CMA3/DAPI fluorochromes: (A) F. doederleini; (B) F. portoi; (C) F. trichocerata; (D) F. longipes. Bar = 10um. 125Karyotype diversity of stingless bees of the genus Frieseomelitta (Hymenoptera, Apidae, Meliponini) supported by a fellowships from CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) pela bolsa concedida durante o Mestrado. We thank Dr. Gabriel Augusto Rodrigues de Melo (UFPR) for spe- cies identification and Dr. Giorgio Cristino Venturieri (Embrapa Amazônia Oriental) for providing samples for cytogenetic analyzes. REFERENCES Barth A, Fernandes A, Pompolo SG, Costa MA (2011) Occurrence of B chromosomes in Tetragonisca Latreille, 1811 (Hymenoptera, Apidae, Meliponini): a new contribution to the cytotaxonomy of the genus. Genetics and Molecular Biology 34(1): 77–79 Caixeiro APA, Pompolo SG. 1999. Caracterização cit- ogenética de heterocromatina constitutiva e sua implicação na evolução do cariótipo de espécies do gênero Plebeia (Hymenoptera, Apidae, Meliponini). Genetics and Molecular Biology 22 (Supplement 3): 31-32. Carvalho AF, Costa MA. 2011. Cytogenetic characteri- zation of two species of Frieseomelitta Ihering, 1912 (Hymenoptera, Apidae, Meliponini). Genetics and Molecular Biology (Impresso), v. 34, p. 237-239. Camargo JMF, Pedro SRM. 2013. Meliponini Lepeletier, 1836. In Moure JS, Urban D and Melo GAR (Orgs). In Catalogue of Bees (Hymenoptera, Apoidea) in the Neotropical Region - online version. Available at http://www.moure.cria.org.br/catalogue. Accessed Agosto/14/2016. Cockerell TDA. 1915. The Real Trigona dorsalis Smith rediscovered (Hym.) Entomological News, 26: 30-32. Costa MA, Pompolo SG, Campos LAO. 1992. Supernu- merary chromosomes in Partamona cupira (Hyme- noptera, Apidae, Meliponinae). Revista Brasileira de Genetica 15, 801–806. Costa KF, Brito RM, Miyazawa CS. 2004. Karyotypic description of four species of Trigona (Jurine, 1807) (Hymenoptera, Apidae, Meliponini) from the State of Mato Grosso, Brazil. Genetics and Molecular Biology. Vol. 27 nº 2 São Paulo. Crozier RH. 1975. Hymenoptera. In: B. John (ed), Ani- mal Cytogenetics 3. Insecta 7. Berlin, Grebuder, Borntraeger. 95p. Domingues AMT, Waldschmidt AM, Andrade SE, Souza VA, Alves RMO, Junior JCS, Costa MA. 2005. Karyo- type characterization of Trigona fulviventris Guérin, 1835 (Hymenoptera, Meliponini) by C banding and fluorochrome staining: Genetics and Molecular Biol- ogy, 28, 3, 390-393. Fabricius JC. 1798. Suplementum Entomologiae System- aticae, Vol. 5 – Hafniae, Apud Proftet Stortch, 572pp. Friese HFAKL. 1900. Neue Arten der Bienengattun- gen Melipona III., und Trigona Jur. Természetrajzi Fuzetek, 23(11): 381-394. Friese HFAKL. 1902. Neue Meliponiden (Hym). Zeitschrift fur Systematische Hymenopterologie und Dipterologie, 2: 382-383. Guerra MS, Souza MJ. 2002. Como Observar Cromo- ssomos: Um Guia de Técnicas em Citogenética Veg- etal, Animal e Humana. Ribeirão Preto, São Paulo, Brazil. Hoshiba H. 1988. Karyological analysis of a stingless bee Melipona favosa (Apidae, Hymenoptera). Cytologia 53:153-156. Hoshiba H, Imai HT. 1993. Chromosome evolution of bees and wasps (Hymenoptera, Apocrita) on the basis of C banding pattern analyses. Japanese Journal of Entomology 61:465-492. Imai HT. 1988. Modes of spontaneous chromosomal mutation and karyotypes evolution in ants with refer- ence to he minimum intraction hypothesis. Japanese Journal of Genetics 63: 159-185. Imai HT. 1991. Mutability of constitutive heterochroma- tin (c-bands) during eukaryotic chromosomal evolu- tion and their cytological meaning. Japanese Journal of Genetics 66: 653-661. Imai HT, Taylor RW, Crosland MWJ, Crozier RH. 1988. Modes of spontaneous evolution in ants with refer- ence to the minimium interaction hypothesis. Japa- nese Journal of Genetics 66: 159-185. Imai HT, Maruyama T, Gojobori T, Inoue Y., Crozier RH. 1986. Theoretical bases for karyotype evolution. The minimum-interaction hypothesis. American Natural- ist 128: 900-920. Imai HT, Taylor RW, Crozier RH. 1994. Experimental bases for the minimium interaction theory. Chromo- some evolution in ants of the Myrmecia pilosula spe- cies complex (Hymenoptera: Formicidae: Myrmeci- nae). Japanese Journal of Genetics 69: 137-182. Imai HT, Brown WL, Kubota JRM, Young HS, Tho YP. 1984. Chromosome observations of tropical ants in western Malasya. Annual Report National Institute of Genetics 63: 66-69. Imai HT, Taylor RW. 1989. Chromosomal polymorphism involving telomere fusion, centromeric inactivation and centromere shipt in the ant Myrmecia (pilosula) n=1. Chromosoma 98: 456-460. Imai HT, Crozier RH, Taylor RW. 1977. Karyotype evolu- tion in Australlian ants. Chromosoma 53: 341-393. Imai HT, Kubota M. 1975. Chromosome polymorphism in the ant, Pheidole nodus. Chromosoma 51: 391-399. 126 Renan Monteiro do Nascimento et al. Imai HT. 1991. Mutability of constitutive heterochroma- tin (c-bands) during eukaryotic chromosomal evolu- tion and their cytological meaning. Japanese Journal of Genetics 66: 653-661. Kerr WE. 1972a. Numbers of Chromosomes in Some Species of Bees. Journal of the Kansas Entomological Society, 45 (1): 111-122. Kerr WE. 1972b. Karyotypic Evolution of Bees and Cor- responding Taxonomic Implications. Evolution, 45 (1): 197-202. Kerr WE, Silveira ZV. 1972. Karyotypc evolution of bees and corresponding taxonomic implications. Evolu- tion 26: 197-202. Lepeletier de Saint-Fargeau A. 1836. Histoire Naturalle des Insects Hyménopteres. nº 10, Bis. Librairie Ency- clopédique de Roret, Paris. 547pp. Mampumbu AR. 2002. Análise citogenética da hetero- cromatina e da NOR em Populações de abelhas sem ferrão Friesella schrottkyi (FRIESE, 1900) (Hymenop- tera: Apidae: Meliponini). Dissertação de Mestrado, Unicamp, Campinas. Martins CCC, Diniz D, Sobrinho-Scudeler PE, Foresti F, Campos LAO, Costa MA. 2012. Investigation of Par- tamona helleri (Apidae, Meliponini) B chromosome origin. An approach by microdissection and whole chromosome painting. Apidologie 44, 75–81 Moure JS. 1946. Contribuição para o Conhecimento dos Meliponinae (Hym. Apoidea). Revista de Entomolo- gia, 17(3): 437-443. Moure JS. 1950. Notas Sobre Alguns Meliponinae Bolivi- anos (Hymenoptera, Apoidea). Dusenia, 2(1): 70-80. Moure JS. 1963. Uma Nova Espécie de “Trigona (Frie- seomelitta)” do Norte do Distrito Tupi (Hymenoptera, Apoidea). Revista Brasileira de Biologia, 23(1): 39-43. Moure JS. 1989. Espécies Novas de Abelhas da Região Central do Estado de Minas Gerais, Brasil (Hyme- noptera, Apoidea). Acta Biológica Paranaense 18(1, 2, 3, 4): 115-127. Moure JS. 1988. Uma Nova Espécie de Frieseomelitta do Oeste da Amazônia (Hymenoptera, Apoidea). Acta Biológica Paranaense 17(1, 2, 3, 4): 141-145. Oliveira FF. 2003. Revisão do gênero Frieseomelitta von Ihering, 1912 (Hymenoptera, Apidae, Meliponinae), com notas bionômicas de algumas Espécies. Tese de Doutorado, Universidade Federal do Paraná, Curiti- ba, 327p. Pompolo SG, Campos LAO. 1995. Karyotypes of two spe- cies of stingless bee, Leurotrigona muelleri and Leuro- trigona pusilla (Hymenoptera, Meliponinae). Revista Brasileira de Genetica 18:181-184. Rasmussen C, Cameron SA. 2010. Global stingless bee phylogeny supports ancient divergence, vicariance, and long-distance dispersal. Biological Journal of the Linnean Society, 99: 206–232. Rocha MP, Pompolo SG. 1998. Karyotypes and hetero- chromatin variation (C-bands) in Melipona species (Hymenoptera, Apidae, Meliponinae). Genetics and Molecular Biology V. 21, p. 41-45. Rocha MP, Pompolo SG, Dergam JÁ, Fernandes A, Cam- pos LAO. 2002. DNA characterization and karyotypic evolution in the bee genus Melipona (Hymenoptera, Meliponini). Hereditas 136:19-37. Rocha MP, Pompolo SG, Campos LAO. 2003. Citogené- tica da tribo Meliponini (Hymenoptera, Apidae). In: Melo, G. A. R. and Santos I. A. (eds) Apoidea Neo- tropica: Homenagem aos 90 anos de Jesus Santiago Moure. UNESC, Criciúma, pp 311-320. Santos JM, Diniz, D, Rodrigues TAS, Cioffi, MB, Wald- schmidt, AM. 2018. Heterochromatin distribution and chromosomal mapping of microsatellite repeats in the genome of Frieseomelitta stingless bees (Hyme- noptera: Apidae: Meliponini). Florida Entomologist. 101(1):33-39. Schweizer D. 1980. Simultaneous fluorescent staining of R bands and specific heterochromatic regions (DA- DAPI bands) in human chromosomes. Cytogenetics and Cell Genetics 27:190-193. Silveira FA, Melo GAR, Almeida EAB. 2002. Abelhas Bra- sileiras – Sistemática e Identificação. Editora Com- posição e Arte, Minas Gerais, Brasil. 1ª edição. Pg 86. Smith F. 1854. Catalogue of Hymenopterous Insects in the Collection of the British Museum: Part II, Api- dae. Printed by Order of the Trustees, London. pp. 199-465. Schwarz HF. 1940. Additional Species and Records of Stingless Bees (Meliponidae) from British Guiana. American Museum Novitates, 1078: 1-12p. Travenzoli, N. M., Barbosa, I. C. de O., Carvalho-Zilse, G. A., Salomão, T. M. F., & Lopes, D. M. (2019) Karyotypic description and repetitive DNA chromo- some mapping of Melipona interrupta Latreille, 1811 (Hymenoptera: Meliponini). Caryologia 72(2), 91-95. Caryologia International Journal of Cytology, Cytosystematics and Cytogenetics Volume 73, Issue 2 - 2020 Firenze University Press The first molecular identification of Egyptian Miocene petrified dicot woods (Egyptians’ dream becomes a reality) Shaimaa S. Sobieh*, Mona H. Darwish Gene flow patterns reinforce the ecological plasticity of Tropidurus hispidus (Squamata: Tropiduridae) Fernanda Ito, Danielle J. Gama-Maia, Diego M. A. Brito, Rodrigo A. Torres* The technique of Plant DNA Barcoding: potential application in floriculture Antonio Giovino1,*, Federico Martinelli2,*, Anna Perrone3 Cytogenetic of Brachyura (Decapoda): testing technical aspects for obtaining metaphase chromosomes in six mangrove crab species Alessio Iannucci1, Stefano Cannicci1,2,*, Zhongyang Lin3, Karen WY Yuen3, Claudio Ciofi1, Roscoe Stanyon1, Sara Fratini1 Comparison of the Evolution of Orchids with that of Bats Antonio Lima-de-Faria Identification of the differentially expressed genes of wheat genotypes in response to powdery mildew infection Mehdi Zahravi1,*, Panthea Vosough-Mohebbi2, Mehdi Changizi3, Shahab Khaghani1, Zahra-Sadat Shobbar4 Populations genetic study of the medicinal species Plantago afra L. (Plantaginaceae) Saeed Mohsenzadeh*, Masoud Sheidai, Fahimeh Koohdar A comparative karyo-morphometric analysis of Indian landraces of Sesamum indicum using EMA-giemsa and fluorochrome banding Timir Baran Jha1,*, Partha Sarathi Saha2, Sumita Jha2 Chromosome count, male meiotic behaviour and pollen fertility analysis in Agropyron thomsonii Hook.f. and Elymus nutans Griseb. (Triticeae: Poaceae) from Western Himalaya, India Harminder Singh2, Jaswant Singh1,*, Puneet Kumar2, Vijay Kumar Singhal1, Bhupendra Singh Kholia2, Lalit Mohan Tewari3 Population genetic and phylogeographic analyses of Ziziphora clinopodioides Lam., (Lamiaceae), “kakuti-e kuhi”: An attempt to delimit its subspecies Raheleh Tabaripour1,*, Masoud Sheidai1, Seyed Mehdi Talebi2, Zahra Noormohammadi3 Induced cytomictic crosstalk behaviour among micro-meiocytes of Cyamopsis tetragonoloba (L.) Taub. (cluster bean): Reasons and repercussions Girjesh Kumar, Shefali Singh* Karyotype diversity of stingless bees of the genus Frieseomelitta (Hymenoptera, Apidae, Meliponini) Renan Monteiro do Nascimento1, Antonio Freire Carvalho1, Weyder Cristiano Santana2, Adriane Barth3, Marco Antonio Costa1,* Karyotype studies on the genus Origanum L. (Lamiaceae) species and some hybrids defining homoploidy Esra Martin1, Tuncay Dirmenci2,*, Turan Arabaci3, Türker Yazici2, Taner Özcan2 Determination of phenolic compounds and evaluation of cytotoxicity in Plectranthus barbatus using the Allium cepa test Kássia Cauana Trapp1, Carmine Aparecida Lenz Hister1, H. Dail Laughinghouse IV2,*, Aline Augusti Boligon1, Solange Bosio Tedesco1