Kent Academic Repository Full text document (pdf) Citation for published version Furo, Ivanete de Oliveira and Kretschmer, Rafael and O’Brien, Patricia Caroline and Pereira, Jorge C. and Garnero, Analía del Valle and Gunski, Ricardo José and O’Connor, Rebecca E. and Griffin, Darren K. and Gomes, Anderson José Baia and Ferguson-Smith, Malcolm Andrew and de Oliveira, Edivaldo Herculano Correa (2020) Chromosomal Evolution in the Phylogenetic DOI https://doi.org/10.3389/fgene.2020.00721 Link to record in KAR https://kar.kent.ac.uk/82591/ Document Version Publisher pdf Copyright & reuse Content in the Kent Academic Repository is made available for research purposes. Unless otherwise stated all content is protected by copyright and in the absence of an open licence (eg Creative Commons), permissions for further reuse of content should be sought from the publisher, author or other copyright holder. Versions of research The version in the Kent Academic Repository may differ from the final published version. Users are advised to check http://kar.kent.ac.uk for the status of the paper. Users should always cite the published version of record. Enquiries For any further enquiries regarding the licence status of this document, please contact: [email protected] If you believe this document infringes copyright then please contact the KAR admin team with the take-down information provided at http://kar.kent.ac.uk/contact.html fgene-11-00721 July 9, 2020 Time: 17:7 # 1 ORIGINAL RESEARCH published: 10 July 2020 doi: 10.3389/fgene.2020.00721 Chromosomal Evolution in the Phylogenetic Context: A Remarkable Karyotype Reorganization in Neotropical Parrot Myiopsitta monachus (Psittacidae) Ivanete de Oliveira Furo1,2,3, Rafael Kretschmer4,5, Patricia Caroline O’Brien3, Jorge C. Pereira6, Analía del Valle Garnero7, Ricardo José Gunski7, Rebecca E. O’Connor5, Darren Karl Griffin5, Anderson José Baia Gomes8, Malcolm Andrew Ferguson-Smith3 and Edivaldo Herculano Correa de Oliveira1,2,9* 1 Instituto de Ciências Biológicas, Universidade Federal do Pará, Belém, Brazil, 2 Laboratório de Cultura de Tecidos e Edited by: Citogenética, Seção de Meio Ambiente, Instituto Evandro Chagas, Ananindeua, Brazil, 3 Department of Veterinary Medicine, Roberto Ferreira Artoni, Cambridge Resource Centre for Comparative Genomics, University of Cambridge, Cambridge, United Kingdom, Universidade Estadual de Ponta 4 Departamento de Genética, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil, 5 School of Biosciences, Grossa, Brazil University of Kent, Canterbury, United Kingdom, 6 Animal and Veterinary Research Centre (CEVAV), University of Tràs-os-Montes and Alto Douro (UTAD), Vila Real, Portugal, 7 Laboratório de Diversidade Genética Animal, Universidade Reviewed by: Federal do Pampa, São Gabriel, Brazil, 8 Instituto Federal do Pará, Abaetetuba, Brazil, 9 Instituto de Ciências Exatas e Daniel Pacheco Bruschi, Naturais, Universidade Federal do Pará, Belém, Brazil Federal University of Paraná, Brazil Alexandr Sember, Academy of Sciences of the Czech Myiopsitta monachus is a small Neotropical parrot (Psittaciformes: Arini Tribe) from Republic (ASCR), Czechia subtropical and temperate regions of South America. It has a diploid chromosome *Correspondence: Edivaldo Herculano Correa de number 2n = 48, different from other members of the Arini Tribe that have usually Oliveira 70 chromosomes. The species has the lowest 2n within the Arini Tribe. In this [email protected] study, we combined comparative chromosome painting with probes generated from Specialty section: chromosomes of Gallus gallus and Leucopternis albicollis, and FISH with bacterial This article was submitted to artificial chromosomes (BACs) selected from the genome library of G. gallus with the Evolutionary and Population Genetics, aim to shed light on the dynamics of genome reorganization in M. monachus in a section of the journal Frontiers in Genetics the phylogenetic context. The homology maps showed a great number of fissions in Received: 02 March 2020 macrochromosomes, and many fusions between microchromosomes and fragments Accepted: 15 June 2020 of macrochromosomes. Our phylogenetic analysis by Maximum Parsimony agree with Published: 10 July 2020 molecular data, placing M. monachus in a basal position within the Arini Tribe, together Citation: Furo IO, Kretschmer R, with Amazona aestiva (short tailed species). In M. monachus many chromosome O’Brien PC, Pereira JC, Garnero AV, rearrangements were found to represent autopomorphic characters, indicating that Gunski RJ, O’Connor RE, Griffin DK, after this species split as an independent branch, an intensive karyotype reorganization Gomes AJB, Ferguson-Smith MA and de Oliveira EHC (2020) Chromosomal took place. In addition, our results show that M. monachus probes generated by Evolution in the Phylogenetic Context: flow cytometry provide novel cytogenetic tools for the detection of avian chromosome A Remarkable Karyotype Reorganization in Neotropical Parrot rearrangements, since this species presents breakpoints that have not been described Myiopsitta monachus (Psittacidae). in other species. Front. Genet. 11:721. doi: 10.3389/fgene.2020.00721 Keywords: Myiopsitta, Arini tribe, phylogenetic, karyotype, rearrangements, breakpoints Frontiers in Genetics| www.frontiersin.org 1 July 2020| Volume 11| Article 721 fgene-11-00721 July 9, 2020 Time: 17:7 # 2 Furo et al. Karyotypic Reorganization in Myiopsitta monachus INTRODUCTION chromosome number among the Arini Tribe (Neotropical Psittacidae) and, together with the African species Agapornis The order Psittaciformes comprises some 330–350 species, roseicollis, has the lowest chromosome complement in the grouped into three families according to a new classification: order (Nanda et al., 2007). Furthermore, M. monachus also Strigopidae, Cacatuidae and Psittacidae (Rheindt et al., atypical sex chromosomes, with an enlarged W chromosome, 2014; Winkler et al., 2015). Within the Psittaciformes, the similar in size and morphology to the Z. These features are earliest divergence was between the New Zealand-restricted uncommon among Neornithes birds, which generally present Strigopidae family (Kakapo, Kea, and Kaka) and other parrots, heteromorphic sex chromosomes. The enlargement of the W is followed by the divergence between the Australasian cockatoos due to the accumulation and amplification of repetitive DNA (Cacatuidae) and remaining parrots (Tavares et al., 2006; (Furo et al., 2017). Wright et al., 2008). Recently, some studies have focused on A study by Tavares et al.(2006) based on mitochondrial and the phylogenetic reconstruction of this order based on ancient nuclear sequences, placed M. monachus in a clade together with geographic considerations. Wright et al.(2008) supported the Amazona, Pionus, Pionopsitta, Triclaria, and Graydidascalus, hypothesis of an Australasian origin, during the cretaceous despite the fact that the M. monachus karyotype has been in Gondwana and afterward the division of the African and characterized only by Giemsa staining and microsatellite India/Madagascar block, with subsequent diversification through mapping (Furo et al., 2017). vicariance and dispersal. Hence, in view of the value of whole chromosome painting in Some studies have proposed the division of Psittacidae family the improved understanding of chromosomal evolution, and the into Old World and New World parrots (Miyaki et al., 1998). fact that M. monachus has an atypical chromosome complement According to Tavares et al.(2006), the New world Parrots in which the homologies with other species is still unknown, the share a much earlier common ancestor with Australian parrots aim of this study has been (a) to investigate the mechanisms (59 million years ago, Mya), prior to the separation of Australia underlining the origin of the low 2n of M. monachus by using from Antarctica and South America. GGA BAC probes and whole chromosome painting probes from Concerning chromosomal data, New World parrots (Arini G. gallus and L. albicollis; (b) to compare the karyotypes of Tribe) show a karyotype homogeneity when compared to Old M. monachus and members of Psittaciformes, to better determine World species, with a constant diploid chromosome number their phylogenetic relationships. Additionally, we show that the (2n = 70) (Caparroz and Duarte, 2004). However, some species use of probes derived from species with highly derived karyotypes have atypical diploid chromosome numbers, as low as 2n = 48 such as M. monachus provides greater resolution in detecting in Myiopsitta monachus (Furo et al., 2017) and 2n = 64 in interchromosomal rearrangements. Graydidascalus brachyurus (Caparroz and Duarte, 2004), or higher, as in Forpus xanthopterygius and Brotogeris versicolurus with 2n = 86 and 82, respectively (de Lucca et al., 1991; MATERIALS AND METHODS de Lucca and Marco, 2014). Advances in molecular cytogenetics have shown that Samples, Cell Culture, and Chromosome ancestral macrochromosomes have been highly rearranged in Preparation Psittaciformes (Nanda et al., 2007; Furo et al., 2015a, 2018). The experiments followed ethical protocols and were approved Hence, although chromosome painting with probes derived by the ethics committee (CEUA-Universidade Federal do Pará) from chicken Gallus gallus (GGA) revealed three associations under no. 170/2013 and SISBIO 68443-1. Metaphases were shared by all species of Psittaciformes analyzed so far – GGA1/4, obtained from fibroblast culture of skin biopsies and feather GGA6/7, and GGA8/9 (Seabury et al.,