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      Is the Karyotype of Neotropical Boid Snakes Really Conserved? Cytotaxonomy, Chromosomal Rearrangements and Karyotype Organization in the Boidae Family

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          Abstract

          Boids are primitive snakes from a basal lineage that is widely distributed in Neotropical region. Many of these species are both morphologically and biogeographically divergent, and the relationship among some species remains uncertain even with evolutionary and phylogenetic studies being proposed for the group. For a better understanding of the evolutionary relationship between these snakes, we cytogenetically analysed 7 species and 3 subspecies of Neotropical snakes from the Boidae family using different chromosomal markers. The karyotypes of Boa constrictor occidentalis, Corallus hortulanus, Eunectes notaeus, Epicrates cenchria and Epicrates assisi are presented here for the first time with the redescriptions of the karyotypes of Boa constrictor constrictor, B. c. amarali, Eunectes murinus and Epicrates crassus. The three subspecies of Boa, two species of Eunectes and three species of Epicrates exhibit 2n = 36 chromosomes. In contrast, C. hortulanus presented a totally different karyotype composition for the Boidae family, showing 2n = 40 chromosomes with a greater number of macrochromosomes. Furthermore, chromosomal mapping of telomeric sequences revealed the presence of interstitial telomeric sites (ITSs) on many chromosomes in addition to the terminal markings on all chromosomes of all taxa analysed, with the exception of E. notaeus. Thus, we demonstrate that the karyotypes of these snakes are not as highly conserved as previously thought. Moreover, we provide an overview of the current cytotaxonomy of the group.

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          Most cited references25

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          Improved telomere detection using a telomere repeat probe (TTAGGG)n generated by PCR.

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            Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization.

            This report describes the use of fluorescence in situ hybridization for chromosome classification and detection of chromosome aberrations. Biotin-labeled DNA was hybridized to target chromosomes and subsequently rendered fluorescent by successive treatments with fluorescein-labeled avidin and biotinylated anti-avidin antibody. Human chromosomes in human-hamster hybrid cell lines were intensely and uniformly stained in metaphase spreads and interphase nuclei when human genomic DNA was used as a probe. Interspecies translocations were detected easily at metaphase. The human-specific fluorescence intensity from cell nuclei and chromosomes was proportional to the amount of target human DNA. Human Y chromosomes were fluorescently stained in metaphase and interphase nuclei by using a 0.8-kilobase DNA probe specific for the Y chromosome. Cells from males were 40 times brighter than those from females. Both Y chromosomal domains were visible in most interphase nuclei of XYY amniocytes. Human 28S ribosomal RNA genes on metaphase chromosomes were distinctly stained by using a 1.5-kilobase DNA probe.
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              Distribution of non-telomeric sites of the (TTAGGG)n telomeric sequence in vertebrate chromosomes.

              The intrachromosomal distribution of non-telomeric sites of the (TTAGGG)n telomeric repeat was determined for 100 vertebrate species. The most common non-telomeric location of this sequence was in the pericentric regions of chromosomes. A variety of species showed relatively large amounts of this sequence present within regions of constitutive heterochromatin. We discuss possible relationships between the non-telomeric distribution of the (TTAGGG)n sequence and the process of karyotype evolution, during which these sites may provide potential new telomeres.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                5 August 2016
                2016
                : 11
                : 8
                : e0160274
                Affiliations
                [1 ]Instituto Nacional de Pesquisas da Amazônia, Coordenação de Biodiversidade, Av. André Araujo 2936, Petrópolis, CEP: 69067-375 Manaus, AM, Brazil
                [2 ]Universidade Federal do Amazonas, Instituto de Ciências Biológicas, Rua General Rodrigo Otávio Num. 3000, Mini-Campus Coroado, CEP: 66077070 Manaus, AM, Brazil
                [3 ]Faculdades Integradas do Tapajós, Laboratório de Pesquisas Zoológicas, Santarém, PA, Brazil
                [4 ]Criadouro Comercial Jiboias Brasil, Belo Horizonte, MG, Brazil
                Institute of Cotton Research of Chinese Academy of Agricultural Sciences, CHINA
                Author notes

                Competing Interests: George Myller Souza is employed by Criadouro Comercial Jiboias Brasil. There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials.

                • Conceptualization: PFV.

                • Methodology: PFV LBR EF GMS HMC MCG.

                • Visualization: PFV LBR EF GMS HMC MCG.

                • Writing - original draft: PFV.

                • Writing - review & editing: PFV LBR EF GMS HMC MCG.

                Article
                PONE-D-16-02302
                10.1371/journal.pone.0160274
                4975421
                27494409
                c7d62bf3-fdb1-4d87-819a-b27a8ab4af88
                © 2016 Viana et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 18 January 2016
                : 15 July 2016
                Page count
                Figures: 7, Tables: 1, Pages: 16
                Funding
                This work was supported by the Pronex/FAPEAM/CNPq 003/2009 (to EF) and MCT/CNPq/MEC/CAPES/FNDCT – Cross Action/FAPs No. 47/2010 – BioPHAM Network, and the CAPES – Pro-Amazon Program: Biodiversity and Sustainability, Public Notice No. 047/2012. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. George Myller Souza is employed by Criadouro Comercial Jiboias Brasil. Criadouro Comercial Jiboias Brasil provided support in the form of salary for author GMS, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific role of this author is articulated in the ‘author contributions’ section.
                Categories
                Research Article
                Biology and Life Sciences
                Genetics
                Cytogenetics
                Karyotypes
                Biology and Life Sciences
                Organisms
                Animals
                Vertebrates
                Amniotes
                Reptiles
                Squamates
                Snakes
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
                Gene Mapping
                Chromosome Mapping
                Research and Analysis Methods
                Molecular Biology Techniques
                Gene Mapping
                Chromosome Mapping
                Biology and Life Sciences
                Cell Biology
                Chromosome Biology
                Chromosomes
                Sex Chromosomes
                Research and Analysis Methods
                Specimen Preparation and Treatment
                Staining
                Chromosome Staining
                Biology and Life Sciences
                Cell Biology
                Chromosome Biology
                Chromatin
                Heterochromatin
                Biology and Life Sciences
                Genetics
                Epigenetics
                Chromatin
                Heterochromatin
                Biology and Life Sciences
                Genetics
                Gene Expression
                Chromatin
                Heterochromatin
                Research and Analysis Methods
                Cytogenetic Techniques
                C Banding
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
                Sequencing Techniques
                Sequence Analysis
                Sequence Motif Analysis
                Research and Analysis Methods
                Molecular Biology Techniques
                Sequencing Techniques
                Sequence Analysis
                Sequence Motif Analysis
                Custom metadata
                All relevant data are within the paper.

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                Uncategorized

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