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ESTUDOS CITOGENÉTICOS EM ESPÉCIES DE PEIXES DE RIACHOS: Hyphessobrycon vinaceus, Bryconamericus aff. iheringii E Odontostilbe pequira (PISCES: CHARACIDAE)

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A família Characidade representa a maior e mais diversificada família de peixes de água doce, na qual se encontram muitos gêneros de pequeno porte. Neste estudo foram analisados, cromossomicamente, exemplares de Hyphessobrycon vinaceus, Bryconamericus aff. iheringii e Odontostilbe pequira. H. vinaceus apresentou 2n=50, sendo 8m, 12sm e 30a, com evidente proporção de ST/A, AgNOR-múltipla e blocos de heterocromatina na porção intersticial. B. aff. iheringii apresentou 2n=52, com 12m, 18sm, 8st e 14a, AgNOR-simples e blocos de heterocromatina pericentroméricos. O. pequira apresentou 2n= 52, sendo 14m, 20sm, 14st e 4a, com determinação cromossômica sexual do tipo ZZ/ZW e sistema de AgNOR-simples, além de blocos de heterocromatina na posição pericentromérica. A diversidade cariotípica, encontrada em peixes de pequeno porte, tem sido explicada através de mecanismos de rearranjos cromossômicos e que, provavelmente, tenham se fixado devido às características biológicas dessas espécies, que ocupam, preferencialmente, cabeceiras de rios e riachos, formando pequenas populações.
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Evolução e Conservação da Biodiversidade Rio Paranaíba, Vol. 6 Nº 1| 13-22| jan-jul 2015
ISSN 2236-3866
DOI: 10.7902/ecb.v6i1.73
Acesso livre em www.simposiodabiodiversidade.com.br/ecb
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ESTUDOS CITOGENÉTICOS EM ESPÉCIES DE PEIXES DE RIACHOS: Hyphessobrycon vinaceus,
Bryconamericus . iheringii E Odontoslbe pequira (CHARACIFORMES: CHARACIDAE)
CYTOGENETIC STUDIES IN SPECIES OF STREAM FISHES: Hyphessobrycon vinaceus, Bryconamericus .
iheringii AND Odontoslbe pequira )
Patricia Belini Nishiyama1; Margarida Maria Vieira Rossi2; Fernanda Errero Porto3, Luciana Andreia Borin3, Ana
Luiza de Brito Portela Castro33







Resumo

      
               
Hyphessobrycon vinaceus, Bryconamericus   iheringii e Odontoslbe pequira, 
             
  Hyphessobrycon vinaceus,
Bryconamericus  iheringii e Odontoslbe pequira. H. vinaceus
Bryconamericus.  iheringii
          
O. pequira
              
              
             

Palavras-chavein situ,
Abstract
 
           
Hyphessobrycon vinaceus, Bryconamericusiheringii and
Odontoslbe pequira
Hyphessobrycon vinaceus, Bryconamericus
 iheringii and Odontoslbe pequira. H. vinaceus 
Bryconamericus  iheringii
    O. pequira 


Evolução e Conservação da Biodiversidade Rio Paranaíba, Vol. 6 Nº 1| 13-22| jan-jul 2015
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


Palavras-chavein situ
Introdução
      
          

        

      
uma enorme diversidade na morfologia, habitats que

 




     
        
Hyphessobrycon, Bryconamericus
e Odontoslbe     
       

    
      

      
    
     
  
     
  

    
        
       
 



      
   
Hyphessobrycon,
Bryconamericus e Odontoslbe.
Considerando a escassez de estudos sobre
       

    
Hyphessobrycon vinaceus, Bryconamericus  iheringii
e Odontoslbe pequira,   
 
contribuindo com melhor entendimento de suas

Material e Métodos
    
  Hyphessobrycon vinaceus 
da nascente do Rio Verruga, Bacia do Rio Pardo, BA
Bryconamericus   iheringii,
        0
540 Odontoslbe pequira, do Córrego do



      

     
       
      
       
      
     
       
Evolução e Conservação da Biodiversidade Rio Paranaíba, Vol. 6 Nº 1| 13-22| jan-jul 2015
15
       
      


       
      
      
     
submetacêntricos (sm), subtelocêntricos (st) e
  
    
  
     
 in situ     
         
  Oreochromis nilocus   
  
     

Resultados
Hyphessobrycon vinaceus  


in situ
      
        
       

     
     
     

Figura 1:in situ com
Hyphessobrycon vinaceus
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Bryconamericus  iheringii mostrou
      



       
      
      



Odontoslbe pequira  
     
        
    
      
      
      

        
 
    
      

 

Figura 2: in situ
Bryconamericus a. iheringii
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
   
  


Discussão
    
Hyphessobrycon vinaceus    

de 50 cromossomos, o qual tem sido o mais frequente

      
       
     
    
       
H. vinaceus
Figura 3: in situ
Odontoslbe pequira 
Evolução e Conservação da Biodiversidade Rio Paranaíba, Vol. 6 Nº 1| 13-22| jan-jul 2015
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 
     




        

      

Em H. vinaceus     

     
     



  

  


      
       
     
    

Bryconamericus iheringii   
   
      

       
       
      
     
        
  Bryconamericus iheringii    
     

       
       
     
      
      
      
       
     
   
       
     Bryconamericus, analisadas


Bryconarericus  iheringii   


     
     
   


    Bryconamericus
  ,    Bryconamericus   
          
Bryconamericus ecai
  

Bryconamericus iheringii 

     


de Bryconamericus inheringii  
       
       
       
   
  
 
      
       

Em Odontoslbe pequira


Evolução e Conservação da Biodiversidade Rio Paranaíba, Vol. 6 Nº 1| 13-22| jan-jul 2015
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  
      
     

foram observadas entre O. pequira   
       
         
     
        
     
      
        
      Serrapinus
heterodon    
     
    

Em Odontoslbe pequira    
     


      
     
     
     
        



 Serrapinus notomelas


      


      





       
bem caracterizado em O. pequira, analisada neste


     
    
  Odontoslbe paranensis e Holoshestes
heterodon     Odontoslbe
microcephala   Serrapinus notomelas e
Serrapinus
    
em O. pequira    
         
       
       
     
    
       

     

     

    
       
     
     
    

Agradecimentos



Referências
     
  Hopliaslacerdae
  , 1:

       
 
Evolução e Conservação da Biodiversidade Rio Paranaíba, Vol. 6 Nº 1| 13-22| jan-jul 2015
20
Journal Fish Biology 76:25712577
 JE,   Menezes NA (1978) Estado

Acta Amazônica
  Portela-Castro ALB, JulioJr  (2008)
     
Bryconamericus iheringii  



       

Caryologia

Castro RMC (1999)   
sul-americanos: padrões gerais e possíveis processos
causais     
       
      

Centofante L, Bertollo AC, Moreira-Filho

  Hyphessobrycon anisitsi (Pisces,
Cytologia
     

     
         Astyanax
alparanae    
       
Molecular Biology
     Peixes da Planície de
Inundação do Alto Rio Paraná e Áreas Adjacentes

    
      
   , 36:

    Pardue ML   
     
Chromosome 53: 25–36

Hereditas.52

       
       
  Parauchenipterus galeatus
       
Micron
 E, Alves AL,    F, 
      
 Astyanaxalparanae,  
2000 and Hyphessobrycon eques  
     


Incertae Sedis


Molecular Research
       
     
      
 
Research
Nelson GJ (2006) Fishes of the word, 4a

       
     
Fish Physiol Biochem, 35:81–100
    
    Bryconamericus exodon
  
Hereditas
     
Evolução e Conservação da Biodiversidade Rio Paranaíba, Vol. 6 Nº 1| 13-22| jan-jul 2015
21
     
of Bryconamericus  iheringii (Characidae,
 ,

      
      
genus Serrapinnus   
       S.
heterodonCaryologia
      
        
     

Gray
   
Proc Natl Acad Sci
      
     
   

     
      
Moenkausiasanctalomenae (Pisces, Characidae) from


Portela-Castro ALB,    
    
   
Cytologia
     IC,
Pavanelli      
Bryconamericus iheringii  
       

      
    
Heredity, 108, 1–3

      
      
Serrapinus (Characidae,
 


     
nucleolus organizer regions in Serrapinnusnotomelas
     
Journal of Fish Biology 

  
chromosome in Bryconamericus ecai

Hereditas
       

Characidae) da Bacia do Rio Paraná  
      


     
Astyanax scabripinnis
Cytologia

,

       
    
     
   

       
Heredity
     
     
of the genus Bryconamericus Brazilian Journal of

Evolução e Conservação da Biodiversidade Rio Paranaíba, Vol. 6 Nº 1| 13-22| jan-jul 2015
22
      
   Bryconamericus (Characidae,
Cytobios
      
     Bryconamericus (Characidae,
Cytologia

      
Chromosome Science
O periódico Evolução e Conservação da Biodiversidade, ISSN 2236-3866, foi licenciado com uma Licença
Creative Commons - Atribuição - NãoComercial - SemDerivados 3.0 Não Adaptada.
Com base no trabalho disponível em www.simposiodabiodiversidade.com.br/ecb.
DOI: 10.7902/issn.2236-3866
Article
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The chromosomal location of 5S rRNA and U2 snRNA genes of Piabina argentea, Piabarchus stramineus and two Bryconamericus species from two different Brazilian river basins were investigated, in order to contribute to the understanding of evolutionary characteristics of these repetitive DNAs in the subfamily Stevardiinae. The diploid chromosome number was 2n = 52 for Bryconamericus cf. iheringii, Bryconamericus turiuba, Piabarchus stramineus and Piabina argentea. The 5S rDNA clusters were located on one chromosome pair in P. stramineus and B. cf. iheringii, and on two pairs in B. turiuba and P. argentea. The U2 snDNA clusters were located on the one pair in all species. Two-color FISH experiments showed that the co-localization between 5S rDNA and U2 snDNA in P. stramineus can represent a marker for this species. Thus, the present study demonstrated that the number of U2 snDNA clusters observed for the four species was conserved, but particular characteristics can be found in the genome of each species.
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Cheirodontinae constitutes a group of 17 genera and 55 species still poorly studied cytogenetically, being that most data available in the literature is for the Alto Paraná basin. In this study Odontostilbe pequira, Serrapinnus kriegi, Serrapinnus calliurus and Serrapinnus microdon from Alto Paraguay basin was characterized cytogenetically. The modal diploid number 2n=52 found for these species, as well as most of the data cited in the literature, is an evidence of a conservadorism in chromosome number for the group. Keywords: Cytotaxonomy. Karyotypic macrostructure. Pantanal
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Fluorescence in situ hybridization (FISH) was undertaken in order to determinate the chromosomal distribution pattern of 18S and 5S ribosomal DNAs (rDNA) in four populations of the characid fish Astyanax altiparanae from the upper Paraná river basin, Brazil. The 18S rDNA probe FISH revealed numerical and positional variations among specimens from the Keçaba stream compared to specimens of the other populations studied. In contrast to the variable 18S rDNA distribution pattern, highly stable chromosomal positioning of the 5S rDNA sites was observed in the four A. altiparanae populations. Divergence in the distribution pattern of 18S and 5S rDNA sites is also discussed.
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Chromosomal data of Hyphessobrycon anisitsi are presented for the first time, comprising 2 populations from adjacent hydrographical basins (Upper Paraná basin and Paraíba do Sul basin), separated by Mantiqueira Hills (water divisor). The same diploid number and karyotype formula were observed in both populations. However, after silver nitrate staining (Ag-NORs) and fluorescent in situ hybridization with 18S rDNA and 5S rDNA probes, it was possible to identify each population by the location of ribosomal sites. Some aspects related to geographical isolates and chromosomal differentiation are approached.
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A remarkable karyotype diversity was reported in five species of the genus Bryconamericus which are still awaiting more precise taxonomic identification. Bryconamericus sp. A, Bryconamericus sp. B, Bryconamericus sp. C and Bryconamericus sp. D had a diploid number equal to 52 chromosomes but their karyotypes varied widely. Bryconamericus sp. E presented a diploid number of 54 chromosomes. Pericentric inversions and heterochromatin variations are the major mechanisms of chromosome diversification among Bryconamericus species. A chromosome fission may have arisen in a more derivative karyotype (2n = 54) observed in Bryconamericus sp. E.
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Cytogenetic studies were performed on two sympatric species of Bryconamericus which are awaiting more precise taxonomic identification. Although both Bryconamericus sp A and Bryconamericus sp B had a chromosome number equal to 52 chromosomes, their karyotypic structures varied widely. The two species presented multiple nucleolar organizer regions (NORs), at times varying in size. Centromeric heterochromatin was the most frequent type in the species studied and C banding showed no significant differences between karyotypes. The data permitted us to propose that non-Robertsonian chromosome rearrangements of the pericentric inversion type led to the differences between the two species of Bryconamericus.
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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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An extensive NOR variability was described in four Bryconamericus species which are still awaiting more precise taxonomic identification. Nine NOR phenotypes were identified by silver nitrate and mithramycin staining, and cytotaxonomy and phylogenetic relationships were inferred. The wide NOR polymorphism probably related to differences in the number of tandemly repeat ribosomic cistrons and rDNA transcriptional regulation thus far may characterize a particular situation of higher level of NOR phenotype diversification in Bryconamericus than in other neotropical genus fish.
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The family Characidae, one of the largest and more diversified fish families in the Neotropical region, is certainly not a monophyletic group, which is supported by the cytogenetic data showing its extensive chromosome variability. In this work, two species of the genus Serrapinnus, S. heterodon and S. piaba, from the São Francisco river basin were analyzed. Both species presented the same diploid number, 2n=52 chromosomes, and the same karyotypic formula, 16m+20sm+14st+2a. The heterochromatin distribution, evidenced by C-banding, was also similar between both species. On the other hand, two NORs were evidenced in S. heterodon, while a multiple NOR system was characterized in S. piaba, both confirmed by silver nitrate staining as well as by fluorescent in situ hybridization with 18S rDNA probe. Furthermore, two additional karyotypes (cytotypes) were observed in S. heterodon, with 15m+20sm+14st+3a and 17m+20sm+14st+la chromosomes, respectively, characterizing a structural chromosome polymorphism in the population. The differences between the three S. heterodon, karyotypic forms are probably due to pericentric inversions related to the 8th and 26th chromosome pairs.
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Studies on different populations of the Characidae fish Astyanax scabripinnis, a species complex, have shown the occurrence of many levels of cytogenetic polymorphisms, such as in the positioning and amount of constitutive heterochromatins, and in the localization of nucleolar rDNA sites. The present paper describes the wide polymorphisms and the differences in location and quantity of heterochromatins in 2 A. scabripinnis populations, which are isolated from each other by waterfalls along the Canta Galo stream (São Paulo State, Brazil). Samples from both populations presented the same karyotype formulae. The specimens from the population located at 980 m of altitude revealed few variations in heterochromatins and NORs sites. On the other hand, those specimens collected from 720 m showed high variability in positioning and number of C-bands and rDNA loci. Events of transposition between telomeric segments, preferential positioning of the nucleolar rDNA clusters and the presence of GC-rich heterochromatins are also discussed.