Genetic diversity of species of the genus Gobio the Tisza River drainage area, Slovakia

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Genetic diversity of species of the genus Gobio
(the Tisza River drainage area, Slovakia)

• 1Mendel, J., 1Lusková, V., 1Halacka, K., 2Koco,
  J., 1Lusk, S.
• 1Academy of Science of the Czech Republic,
  Institut of Vertebrate Biology, Department of
  Ichthyology, Kvetná 8, 603 65, Brno, Czech
  Republic
• 2Department of Ekology FHPV, Preov University,
  17. Novembra 1 081 16 Slovakia

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Aims

• actual distribution of the Gobio species in
  eastern Slovakia
• detection of intraspecific diversity
• relationship of species genus Gobio
• conservation
• council directive No 92/43/EEC annex II
  Gobio albipinnatus, Gobio uranoscopus, Gobio
  kesslerii NATURA 2000 network

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Methods

• electrofishing
• standard karyological methods (kidney tissue)
• starch gel electrophoresis (skeletal muscle,
  
• liver tissue)
• direct sequencing of D-loop mtDNA
• software packages (Genetix, Mega, DnaSP,
• Bioedit etc)

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G.albipinnatus
recent
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G.kesslerii
recent
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G.uranoscopus
recent
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Summary table partial differences exist between
single species (the results more or less
correspond with the literature dates)
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G.gobio
M 22 SM 24 ST 2 A 2
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G.albipinnatus
M 24 SM 24 ST 0 A 2
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One asymmetrical pair of the chromosomes -
cytotype polymorphism SM/A
G.kesslerii
M 24 SM 24 1 ST 0 A 0 1
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G.uranoscopus
M 22 SM 26 ST 2 A 0
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• for allozyme characteristics of the gudgeon
  species was used starch gel electrophoresis of
  tissue extracts
• four buffer systems
• enzyme specific staining of gels
• 14 enzyme systems
• products of 20 loci
• basic genetic characterization of populations
• genetic similarities

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Genetic variability of Slovak populations of
genus Gobio
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Distinguishing and rare alleles in four species
of gudgeons (part 1.)
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Distinguishing and rare alleles in four species
of gudgeons (part 2.)
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Gobio kesslerii
Gobio uranoscopus
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Method of direct sequencing

• Purification of PCR product
• Sequencing PCR
• Four terminators (ddNTPs-dideoxynucleotidtriphosph
  ate) carry other fluorescence mark on 3end
• terminators incorporated currently into growing
  chains chain terminated and given mark
  respective to given base
• Created fragments are divided using capillary
  electrophoresis in genetic analyser
• CCD camera of analyser takes then the
  fluorescence, brought about by laser ray, from
  all four marks simultaneously because emission
  maxima are of defferent wave lengths, all
  fragments can be analysed simultaneously.

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The localities of samplings
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Variable nucleotide and amino acid positions for
of D-loop
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Intrapopulation and interpopulation variability

• Intrapopulation diversity (DNAdist,Kimura -2
  parameter)
• G. uranoscopus Laborec (0,001)
• G. Kesslerii Laborec (0,001), Morava-Dub
  (0,005), Topla (0,013)
• G. albipinnatus Dnieper (0,002), Topla
  (0,001), Dniester (0,001), Revitia (0,001)
• G. gobio Topla (0,000), Revitia (0,017),
  Belanský p. (0,002), Laborec (0,006), S. Bug
  (0,002)

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Haplotypes and intrapopulation variability
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Phylogenetic tree I
UPGMA method (Kimura -2-parameter)
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Phylogenetic tree II
Neighbor-joining method (Kimura -2-parameter )
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The mean interspecific divergence of D-loop
sequences was G. gobio vs. G. albipinnatus 4.5
(excluding pop. Dniester) G. gobio vs G.
uranoscopus 8.1 G. gobio vs. G. kesslerii 6.0
G. albipinnatus vs. G. uranoscopus 5.6 G.
albipinnatus vs. G. kesslerii 5.0 G.
uranoscopus vs. G. kesslerii 7.0
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