Genetic characteristics of Perccottus glenii from the River Tisza Slovakia

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Genetic characteristics of Perccottus glenii from
the River Tisza (Slovakia)

• 1Lusková, V., 1Mendel, J., 1Halacka, K., 2Koco,
  J.
• 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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Introduction of P.g. from the native area to
Europe
1912
1948
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Distribution of P.g. in east Slovakia
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S
St.Maria - Somotor locality, channel
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St.Maria - Somotor locality, backwater
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Cíle

• Research of the population diversity in P.g. by
• classic karyology
• allozyme analyses by starch gel electrophoresis
• direct sequencing of cytb for molecular genetic
  study

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Lokalities of the karyological observation of P.g.
Khabarovsk 14 spec.
Moskow 13 spec.
Slovakia 4 spec.
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KHABAROVSK
2n 44 44 subtelo- and akrocentric chromosomes
MOSKOW

• 2n 44 (two types)
• 44 subtelo- and akrocentric chromosomes
• 43 subtelo- and akrocentric
• 1 submetacentric chromosomes

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St.Maria-Somotor
2n 44 2 submetacentric 32 subtelocentric 10
acrocentric chromosomes
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Khabarovsk
Moskow
St.Maria
Original state
Genesis of the chromosome aberations
Expansion of this aberation and fixation in
populations by their pairing
Problems
- too little dates - accurate classification
chromosomes
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Allozyme patterns of polymorphic loci
(electrophoresis) in P.g.
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Electrophoretic analysis of Amur sleeper - enzyme
loci, condition
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Fis parameter and its significance in polymorphic
loci
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Allelic frequences and Fis index
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Molecular method

• isolation of DNA from pectoral fins by the
  phenol-chloroform-isoamylalcohol method
• selection of the sequence of mtDNA cytb 545 bp
  in 5 specimens
• designing of primers by OLIGO program - for
  define of the length of fragment analysed
• primer sequences
• Cyt bL 5- TAA TGG CCA GCC TAC GAA A- 3 (19
  bp)
• Cyt bU 5- GGA AGG TAA AGA AAC GGG TAA- 3 (21
  bp)
• analyse of sequence on the genetic analyser ABI
  PRISM 310
• statistical evaluation (MEGA,DnaSP 3.99, BioEdit,
  Lasergene)

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Table DNA distances
1 2 3
4 5 6 7 8 9 10 11 1
Perc5 2 Perc3 0.009 3 Perc1
0.000 0.009 4 Perc2 0.009 0.000 0.009 5
Perc4 0.000 0.009 0.000 0.009 6
O.obscura 0.218 0.220 0.218 0.220 0.218 7
M.swinhonis 0.211 0.213 0.211 0.213 0.211
0.266 8 E.acanthopoma 0.273 0.273 0.273 0.273
0.273 0.259 0.307 9 H.temporalis 0.184 0.196
0.184 0.196 0.184 0.238 0.240 0.238 10 G.gobio
0.302 0.305 0.302 0.305 0.302 0.330 0.287
0.284 0.250 11 P.fluviatilis 0.251 0.251 0.251
0.251 0.251 0.281 0.240 0.279 0.240 0.248

• Distance method Nucleotide Kimura 2-parameter
  Between group average

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Phylogenetic tree I

• constructed by Neighbor-joining method.
  Separation of samples P. glenii (bootstraping
  100) with agreement their haplotypes.
  Quantification of relationship of different
  families.

Neighbor-joining method (Kimura 2 -parameter)
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Phylogenetic tree II

• constructed by the UPGMA method. Difference
  groups of the Cichlidae and Odontobutidae
  families was broken in the agreement of the table
  DNAdist. More affinity of P. glenii to Cichlidae
  family than Odontobutidae.

UPGMA method (Kimura 2 -parameter)
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Conclusions

• karyology 2n44, one pair sm (differs from the
  native eastern population),16 pairs st,5 pairs
  acrocentric chromosomes
• allozyme analysis has been carried out for the
  first time. Low heterozigosity level in loci and
  uniform allozyme pattern
• by molecular analyses observed two haplotypes
  differed in 5 nucleotides (i.e. 0,92 of all
  sequence analysed)
• variable place characterized 5 transitions, i.e.
  reciprocal exchange of purine/purine or
  pyrimidine/pyrimidine bases
• mean intrapopulation similarity in sequence of
  cytb mtDNA in population was high - 99,6 which
  indicate of origin from small count of specimens

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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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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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G.kesslerii
M 24 SM 24 1 ST 0 A 0 1
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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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Table 1. Electrophoretic analysis of Amur sleeper
enzyme loci, conditions
1 muscle (m), liver (l) 2 Valenta
(V) Ferguson (F) Citric Phosphate
(CP) Morpholine citric (MC2)
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Table 2. Fis parameter calculated according to
Weir and Cockerham (1984) in
Perccottus glenii from Sv. Maria - Somotor and
parameters its significance in
polymorphic loci
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Table 3. Allelic frequences and Fis index in the
population of Perccottus glenii in
Sv.Maria-Somotor
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