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Genetic Polymorphism Of Zatorska Geese Flock
Estimated Through RAPD Markers Krzysztof
Andres1, Ewa Kapkowska1, Urszula
Kaczor2 1Department of Poultry and Fur Animals
Breeding and Animal Hygiene, 2Department of
Sheep and Goat Breeding, Agricultural
University, al. Mickiewicza 24/28, 30-059
Kraków, Poland
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• Zatorska goose
• one of the fourteen breeds of geese belonging to
  the protected waterfowl genetic resources in
  Poland
• Scherf B. D. (ed.) (2000). World Watch List for
  Domestic Animal Diversity 3rd edn., FAO Rome
  Italy (page 352).

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History of the Zatorska goose flock

• breed was created during 1956-61 in the Research
  Institute of Animal Production at Zator, by the
  breeder team coordinated by Prof. dr Helena
  Baczkowska.
• originated from four indigenous Polish goose
  varieties Podkarpacka from Nowy Targ area,
  Garbonosa, Suwalska, and Pomorska (Baczkowska,
  1967).
• 1961, flock was closed and transferred to the
  Experimental Farm of Research Institute of Animal
  Production in Koluda Wielka, where the breeding
  work was continued for 6 years.

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History of the Zatorska goose flock

• 1967 - moved again to the Ostrów Szlachecki farm
  of Agricultural University in Krakow. At that
  time the flock belonged to the national pedigree
  breeding stock.
• 1984 - classified as a gene pool flock.
• From 1996 the Zatorska geese is kept at the
  Research Farm in Rzaska of the Agricultural
  University in Kraków. At first it was named a
  conservation flock and later as a flock belonging
  to animal genetic resources.
• Whole breeding documentation, including Pedigree
  Books, for the period of 50 years are kept in our
  Department.

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Studies on Zatorska goose
Several studies on the production traits and
genetic parameters of Zatorska goose were
undertaken over the years and are well documented
(Rabsztyn 2006). Molecular analysis in Zatorska
geese stock were provided only to some extent by
Zawadzka (1999) by the use of minisatellite DNA
fingerprinting.
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Reproduction scheme Mating for flock renovation
Yearly to 1996, then in
1998
1999
2000
2001
2002
2003
2004
2005
2006
2007
1997
- flock renovation
Number of mating pens 20 One pen 1
sire
6 dams Inbreeding rate has been reduced
through avoiding first cousins matings.
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The purpose of the present study was to estimate
the genetic polymorphism of the Zatorska geese
flock through the RAPD analysis
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Technique of RAPD

• RAPD is a PCR based molecular method which is
  useful for diversity studies.
• Suitable for genome analysis of species that are
  poorly studied (like goose etc.).
• RAPD loci are inherited in Mendelian fashion and
  can be used as genetic markers.
• Employs single short primers of random sequence,
  usually 10-mers, which produce multiband patterns
  detected by gel electrophoresis.
• Limitations sensitiveness to reaction conditions
  and dominance character of markers.

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Material and methods

• The experiment was carried out on 20 sires and
  120 dams mated for renovation of the Zatorska
  geese flock in 2004.
• RAPD-PCR was performed with the individual
  genomic DNA samples.
• DNA was isolated from blood using
  phenol-chlorophorm standard procedure .
• Four published primers OPP-17, PBG-3, PBG-7,
  PBG-13 (Ahlavat et al. 2004 Singh and Sharma
  2002) were used from the set of ten primers
  giving the most reproducible banding patterns.

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• Amplification conditions
• Reagents
• 40 ng of genomic DNA,
• 0.5 µM of primer,
• 200 µM of each dNTP, 2 mM MgCl2,
• 1 U Taq DNA Polymerase (Fermentas) and 1? Taq
  buffer.
• Thermal profile
• 15 s at 94C
• 30 s at 31C x 45
• 1 min at 72C
• final extension at 72C for 8 min /Eppendorf
  Mastercycler/
• Electrophoresis
• 3 Nusieve GTG agarose gel with ethidium
  bromide.

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Statistical analysis

• Within-population genetic similarity based on
  band sharing (BS) formula
• where
• Na and Nb are the numbers of amplified fragments
  in samples a and b
• Nab is the number of bands in common.
• Genetic similarity based on RAPD band frequency
  (BF)
• where Vi is the proportion of individuals
  possessing ith band across all the individuals
  and N is the total number of bands scored.

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Results
Example of RAPD polymorphisms amplified by primer
OPP-17 from 11 individuals of Zatorska geese
M
M
-3000bp
-1000bp
-500bp
M - GeneRuler 100bp DNA Ladder Plus, Fermentas
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Total number of bands and polymorphic bands per
primer, average band sharing (BS) and band
frequency (BF) estimated from RAPD of Zatorska
geese flock
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• Conclusions
• This is the first report estimating genetic
  similarity in Zatorska geese based on RAPD.
• Our results suggest that breeding strategy is
  effective in maintenance of genetic diversity in
  the flock.
• The received data informed about the current
  condition of genetic polymorphism in this
  population which could be the starting point for
  tracing the changes of genetic diversity in
  future generations.

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Zatorska geese flock
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