Agricultural University, al. Mickiewicza 24/28, Kraków, Poland

Transcription

1 Genetic Polymorphism Of Zatorska Geese Flock Estimated Through RAPD Markers Krzysztof Andres 1, Ewa Kapkowska 1, Urszula Kaczor 2 1 Department of Poultry and Fur Animals Breeding and Animal Hygiene, 2 Department of Sheep and Goat Breeding, Agricultural University, al. Mickiewicza 24/28, Kraków, Poland

2 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).

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

4 History of the Zatorska goose flock moved again to the Ostrów Szlachecki farm of Agricultural University in Krakow. At that time the flock belonged to the national pedigree breeding stock classified as a gene pool flock. From 1996 the Zatorska geese is kept at the Research Farm in Rząska 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.

5 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.

6 Mating for flock renovation: Reproduction scheme Yearly: to 1996, then in: flock renovation Number of mating pens: 20 One pen: 1 sire 6 dams Inbreeding rate has been reduced through avoiding first cousins matings.

7 The purpose of the present study was to estimate the genetic polymorphism of the Zatorska geese flock through the RAPD analysis

8 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.

9 Material and methods The experiment was carried out on 20 sires and 120 dams mated for renovation of the Zatorska geese flock in 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.

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

11 Within-population genetic similarity based on band sharing (BS) formula: where: Statistical analysis BS ab = 2Nab ( Na + N N a and N b are the numbers of amplified fragments in samples a and b N ab is the number of bands in common. b ) Genetic similarity based on RAPD band frequency (BF): BF = 1/ where V i is the proportion of individuals possessing ith band across all the individuals and N is the total number of bands scored. N N i Vi i= 1

12 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

13 Total number of bands and polymorphic bands per primer, average band sharing (BS) and band frequency (BF) Primer estimated from RAPD of Zatorska geese flock Within population genetic similarity based on: Total number of bands (polymorphic bands) Band Sharing (BS) Band Frequency (BF) OPP-17 8 (5) PBG-3 10 (5) PBG-7 9 (8) PBG-13 9 (6) Mean 9 (6) ± ± 0.121

14 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.

15 THANK YOU FOR YOUR ATTENTION Zatorska geese flock