Kullaj, E. , S. imon, S. , A. akalli & I.

Peji
1 2 3
1 2 3

Department of Horticulture, Faculty of Agriculture and Environment, Agricultural University of Tirana, Albania Genetic Resources Centre, Agricultural University of Tirana, Tirana, Albania

Department of Plant Breeding, Genetics and Biometrics, Faculty of Agriculture University of Zagreb, University of Zagreb, Croatia

Introduction
The wild grapevine [Vitis vinifera ssp. sylvestris (G.C. Gmel.) Hegi] is one of the two forms co-existing in Eurasia and North Africa, which conservation and characterization are under the high priority. Since 2003, research has been launched in order to identify, map, describe and characterize the populations of wild grapevine in northern Albania. In this study, three populations found in Mat, Drin and Shkreli valleys represented with total of 19 individual plants have been the subject of molecular characterization using 9 SSR markers.

Materials & Methods

DNA was extracted from young leaves using Qiagen DNeasy Plant mini kit following the protocol of manufacturer. PCR products were separated using capillary electrophoresis with ABI 3130 Genetic Analyzer (Applied Biosystems). Allele sizes were detected using GeneMapper 4.0. with the sizing algorithm second order least square. Relevant population statistics parameters such as average number of alleles, allele frequencies, He, Ho and PIC, as well as F-Statistics measures (FIS and FST) were calculated according to Nei (1987) for the 9 SSR loci using the POPGENE V3.2 (Yeh and Boyle 1997). Also, cluster analysis was carried out based on Dice similarity indices (Dice, 1945) calculated for each possible pair-wise combination of analyzed genotypes. The similarity matrix was subjected to sequential agglomerative hierarchical nested (SAHN) clustering using unweighted pair-group method analysis (UPGMA) as implemented in NTSYS V2.2 software (Rholf, 1998).

Results & Conclusions

Cluster analysis
Substantial genetic polymorphism was observed with 5 to 9 alleles per locus. The most polymorphic loci were VVS2 and VVMD7 with majority of genotypes being heterozygous. Marker PIC values varied from 0.452 0.753. The level of heterozygosity (Ho=0.696; He=0.656) was rather high compared to similar studies. In the same time, little inbreeding was observed (FIS=0.042). In spite of geographical distance among populations, overall population differentiation was very small (FST = 0.044) suggesting common origin and absence of gene flow barriers. However, results might be influenced by insufficient number of plants per population and markers. This study will continue analyzing more genotypes and using additional markers with higher PIC values such as AFLP or RAPD markers. REFERENCES Dice (1945) Nei M (1978) Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 89:583590 Rholf (1998) Yeh FC, Boyle T (1997) Population genetic analysis of codominant and dominant markers and quantitative traits. Belg J Bot 129:157

Populations polymorphism
Expected Heterozigocity Observed Heterozigocity (He) (Ho) Locus 3 popul. Locus 3 popul. VVS2 0.804 VVS2 0.789 MD7 MD27 ZAG62 ZAG79 MD5 MD28 MD32 MD25 Average 0.789 0.684 0.486 0.647 0.720 0.703 0.670 0.764 0.656 MD7 MD27 ZAG62 ZAG79 MD5 MD28 MD32 MD25 Average 0.895 0.684 0.474 0.474 0.588 0.632 0.632 0.737 0.696 PIC values Locus VVS2 MD7 MD27 ZAG62 ZAG79 MD5 MD28 MD32 MD25 3 popul. 0.753 0.735 0.633 0.452 0.562 0.652 0.639 0.596 0.711

Population statistics
Locus VVS2 MD7 MD27 ZAG62 ZAG79 MD5 MD28 MD32 MD25 Average FIS 0.005950 -0.127795 -0.166550 -0.001677 0.244413 0.183163 0.112249 0.083493 0.046772 0.042 FST 0.034966 -0.012799 0.197121 0.041676 0.055811 0.008223 0.014203 0.040192 0.016235 0.044

Contact: ekullaj@ubt.edu.al