In the thesis, we focused on the genotyping of Tartary buckwheat (Fagopyrum tataricum Gaertn.), analyzing 14 different genetic resources. Of these, 7 were local and 7 were foreign, all stored in the gene bank of the Biotechnical Faculty, University of Ljubljana. Using the isolation procedure, we extracted high-quality DNA from leaves, which was then amplified with 8 microsatellite markers using polymerase chain reaction (PCR). To verify and visualize the amplified DNA fragments, we used agarose gel electrophoresis, which enables the analysis of fragment presence and size. All 8 loci were successfully amplified. Subsequently, capillary electrophoresis was used to obtain data, which were processed with the GeneMapper 6 software. Genetic variability parameters were calculated using Cervus software. A total of 20 different alleles were successfully amplified, with an average of 2.5 alleles per locus. The average polymorphism information content (PIC) was 0.341. The shortest allele measured at 141 bp, while the longest was 267 bp. The average observed heterozygosity (Ho) was 0.5, slightly higher than the average expected heterozygosity (He) of 0.4. Three out of the 8 SSR markers were monomorphic, indicating a high level of homozygosity in the studied genotypes. A well-defined group, strongly supported by a high bootstrap value, was formed by 4 genotypes: 122, B, 105, and 132.
|
