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A codominant marker for homozygosity testing and species discrimination needed in breeding programs was developed and applied to different Mimulus L. species and cultivars. Degenerative primers used to amplify intron 10 of topoisomerase 6 subunit B (top6B) in distant species also amplified the locus in all analyzed Mimulus species. The sequences obtained revealed the presence of a microsatellite motif and were used to design a specific microsatellite primer pair, Mim-top6B, for Mimulus species. The microsatellite marker showed a high degree of polymorphism in Mimulus species, and the heterozygous nature of most M. aurantiacus Curtis cultivars. The marker was further used to analyze putative doubled haploids of M. aurantiacus and showed that all but one was heterozygous, indicating their hybrid origin.
A sample of 94 pear (Pyrus L.) genotypes, traditionally present in the Slovenian landscape, was analyzed by amplified fragment length polymorphism (AFLP) and SSR (SSR), focusing on the assessment of genetic relationships. The analyzed samples involved germplasm of Pyrus communis L., P. nivalis Jacq., and P. pyraster L. The AFLP technique, using five EcoRI and MseI primer combinations, revealed molecular polymorphism at a level of 65.95%, representing 93 polymorphic bands among the total of 141 scored. With SSR analysis, 64 polymorphic alleles were found at seven microsatellite loci, with an average of 9.14 alleles per locus. Genetic distances among the individuals being studied were calculated using the Dice coefficient of similarity, and a dendrogram supported by AFLP and SSR data was constructed using the neighbor-joining method. The clustering method grouped the analyzed genotypes into three main clusters. The first cluster included the P. communis germplasm. The genotypes resembling P. nivalis were grouped in the second largest cluster, which could be divided in to four subclusters. The germplasm of P. pyraster, in this cluster, was found to be much less distinct than we had assumed. The most typical cultivar group in the third cluster was ‘Vinska Mostnica’. The study indicated that P. nivalis germplasm is frequently present in Slovenia, but not as a pure species.
There is a long tradition of common bean cultivation in Slovenia, which has resulted in the development of numerous landraces in addition to newly established cultivars. The genetic diversity of 100 accessions from the Genebank of the Agricultural Institute of Slovenia (AIS) were evaluated with amplified fragment length polymorphism (AFLP) markers and phaseolin seed protein. Twenty-seven standard accessions of known Mesoamerican and Andean origin, 10 wild Phaseolus vulgaris accessions and two related species, P. coccineus L. and P. lunatus L., were also included. Ten AFLP primer combinations produced 303 polymorphic bands, indicating a relatively high level of genetic diversity. Based on the marker data, unweighted pair group method with arithmethic mean (UPGMA) analysis and principal coordinate analysis (PCoA) all P. vulgaris accessions were separated into three well-defined groups. Two groups consisted of accessions of Mesoamerican and Andean origin, while the third was comprised of only four wild P. vulgaris accessions. A set of Slovene accessions formed a well-defined sub-group within the Andean cluster, showing their unique genetic structure. These data were supported by phaseolin analysis, which also revealed additional variants of “C” and “T” phaseolin types. The results are in agreement with previous findings concerning diversification of common bean germplasm introduced in Europe.