Sweet cherries are self-incompatible, which is determined by a gametophytic self-incompatibility system (GSI). The self-incompatibility is controlled by a multi-allelic S-locus. Knowledge about the S-allele constitution of the cultivars is essential for fruit growers and breeders. Recently, molecular PCR-based methods have been developed to distinguish all S-alleles in sweet cherries. In our work, we analysed S-locus genotypes by 13 universal and allele-specific PCR primer combinations within 117 registered, old and local sweet cherry cultivars from the Czech genetic resources of the Research and Breeding Institute of Pomology in Holovousy, the Czech Republic. We confirmed the previous S-genotyping for 66 accessions except for Drogans Gelbe, Hedelfinger, Erika, Meckenheimer Frühe, Badeborner, Bing, Alfa, Gamma, Huldra, Rivan, Valerij Tschkalov, Viola and Winkler’s Frühe. It could be due to either mislabelling or mistakes in the previous analyses. Newly, S-genotyping was determined for 51 accessions in which we found 4 new S-loci combinations. We detected the S-locus combinations in 19 incompatibility groups. The most frequent incompatibility groups were III (S3S4), II (S1S3), IV (S2S3), and VI (S3S6) with 22, 20, 12 and 12 genotypes, respectively.
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