Gladiolus as economically important flowering plants reflect the wide range of morphological variability of the inflorescences. The aim of current gladiolus breeders is to create interesting new genotypes by different breeding techniques, carrying a variety of colour, size, texture and shapes of flowers and inflorescences. Despite the well-established morphological characterization of gladiolus germplasm, genomic screening by functional markers can provide useful information for breeders. The miRNA-based assay was conducted on 9 gladiolus (Gladiolus × gandavensis Van Houtte) hybrids by 9 individual markers of following families: miR156, miR160, miR398, miR408, miRr414 and miR482. A total of 291 loci were amplified, of which loci of marker miR408 were the most abundant (25%), with the following representation of other marker types: 21% – miR414; 19% miR156; 16% – miR160; 12% – miR482 and 7% – miR398. Genetic diversity of selected gladiolus cultivars was assessed by two markers, lus-miR408 and hyp-miR414, which provided genotype-specific marker profiling in the form of molecular fingerprinting. Stress-sensitive marker miR398 specifically amplified loci in genotype Správna Susane, which is the most susceptible to water deficit of all analysed genotypes. Marker miR141 was able to distinguish cultivars Athos and Dandy among themselves, but also from other cultivars.
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