Features of varietal DNA identification in Lotus corniculatus L. samples

This study focuses on molecular-genetic identification of encrypted samples of birdsfoot trefoil (Lotus corniculatus L.) using Inter-Primer Binding Site (iPBS) and Sequence-Related Amplified Polymorphism (SRAP) marker systems. The research aimed to verify whether the encrypted samples genetically match the reference varieties Smolensky 1, Luch, and Cheprasovsky. The analysis involved a collection of known reference samples and encrypted accessions of the same varieties but from different harvest years. Genomic DNA was isolated from seven-day-old seedlings using a pooled sample of 30 genotypes per variety. Following preliminary screening, three informative SRAP primer combinations (ME3-EM3, F9-R14, F11-R14) and three iPBS primers (2217, 2257, 2239) revealing interspecific polymorphism were selected for analysis. The PCR products were separated by agarose gel electrophoresis, and the resulting binary matrices were processed using principal coordinate analysis (PCoA) in GenAlEx 6.5. The first two PCoA coordinates explained 57.44% of the total molecular variance. The results demonstrated that the DNA profiles of the encrypted samples did not form clear clusters with the reference varieties, preventing their unambiguous identification. The observed genetic drift between biological replicates of the known varieties underscores the inherent variability in this cross-pollinating species and validates the legislative requirement for periodic updates of genetic reference passports every 3-5 years to ensure reliable cultivar identification