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Assessment of intervarietal polymorphism of raspberry using SSR-PCR analysis

Raspberry (Rubus idaeus L.) is one of the leading berry crops in Ukraine. The necessity to improve its varietal composition in several directions requires the use of DNA markers as a modern approach to working with genetic resources. For the first time, microsatellite DNA markers (SSR-PCR) were applied to study the genetic structure of 12 raspberry cultivars cultivated in Ukraine and to evaluate their effectiveness in determining the genetic relatedness among them. Genotyping was carried out using 10 microsatellite loci: RiM017, RiM019, RhM003, RhM011, RhM043, Rub1a, Rub4a, Rub223a, Rub228a, and Rub262b. The analysis of microsatellite locus polymorphism was performed by electrophoretic separation of PCR products in an 8 % denaturing polyacrylamide gel (PAGE), followed by silver nitrate staining. A high level of polymorphism of the SSR markers in raspberry was revealed. The average number of alleles per locus was 4.61. The effective number of alleles ranged from 1.291 (RiM017) to 5.053 (Rub228a). Shannon index values varied from 0.456 (Rub228a) to 1.770 (RhM043). The highest observed heterozygosity was recorded for locus RiM019 (83.3 %), and the lowest for locus RhM011 (8.3 %). Based on the allele frequencies, the polymorphism information content (PIC) values were calculated and ranged from 0.212 to 0.777 across the studied markers. The genetic distances among the cultivars varied from 0.0093 to 2.0127, while genetic identity indices ranged from 0.0056 to 0.9398. The obtained results demonstrate the potential of the tested SSR markers for assessing the genetic diversity of the national raspberry gene pool and for the identification and certification (passportization) of raspberry cultivars.

Key words: raspberry, Rubus idaeus L., polymorphism, SSR markers, microsatellites, cultivar identification.

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