Using RAPD-markers in studying molecular genetic polymorphism in the genus Miscanthus species

Abstract

Aim. The study aims to investigate molecular genetic polymorphism in the representatives of different populations and bioenergy crops plant groups of the genus Miscanthus of the collection of the Institute of Bioenergy Crops and Sugar Beet of the NAAS of Ukraine as well as to search and select the molecular markers to differentiate them using RAPD PCR methods. Methods. The studies used the molecular genetic method for detecting polymorphism by analyzing the lengths of amplification fragments and the method of electrophoretic distribution of amplification products in agarose gel. Results. In the course of amplification with 7 RAPD primers, 28 loci were obtained, of which 23 were polymorphic. The polymorphism degree averaged 65 %. The index of polymorphism locus ranged from 0.17 to 0. 56. There were identified six alleles by marker P822 -with sizes ranging from 230 to 613 bp. P820 primer amplified two loci of 311 and 482 bp, three alleles sized from 219 to 530 bp were identified by P816 and P817 markers. The spectra of the amplicons obtained using the above-mentioned primers make it possible to differentiate representatives of the Miscanthus genus of different species, since the difference in the number of loci for each species representative is established. A unique allele with the frequency of 0.35 and the size of 605 bp was obtained using the RAPD2 primer. Conclusions. According to the results it was found that the use of primers P816 and RAPD2 allowed to separate genotypes of M. sacchariflorus, M. gigantheus, M. sinensis. A large proportion of polymorphic loci confirm the population composition of the model sample which resulted in high level of polymorphism. Key words: genus Miscanthus, RAPD primers, PCR, polymorphism, allele frequencies.