STUDY OF GENETIC POLYMORPHISM OF RUSSIAN ORIGIN HEMP CULTIVARS WITH THE USE OF ISSR–MARKERS

Abstract

Nowadays, hemp production places high demands on the efficiency of breeding and the reliability of seed multiplication of hemp (Cannabis sativa L. subsp. Sativa). Therefore, a detailed study of genetic polymorphism, intervarietal and intravarietal variability, and the peculiarities of the formation of the fund of available genetic diversity of culture becomes urgent. The aim was to study the genetic polymorphism of some modern Russian species of industrial hemp using molecular ISSR markers. Experimental data were obtained in 2021 in the laboratory of molecular-genetic researching and cell selection of the Federal Research Center of Bast Crops. We studied ten varieties of hemp of Russian breeding from the collection of Penza Unit of Federal Research Center for Fiber Crops: ‘Milena’, ‘Vera’, ‘Surskaya’, ‘Nadezhda’, ‘Ingreda’, ‘Diana’, ‘Juliana’, ‘Gentus’, ‘Dimra’, ‘Margo’. DNA was isolated from individual seeds by the CTAB method. When setting up PCR, 20 ISSR primers were used; amplification products were separated in agarose gel. A total of 99 alleles of 430–1500 bp were identified, the genetic profile of each studied sample turned out to be individual. Cluster analysis and genetic similarity dendrogram construction made it possible to reveal the presence of intra- and intervarietal genetic polymorphism, visualize the phylogenetic relationships of the studied species. The studied samples formed dense intravarietal groups and were divided into three intervarietal clusters. The presence of similar components of crossing in the pedigrees of the studied varieties is associated with the peculiarity of the identified clustering and determines the prerequisites for the formation of the genetic narrowness of modern varieties of cannabis. The set of 20 used ISSR-markers is distinguished by good resolution for studying the genetic polymorphism of hemp. Intravarietal polymorphism with an average genetic distance of 0.14 and intervarietal polymorphism with a distance of 0.61 were revealed in 10 cultivars of hemp of the Central Russian type. According to the results of cluster and factor analyzes, which showed the similarity of conclusions, the samples were divided into three main clusters, mainly differing in the originator of the variety. The study of DNA polymorphism of seeds of the ‘Diana’, obtained after reproduction of two years, indicates its genetic stability.