Aim. To investigate the polymorphism of microsatellite loci of the 5HL chromosome of barley, including those which
are location in the area of LT-resistance key genes and close to them, to determine and evaluate the frequence of
microsatellite alleles in the genetic material of autumn-sown barley of different origin. Methods. The isolation of
genomic DNA, microsatellite analysis, PCR amplification, gel electrophoresis, and statistical analysis. We studied 46
barley genotypes of different origin, including 33 winter barley varieties, and 13 varieties of alternative (facultative)
growth habit; among these 46 barley genotypes, 21 varieties were developed by PBGI-NCSCI, 25 – of other origin,
including 21 from the countries of Central Europe (Czech Republic/CZ – 2; Grabe, Luran; Germany/DE – 4: Skarpia,
Majbrit, Сinderella, Highlight), Western Europe (France/FR – 1: Anzhelika; the Netherlands/NL – 1: Gerlach) and
Eastern Europe (NGC named after P.P. Lukianenko/RU – 13, 11 winter varieties: Derzhavnyi, Espada, Zhavoronok,
Kondrat, Kumach, Meteor, Metaksa, Mikhailo, Platon, Tigr, Khutorok, and 2 winter-and-spring varieties: Putnik,
Timofei) and 4 Western-Asian varieties (Syria/SYR – 4: Pamir013/Sonata, Pamir065/Pamir149, CWB-117-77-97,
ROHO). Results. The allelic polymorphism was studied by 14 microsatellite loci of the 5HL chromosome of barley,
including those in the area of genes Fr-H1, Fr-H2 and close to these regions, the distribution of the identified micro-
satellite alleles was studied along with their frequencies and genetic diversity in the sampling of 46 collection varieties
of autumn-sown barley of different origin. Among the investigated microsatellite areas of the 5HL chromosome of
barley, only 50 % of loci in the selected samples of varieties were found to be polymorphic. These were microsatellite
loci Bmag0760, GMS061, Bmag0337, UMB702, Bmag0323, Bmag0223, and Bmag0222. We found the dominating
alleles and those with reliably lower frequency, and the alleles specific only for some regions. The estimated values
of the polymorphism information content (PIC) for the investigated polymorphic microsatellite loci varied between
0.29 (UMB702) – 0.77 (Bmag0223). The diversity index was 0.54 on average. Conclusions. The results of the study
demonstrated moderate allelic diversity of the investigated microsatellite loci of the 5HL chromosome of barley,
which indicates the presence of potential genetic variability of some loci, the possibility of applying them in further
studies on determining the effects of specific alleles of each locus and their associations with required economically
valuable traits of barley, including resistance to low temperatures (LT-resistance). The potential of polymorphic alleles
as markers of frost-resistance traits of autumn-sown barley genotypes is discussed.