Molecular Genetic Analysis of a Rare Plant Species Pulsatilla patens (L.) Mill. of Northern Kazakhstan

The genetic diversity of 5 cenopopulations of the rare plant species Pulsatilla patens (L.) Mill, located on the territories of Pavlodar, Akmola and Kostanay regions, Kazakhstan Republic, was studied. Intermicrosatellite DNA analysis (ISSR — Inter Simple Sequence Repeats) was used to determine the indicators of genetic diversity. Polymerase chain reaction with 5 primers: ISSR-1, ISSR-3, M1, M27, X11 was performed. The number of polymorphic DNA fragments in the total plant samples ranged from 9 to 28. Fragment’s size ranged from 200 to 1420 bp. The percentage of polymorphic loci in the total sample P. patens was 0.965. Expected heterozygosity is 0.162, and the number of effective alleles is 1.361. Indicators of genetic diversity are higher in the cenopopulation from the Kostanay region (Borovskoe village): HE =0.209, ne =1.743, and lower in the population from the Pavlodar region: HE =0.131, ne =1.597. Two rare DNA fragments were found in the studied P. patens cenopopulations: in cenopopulation from the Pavlodar region and in cenopopulation from the Akmola region (village Yerementau). Cenopopulation from the Kostanay region (village Borovskoe) has the most balanced diversity structure: h=0.139, and cenopopulation of the Kostanay region (village Kamensk-Uralsk) is the least balanced: h = 0.188. The Shannon Index revealed the greatest diversity in the cenopopulation from the Kostanay region (village Borovskoe) I=0.327, and the lowest diversity in cenopopulation from the Pavlodar region I=0.215.