Evaluation of the Effectiveness of ISSR-Marker Informativeness Indicators for the Analysis of Plant Genetic Polymorphism
Authors: Prokopiak M.  Z. , Fliachok A.  I. , Maiorova O. Yu. , Hrytsak L.  R. , Drobyk N.  M. 
Published: September 2022

Introduction. A number of generations of different types of molecular genetic markers have changed to the present day.The DNA nucleotide sequence marker system allows testing genetic diversity at the gene level. The creation of molecular markers and their use in biological researches made it possible to study genetic polymorphism in more detail, as well as to investigate the level of kinship at the inter- and intra-species levels. The use of various DNA markers that are associated with different sequences of the genome is effective at the time of studying genetic polymorphism. At the same time, it is also important to reduce the cost of polymerase chain reaction analysis, which is associated with the use of a small number of primers, and to choose the most informative primers.Purpose. The purpose of the research is to select the most informative ISSR primers for assessing the genetic polymorphism of plant populations using the example of representatives of the Gentiana L. genus, as well as to determine the most effective indicators of the informativeness of DNA markers.Methods. Molecular genetic markers, statistical methods, calculation of indicators of informativeness of primers.Results. It was established that all the used indicators of the informativeness of the primers to a certain extent characterize the informativeness of the ISSR-markers, which are used for learning of the genetic polymorphism and genetic structure of G. lutea.Three ISSR primers (UBC#807, UBC#811 and UBC#840) with the highest indicators of discriminating(D) and resolving power (Rp) were selected by us. It was established that in general the value of the percentageof polymorphic amplicons was similar, and the values of the indicators of expected heterozygosity and the Shannon index were higher.This was established as a result of comparing the genetic polymorphism indices of the G. luteapopulations obtained using three ISSR primers (UBC#807, UBC#811 and UBC#840) with the genetic polymorphism indices estimated by nine ISSR primers (UBC #807, UBC#809, UBC#810, UBC#811, UBC#827, UBC#835, UBC#840, UBC#857, UBC#889). The increase in genetic diversity is probably caused by an increase in the polymorphic primers due to the removal from the sample of those that form monomorphic amplicons.Originality. For the first time, an effective indicator of the informativeness of primers was selected and the most effective ISSR markers were selected for the assessment of the genetic diversity of plants of the genus Gentiana L. on the example of seven populations of G. lutea from the Ukrainian Carpathians.Conclusions. The most informative primers were selected and their effectiveness was demonstrated for assessing the genetic diversity of a sample of plants from seven populations located in the mountain ranges of the Ukrainian Carpathians. In general, UBC#811 and UBC#807 were identified as the most informative, and UBC#827 and UBC#889 were the least effective in evaluating the genetic polymorphism of G. lutea. This conclusion was made by evaluating indicators(total number of amplicons, number of polymorphic amplicons, PIC (polymorphism information content), MI (marker index), Rp, D). The relationship between all the parameters of the evaluation of the effectiveness of the primers was characterized by a high level of reliability (р<0.001, р=0.05), only in some cases it was higher than 5%. The level of genetic polymorphism of G. lutea for UBC#807, UBC#811 and UBC#840 was evaluated.Key words: Gentianalutea L.; genetic diversity; resolving power; discriminating power, number of polymorphic amplicons