Molecular markers to assess genetic diversity of Gentiana lutea L. from the Ukrainian Carpathians

Abstract

The aim of the study was to develop the system of polymerase chain reaction (PCR)-based markers for the assessment of genetic diversity and population genetic studies of Gentiana lutea L. as well as to determine the utility of two indices of marker informativeness. The informativeness was determined for 40 PCR primers of different types (random amplified polymorphic DNA, inter simple sequence repeat, inter-retrotransposon amplified polymorphism, resistance gene analog polymorphism and conserved DNA-derived polymorphism markers) by evaluating discriminating power (D L ) and resolving power (R p ) in a sample of 30 plants from two populations. Analysis of correlation between the index value and the number of differentiated pairs of genotypes in the given sample revealed that D L is more efficient than R p ; therefore, we selected primers based on the D L value. In total, 12 primers with the largest values of D L were chosen. Analysis of genetic relationship among 86 plants from six populations showed that the number of bands produced by the three of selected primers was sufficient to give average bootstrap support across six key nodes in the dendrogram higher than 85%, while using six of the primers resulted in average bootstrap value exceeding 99%. Thus, a minimal set of three to six selected primers are sufficient for a quick assessment of genetic diversity of G. lutea populations, depending on the sample size and degree of differentiation between populations, while the rest of the primers with D L values above 0.8 may be used for ecogenetic surveys. Preliminary results obtained with selected primers indicate the moderate level of genetic variation within the species and significant differentiation among individual populations.

The assessment of the current status of Gentiana lutea L. populations of the Ukrainian Carpathians: Ecological and genetic approaches

“…All these populations grow on the territory of the Carpathian Biosphere Reserve. In addition, we studied the artificially created (in the 1970s) agropopulation located on Pozhyzhevska Mountain (Chornogora ridge, Ukrainian Carpathians, Carpathian National Nature Park) (Mosula et al, 2015). The material for introduction of Pozhyzhevska population was taken from Sheshul-Pavlyk population (Bedej et al, 2010).…”