Testing validity of cpDNA, rnDNA and SSR markers for evaluation of Lythrum salicaria L. populations

Abstract

Higher soil concentrations of sodium chloride might be reflection of natural edaphic features and consequence of man-related activities. Near the rivers with elevated soil salinity, more intensive growth of Lythrum salicaria L. is well-known phenomenon. Anthropogenic activities, such as species introduction to the new areas, might contribute to the shifts of genetic diversity of L. salicaria. In this context, L. salicaria as halophyte and invasive species, is an important model for evaluation of anthropogenic effects. Till now data concerning genetic diversity of this species in Europe are very limited. Thereby our study aimed at molecular evaluation of populations of Lythrum salicaria growing in the riverbanks of Lithuania. Successes of comparison of populations firstly depends on the DNA marker selection. There are many cases when markers applied for one species might work for the other species of the same family. Among Lythraceae family representatives, the widest scope genetic evaluations were done for Punica granatum. Simple sequence repeat, SSR markers have been created for the accessions of horticultural importance of this species. SSR primers of Punica granatum failed to amplify DNA of Lythrum salicaria populations of Lithuania. It was true for the conditions described in Punica granatum protocol also for the set of buffer type, cycle number and temperature-related modifications of polymerase chain reactions (PCR). In addition to SSR markers, attempts were made to select primers for sequencing some nrDNA and cpDNA fragments. PCR reaction failed for primer combinations like rps16.5R+rps16.50F, trnK.UUU.3R+trnK.UUU.3F, 126614_F+127383_R (55 °C), although some primers created on the bases of chloroplast DNA, showed amplification of Lythrum salicaria DNA. Furthermore, several pairs of nrDNA primers, ITS4_R, ITS5_F and ITS7_F, demonstrated positive results of PCR. [...]