Modern Phytomorphology 6: 87–88, 2014

SEARCHING FOR SCAR MARKERS NEEDED FOR DIVERSIFICATION OF THREE GROUPS WITHIN TRICHOPHYLLUS (, SUBGEN.BATRACHIUM )

Magdalena Jopek * & Magdalena Dąbrowska

Key words: Ranunculus, Batrachium, aquatic , diversity, molecular identification, SCAR markers, , Poland

Institute of Botany, Jagiellonian University, Kopernika 27, PL-31–501 Krakow, Poland; * [email protected]

Batrachium is a group of aquatic plants allowing for convenient and fast way to identify classified within the genus Ranunculus L. groups. (Ranunculaeceae). It is a widely distributed and one of the most difficult group of aquatic plants. For the analysis there were selected 8 During the analysis of the ITS sequences species of the genus Ranunculus and 3 groups of Ranunculus trichophyllus Chaix ex Vill. of R. trichophyllus. Other taxa were treated as specimens from different parts of a reference point that allows the exclusion of significant differences were detected, and as products varying within R. trichophyllus, but a result intraspecific diversification of this being alike with other species. In each species, taxon into three groups was made. Specimens a different number of samples were taken. forming different groups have different The differences in numbers of samples were ecological preferences, partly also differ in determined by the availability of previously range. Most likely, these groups correspond defined species based on the analysis of ITS to a separate species but the full picture still sequences. needs morphological analysis. However one of Samples were combined and treated as a the adaptations to the aquatic environment is single sample during the primers test. From their plastic vegetation response to changing 26 tested RAPD primers 11 were selected for environmental conditions, manifested in their further analysis. It was these primers which morphological variability. This phenotypic gave profiles showing the specific products for variability is taxonomically insignificant but each group. Next, primers were tested whether impedes the identification of individual species. specific products were present while using That is why molecular analysis can be useful. DNA from individual samples within a group. Although amplification and direct sequencing of From analyzed profiles five products were ITS (Internal Transcribed Spacers) is a reliable chosen to be cloned into plasmids and few method to determine species but this method of them sequenced. On the basis of obtained is relatively time-consuming and expensive. As sequences there were designed at least one pair an alternative to sequencing, RAPD (Random of primers for each sequence. From four pairs Amplified Polymorphic DNA) and ISSR (Inter- of starters designed for trichophyllus type I one Simple Sequence Repeats) are often chosen as gave products for individuals from this group cheap and simple methods. It is because they and didn’t for individuals from other two. For do not require an information sequence and are trichophyllus type II two pairs of starters were effective in obtaining the characteristic DNA tried and one of them worked as expected. Also profile. The main drawback of these methods is one pair of starter gave products for individuals low repeatability. Conversion RAPD markers to from for trichophyllus type I group without the SCAR increase the specificity and stability, products for DNA from other groups. These © The Author(s), 2014 88 Modern Phytomorphology 6 (2014) markers however work well for deliberate as provide new information about their habitat population but usually didn’t mark hybrids. preferences and morphological variation and This is the worse flaw however shouldn’t impact shed a new light on taxonomical aspects of this the significance of their role. group. These markers will give the possibility to examine a much larger number of samples than Acknowledgements it would be possible by direct sequencing. This can lead to obtain knowledge of distribution of The study was financially supported by the particular groups within R. trichophyllus, as well Jagiellonian University no. K/DSC/001677.