Selection of nrDNA and cpDNA markers for comparison of Echinocystis lobata (michx.) Torr. et A. gray populations

Abstract

Wild cucumber (Echinocystis lobata), Cucurbitaceae representative, is vine, climbing with the help of branched tendrils, forming big capsules with spines. This annual herb is cultivated in the gardens due to numerous fragrant whitish flowers, attention attracting fruits, hanging on the fences. E. lobata spread, as introduced to Europe species, is related to ornamental features. Romania is proposed as the first country of E. lobata settlement in Europe, where it has arrived from North America. E. lobata has escaped from the sites of anthropogenic origin to overmoistured natural habitats of temperate climate zone. Within last decades it is recognized as an intensively spreading invasive species along riverbanks of Lithuania. For invasion process elucidation and eradication of this plant it is important to get comprehensive information about alien species effects on natural ecosystems. Genetic tools are important for selection of management strategies concerning invaders. Till now molecular data about E. lobata is very scarce. The aim of this study was to find out chloroplast and ribosomal DNA markers for analyses of E. lobata populations. For selected individuals the best conditions (reaction mix, temperature, number of cycles) of polymerase chain reaction were determined. Ribosomal markers ITS4, ITS5, ITS7 and chloroplast DNA markers such as rps16.50, trnK.UUU.3, ycf1.59, ycf1.70, 127383, 127647, 126614, 126489, atpB, rbcL, trnH (GUG), psbA, petA, psbJ, atpI, atpH, F71, R1516, R1661 were tested. Amplified DNA fragments were visualized on 1.5 % agarose gel, cut out and cleaned with GeneJET PCR Purification Kit. Finally, one (ITS4, ITS7) out of two nrDNA primer pairs and five (ycf1.59-ycf1.70, atpB-rbcL, atpI-atpH, F71- R1516, F71-R1661) out of thirteen cpDNA primer pairs have generated pure enough and readable DNA fragments for sequencing.