DNA barcoding and phylogenetic analysis of endemic Astragalus nezaketiae and Vicia alpestris subsp. hypoleuca (Fabaceae): Evidence from nrDNA ITS and cpDNA matK and rbcL sequences

Abstract

In this study, we performed DNA barcoding and phylogenetic analysis using one nuclear (ITS) and two chloroplast DNA regions (matK and rbcL) of endemic Astragalus nezaketiae A. Duran & Aytaçand Vicia alpestris Stev. subsp. hypoleuca (Boiss.) Davis taxa in Turkey. PCR reactions were performed using universal primers. Sequences of the PCR products were edited using BioEdit and FinchTV software and contigs were obtained. All contigs were Blasted at NCBI and similarities were analysed. Using the MEGA 6.0 program, maximum likelihood trees were constructed including some sequences retrieved from NCBI. For Astragalus nezaketiae; in the ITS analysis, Astragalus nezaketiae appeared separately from other species, and for matK, Astragalus nezaketiae appeared together with Astragalus cicer L. However, rbcL tree was polytomic. For Vicia alpestris subsp. hypoleuca; in ITS, rbcL and matK results Vicia alpestris subsp. hypoleuca were found together with Vicia cracca L., Vicia benghalensis L. and Vicia villosa Roth species. Analysis of the combined data revealed similar results with all barcode regions for Vicia alpestris subsp. hypoleuca while different phylogenetic results were obtained for Astragalus nezaketiae.