Abstract
Recently, within the Fabaceae family, the Vicia genus has been recognized for its vital role in sustainable agriculture. Vicia species are economically important grain and forage crops. However, the presence of complex morphological characteristics makes identification and recognition of native species difficult. In this study, the possibility of using DNA barcoding regions (ITS2, matK, and rbcL) to distinguish among 19 Vicia taxa (59 accessions) found in South Korea was evaluated. The sequence alignment analysis revealed considerable nucleotide diversity (π) between the loci, in which ITS2 showed the highest mean interspecific distance, whereas there was no intraspecific variability among the barcode regions in 12 of the 19 taxa. Phylogenetic analysis of combined barcoding regions revealed well-resolved phylogeny with the highest species level discrimination. Combinations of barcode loci were also used in classification at the subgenera and section levels. The results revealed that the combined barcoding regions can be used effectively to differentiate the following species: Vicia angustifolia var. segetilis, Vicia bungei, Vicia villosa, Vicia cracca, Vicia dasycarpa, Vicia hirsuta, Vicia tetrasperma, Vicia amurensis, Vicia hirticalycina, and Vicia chosenensis. However, it is difficult to differentiate the species of Vicia unijuga, Vicia unijuga var. kaussanensis, Vicia linearifolia, Vicia unijuga f. angustifolia, Vicia nipponica, Vicia amoena, Vicia venosa var. cuspidata, Vicia pseudo-orobus, and Vicia japonica with the tested barcode regions. These species come under sect. Vicilla and are found to be closely related or species that have recently undergone speciation; thus, it has limitation to distinguish with recommended barcodes. Hence, to differentiate the unclassified species, 39 morphological characteristics were investigated, in which 16 useful characteristics were selected for efficient classification. Finally, the 16 selected morphological useful traits efficiently differentiated all the Vicia species. In conclusion, a combination of barcoding loci together with morphological characteristics of this study efficiently discriminated all the Korean Vicia species.
Highlights
The genus Vicia L. belongs to the Fabaceae (Leguminosae) family and is the third largest family of flowering plants worldwide
Barcode sequence of ITS2, matK, and rbcL for 33 Vicia species conserved at the National Agrobiodiversity Center, South Korea were downloaded from National Center for Biotechnology Information (NCBI) GenBank
As a result of amplification with universal primers (ITS2, matK, and rbcL) using template Genomic DNA (gDNA) extracted from each sample, the sequence length of ITS2 varied from 339 to 355 bp, whereas matK and rbcL measured 705 and 542 bp, respectively (Table 3)
Summary
The genus Vicia L. belongs to the Fabaceae (Leguminosae) family and is the third largest family of flowering plants worldwide. 150–210 species are distributed across Europe, Asia, and North America, and the majority of species is found in the Mediterranean (Cacan et al, 2016; Al-Joboury, 2017). Approximately 17–21 Vicia species have been reported in Korea (Lee, 1996; Choi, 2007; Nam et al, 2012). (Kupicha, 1976). [=Vicilla (SCHUR) ROUY, five sections], based on the presence of stipule nectaries, relative peduncle length, subtending leaves, and number of flowers per inflorescence (Kupicha, 1976; Leht, 2009) The Vicia genus is divided into two subgenera, Vicia L. (22 sections) and Cracca Peterm. [=Vicilla (SCHUR) ROUY, five sections], based on the presence of stipule nectaries, relative peduncle length, subtending leaves, and number of flowers per inflorescence (Kupicha, 1976; Leht, 2009)
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