Identification of species of Vicia subgenus Vicia (Fabaceae) using chloroplast DNA data

Vicia subgenus Vicia (Fabaceae) contains several economically important food and forage legume species. Generally, the taxonomy and identification of these species have relied heavily on a few diagnostic morphological characters (e.g., the number of flowers per inflorescence). However, there are several specimens with intermediate morphological characters that make diagnosis of these species using those characters a difficult task for nonspecialists and render results that are not always accurate. The use of molecular markers, therefore, is the most attractive alternative strategy for a more accurate identification of species of the subgenus Vicia. In the present study, an attempt was made to investigate polymorphism in the chloroplast DNA of 22 species of the subgenus Vicia using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) on 9 chloroplast regions and a set of 12 consensus chloroplast simple sequence repeat (ccSSR) primer pairs for the diagnosis of the targeted species. Based on the data generated, it was possible to identify all of the species analyzed, except for 4 species that were placed in 2 groups (1, V noeana and V dionysiensis; and 2, V serratifolia and V hyaeniscyamus), each of which had a unique chloroplast type. Species-specific markers developed here are useful for the early diagnosis of targeted Vicia species and can provide essential baseline data for the evolution of systematic breeding and conservation strategies, as well as guidance for germplasm collections.