Comparative analysis of genus Vigna seeds using antiserum against a synthesized multiple antigenic peptide

Antiserum for 33 kDa vicilin‐like seed proteins in V. angularis was prepared using a synthesized multiple antigenic peptide (MAP). The anti‐MAP antiserum was applied to the protein analysis of all of genus Vigna , seeds which are stored in the Gene Bank at the National Institute of Agrobiological Resources, Tsukuba, Japan. The anti‐MAP antiserum specifically reacted with 33 kDa vicilin‐like proteins and weakly with 55 kDa vicilin‐like proteins of V. angularis by immunoblotting and could distinguish between these two types of vicilin‐like 7 S proteins. Anti‐MAP antiserum reacted with 33 kDa band both in wild and cultivated types of V. angularis (4 spots) and half species of V. radiata (2 spots). The N ‐terminal amino acid sequence of the major immunoreacting spot in V. radiata seeds was analyzed. A partially homologous amino acid sequence with MAP was found in immunoreacted protein and the anti‐MAP antiserum was able to be applied as a probe to identify homologous amino acid sequence in other proteomes. V. radiata species could be divided by their immunoreactivities into two groups: the group from Southeast Asia and Australia, which reacted with the anti‐MAP antiserum, and the group from West Asia and Madagascar, which did not. The abundant proteins in V. mungo , seeds at 55 kDa showed strong reactivity signals with anti‐MAP antiserum. The existance of a homologous amino acid sequence with MAP was suggested in the 55 kDa proteins. The seed proteins in V. aconitifolia, V. umbellata, V. vexillata, V. marina, V. unguiculata , and V. oblongifolia did not show any reactions and they do not possess the homologous amino acid sequence with MAP in their seed proteins.