Development and Characterization of a Soybean Experimental Line Lacking the α′ Subunit of β‑Conglycinin and G1, G2, and G4 Glycinin

A soybean experimental line (BSH-3) devoid of a subset of seed storage proteins was developed by crossing a mutant donor line “HS99B” with a Chinese cultivar “Dongnong47” (DN47). One-dimensional and high-resolution 2-D gel electrophoresis revealed the absence of G1 (A1<sub>a</sub>B<sub>2)</sub>, G2 (A<sub>2</sub>B1<sub>a)</sub>, and G4 (A<sub>5</sub>A<sub>4</sub>B<sub>3)</sub> glycinin and the α′ subunit of β-conglycinin in BSH-3 seeds. Despite the lack of these abundant seed proteins, BSH-3 seeds still accumulated 38% protein. BSH-3 seeds also accumulated high levels of free amino acids as compared with DN47 seeds, particularly arginine, and the amount of several essential amino acids were significantly elevated in BSH-3 seeds. Elevated accumulation of α and β-subunit of β-conglycinin, G5 glycinin, Kunitz trypsin inhibitor, and Bowman-Birk protease inhibitor indicates seed proteome rebalancing in BSH-3 seeds. Immunoblot analysis using sera from soybean allergic patients demonstrated the complete lack of a major allergen (α′ subunit of β-conglycinin) in BSH-3 seeds. However, elevated levels of other allergens were found in BSH-3 seeds due to proteome rebalancing. Transmission electron microscopy observation of mature seeds of BSH-3 revealed striking differences in the appearance of the protein storage vacuoles when compared with DN47.