Analysis of lectin binding by Bradyrhizobium japonicum strains grown on nitrocellulose filters using peroxidase-labeled lectin

• Published in: Analytical biochemistry Vol. 164; no. 2; pp. 488 - 493
• Main Authors: Liang, Rongti, Emerich, David W.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.08.1987; Elsevier
• Subjects: Bacteriological methods and techniques used in bacteriology; Bacteriology; binding assays; Binding Sites; Biological and medical sciences; Bradyrhizobium japonicum; cell adhesion; cell surface; Collodion; defective mutant; Fluorescein-5-isothiocyanate; Fluoresceins; Fundamental and applied biological sciences. Psychology; Glycine max; interspecific relationships; lectin; lectins; Lectins - metabolism; Microbiology; Mutation; Peroxidases; Plant Lectins; polysaccharide; receptors; Rhizobium - genetics; Rhizobium - metabolism; Soybean Proteins; symbiosis; Thiocyanates; cell surface; binding assays; receptors; polysaccharide; lectin; cell adhesion; Screening; Lectin; Binding capacity; Analysis; Bradyrhizobium japonicum; Bacteria; Rhizobiaceae
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/0003-2697(87)90523-9

A procedure was developed to assess the ability of wild-type and mutant strains of Bradyrhizobium japonicum to bind soybean lectin. The lectin-binding ability of bacteria grown on nitrocellulose filters was determined using peroxidase-labeled soybean lectin. The assay produced clear differences between strains known to be unable to bind soybean lectin and those which can. The assay gave results identical to those of the fluorescein isothiocyanate-soybean lectin-binding assay of T. V. Bhuvaneswari, S. G. Pueppke, and W. D. Bauer (1977, Plant Physiol. 60, 486–491) with regard both to the ability of particular B. japonicum strains to bind lectin and to the inhibition caused by N-acetyl- d-galactosamine. The method was used to screen Tn5-induced mutants of B. japonicum 2143 for their inability to bind soybean lectin. The procedure provides a sensitive and convenient method to screen Bradyrhizobium strains for the ability to bind soybean lectin.