An ELISA Procedure for Quantification of Relative Amounts of Intercellular Glycoprotein in Legume Nodules

An enzyme-linked immunosorbant assay (ELISA) method based ona monoclonal antibody (MAC236) is described in which relativeamounts of an intercellular glycoprotein were quantified inextracts of whole legume nodules. This glycoprotein has recentlybeen shown to be an important component of the cortical oxygendiffusion barrier. The ELISA method is demonstrated on threeexamples of soybean (Glycine max L. Merr.) nodule systems whichhave been the subject of previously published investigations:(a), cv. Clarke inoculated with Bradyrhizobium japonicum RCR3442,nodulated root systems of which were subject to 10, 21 or 40%oxygen continuously for 28 d; (b), cv. Bragg and its supernodulatingmutant derivative (nts382) inoculated with Bradyrhizobium japonicumUSDA110; (c), cv. Clarke inoculated with Bradyrhizobium japonicumRCR3442 or RCR3407. ELISA results are related to oxygen diffusioncharacteristics defined in previous publications and show thatincreases in the amount of glycoprotein present correlated withincreases in supra-ambient (40%) levels of rhizosphere pO₂,in minimum gas diffusion resistance and in speed of diffusionbarrier response. Area data of component parts of nodule inner cortices suggestthat diffusion resistance control under sub-ambient (10%) oxygenlevels also involves cell expansion. The amount of MAC236 antigen in nodules is affected by bothhost plant genotype and rhizobial strain and the latter alsoappears to be involved in determining the morphological developmentof the nodule inner cortex.Copyright 1993, 1999 Academic Press Oxygen diffusion resistance, glycoprotein, nodules, Glycine max, ELISA