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Polyacrylamide gel electrophoresis and electron microscopy revealed that accumulation of iron-protein in soybean nodules is influenced by nodule age, mutation in bradyrhizobia, and rhizobial/bradyrhizobial strain-soybean cultivar interactions. Iron-protein concentrations (micrograms per milligram protein) were inversely related to heme concentrations (nanomoles per milligram protein), with correlation coefficients (r values) ranging from -0.98 in young nodules to -0.83 in mature ones. Bradyrhizobium japonicum symbiotic mutants HS 129 and HS 145 (Nod(+) Fix(-)) produced nodules high in iron-protein. Electrophoresis of homogenate prepared from nodules on Lee 68 produced by B. japonicum HS 129 yielded two different forms of the iron-proteins, 570 and 600 kilodaltons. The 570 kilodalton iron-protein isolated by preparative polyacrylamide gel electrophoresis behaved like horse-spleen-ferritin in responses to iron-stains, heat stability, ultraviolet absorption spectrum, iron unloading and reloading, and characteristic appearance in electron micrographs. These properties led to the conclusion that the 570 kilodalton iron-protein is phytoferritin. The nodule phytoferritin differed from horse-spleen-ferritin in electrophoretic mobility, serological properties, and molecular size and was distinct from most other known phytoferritins in that it was composed of different subunit types.

The Occurrence of Phytoferritin and Its Relationship to Effectiveness of Soybean Nodules