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Proteomic Analysis of Soybean Root Hairs After Infection by Bradyrhizobium japonicum

¹National Center for Soybean Biotechnology, Department of Plant Microbiology and Pathology, University of Missouri, Columbia, MO 65211, U.S.A.; ²Department of Biology, Maryville College, Maryville, TN 37804, U.S.A.; ³Department of Computer Science, University of Missouri, Columbia, MO 65211, U.S.A.; ⁴Department of Biochemistry, University of Missouri, Columbia, MO 65211, U.S.A.; ⁵Proteomics Center, University of Missouri, Columbia, MO 65211, U.S.A.

Infection of soybean root hairs by Bradyrhizobium japonicum is the first of several complex events leading to nodulation. In the current proteomic study, soybean root hairs after inoculation with B. japonicum were separated from roots. Total proteins were analyzed by two-dimensional (2-D) polyacrylamide gel electrophoresis. In one experiment, 96 protein spots were analyzed by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) to compare protein profiles between uninoculated roots and root hairs. Another 37 spots, derived from inoculated root hairs over different timepoints, were also analyzed by tandem MS (MS/MS). As expected, some proteins were differentially expressed in root hairs compared with roots (e.g., a chitinase and phosphoenolpyruvate carboxylase). Out of 37 spots analyzed by MS/MS, 27 candidate proteins were identified by database comparisons. These included several proteins known to respond to rhizobial inoculation (e.g., peroxidase and phenylalanine-ammonia lyase). However, novel proteins were also identified (e.g., phospholipase D and phosphoglucomutase). This research establishes an excellent system for the study of root-hair infection by rhizobia and, in a more general sense, the functional genomics of a single, plant cell type. The results obtained also indicate that proteomic studies with soybean, lacking a complete genome sequence, are practical.

Additional keywords: legume , rhizobium , symbiosis .