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Plant and Cell Physiology, 1992, Vol. 33, No. 2 151-156
© 1992

Development and Characterization of a Monoclonal Antibody to Phytosiderophores

Shuichi Mihashi1, Lawrence J. Sikora2, Jeffrey S. Buyer2, Shinji Fushiya3 and Satoshi Mori1

1Department of Agricultural Chemistry, The University of Tokyo Bunkyo-ku, Tokyo, 113 Japan
2Soil-Microbial Systems Laboratory, USDA-ARS, Beltsville Agricultural Research Center Beltsville, MD 20705, U.S.A.
3Pharmaceutical Institute, Tohoku University Aobayama, Sendai, 980 Japan

Mugineic acid-family phytosiderophores (MAs) are low molecular weight chelators that are secreted by graminaceous plants, form complexes with soil Fe(III) and are essential for plant growth. Methods to detect MAs which include HPLC and radio-immunoassay with polyclonal antibody require sophisticated equipment or radio-labelled MAs which are difficult to synthesize. Our objective was to develop a detection and quantitation system for MAs based on monoclonal antibody specificity and technology. A monoclonal antibody was produced which reacts with nicotianamine (NA), deoxymugineic acid (DMA), mugineic acid (MA) and epi-hydroxymugineic acid (epi-HMA) in a competitive ELISA. Azetidine-2-carboxylic acid (A-2-C) was not reactive while N-(3-amino-3-carboxypropyl) azetidine-2-carboxylic acid (A-2-C dimer) was partially reactive. The range of detection using the competitive ELISA is from 2 x 10–6 to 2 x 10–7 M MAs. Besides detection and quantification of MAs, the potential uses for the monoclonal antibody are numerous and include affinity chromatography and immunocytochemistry.

(Received September 26, 1991; Accepted December 16, 1991)
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