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Differentiation of Clavibacter michiganensis subsp. michiganensis from seed-borne saprophytes using ELISA, Biolog and 16S rDNA sequencing

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Abstract

Specificity of a monoclonal antibody (MAb), Cmm1, to geographically diverse strains of the seed-borne tomato pathogen, Clavibacter michiganensis subsp. michiganensis (Cmm), was assessed and the MAb was tested for its usefulness as a tool to separate the pathogen from saprophytes in naturally infested tomato seed. Of the 236 international Cmm strains tested, 99% reacted with MAb Cmm1. MAb Cmm1 was also strongly reactive with an additional 32 strains isolated from seed that were later identified as Cmm by the Biolog MicroLog™ microbial identification system (Biolog, Inc., Hayward, CA) and 16S rDNA sequence analysis. It correctly differentiated these strains from 12 MAb Cmm1-negative seed strains that possessed similar colony morphology but were later identified as other Gram-positive genera and species. The specificity of MAb Cmm1 to the pathogen and the near universality of the MAb Cmm1-reactive antigen among diverse Cmm strains make this antibody a useful detection and identification tool. The finding that a large proportion of the Cmm strains associated with naturally infested tomato seed were putatively hypovirulent or non-virulent indicates that such populations cannot be ignored and points to a need for studies to determine their significance in host-pathogen interactions.

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Acknowledgements

This paper is dedicated to the late Dr. Albert Benedict, who contributed greatly to the production and characterization of these and other monoclonal antibodies for numerous bacterial plant pathogens. Many thanks go to German Hoyos (Syngenta Seeds), Mark Ricker and Bert Woudt for generously providing the infested seed lots used in this study. This work was supported by grants from the United States Department of Agriculture Tropical and Subtropical Agricultural Research Grants Programme (Award No. 2003-34135-13982) and the American Seed Research Foundation Vegetable and Flower Seed Division Permanent Research Fund.

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Author notes

  1. Carla Y. Mizumoto

    Present address: Department of Tropical Medicine, John A. Burns School of Medicine, University of Hawaii, 651 Ilalo Street, Honolulu, HI, 96813, USA

Authors and Affiliations

  1. Department of Plant and Environmental Protection Sciences, University of Hawaii, 3190 Maile Way, St. John Plant Science Building 307, Honolulu, HI, 96822, USA

    Wendy S. Kaneshiro, Carla Y. Mizumoto & Anne M. Alvarez

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  1. Wendy S. Kaneshiro
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  2. Carla Y. Mizumoto
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  3. Anne M. Alvarez
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Correspondence to Anne M. Alvarez.

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Kaneshiro, W.S., Mizumoto, C.Y. & Alvarez, A.M. Differentiation of Clavibacter michiganensis subsp. michiganensis from seed-borne saprophytes using ELISA, Biolog and 16S rDNA sequencing. Eur J Plant Pathol 116, 45–56 (2006). https://doi.org/10.1007/s10658-006-9036-1

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