Tomato Plants Overexpressing a Celery Mannitol Dehydrogenase (MTD) Have Decreased Susceptibility to Botrytis cinerea


The oxidative burst is a critical early event in plant-pathogen interactions that leads to a localized, programmed cell death (PCD) called the hypersensitive response (HR). The HR and associated PCD retard infection by biotrophic pathogens, but can, in fact, enhance infection by necrotrophic pathogens like Botrytis cinerea. In addition to signaling the induction of the HR, reactive oxygen species (ROS) produced during the oxidative burst are antimicrobial. We hypothesize that pathogens such as B. cinerea survive the antimicrobial effects of ROS, at least partially by secreting the antioxidant mannitol during infection. This is supported by the previous observation that overexpression of the catabolic enzyme mannitol dehydrogenase (MTD) can decrease a plants susceptibility to mannitol-secreting pathogens like B. cinerea. To extend the above hypothesis, and test the general utility of this approach in an important horticultural crop, we overexpressed celery MTD in tomato (Solanum lycopersicum cv. “Moneymaker”). In these studies, we observed a significant increase (up to 90%) in resistance to B. cinerea in transgenic tomatoes expressing high amounts of MTD.

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Patel, T. , Krasnyanski, S. , Allen, G. , Louws, F. , Panthee, D. and Williamson, J. (2015) Tomato Plants Overexpressing a Celery Mannitol Dehydrogenase (MTD) Have Decreased Susceptibility to Botrytis cinerea. American Journal of Plant Sciences, 6, 1116-1125. doi: 10.4236/ajps.2015.68116.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] van Kan, J.(2005) Infection Strategies of Botrytis cinerea. Acta Horticulturae, 669, 77-89.
[2] Nakajima, M. and Akutsu, K. (2014) Virulence Factors of Botrytis cinerea. Journal of General Plant Pathology, 80, 15-23.
[3] Levine, A., Tenhaken, R., Dixon, R. and Lamb, C. (1994) H2O2 from the Oxidative Burst Orchestrates the Plant Hy- persensitive Disease Resistance Response. Cell, 79, 583-593.
[4] Torres, M. (2010) ROS in Biotic Interactions. Physiologia Plantarum, 178, 414-429.
[5] Gorvin, M. and Levine, A. (2000) The Hypersensitive Response Facilitates Plant Infection by the Necrotrophic Pathogen Botrytis cinerea. Current Biology, 10, 751-757.
[6] Yu, I., Parker, J. and Bent, A. (1998) Gene for Gene Resistance without the Hypersensitive Response in Arabidopsis dnd1 Mutant. Proceedings of the National Academy of Sciences of the United States of America, 95, 7819-7824.
[7] Rolke, Y., Liu, S., Quidde, T., Williamson, B., Schouten, A., Weltring, K., Siewers, V., Tenberge, K., Tudzynski, B. and Tudzynski, P. (2004) Functional Analysis of H2O2-Generating Systems in Botrytis cinerea: The Major Cu-Zn-Superoxide Dismutase (BcSOD1) Contributes to Virulence on French Bean, Whereas a Glucose Oxidase (BcGOD1) Is Dispensable. Molecular Plant Pathology, 5, 17-27.
[8] Segmuüller, N., Kokkelink, L., Giesbert, S., Odinius, D., Kan, J. and Tudzynski, P. (2008) NADPH Oxidases Are involved in Differentiation and Pathogenicity in Botrytis cinerea. Molecular Plant-Microbe Interactions, 21, 808-819.
[9] Williamson, J., Desai, A., Krasnyanski, S., Ding, F., Guo, W., Nguyen, T., Olson, H., Dole, J. and Allen, G. (2013) Over-expression of Mannitol Dehydrogenase in Zonal Geranium Confers Increased Resistance to the Mannitol Secreting Fungal Pathogen Botrytis cinerea. Plant Cell, Tissue and Organ Culture (PCTOC), 115, 367-375.
[10] Jennings, D., Daub, M., Pharr, D. and Williamson, J. (2002) Constitutive Expression of a Celery Mannitol Dehydrogenase in Tobacco Enhances Resistance to the Mannitol-Secreting Fungal Pathogen Alternaria alternata. The Plant Journal, 32, 41-49.
[11] Voegele, R., Hahn, M. and Mendgen, K. (2005) Possible Roles for Mannitol and Mannitol Dehydrogenase in the Biotrophic Plant Pathogen Uromyces fabae. Plant Physiology, 137, 190-198.
[12] Joosten, M., Hendrickx, L. and de Wit, P. (1990) Carbohydrate Composition of Apoplastic Fluids Isolated from Tomato Leaves Inoculated with Virulent or Avirulent Races of Cladosporium fulvum (Syn. Fulvia fulva). Netherlands Journal of Plant Pathology, 96, 103-112.
[13] Smirnoff, N. and Cumbes, Q.J. (1989) Hydroxyl Radical Scavenging Activity of Compatible Solutes. Phytochemistry, 28, 1057-1060.
[14] Chaturvedi, V., Bartiss, A. and Wong, B. (1997) Expression of Bacterial mtlD in Saccharomyces cerevisiae Results in Mannitol Synthesis and Protects a Glycerol-Defective Mutant from High-Salt and Oxidative Stress. Journal of Bacteriology, 179, 157-162.
[15] Vélez, H., Glassbrook, N. and Daub, M. (2008) Mannitol Biosynthesis Is Required for Plant Pathogenicity by Alternaria alternata. FEMS Microbiology Letters, 285, 122-129.
[16] Stoop, J., Williamson, J. and Pharr, D. (1996) Mannitol Metabolism in Plants: A Method for Coping with Stress. Trends in Plant Science, 1, 139-144.
[17] Stoop, J. and Pharr, D. (1992) Partial Purification and Characterization of Mannitol: Mannose 1-Oxidoreductase from Celeriac (Apium graveolens var. rapaceum) Roots. Archives of Biochemistry and Biophysics, 298, 612-619.
[18] Williamson, J., Massel, M., Conkling, M. and Pharr, D. (1995) Sequence Analysis of a Mannitol Dehydrogenase cDNA from Plants Reveals a Function for the Pathogenesis-Related Protein ELI3. Proceedings of the National Academy of Sciences of the United States of America, 92, 7148-7152.
[19] Jennings, D., Ehrenshaft, M., Pharr, M. and Williamson, J. (1998) Roles for Mannitol and Mannitol Dehydrogenase in Active Oxygen-Mediated Plant Defense. Proceedings of the National Academy of Sciences of the United States of America, 95, 15129-15133.
[20] Lauter, F. (1996) Root-Specific Expression of the LeRse-1 Gene in Tomato Is Induced by Exposure of the Shoot to Light. Molecular Genetics and Genomics, 252, 751-754.
[21] Kiedrowski, S., Kawalleck, P., Hahlbrock, K., Somssich, I. and Dangl, J. (1992) Rapid Activation of a Novel Plant Defense Gene Is Strictly Dependent on the Arabidopsis RPM1 Disease Resistance Locus. The EMBO Journal, 11, 4677-4684.
[22] Cheng, F.-Y., Zamski, E., Guo, W.-W., Pharr, D. and Williamson, J. (2009) Salicylic Acid Stimulates Secretion of the Normally Symplastic Enzyme Mannitol Dehydrogenase (MTD): A Possible Defense against Mannitol-Secreting Fungal Pathogens. Planta, 230, 1093-1103.
[23] Blackburn, K., Cheng, F., Williamson, J. and Goshe, M. (2010) Data-Independent Liquid Chromatography/Mass Spectrometry (LC/MSE) Detection and Quantification of the Secreted Apium graveolens Pathogen Defense Protein Mannitol Dehydrogenase. Rapid Communications in Mass Spectrometry, 24, 1009-1016.
[24] Murashige, T. and Skoog, F. (1962) A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiologia Plantarum, 15, 473-497.
[25] Gamborg, O., Miller, R. and Ojima, K. (1968) Nutrient Requirements of Suspension Cultures of Soybean Root Cells. Experimental Cell Research, 50, 151-158.
[26] Bradford, M. (1976) A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding. Analytical Biochemistry, 72, 248-252.
[27] Laemmli, U. (1970) Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature, 227, 680-685.
[28] Stoop, J., Williamson, J., Conkling, M. and Pharr, D. (1995) Purification of NAD-Dependent Mannitol Dehydrogenase from Celery Suspension Cultures. Plant Physiology, 108, 1219-1225.
[29] Spurr Jr., H. (1973) An Efficient Method for Producing and Studying Tobacco Brown-Spot Disease in the Laboratory. Tobacco Science, 17, 145-148.
[30] Floryszak-Wieczorek, J., Arasimowicz, M., Milczarek, G., Jelen, H. and Jackowiak, H. (2007) Only an Early Nitric Oxide Burst and the Following Wave of Secondary Nitric Oxide Generation Enhanced Effective Defense Responses of Pelargonium to a Necrotrophic Pathogen. New Phytologist, 175, 718-730.
[31] Barna, B. and Györgyi, B. (1992) Resistance of Young versus Old Tobacco Leaves to Necrotrophs, Fusaric Acid, Cell Wall Degrading Enzymes and Autolysis of Membrane Lipids. Physiological and Molecular Plant Pathology, 40, 247-257.
[32] Dulermo, T., Rascle, C., Billon-Grand, G., Gout, E., Bligny, R. and Cotton, P. (2010) Novel Insights into Mannitol Metabolism in the Fungal Plant Pathogen Botrytis cinerea. Biochemical Journal, 427, 323-332.
[33] Ceccaroli, P., Saltarelli, R., Guescini, M., Polidori, E., Buffalini, M., Menotta, M., Pierleoni, R., Barbieri, E. and Stocchi, V. (2007) Identification and Characterization of the Tuber borchii D-Mannitol Dehydrogenase Which Defines a New Subfamily within the Polyol-Specific Medium Chain Dehydrogenases. Fungal Genetics and Biology, 44, 965-978.
[34] Glick, B. (1995) Metabolic Load and Heterologous Gene Expression. Biotechnology Advances, 13, 247-261.
[35] Howell, S. (2013) Endoplasmic Reticulum Stress Responses in Plants. Annual Review of Plant Biology, 64, 477-499.

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