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Salicylic acid stimulates secretion of the normally symplastic enzyme mannitol dehydrogenase: a possible defense against mannitol-secreting fungal pathogens

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Abstract

The sugar alcohol mannitol is an important carbohydrate with well-documented roles in both metabolism and osmoprotection in many plants and fungi. In addition to these traditionally recognized roles, mannitol is reported to be an antioxidant and as such may play a role in host–pathogen interactions. Current research suggests that pathogenic fungi can secrete mannitol into the apoplast to suppress reactive oxygen-mediated host defenses. Immunoelectron microscopy, immunoblot, and biochemical data reported here show that the normally symplastic plant enzyme, mannitol dehydrogenase (MTD), is secreted into the apoplast after treatment with the endogenous inducer of plant defense responses salicylic acid (SA). In contrast, a cytoplasmic marker protein, hexokinase, remained cytoplasmic after SA-treatment. Secreted MTD retained activity after export to the apoplast. Given that MTD converts mannitol to the sugar mannose, MTD secretion may be an important component of plant defense against mannitol-secreting fungal pathogens such as Alternaria. After SA treatment, MTD was not detected in the Golgi apparatus, and its SA-induced secretion was resistant to brefeldin A, an inhibitor of Golgi-mediated protein transport. Together with the absence of a known extracellular targeting sequence on the MTD protein, these data suggest that a plant’s response to pathogen challenge may include secretion of selected defensive proteins by as yet uncharacterized, non-Golgi mechanisms.

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Abbreviations

BFA:

Brefeldin A

HK:

Hexokinase

JA:

Jasmonic acid

MTD:

Mannitol dehydrogenase

PCR:

Polymerase chain reaction

PR:

Pathogenesis related

ROS:

Reactive oxygen species

SA:

Salicylic acid

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Acknowledgments

We thank Drs. Steven Spiker (NC State University, Genetics) for NT1 cultures, Jen Sheen (Harvard Medical School) for HK antiserum, Dan Klessig (Boyce Thompson Institute) for PR1a antiserum, and Drs. Candace Haigler (NC State University, Crop Science and Plant Biology) and Marcela Rojas-Pierce (NC State University, Plant Biology) for critical discussion and helpful comments. TEM analyses were conducted at the NCSU Center for Electron Microscopy. This work was supported in part by US Department of Agriculture—National Research Initiative grant no. 2000-0884 to JDW and DMP.

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Authors and Affiliations

  1. Department of Horticultural Science, North Carolina State University, Raleigh, NC, 27695-7609, USA

    Fang-yi Cheng, Wei-wen Guo, D. Mason Pharr & John D. Williamson

  2. Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76-100, Rehovot, Israel

    Eli Zamski

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  1. Fang-yi Cheng
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  2. Eli Zamski
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  3. Wei-wen Guo
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Correspondence to John D. Williamson.

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Cheng, Fy., Zamski, E., Guo, Ww. et al. Salicylic acid stimulates secretion of the normally symplastic enzyme mannitol dehydrogenase: a possible defense against mannitol-secreting fungal pathogens. Planta 230, 1093–1103 (2009). https://doi.org/10.1007/s00425-009-1006-3

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