Plant Molecular Biology

, Volume 47, Issue 5, pp 621–631 | Cite as

Analysis of celery (Apium graveolens) mannitol dehydrogenase (Mtd) promoter regulation in Arabidopsis suggests roles for MTD in key environmental and metabolic responses

  • Eli Zamski
  • Wei-Wen Guo
  • Yuri T. Yamamoto
  • D. Mason Pharr
  • John D. Williamson


Of the growing list of promising genes for plant improvement, some of the most versatile appear to be those involved in sugar alcohol metabolism. Mannitol, one of the best characterized sugar alcohols, is a significant photosynthetic product in many higher plants. The roles of mannitol as both a metabolite and an osmoprotectant in celery (Apium graveolens) are well documented. However, there is growing evidence that `metabolites' can also have key roles in other environmental and developmental responses in plants. For instance, in addition to its other properties, mannitol is an antioxidant and may have significant roles in plant-pathogen interactions. The mannitol catabolic enzyme mannitol dehydrogenase (MTD) is a prime modulator of mannitol accumulation in plants. Because the complex regulation of MTD is central to the balanced integration of mannitol metabolism in celery, its study is crucial in clarifying the physiological role(s) of mannitol metabolism in environmental and metabolic responses. In this study we used transformed Arabidopsis to analyze the multiple environmental and metabolic responses of the Mtd promoter. Our data show that all previously described changes in Mtd RNA accumulation in celery cells mirrored changes in Mtd transcription in Arabidopsis. These include up-regulation by salicylic acid, hexokinase-mediated sugar down-regulation, and down-regulation by salt, osmotic stress and ABA. In contrast, the massive up-regulation of Mtd expression in the vascular tissues of salt-stressed Arabidopsis roots suggests a possible role for MTD in mannitol translocation and unloading and its interrelation with sugar metabolism.

carbohydrate translocation and regulation gene regulation mannitol metabolism plant-pathogen interaction salt and osmotic stress 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. An, G., Ebert, P.R., Mitra, A. and Ha, S.B. 1988. Binary vectors, In: S. Gelvin and R. Schilperoort (Eds.) Plant Molecular Biology Manual (Supplement 4, 1992), Kluwer Academic Publishers, Dordrecht, Netherlands, pp. A3: 1–19.Google Scholar
  2. Chiu, W.-L., Niwa, Y., Zeng, W., Hirano, T., Kobayashi, H. and Sheen, J. 1996. Engineered GFP as a vital reporter in plants. Curr. Biol. 63: 325–330.Google Scholar
  3. Clough, S.J. and Bent, A.F. 1998. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16: 735–743.Google Scholar
  4. Creelman, R.A. and Mullet, J.E. 1997. Biosynthesis and action of jasmonates in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48: 355–381.Google Scholar
  5. Doares, S.H., Narvaez-Vasquez, J., Conconi, A. and Ryan, C.A. 1995. Salicylic acid inhibits synthesis of proteinase inhibitors in tomato leaves induced by systemin and jasmonic acid. Plant Physiol. 108: 1741–1746.Google Scholar
  6. Durner, J. and Klessig, D.F. 1996. Salicylic acid is a modulator of tobacco and mammalian catalases. J. Biol. Chem. 271: 28492–28501.Google Scholar
  7. Everard, J.D., Gucci, R., Kann, S.C., Flore, J.A. and Loescher, W.H. 1994. Gas exchange and carbon partitioning in the leaves of celery (Apium graveolens L.) at various levels of root zone salinity. Plant Physiol. 106: 281–292.Google Scholar
  8. Fellman, J.K. and Loescher, W.H. 1987. Comparative studies on sucrose and mannitol utilization in celery (Apium graveolens L.). Physiol. Plant. 69: 337–341.Google Scholar
  9. Geigenberger, P., Langenberger, S., Wilke, I., Heineke, D., Heldt, H.W. and Stitt, M. 1993. Sucrose is metabolized by sucrose synthase and glycolysis within the phloem complex of Ricinis communis L. seedlings. Planta 190: 446–453.Google Scholar
  10. Hause, B., Demus, U., Teichmann, C., Parthier B. and Wasternack C. 1996. Developmental and tissue-specific expression of JIP-23, a jasmonate-inducible protein of barley. Plant Cell Physiol. 37: 641–649.Google Scholar
  11. Hood, E.E., Gelvin, S.B., Melchers, L.S. and Hoekema, A. 1993. New Agrobacterium helper plasmids for gene transfer to plants. Transgenic Res. 2: 208–218.Google Scholar
  12. Huijser, C., Kortstee, A., Pego, J.V., Wisman, E., Weisbeek, P. and Smeekens, S.C.M. 2000. The Arabidopsis SUN6 gene is identical to ABI4. Plant J. 23: 577–585.Google Scholar
  13. Iwasaki, T., Yamaguchi-Shinozaki, K. and Shinozaki, K. 1995. Identification of a cis-regulatory region of a gene in Arabidopsis thaliana whose induction by dehydration is mediated by abscisic acid and requires protein synthesis. Mol. Gen. Genet. 247: 391–398.Google Scholar
  14. Jennings, D.B., Eherenshaft, M., Pharr, D.M. and Williamson, J.D. 1998. Roles for mannitol and mannitol dehydrogenase in active-oxygen-mediated plant defense. Proc. Natl. Acad. Sci. USA 95: 15129–15133.Google Scholar
  15. Koch, K.E. 1996. Carbohydrate-modulated gene expression in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47: 509–540.Google Scholar
  16. Korfhage, U., Trezzini, G.F., Meier, I., Hahlbrock, K. and Somssich, I.E. 1994. Plant homeodomain protein involved in transcriptional regulation of a pathogen defense-related gene. Plant Cell 6: 695–708.Google Scholar
  17. Korsmeyer, S.J. 1995. Regulators of cell death. Trends Genet. 11: 101–105.Google Scholar
  18. Leung, J. and Giraudat, J. 1998. Abscisic acid signal transduction. Annu. Rev. Plant Physiol. Plant Mol. Biol. 49: 199–222.Google Scholar
  19. Lew, R.R. 1996. Pressure regulation of the electrical properties of growing Arabidopsis thaliana L. root hairs. Plant Physiol. 112: 1089–1100.Google Scholar
  20. Lu, C.A., Lim, E.K. and Yu, S.M. 1998. Sugar response sequence in the promoter of a rice α-amylase gene serves as a transcriptional enhancer. J. Biol. Chem. 273: 10120–10131.Google Scholar
  21. Mathur, J. and Koncz, C. 1998. Establishment and maintenance of cell suspension cultures. In: J. Martinez and J. Salinas (Eds.) Methods in Molecular Biology, Vol. 82: Arabidopsis Protocols, Humana Press, Totowa, NJ, pp. 27–30.Google Scholar
  22. Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497.Google Scholar
  23. Noiraud, N., Delrot, S. and Lemoine, R. 2000. The sucrose transporter of celery. Identification and expression during salt stress. Plant Physiol. 122: 1447–1456.Google Scholar
  24. Nolte, K.D. and Koch, K.E. 1993. Companion-cell-specific localization of sucrose synthase in zones of phloem loading and unloading. Plant Physiol. 101: 899–905.Google Scholar
  25. Obaton, M.F. 1929. Evolution de la mannite chez les végétaux. Rev. Gén. Bot. 41: 622–633.Google Scholar
  26. PeÑa-Cortés, H., Albrecht, T., Prat, S., Weiler, E.W. and Willmitzer, L. 1993. Aspirin prevents wound-induced gene expression in. tomato leaves by blocking jasmonic acid biosynthesis. Planta 191: 123–128.Google Scholar
  27. Prata, R.T.N., Williamson, J.D., Conkling, M.A. and Pharr, D.M. 1997. Sugar repression of mannitol dehydrogenase activity in celery cells. Plant Physiol. 114: 307–314.Google Scholar
  28. Riesmeier, J.W., Hirner, B. and Frommer, W.B. 1993. Potato sucrose transporter expression in minor veins indicate a role in phloem loading. Plant Cell 5: 1591–1598.Google Scholar
  29. Salmon, S., Lemoine, R., Jamai, A., Bouché-Pillon, S. and Fromont, J.C. 1995. Study of sucrose and mannitol transport in plasma-membrane vesicles from phloem and non-phloem tissues of celery (Apium graveolens L.) petioles. Planta 197: 76–83.Google Scholar
  30. Schindler, U., Terzaghi, W., Beckmann, H., Kadesch, T. and Cashmore, A.R. 1992. DNA binding site preferences and transcriptional activation properties of the Arabidopsis transcription factor GBF1. EMBO J. 11: 1275–1289.Google Scholar
  31. Shah, J. and Klessig, D.F. 1996. Identification of a salicylic acid-responsive element in the promoter of the tobacco pathogenesis-related β-1,3-glucanase gene, PR-2d. Plant J. 10: 1089–1101.Google Scholar
  32. Sheen, J., Hwang, S., Niwa, Y., Kobayashi, H. and Galbraith, D.W. 1995. Green-fluorescent protein as a new vital marker in plant cells. Plant J. 8: 777–784.Google Scholar
  33. Shen, B., Jensen, R. and Bohnert, H.J. 1997. Increased resistance to oxidative stress in transgenic plants by targeting mannitol biosynthesis to chloroplasts. Plant Physiol. 113: 1177–1183.Google Scholar
  34. Smeekens, S. 2000. Sugar-induced signal transduction in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 51: 49–81.Google Scholar
  35. Smirnoff, N. and Cumbes, Q.J. 1989. Hydroxyl radical scavenging activity of compatible solutes. Phytochemistry 28: 1057–1060.Google Scholar
  36. Stoop, J.M.H. and Pharr, D.M. 1992. Partial purification and characterization of mannitol:mannose 1-oxidoreductase from celeriac (Apium graveolens var. rapaceum) roots. Arch. Biochem. Biophys. 298: 612–619.Google Scholar
  37. Stoop, J.M.H. and Pharr, D.M. 1994. Mannitol metabolism in celery stressed by excess macronutrients. Plant Physiol. 106: 503–511.Google Scholar
  38. Stoop, J.M.H., Williamson, J.D., Conkling, M.A. and Pharr, D.M. 1995. Purification of NAD-dependent mannitol dehydrogenase from celery suspension cultures. Plant Physiol. 108: 1219–1225.Google Scholar
  39. Stoop, J.M.H., Williamson, J.D. and Pharr, D.M. 1996. Mannitol metabolism in plants: a method for coping with stress. Trends Plant Sci. 1: 139–144.Google Scholar
  40. Sutherland, M.W. 1991. The generation of oxygen radicals during host plant responses to infection. Physiol. Mol. Plant Path. 39: 79–93.Google Scholar
  41. Truernit, E. and Sauer, N. 1995. The promoter of the Arabidopsis thaliana SUC2 sucrose-H+symporter gene directs expression of β-glucuronidase to the phloem: evidence for phloem loading and unloading by SUC2. Planta 196: 564–570.Google Scholar
  42. Urwin, N.A. and Jenkins, G.I. 1997. A sucrose repression element in the Phaseolus vulgaris rbcS2 gene promoter resembles elements responsible for sugar stimulation of plant and mammalian genes. Plant Mol. Biol. 35: 929–942.Google Scholar
  43. Vreugdenhil, D. 1983. Abscisic acid inhibits phloem loading of sucrose. Physiol. Plant. 57: 463–467.Google Scholar
  44. Williamson, J.D. and Scandalios J.G. 1992. Differential response of maize catalases to abscisic acid: Vp1 transcriptional activator is not required for ABA-regulated Cat1 expression. Proc. Natl. Acad. Sci. USA 89: 8842–8846.Google Scholar
  45. Williamson, J.D. and Scandalios, J.G. 1994. The maize (Zea mays L.) Cat1 catalase promoter displays differential binding of nuclear proteins isolated from germinated and developing embryos and from embryos grown in the presence and absence of abscisic acid. Plant Physiol. 106: 1373–1380.Google Scholar
  46. Williamson, J.D., Stoop, J.M.H., Massel, M.O., Conkling, M.A. and Pharr, D.M. 1995. Sequence analysis of a mannitol dehydrogenase cDNA from plants reveals a function for the PR protein ELI3. Proc. Natl. Acad. Sci. USA 92: 7148–7152.Google Scholar
  47. Williamson, J.D., Guo, W-W. and Pharr, D. M. 1998. Cloning and characterization of a genomic clone (Accession No. AF067082) encoding mannitol dehydrogenase from celery (Apium grave-olens) (PGR98-137). Plant Physiol. 118: 329.Google Scholar
  48. Yamamoto, Y.T., Zamski, E., Williamson, J.D., Conkling, M.A. and Pharr, D.M. 1997. Subcellular localization of celery mannitol dehydrogenase: a cytoplasmic metabolic enzyme in nuclei. Plant Physiol. 115: 1397–1403.Google Scholar
  49. Zamski, E., Yamamoto, Y.T., Williamson, J.D., Conkling, M.A. and Pharr, D.M. 1996. Immunolocalization of mannitol dehydrogenase in celery plants and cells. Plant Physiol. 112: 931–938.Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Eli Zamski
    • 1
  • Wei-Wen Guo
    • 1
  • Yuri T. Yamamoto
    • 2
  • D. Mason Pharr
    • 1
  • John D. Williamson
    • 1
  1. 1.Departments of Horticultural ScienceUSA
  2. 2.Forestry, North Carolina State UniversityRaleighUSA

Personalised recommendations