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Enhanced NaCl Stress Tolerance in Transgenic Tobacco Expressing Bacterial Choline Dehydrogenase

Abstract

The intracellular accumulation of osmoprotectants, such as glycine betaine and other low molecular weight compounds, is a well investigated response of environmental stress occurring in a wide range of organisms. By introducing the bacterial bet A gene, encoding choline dehydrogenase (CDH), into tobacco both a salt and choline resistant phenotype was achieved. As measured by dried weights, there was an 80% increase in salt tolerance between the transgenic and wild-type plants at 300 mM NaCl.

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References

  1. Boyer, J.S. 1982. Plant productivity and environment. Science 218: 443–448.

    Article  CAS  Google Scholar 

  2. Yancey, P.H., Clark, M.E., Hand, S.C., Bowlus, R.D. and Somero, G.N. 1982. Living with water stress: Evolution of osmolyte systems. Science 217: 1214–1222.

    Article  CAS  Google Scholar 

  3. McCue, K.F. and Hanson, A.D. 1990. Drought and salt tolerance: towards understanding and application. Tibtech 8: 358–362.

    Article  CAS  Google Scholar 

  4. Incharoensakdi, A., Takabe, T. and Akazawa, T. 1986. Effect of betaine on enzyme activity and subunit interaction of ribulose-l,5-bisphosphate carboxy-lase/oxygenase fromAphanothece halophytica . Plant Physiol. 81: 1044–1049.

    Article  CAS  Google Scholar 

  5. Wyn Jones, R.G. and Storey, R. 1981. Physiology and biochemistry of drought resistance in plants, Betaines, p. 172–204. Academica Press Australia.

    Google Scholar 

  6. Kishitani, S., Watanabe, K., Yasuda, S., Arakawa, K. and Takabe, T. 1994. Accumulation of glycinebetaine during cold acclimation and freezing in leaves of winter and spring barley plants. Plant, Cell and Environment. 17: 89–95.

    Article  CAS  Google Scholar 

  7. McCue, K.F. and Hanson, A.D. 1992. Salt-inducible betaine aldehyde dehydrogenase from sugar beet: cDNA cloning and expression. Plant Mol. Biol. 18: 1–11.

    Article  CAS  Google Scholar 

  8. Weretilnyk, E.A., Bednarek, S., McCue, K.F., Rhodes, D. and Hanson, A.D. 1989. Comparative biochemical and immunological studies of the glycine betaine synthesis pathway in diverse families of dicotyledons. Planta 178: 342–352.

    Article  CAS  Google Scholar 

  9. Hanson, A.D. and Rhodes, D. 1983. 14C Tracer evidence for synthesis of choline and betaine via phosphoryl base intermediates in salinized sugarbeet leaves. Plant Physiol. 71: 692–700.

    Article  CAS  Google Scholar 

  10. Brouquisse, R., Weigel, P., Rhodes, D., Yocum, C.F. and Hason, A.D. 1988. Evidence for a ferredoxin-dependent choline monooxygenase from spinach chloroplast stroma. Plant Physiol. 90: 322–329.

    Article  Google Scholar 

  11. Landfald, B. and Ström, A.R. 1986. Choline-glycine betaine pathway confers a high level of osmotic tolerance in Escherichia coli J . Bacteriol. 165: 849–855.

    Article  CAS  Google Scholar 

  12. Lamark, T. et al. 1991. DNA sequence and analysis of the bet genes encoding the osmoregulatory choline-glycine betaine pathway of Escherichia coli . Mol. Microbiol. 5: 1049–1064.

    Article  CAS  Google Scholar 

  13. Boyd, L.A. et al. 1991. Characterization of an Escherichia coli gene encoding betaine aldehyde dehydrogenase (BADH): structural similarity to mammalian ALDHs and a plant BADH. Gene 103: 45–52.

    Article  CAS  Google Scholar 

  14. Rathinasabapathi, B., McCue, K.F., Gage, D.A. and Hanson, A.D. 1994. Metabolic engineering of glycine betaine synthesis: plant betaine aldehyde dehydrogenase lacking typical transit peptide are targeted to tobacco chloro-plasts where they confer betaine aldehyde resistance. Planta 193: 155–162.

    Article  CAS  Google Scholar 

  15. Holmström, K. et al.1994. Production of Escherichia coli betaine-aldehyde dehydrogenase, an enzyme required for the synthesis of the osmoprotectant glycine betaine, in transgenic plants. Plant J. 6: 749–758.

    Article  Google Scholar 

  16. Giannoni, F., Field, J.F. and Davidson, N.O. 1994. An improved reverse transcription polymerase chain reaction method to study a lipoprotein gene expression in Caco-2 cells. J. Lip. Res. 35: 340–350.

    CAS  Google Scholar 

  17. Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W. 1987. GUS fusions: β-glu-curonidase as a sensitive and versatile gene fusion marker in higher plants.EMBO J. 6: 3901–3907.

    Article  CAS  Google Scholar 

  18. Bartels, D. and Nelson, D. 1994. Approaches to improve stress tolerance using molecular genetics. Plant, Cell and Environment 17: 659–667.

    Article  CAS  Google Scholar 

  19. Tarszynski, M.C., Jensen, R.G. and Bohnert, H.J. 1993. Stress protection of tobacco by production of the osmolyte mannitol. Science 259: 508–510.

    Article  Google Scholar 

  20. Le Rudlier, D., Ström, A.R., Dandekar, A.M., Smith, L.T. and Valentine, R.C. 1984. Molecular biology of osmoregulation. Science 224: 1064–1068.

    Article  Google Scholar 

  21. Ishitani, M., Arakawa, K., Mizuno, K., Kishitani, S. and Takabe, T. 1993. Betaine aldehyde dehydrogenase in Gramineae: levels in leaves of both betaine accumulating and nonaccumulating cereal plants. Plant Cell Physiol. 34: 493–495.

    CAS  Google Scholar 

  22. Saki, R.K. et al. 1988. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239: 487–91.

    Article  Google Scholar 

  23. Walden, R., Koncz, C. and Schell, J. 1990. The use of gene vectors in plant molecular biology. MMCB 1: 175–194.

    CAS  Google Scholar 

  24. Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

  25. Figurski, D.H. and Helinski, D.R. 1979. Replication of an origin-containing derivate of plasmid RK2 dependent on a plasmid function provided in trans . Proc. Natl. Acad. Sci. USA 76: 1648–1652.

  26. Edwards, K., Johnstone, C. and Thomson, C. 1991. A simple and rapid method for the preparation of plant genomic DNA for PCR analysis.Nucl. Acids Res. 19: 1349.

    Article  CAS  Google Scholar 

  27. Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassay with tobacco tissue cultures.Physiol. Plant 15: 473–497.

    Article  CAS  Google Scholar 

  28. Rogers, S.G. and Fraley, R.T. 1986. Gene transfer to plants: production of transformed plants using Ti plasmid vectors. Methods in Enzymology 118: 627–684.

    Article  CAS  Google Scholar 

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Lilius, G., Holmberg, N. & Bülow, L. Enhanced NaCl Stress Tolerance in Transgenic Tobacco Expressing Bacterial Choline Dehydrogenase. Nat Biotechnol 14, 177–180 (1996). https://doi.org/10.1038/nbt0296-177

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