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Expression of the Talaromyces flavus glucose oxidase gene in cotton and tobacco reduces fungal infection, but is also phytotoxic

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Abstract

Glucose oxidase secreted by the fungus Talaromyces flavus generates, in the presence of glucose, hydrogen peroxide that is toxic to phytopathogenic fungi responsible for economically important diseases in many crops. A glucose oxidase gene from T. flavus, was modified with a carrot extensin signal peptide and fused to either a constitutive or root-specific plant promoter. T1 tobacco plants expressing the enzyme constitutively were protected against infection by the seedling pathogen Rhizoctonia solani. Constitutive expression in tobacco was associated with reduced root growth, and slow germination on culture medium, and with reduced seed set in glasshouse conditions. Several independent transformed cotton plants with a root-specific construct expressed high glucose oxidase activity in the roots, excluding the root tip. Selected T3 homozygous lines showed some protection against the root pathogen, Verticillium dahliae, but not against Fusarium oxysporum. High levels of glucose oxidase expression in cotton roots were associated with reduced height, seed set and seedling germination and reduced lateral root formation. If this gene is to be of value for crop protection against pathogens it will require precise control of its expression to remove the deleterious phenotypes.

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References

  1. Anderson CR, Llewellyn DJ, Peacock WJ, Dennis ES: Cellspecific expression of the promoters of two nonlegume hemoglobin genes in a transgenic legume, Lotus corniculatus. Plant Physiol. 113: 45–57 (1997).

    Article  Google Scholar 

  2. Avigard G: Sucrose and other disaccharides. In: Loewus FA, Tanner W (eds) Encyclopedia of Plant Physiology 13A, pp. 217–347. Springer-Verlag, Berlin (1982).

    Google Scholar 

  3. Bent AF: Plant disease resistance genes: function meets structure. Plant Cell 8: 1757–1771 (1996).

    Article  PubMed  CAS  Google Scholar 

  4. Bogusz D, Llewellyn DJ, Craig S, Dennis ES, Appleby CA, Peacock WJ: Nonlegume haemoglobin genes retain organspecific expression in heterologous transgenic plants. Plant Cell 2: 633–641 (1990).

    Article  PubMed  CAS  Google Scholar 

  5. Boosalis MG: Effect of soil temperature and green-manure amendment of unsterilized soil on parasitism of Rhizoctonia solani by Penicillium vermiculatum and Trichoderma sp. Phytopathol. 46: 473–478 (1956).

    Google Scholar 

  6. Broglie K, Chet I, Holliday M, Cressman R, Biddle P, Knowlton S, Mauvais CJ, Broglie R: Transgenic plants with enhanced resistance to the fungal pathogen Rhizoctonia solani. Science 254: 1194–1197 (1991).

    CAS  PubMed  Google Scholar 

  7. Burke TJ, Callis J, Vierstra RD: Characterisation of a polyubiquitin gene from Arabidopsis thaliana. Mol. Gen. Genet. 213: 435–443 (1988).

    Article  PubMed  CAS  Google Scholar 

  8. Camp WV, Montagu MV, Inzé D: H2O2 and NO: redox signals in disease resistance. Trends in Plant Sci. 3: 330–334 (1998).

    Article  Google Scholar 

  9. Chen J, Varner JE: An extracellular matrix protein in plants: characterisation of a genomic clone for carrot extensin. EMBO J. 4: 2145–2151 (1985).

    PubMed  CAS  Google Scholar 

  10. Cook SA, Stanley PG: Tetrazolium chloride as an indicator of pine pollen germinability. Silvacult Genet. 9: 134–136 (1960).

    CAS  Google Scholar 

  11. Cousins YL, Lyon BR, Llewellyn DJ: Transformation of an Australian cotton cultivar: Prospects for cotton improvement through genetic engineering. Aust J Plant Physiol 18: 481–494 (1991).

    Article  CAS  Google Scholar 

  12. Dolferus R, Jacobs M, Peacock WJ, Dennis ES: Differential interactions of promoter elements in stress responses of the Arabidopsis Adh gene. Plant Physiol 105: 1075–1087 (1994).

    Article  PubMed  CAS  Google Scholar 

  13. Ellis JG, Llewellyn DJ, Dennis ES, Peacock WJ: Maize Adh-1 promoter sequences control anaerobic regulation: addition of upstream promoter elements from constitutive genes is necessary for expression in tobacco. EMBO J 6: 11–16 (1987).

    PubMed  CAS  Google Scholar 

  14. Fiedurek J, Rogalski J, Ilczuk Z, Leonowicz A: Screening and mutagenesis of moulds for the improvement of glucose oxidase production. Enzyme Microb Technol 8: 734–736 (1986).

    Article  CAS  Google Scholar 

  15. Fravel DR, Kim, KK, Papavizas GC: Viability of microsclerotia of Verticillium dahliae reduced by a metabolite produced by Talaromyces flavus. Phytopathol 77: 616–619 (1987).

    Google Scholar 

  16. Grignon C, Sentenac H: pH and ionic conditions in the apoplast. Ann. Rev. Plant Physiol Plant Mol Biol 42: 103–128 (1991).

    Article  CAS  Google Scholar 

  17. Heineke D, Sonnewald U, Büssis D, Günter G, Leidreiter K, Wilke I, Raschke K, Willmitzer L, Heldt HW: Apoplastic expression of yeast-derived invertase in potato. Plant Physiol 100: 301–308 (1992).

    Article  PubMed  CAS  Google Scholar 

  18. Hewitt EJ: Sand and water culture methods used in the study of plant nutrition. pp. 187–193. 2nd edition (1966).

  19. Hughes DW, Galau G: Preparation of RNA from cotton leaves and pollen. Plant Mol Biol Rep 6: 253–257. (1988).

    CAS  Google Scholar 

  20. Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).

    CAS  Google Scholar 

  21. Kim KK, Fravel DR, Papavizas GC: Production, purification and properties of glucose oxidase from the biocontrol fungus Talaromyces flavus. Can J Microbiol 36: 199–205 (1990).

    CAS  Google Scholar 

  22. Kim KK, Fravel DR, Papavizas GC: Glucose oxidase as the antifungal principle of talaron from Talaromyces flavus. Can J Microbiol 36: 760–764 (1990).

    Article  PubMed  CAS  Google Scholar 

  23. Kirschman JA, Cramer JH: Two new tools: multipurpose cloning vectors that carry kanamycin or spectinomycin/ streptomycin resistance markers. Gene 68: 163–165 (1988).

    Article  PubMed  CAS  Google Scholar 

  24. Lazo GR, Stein PA, Ludwig RA: A DNA transformation competent Arabidopsis library in Agrobacterium. Bio/technology 9: 963–967 (1991).

    Article  PubMed  CAS  Google Scholar 

  25. Lin W, Anuratha CS, Datta K, Potrykus I, Muthukrishnan S, Datta SK: Genetic engineering of rice for resistance to sheath blight. Bio/technology 13: 686–690 (1995).

    Article  CAS  Google Scholar 

  26. Lucus W, Madore MA: Recent advances in sugar transport. In: Stumpf PK, Conn EE (eds) The Biochemistry of Plants: A Comprehensive Treatise, vol 14, pp. 35–84. Academic Press, San Diego (1988).

    Google Scholar 

  27. Marois JJ, Dunn MT, Papavizas GC: Biological control of Verticillium wilt of eggplant in the field. Plant Dis. 66: 1166–1168 (1982).

    Article  Google Scholar 

  28. Mitchell LE, Dennis ES, Peacock WJ: Molecular analysis of an alcohol deydrogenase (Adh) gene from chromosome 1 of wheat. Genome 32: 349–358 (1989).

    PubMed  CAS  Google Scholar 

  29. Murray FR, Llewellyn DJ, Peacock WJ, Dennis ES: Isolation of the glucose oxidase gene from Talaromyces flavus and characterisation of its role in the biocontrol of Verticillium dahliae. Current Genetics 32: 367–375 (1997).

    Article  PubMed  CAS  Google Scholar 

  30. Odell JT, Nagy F, Chua N: Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313: 810–812 (1985).

    Article  PubMed  CAS  Google Scholar 

  31. Olson PD, Varner JE: Hydrogen peroxide and lignification. Plant J 4: 887–892 (1993).

    Article  CAS  Google Scholar 

  32. Roitsch T, Tanner W: Cell wall invertase: bridging the gap. Bot. Acta 109: 90–93 (1996).

    CAS  Google Scholar 

  33. Sanger M, Daubert S, Goodman RM: Characteristics of a strong promoter from figwort mosaic virus: comparison with the analogous 35S promoter from cauliflower mosaic virus and the regulated mannopine synthase promoter. Plant Mol Biol 14: 433–443 (1990).

    Article  PubMed  CAS  Google Scholar 

  34. Sonnewald U, Brauer M, Schaewen A, Stitt M, Willmitzer L: Transgenic tobacco plants expressing yeast-derived invertase in either the cytosol, vacuole or apoplast: a powerful tool for studying sucrose metabolism and sink/source interactions. Plant J 1: 95–106 (1991).

    PubMed  CAS  Google Scholar 

  35. Su SJ, Leu LS: Three parasitic fungi on Sclerotinia sclerotiorum (Lib) de Bary. Plant Prot Bull (Taipei) 22: 253–262 (1980).

    Google Scholar 

  36. Tabe LM, Wardley-Richardson T, Ceriotti A, Aryan AP, McNabb W, Moore AE, Higgins TJV: A biotechnological approach to improving the nutritive value of alfalfa. J Anim Sci 73: 2752–2759 (1995).

    PubMed  CAS  Google Scholar 

  37. Terry ME, Bonner BA: An examination of centrifugation as a method of extracting an extracellular solution from peas, and its use for the study of indoleacetic acid-induced growth. Plant Physiol 66: 321–325 (1980).

    PubMed  CAS  Google Scholar 

  38. Thordal-Christensen H, Zhang Z, Wei Y, Collinge DB: Subcellular localisation of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barleypowdery mildew interaction. Plant J 11: 1187–1194 (1997).

    Article  CAS  Google Scholar 

  39. Weber H, Borisjuk L, Wobus U: Sugar import and metabolism during seed development. Trends Plant Sci 2: 169–174 (1997).

    Article  Google Scholar 

  40. Wu G, Shortt BJ, Lawrence EB, Levine EB, Fitzsimmons KC, Shah DM: Disease resistance conferred by expression of a gene encoding H2O2-generating glucose oxidase in transgenic potato plants. Plant Cell 7: 1357–1368 (1995).

    Article  PubMed  CAS  Google Scholar 

  41. Yamamoto YT, Taylor CG, Acedo GN, Cheng CL, Conkling MA: Characterisation of cis-acting sequences regulating rootspecific gene expression in tobacco. Plant Cell 3: 371–382 (1991).

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. CSIRO Plant Industry, P.O. Box 1600, Canberra City, ACT, 2601, Australia

    Fiona Murray, Danny Llewellyn, Helen McFadden, David Last, Elizabeth S. Dennis & W. James Peacock

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  1. Fiona Murray
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  2. Danny Llewellyn
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  3. Helen McFadden
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  4. David Last
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  5. Elizabeth S. Dennis
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  6. W. James Peacock
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Murray, F., Llewellyn, D., McFadden, H. et al. Expression of the Talaromyces flavus glucose oxidase gene in cotton and tobacco reduces fungal infection, but is also phytotoxic. Molecular Breeding 5, 219–232 (1999). https://doi.org/10.1023/A:1009625801909

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