Summary
Simultaneous expression of a tobacco class I chitinase and a class I β-1,3-glucanase gene in tomato resulted in increased fungal resistance, whereas transgenic tomato plants expressing either one of these genes were not protected against fungal infection. After infection with Fusarium oxysporum f.sp. lycopersici, a 36% to 58% reduction in disease severity was observed in resistant tomato lines. Two transgenic lines largely recovered from the initial infection by the time wild-type tomato plants had died.
The overall results are consistent with the observation that class I chitinases and class I β-1,3-glucanases synergistically inhibit the growth of fungi in vitro and provide the first experimental support to the hypothesis that such synergy can contribute to enhanced fungal resistance in planta.
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Alexander D., R.M. Goodman, M. Gut-Rella, C. Glascock, K. Weymann, L. Friedrich, D. Maddox, P. Ahl-Goy, T. Luntz, E. Ward & J.A. Ryals, 1993. Increased tolerance to two oomycete pathogens in transgenic tobacco expressing pathogenesis-related protein 1 a. Proc. Natl. Acad. Sci. USA 90: 7327–7331.
Broglie K., I. Chet, M. Holliday, R. Cressman, P. Biddle, S Knowlton, C.J. Mauvais & R. Broglie, 1991. Transgenic plants with enhanced resistance to the fungal pathogen Rhizoctonia solani. Science 254: 1194–1197.
Collinge D.B., K.M. Kragh, J.D. Mikkelsen, K.K. Nielsen, U. Rasmussen & K. Ved, 1993. Plant chitinases. The Plant Journal 3: 31–40.
Cornelissen B.J.C. & L.S. Melchers, 1993. Strategies for control of fungal diseases in plants. Plant Physiol. 101: 709–712.
Hain R., H.J. Reif, E. Krause, R. Langebartels, H. Kindl, B. Vornam, W. Wiese, E. Schmeltzer, P.H. Schreier, R.H. Stöcker & K. Stenzel, 1993. Disease resistance results from foreign phytoalexin expression in a novel plant. Nature 361: 153–156.
Hood E.E., S.B. Gelvin, L.S. Melchers & A. Hoekema, 1993. New Agrobacterium helper plasmids for gene transfer to plants. Transgenic Research 2: 208–218.
Joosten M.H.A.J. & P.J.G.M.de Wit, 1989. Identification of several pathogenesis-related proteins in tomato leaves inoculated with Cladosporium fulvum (syn. Fulvia fulva) as 1,3-β-glucanases and chitinases. Plant Physiology 89: 945–951.
Kalloo G. & B.O. Bergh, 1993. Genetic Improvement of Vegetable Crops. Pergamon Press Ltd., Oxford.
Koorneef M., J.A.M.van Diepen, C.J. Hanhart, A.C. Kieboomde Waard, L. Martinelli, H.C.H. Schoenmakers & J. Wijbrandi, 1989. Chromosomal instability in cell and tissue cultures of tomato haploids and diploids. Euphytica 43: 179–186.
Lamb C.J., J.A. Ryals, E.R. Ward & R.A. Dixon, 1992. Emerging strategies for enhancing crop resistance to microbial pathogens. Bio/Technology 10: 1436–1445.
Leah R., H. Tommerup, I. Svendsen & J. Mundy, 1991. Biochemical and molecular characterization of three barley seed proteins with antifungal properties. J. Biol. Chem. 266: 1464–1573.
Logemann J., G. Jach, H. Tommerup, J. Mundy & J. Schell, 1992. Expression of a barley ribosome-inactivating protein leads to increased fungal protection in transgenic tobacco plants. Bio/Technology 10: 305–308.
Logemann J., G. Jach, S. Logemann, R. Leah, G. Wolf, J. Mundy, A. Oppenheim, I. Chet & J. Schell, 1993. Expression of a ribosome inhibiting protein (RIP) or a bacterial chitinase leads to fungal resistance in transgenic plants. In: B. Fritig & M. Legrands (Eds) Mechanisms of plant defense responses. pp. 446–448. Kluwer Academic Publishers, Dordrecht.
Mauch F., B. Mauch-Mani & T. Boller, 1988. Antifungal hydrolases in pea tissue. II. Inhibition of fungal growth by combinations of chitinase and β-1,3-glucanase. Plant Physiol. 88: 936–942.
Melchers L.S., M.B. Sela-Buurlage, S.A. Vloemans, C.P. Woloshuk, J.S.C.van Roekel, J. Pen, P.J.M.van den Elzen & B.J.C. Comelissen, 1993. Extracellular targeting of vacuolar tobacco proteins AP24, chitinase and β-1,3-glucanase in transgenic plants. Plant Molec. Biol. 21: 583–593.
Melchers L.S., M. Aphotheker-de Groot, J.A.van der Knaap, A.s. Ponstein, M.B. Sela-Buurlage, J.F. Bol, B.J.C. Cornelissen, P.J.M.van der Elzen & H.J.M. Linthorst, 1994. A new class of tobacco chitinases homologous to bacterial exo-chitinases displays antifungal activity. The Plant Journal 5: 469–480.
Neuhaus J.M., P. Ahl-Goy, U. Hinz, S. Flores & F. Meins, 1991. High level expression of a tobacco chitinase gene in Nicotiana sylvestris. Susceptibility of transgenic plants to Cercospora nicotianae infection. Plant Molec. Biol. 16: 141–151.
Poehlman J.M., 1987. Breeding Field Crops. Van Nostrand Reinhold Publishers, New York, third edition.
Ponstein A.P., S.A. Bres-Vloemans, M.B. Sela-Buurlage, P.J.M.van den Elzen, L.S. Melchers & B.J.C. Comelissen, 1993. A novel pathogen and wound-inducible tobacco protein with antifungal activity. Plant Physiol. 104: 109–118.
Sela-Buurlage M.B., A.S. Ponstein, S.A. Bres-Vloemans, L.S. Melchers, P.J.M.van den Elzen & B.J.C. Cornelissen, 1993. Only specific tobacco (Nicotiana tabacum) chitinases and β-1,3-glucanases exhibit antifungal activity. Plant Physiol. 101: 857–863.
Sijmons P.C., B.M.M. Dekker, B. Schrammeijer, T.C. Verwoerd, P.J.M.van den Elzen & A. Hoekema, 1990. Production of correctly processed human serum albumin in transgenic plants. Bio/Technology 8: 217–221.
Strittmatter G. & D. Wegener, 1993. Genetic engineering of disease and pest resistance in plants: present state of the art. Z. Naturforsch. 48c: 673–688.
Tuzun S., M.N. Rao, U Vogeli, C.L. Schardl & J. Kuc, 1989. Induced systemic resistance to blue mold: early induction and accumulation of β-1,3-glucanases, chitinases and other pathogenesis-related proteins in immunized tobacco. Phytopathology 79: 979–983.
Van den Elzen P.J.M., E. Jongedijk, L.S. Melchers & B.J.C. Cornelissen, 1993. Virus and fungal resistance: from laboratory to field. Phil. Trans. R. Soc. Lond. B 342: 271–278.
Van Roekel J.S.C., B. Damm, L.S. Melchers & A. Hoekema, 1993. Factors influencing transformation frequency of tomato (Lycopersicon esculentum). Plant Cell Reports 12: 644–647.
Vogelsang R. & W. Barz, 1993. Purification, characterization and differential hormonal regulation of a β-1,3-glucanase and two chitinases from chickpea (Cicer arientinum L.). Planta 189: 60–69.
Ward E.R., G.B. Payne, M.B. Moyer, S.C. Williams, S.S. Dincher, K.C. Sharkey, J.H. Beck, H.T. Taylor, P. Ahl-Goy, F. Meins & J.A. Ryals, 1991. Differential regulation of β-1,3-glucanse messenger RNAs in response to pathogen infection. Plant Physiol. 96: 390–397.
Weide R., M. Koorneef & P. Zabel, 1989. A simple nondestructive spraying assay for the detection of an active kanamycin resistance gene in transgenic tomato plants. Theor. Appl. Genet. 78: 169–172.
Wessels J.G.H. & J.H. Sietsma, 1981. Fungal cell walls: a survey. In: W. Tanner & F.A. Loewus (Eds) Encyclopedia of Plant Physiology, new series, vol. 13B: Plant carbohydrates, pp. 352–394. Springer Verlag, Berlin.
Yenofsky R.L., M. Fine & J.W. Pellow, 1990. A mutant neomycin phosphotransferase II gene reduces the resistance of transformants to antibiotic selection pressure. Proc. Natl. Acad. Sci. USA 87: 3435–3439.
Zu Q., E.A. Maher, S. Masoud, R.A. Dixon & C. Lamb, 1994. Enhanced protection against fungal attack by constitutive co-expression of chitinase and glucanase genes in transgenic tobacco. Bio/Technology 12: 807–812.
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Jongedijk, E., Tigelaar, H., van Roekel, J.S.C. et al. Synergistic activity of chitinases and β-1,3-glucanases enhances fungal resistance in transgenic tomato plants. Euphytica 85, 173–180 (1995). https://doi.org/10.1007/BF00023946
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DOI: https://doi.org/10.1007/BF00023946