Summary
We used in vitro growth inhibition assays to demonstrate that synthetic cecropin protein has potent activity against a range of plant pathogenic bacteria. We then prepared transgenic tobacco plants which express cecropin mRNA and protein. We have used Pseudomonas syringae pv tabaci infection of these transgenic tobacco as a model system to evaluate whether the plants which express cecropin protein also have increased tolerance to infection. We found no dramatic difference in disease response between plants which are expressing cecropin protein and control plants which were derived from the transformation with a binary vector which did not carry the gene encoding cecropin protein.
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References
Agrios, G.N. (1988). In Plant Pathology, (eds.). New York.: Academic Press, pp.
Andreu, D., Merrifield, R., Steiner, H. and Boman, H. (1983). Solidphase synthesis of cecropin A and related peptides. Proc Natl Acad Sci USA. 80: 6475–6479.
Andreu, D., Merrifield, R., Steiner, H. and Boman, H. (1985). Nterminal analogues of cecropin A: synthesis, antibacterial activity, and conformational properties. Biochemistry. 24: 1683–1688.
Boman, J. and Hultmark, D. (1987). Cell-free immunity in insects. Ann Rev Microbiol 41: 103–126.
Bradford, M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Anal Biochem. 72: 248–254.
Christensen, B., Fink, J., Merrifield, R. and Mauzerall, D. (1988). Channel-forming properties of cecropins and related model compounds incorporated into planar lipid membranes. Proc Natl Acad Sci USA. 85: 5072–5076.
Denecke, J., Botterman, J. and Deblaere, R. (1990). Protein secretion in plant cells can occur via a default pathway. The Plant Cell. 2: 51–59.
Destefano-Beltran, L., Nagpala, P., Kim, J., Dodds, J. and Jaynes, J. (1991). Genetic transformation of potato to enhance nutritional value and confer disease resistance. Plant Gene Research; Molecular approaches to crop improvement. 17–32.
Diachun, S. and Troutman, J. (1954). Multiplication of Pseudomonas tabaci in leaves of burley tobacco, Nicotiana longiflora, and hybrids. Phytopathology. 44:186–187.
Figurski, D. and Helinski, D. (1979). Replication of an origincontaining derivative of plasmid RK2 dependent on a plasmid function in rans. Proc Natl Acad Sci USA. 76: 1648–1652.
Green, N., Alexander, H., Olson, A., Alexander, S., Shinnick, T., Sutcliffe, J. and Lerner, R. (1982). Immunogenic structure of the influenza virus hemagglutimn. Cell. 28: 477–487.
Harlow, E. and Lane, D. (1988). Antibodies, A Laboratory Manual. In (eds.). New York: Cold Spring Harbor, pp.
Hoekema, A., Hirsch, P., Hooykas, P. and Schilperoort, R. (1983). A binary vector plant strategy based on separation of vir and T-region of Agrobacterium tumefaciens T-plasmid. Nature. 303: 179–180.
Jaynes, J., Nagpala, P., Destefano-Beltran, L., Huang, J., Kim, J., Denny, T. and Cetiner, S. (1993). Expression of a cecropin B lytic peptide analog in transgenic tobacco confers enhanced resistance to bacterial wilt caused by Pseadomonas solanacearum. Plant Science. 89: 43–53.
Jaynes, J., Xanthopoulos, K., Destéfano-Beltrán, L. and Dodds, J. (1987). Increasing bacterial disease resistance in plants utilizing antibacterial genes from insects. BioEssays. 6: 263–270.
Jones, R., Barry, A., Gavan, T. and Washington II, J. (1985). Susceptibility tests: microdilution and macrodilution broth procedures. Manual of Clinical Microbiology. 972–977.
Mauch, F. and Staehelin, L. (1989). Functional implications of the subcellular localization of ethylene-induced chitinase and beta-1,3-glucanase in bean leaves. The Plant Cell. 1: 447–457.
Perlak, F., Fuchs, R., Dean, D., McPherson, S. and Fischoff, D. (1991). Modification of the coding sequence enhances plant expression of insect control protein genes. Proc Natl Acad Sci USA. 88: 3324–3328.
Reichlin, M. (1980). Use of glutaraldehyde as a coupling agent for proteins and peptides. Methods in Enzymology. 70: 159–165.
Salunkhe, D. and Desai, R. (1984). Postharvest Biotechnology of Vegetables. In (eds.). New York,: CRC Press, pp.
Sherf, A. and Macnab, A. (1986). Vegetable Diseases and Their Control. In (eds.). New York: Wiley and Sons, pp.
Steiner, H., Hultmark, D., Engström, Å., Bennich, H. and Boman, H. (1981). Sequence and specificity of two antibacterial proteins involved in insect immunity. Nature. 292: 246–248.
van den Elzen, P., Lee, K., Townsend, J. and Bedbrook, J. (1985). Simple binary vectors for DNA transfer to plant cells. Plant Mol Biol. 5: 249–254.
van Hofsten, P., Faye, I., Kockum, K., Lee, J.-Y., Xanthopoulos, K., Boman, I., Boman, H., Engström, Å., Andreu, D. and Merrifield, R. (1985). Molecular cloning, cDNA sequencing, and chemical synthesis of cecropin B from Hyalophora cecropia. Proc Natl Acad Sci USA. 82: 2240–2243.
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Hightower, R., Baden, C., Penzes, E. et al. The expression of cecropin peptide in transgenic tobacco does not confer resistance to Pseudomonas syringae pv tabaci . Plant Cell Reports 13, 295–299 (1994). https://doi.org/10.1007/BF00233324
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DOI: https://doi.org/10.1007/BF00233324