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Enhanced Resistance to Phytopathogenic Bacteria in Transgenic Tobacco Plants with Synthetic Gene of Antimicrobial Peptide Cecropin P1

  • Molecular Genetics
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Abstract

Plasmids with a synthetic gene of the mammalian antimicrobial peptide cecropin P1 (cecP1) controlled by the constitutive promoter 35S RNA of cauliflower mosaic virus were constructed. Agrobacterial transformation of tobacco plants was conducted using the obtained recombinant binary vector. The presence of gene cecP1 in the plant genome was confirmed by PCR. The expression of gene cecP1 in transgenic plants was shown by Northern blot analysis. The obtained transgenic plants exhibit enhanced resistance to phytopathogenic bacteria Pseudomonas syringae, P. marginata, and Erwinia carotovora. The ability of transgenic plants to express cecropin P1 was transmitted to the progeny. F0 and F1 plants had the normal phenotype (except for a changed coloration of flowers) and retained the ability to produce normal viable seeds upon self-pollination. Lines of F1 plants with Mendelian segregation of transgenic traits were selected.

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REFERENCES

  1. Boman, H., Peptide Antibiotics and Their Role in Innate Immunity, Annu. Rev. Immunol., 1995, vol. 13, pp. 61–92.

    Article  PubMed  CAS  Google Scholar 

  2. Terras, F.R.G., Eggermont, K., Kovaleva, V., et al., Small Cystein-Rich Antifungal Proteins from Radish: Their Role in Host Defense, Plant Cell, 1995, vol. 7, pp. 573–588.

    Article  PubMed  CAS  ISI  Google Scholar 

  3. Hyltmark, D., Engstrom, A., Bennich, H., et al., Insect Immunity: Isolation and Structure of Cecropin D and Four Minor Antibacterial Components from Cecropia pupa, Eur. J. Biochem., 1982, vol. 127, pp. 207–217.

    Google Scholar 

  4. Mills, D. and Hammerschlag, F.A., Effect of Cecropin B on Peach Pathogens, Protoplasts, and Cells, Plant Sci., 1993, vol. 93, pp. 143–150.

    Article  CAS  Google Scholar 

  5. Jaynes, J.M., Nagpala, P., Destefano-Beltran, L., et al., Expression of a Cecropin B Lytic Peptide Analog in Transgenic Tobacco Confers Enhanced Resistance to Bacterial Wilt Caused by Pseudomonas solanacearum, Plant Sci., 1993, vol. 89, pp. 43–53.

    Article  CAS  Google Scholar 

  6. Ohshima, M., Mitruhara, I., Okamoto, M., et al., Enhanced Resistance to Bacterial Diseases of Transgenic Tobacco Plants Overexpressing Sarcotoxin IA, a Bactericidal Peptide of Insect, J. Biochem., 1999, vol. 125, pp. 431–435.

    PubMed  CAS  Google Scholar 

  7. Parashina, E.V., Shadenkov, A.A., Lavrova, N.V., and Avetisov, V.A., Use of a Defensive Peptide (Defensin) Gene from Radish Seeds to Improve the Tomato Resistance to Fungal Diseases, Biotekhnologiya, 1999, vol. 6, pp. 35–41.

    Google Scholar 

  8. Bur'yanov, Ya.I. and Kado, K.I., Strategies of Constructing Transgenic Plants Resistant to Phytopathogens and Pests, Bioorg. Khim., 1999, vol. 25, pp. 903–910.

    Google Scholar 

  9. Allefs, S., Florask, D., Hoogendoorn, C., and Stiekema, W., Erwinia Soft Rot Resistance of Potato Cultivars Transformed with a Gene Construct Coding for Antimicrobial Peptide Cecropin B Is Not Altered, Am. Potato J., 1995, vol. 72, pp. 437–445.

    CAS  Google Scholar 

  10. Martemyanov, K.A., Spirin, A.S., and Gudkov, A.T., Synthesis, Cloning and Expression of Genes for Antibacterial Peptides: Cecropin, Magainin, and Bombinin, Biotechnol. Lett., 1996, vol. 18, pp. 1357–1362.

    Article  CAS  Google Scholar 

  11. Murashige, T. and Skoog, F., A Revised Medium for Rapid Growth and Bioassays with Tobacco Cultures, Physiol. Plant., 1962, vol. 15, pp. 473–497.

    CAS  Google Scholar 

  12. Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, New York: Cold Spring Harbor Lab., 1989.

    Google Scholar 

  13. Topfer, R., Matzeit, V., Gronenborn, B., et al., A Set of Plant Expression Vectors for Transcriptional and Translational Fusions, Nucleic Acids Res., 1987, vol. 15, p. 5890.

    PubMed  CAS  Google Scholar 

  14. An, G., Ebert, P., Mitra, A., and Ha, S.B., Binary Vectors, Plant Molecular Biology Manual, Gelvin, S.B., Schilperoort, R.A., and Verma, D.P.S., Eds., Dordrecht: Kluwer Academic, 1988, vol. A3, pp. 1–19.

    Google Scholar 

  15. Koncz, C. and Schell, J., The Promoter of TL-DNA Gene 5 Controls the Tissue Specific Expression of Chimeric Genes Carried by a Novel Type of Agrobacterium Binary Vector, Mol. Gen. Genet., 1986, vol. 204, pp. 383–396.

    Article  CAS  Google Scholar 

  16. Van Haute, E., Joos, H., Maes, M., et al., Intergeneric Transfer and Exchange Recombination of Restriction Fragments Cloned in pBR322: A Novel Strategy for the Reversed Genetics of the Ti Plasmid of Agrobacterium tumefaciens, EMBO J., 1983, vol. 2, pp. 291–299.

    Google Scholar 

  17. Draper, J., Scott, R., and Hamil, J., Transformation of Dicotyledonous Plant Cells Using the Ti Plasmid of Agrobacterium tumefaciens and the Ri Plasmid of A. rhizogenes, Plant Genetic Transformation and Gene Expression: A Laboratory Manual, Draper, J., Scott, R., Armitage, P., and Walden, R., Eds., Oxford: Blackwell Sci., 1988, pp. 69–160.

    Google Scholar 

  18. Edwards, K., Johnstone, C., and Thompson, C., A Simple and Rapid Method for the Preparation of Plant Genomic DNA for PCR Analysis, Nucleic Acids Res., 1991, vol. 19, p. 1349.

    PubMed  CAS  Google Scholar 

  19. Pawlowski, K., Kunze, R., Vries, S., and Bisseling, T., Isolation of Total, Poly(A) and Polysomal RNA from Plant Tissues, Plant Molecular Biology Manual, Gelvin, S.B., Schilperoort, R.A., and Verma, D.P.S., Eds., Dordrecht: Kluwer Academic, 1994, vol. D5, pp. 1–13.

    Google Scholar 

  20. Nagy, F., Kay, S.A., and Chua, N.-H., Analysis of Gene Expression in Transgenic Plants, Plant Molecular Biology Manual, Gelvin, S.B., Schilperoort, R.A., and Verma, D.P.S., Eds., Dordrecht: Kluwer Academic, 1988, vol. B4, pp. 1–29.

    Google Scholar 

  21. Bradford, M.M., A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding, Anal. Biochem., 1976, vol. 72, pp. 248–254.

    Article  PubMed  CAS  Google Scholar 

  22. Matzke, A.J.M., Neuhuber, F., Park, Y.-D., et al., Homology-Dependent Gene Silencing in Transgenic Plants: Epistatic Silencing Loci Contain Multiple Copies of Methylated Transgenes, Mol. Gen. Genet., 1994, vol. 244, pp. 219–229.

    Article  PubMed  CAS  Google Scholar 

  23. Owens, L.D. and Heutte, T.M., A Single Amino Acid Substitution in the Antimicrobial Defense Protein Cecropin B Is Associated with Diminished Degradation by Leaf Intercellular Fluid, Mol. Plant-Microbe Interact., 1997, vol. 10, pp. 525–528.

    PubMed  CAS  Google Scholar 

  24. Sipos, D., Andersson, M., and Ehrenberg, A., The Structure of the Mammalian Antibacterial Peptide 3 Cecropin P1 in Solution, Determined by Proton-NMR, Eur. J. Biochem., 1992, vol. 209, pp. 163–169.

    Article  PubMed  CAS  Google Scholar 

  25. Fischer, R., Budde, I., and Hain, R., Stilbene Synthase Gene Expression Causes Changes in Flower Color and Male Sterility in Tobacco, Plant J., 1997, vol. 11, pp. 489–498.

    Article  CAS  Google Scholar 

  26. Poroiko, V.A., Rukavtsova, E.B., Orlova, I.V., and Bur'yanov, Ya.I., Phenotypic Changes in Transgenic Tobacco Plants Caused by Expression of the Antisense Form for the hmg1 Gene, Russ. J. Genet., 2000, vol. 36, no.9, pp. 998–1002.

    CAS  Google Scholar 

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Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Pushchino Branch), Russian Academy of Sciences, Pushchino, Moscow oblast, 142290, Russia

    N. S. Zakharchenko, E. B. Rukavtsova & Ya. I. Buryanov

  2. Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow oblast, 142290, Russia

    A. T. Gudkov

Authors
  1. N. S. Zakharchenko
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  2. E. B. Rukavtsova
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  3. A. T. Gudkov
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  4. Ya. I. Buryanov
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__________

Translated from Genetika, Vol. 41, No. 11, 2005, pp. 1445–1452.

Original Russian Text Copyright © 2005 by Zakharchenko, Rukavtsova, Gudkov, Buryanov.

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Zakharchenko, N.S., Rukavtsova, E.B., Gudkov, A.T. et al. Enhanced Resistance to Phytopathogenic Bacteria in Transgenic Tobacco Plants with Synthetic Gene of Antimicrobial Peptide Cecropin P1. Russ J Genet 41, 1187–1193 (2005). https://doi.org/10.1007/s11177-005-0218-2

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  • DOI: https://doi.org/10.1007/s11177-005-0218-2

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