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Cytokinin-mediated insect resistance in Nicotiana plants transformed with the ipt gene

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Abstract

The bacterial isopentenyl transferase (ipt) gene involved in cytokinin biosynthesis was fused with a promoter from the proteinase inhibitor II (PI-IIK) gene and introduced into Nicotiana plumbaginifolia. Transcripts of the ipt gene were wound-inducible in leaves of transgenic PI-II-ipt plants. In leaf disks excised from fully expanded leaves, transcript levels increased 25- to 35-fold within 24 h and by 48 h were reduced by about 50%. In flowering plants, message levels were 2- to 5-fold higher than in preflowering plants. These plants were used to test for defensive properties of cytokinins against insects. Manduca sexta larvae consumed up to 70% less of the PI-II-ipt leaf material on flowering plants than larvae feeding on controls. Normal development of Myzus persicae nymphs was also delayed. Approximately half as many nymphs reached adulthood on PI-II-ipt leaves than on controls. Zeatin and zeatinriboside levels in leaves remaining on PI-II-ipt plants after hornworm feeding were elevated by about 70-fold and the chlorophyll a/b content was double that of controls. Exogenous applications of zeatin to the PI-II-ipt leaves enhanced the level of resistance to the tobacco hornworm and almost completely inhibited normal development of the green peach aphid nymphs. Transcript levels of an acidic chitinase gene were low and minimally inducible in PI-II-ipt leaves. The mode of action of the cytokinin gene product on enhanced insect resistance is not clear but may involve the products of secondary metabolic pathways.

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References

  1. Akiyoshi DE, Morris RO, Hinz R, Mischke BS, Kosuge T, Garfinkel D, Gordon MP, Nester EW: Cytokinin/auxin balance in crown gall tumors is regulated by specific loci in the T-DNA. Proc Natl Acad Sci USA 80: 407–411 (1983).

    Google Scholar 

  2. An G, Mita A, Choi HK, Costa MA, An K, Thornburg RW, Ryan CA: Functional analysis of the 3′-control region of the potato wound-inducible proteinase inhibitor II gene. Plant Cell 1: 115–122 (1989).

    Article  PubMed  Google Scholar 

  3. Bailiss KW, Cocker FM, Cassels AC: The effect of Benlate and cytokinins on the content of tobacco mosaic virus in tomato leaf disks and cucumber mosaic virus in cucumber cotyledon disks and seedlings. Ann Appl Biol 87: 383–392 (1977).

    Google Scholar 

  4. Balazs E, Kiraly Z: Virus content and sympton expression in Samsun tobacco treated with kinetin and a benzimidazole derivative. Phytopath Z 100: 356–360 (1981).

    Google Scholar 

  5. Barker RF, Idler KB, Thompson DV, Kemp TD: Nucleotide sequence of the T-DNA region from the Agrobacterium tumefaciens octopine Ti plasmid pTil5955. Plant Mol Biol 2: 335–350 (1983).

    Google Scholar 

  6. Binns AN, Chen RH, Wood HN, Wood HN, Lynn DG: Cell division promoting activity of naturally occurring dehydrodiconiferyl glucosides: do cell wall components control cell division. Proc Natl Acad Sci USA 84: 980–984 (1987).

    PubMed  Google Scholar 

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

    Google Scholar 

  8. Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ: Isolation of biologically active RNA from sources enriched in RNase. Biochemistry 18: 5294–5299 (1979).

    PubMed  Google Scholar 

  9. Davis PJ. Plant Hormones and their Role in Plant Growth and Development. Kluwer Academic Publishers, Boston, MA (1987).

    Google Scholar 

  10. Dellaporta SL, Wood J, Hicks JB: Maize DNA miniprep. In: Malmberg R, Messing J, Sussex I (eds) Molecular Biology of Plants: A Laboratory Course Manual, pp. 36–37. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1984).

    Google Scholar 

  11. Gatehouse JA: Breeding for resistance to insects. In: Murray DR (ed) Advanced Methods in Plant Breeding and Biotechnology, pp. 250–276. Redwood Press, Melksham, UK (1991).

    Google Scholar 

  12. Gatehouse AMR, Boulter D, Hilder VA: Potential of plant derived genes in the genetic manipulation of crops for insect resistance. In: Gatehouse AMR, Hilder VA, Boulter D (eds) Plant Genetic Manipulation for Crop Protection, pp. 155–181. Redwood Press, Melksham, UK (1992).

    Google Scholar 

  13. Hallahan DL, Pickett JA, Wadhams LJ, Wallsgrove RM, Woodcock C: Potential of secondary metabolites in genetic engineering of crops for resistance. In: Gatehouse AMR, Hilder VA, Boulter D, (eds) Plant Genetic Manipulation for Crop Protection, pp. 212–248. Redwood Press, Melksham, UK (1992).

    Google Scholar 

  14. Hedin PA, McCarty JC, Thompson AC, Jenkins JN, Smith DH, Shepherd RL, Parrott WL: Plant bioregulator induced increases in the protein content of cotton plant tissues. J Agric Food Chem 36: 742–745 (1988).

    Google Scholar 

  15. Hedin PA, Williams WP, Davis FM, Thompson AC: Effects of plant bioregulators on nutrients, insect resistance, and yield of corn (Zea mays L.). J Agric Food Chem 36: 746–748 (1988).

    Google Scholar 

  16. Holden M: Chlorophylls. In: Goodwin TW (ed) Chemistry and Biochemistry of Plant Pigments, pp 1–32. Academic Press, New York (1976).

    Google Scholar 

  17. Hood EE, Helmer GL, Fraley RT, Chilton MD: The hypervirulence of Agrobacterium tumefaciens A 281 is encoded in a region of pTIBo542 outside of T-DNA. J Bact 168: 1291–1301 (1986).

    PubMed  Google Scholar 

  18. Horsch RB, Fry JE, Hoffmann N, Eichholtz D, Rogers SG, Fraley RT. A simple and general method for transferring genes into plants. Science 227: 1229–1231 (1985).

    Google Scholar 

  19. Johnson R, Narvaez J, An G, Ryan C: Expression of proteinase inhibitors I and II in transgenic tobacco plants: effects on natural defense against Manduca sexta larvae. Proc Natl Acad Sci USA 86: 9871–9875 (1989).

    PubMed  Google Scholar 

  20. Lorberth R, Dammann C, Ebneth M, Amati S, Sanchez-Serrano J: Promoter elements involved in environmental and developmental control of potato proteinase inhibitor II expression. Plant J 2: 477–486 (1992).

    Article  PubMed  Google Scholar 

  21. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  22. Memelink J, Hoge JHC, Schilperoort RA: Cytokinin stress changes the developmental regulation of several defence-related genes in tobacco. EMBO J 6: 3579–3583 (1987).

    Google Scholar 

  23. Moffat AS: Improving plant disease resistance. Science 257: 482–483 (1992).

    Google Scholar 

  24. Nicholson RL: Phenolic compounds and their role in disease resistance. Annu Rev Phytopath 30: 369–389 (1992).

    Article  Google Scholar 

  25. Orr JD, Lynn DG: Biosynthesis of dehydrodiconiferyl alcohol glucosides: implications for the control of tobacco cell growth. Plant Physiol 98: 343–352 (1992).

    Google Scholar 

  26. Payne G, Ahl P, Moyer M, Harper A, Beck J, Meins F, Ryals J: Isolation of complementary DNA clones encoding pathogenesis-related proteins P and Q, two acidic chitinases from tobacco. Proc Natl Acad Sci USA 87: 98–102 (1990).

    PubMed  Google Scholar 

  27. Sanchez-Serrano JJ, Keil M, O'Connor A, Schell J, Willmitzer L: Wound-induced expression of a potato proteinase inhibitor II gene in transgenic tobacco plants. EMBO J 6: 303–306 (1987).

    Google Scholar 

  28. Skoog F, Miller CO: Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp Soc Exp Biol 11: 118–130 (1957).

    Google Scholar 

  29. Smigocki AC: Cytokinin content and tissue distribution in plants transformed by a reconstructed isopentenyl transferase gene. Plant Mol Biol 16: 105–115 (1991).

    PubMed  Google Scholar 

  30. Smigocki AC, Owens LD: Cytokinin gene fused with a strong promoter enhances shoot organogenesis and zeatin levels in transformed plant cells. Proc Natl Acad Sci USA 85: 5131–5135 (1988).

    Google Scholar 

  31. Smigocki AC, Owens LD: Cytokinin-to-auxin ratios and morphology of shoots and tissues transformed by a chimeric isopentenyl transferase gene. Plant Physiol 91: 808–811 (1989).

    Google Scholar 

  32. Sollner-Webb D, Reeder RH: The nucleotide sequence of the initiation and termination sites for ribosomal RNA transcription in X. laevis. Cell 18: 485–499 (1979).

    Article  PubMed  Google Scholar 

  33. Teutonico RA, Dudley MW, Orr JD, Lynn DG, Binns AN: Activity and accumulation of cell division-promoting phenolics in tobacco tissue cultures. Plant Physiol 97: 288–297 (1991).

    Google Scholar 

  34. Thomas TH, Balkesley D: Cytokinins-plant hormones in search of a role-practical and potential uses of cytokinins in agriculture and horticulture. Monogr Br Growth Regulator Group 14: 69–83 (1987).

    Google Scholar 

  35. Thornburg RW, Kernana A, Molin L: Cj-hloramphenicol acetyl transferase (CAT) protein is expressed in transgenic tobacco in field tests following attack by insects. Plant Physiol 92: 500–505 (1990).

    Google Scholar 

  36. Thornburg RW, An G, Cleveland TE, Johnson R, Ryan CA: Wound-inducible expression of a potato inhibitor II-CAT gene fusion in transgenic tobacco plants. Proc Natl Acad Sci USA 84: 744–748 (1987).

    Google Scholar 

  37. Ward ER, Uknes JJ, Williams SC, Dincher SS, Wiederhold DL, Alexander DC, Ahl-Goy P, Metraux J-P, Ryals JA: Coordinate gene activity in response to agents that induce systemic acquired resistance. Plant Cell 3: 1085–1094 (1991).

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. Plant Molecular Biology Laboratory Agricultural Research Service, U.S. Department of Agriculture, 20705, Beltsville, MD, USA

    A. Smigocki & I. McCanna

  2. Florist and Nursery Crops Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 20705, Beltsville, MD, USA

    J. W. Neal Jr.

  3. Animal Sciences Department, University of Maryland, 20742, College Park, MD, USA

    L. Douglass

Authors
  1. A. Smigocki
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  2. J. W. Neal Jr.
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  3. I. McCanna
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  4. L. Douglass
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Smigocki, A., Neal, J.W., McCanna, I. et al. Cytokinin-mediated insect resistance in Nicotiana plants transformed with the ipt gene. Plant Mol Biol 23, 325–335 (1993). https://doi.org/10.1007/BF00029008

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  • DOI: https://doi.org/10.1007/BF00029008

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