Skip to main content
SpringerLink
Log in

Phytodecta fornicata Brüggemann resistance mediated by oryzacystatin II proteinase inhibitor transgene

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

Phytodecta fornicata Brüggemann is a serious pest of alfalfa (Medicago sativa L.) that causes significant crop loss in the Balkan peninsula of Europe. We introduced a wound-inducible oryzacystatin II (OCII) gene to alfalfa to evaluate its effect on survival of P. fornicata larvae. Feeding bioassays with second, third and fourth instars were carried out using transgenic plants that were shown to express OCII at 24 and 48 h after wounding. Second and third instars were the most sensitive to the ingestion of OCII, whereas no effects were observed with fourth instars. About 80% of the second and third instars died after 2 days of feeding on the transgenic plants as compared to 0–40% on the controls. This is the first report that demonstrates significant increase in mortality of P. fornicata on transgenic plants that express a cysteine proteinase inhibitor gene, and this knowledge should lead to the development of effective management strategies for this devastating pest of alfalfa.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Abe K, Arai S (1985) Purification of a cysteine proteinase inhibitor from rice, Oryza sativa L. japonica. Agric Biol Chem 49:3349–3350

    CAS  Google Scholar 

  • Applebaum SW (1985) Biochemistry of digestion. In: GA Kerkut, LI Gilbert (eds) Comparative physiology, biochemistry and pharmacology of insects, vol 4. Pergamon Press, London, pp 279–311

    Google Scholar 

  • Barrett AJ (1986) The classes of proteolytic enzymes. In: MJ Dalling (ed) Plant proteolytic. Enzymes CRC, Boca Raton, FL, pp 1–16

    Google Scholar 

  • Blake MS, Johnston KH, Russel-Jones GJ, Gotschlich EC (1984) A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on western blots. Anal Biochem 136:175–179

    Article  PubMed  CAS  Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein using the principle of dyebinding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Brzin J, Popovic T, Ritonja A, Puizdar V, Kidric M (1998) Related cystatin inhibitors from leaf and from sead Phaseolus vulgaris L. Plant Sci 138:17–26

    Article  CAS  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cels. Exp Cell Res 50:151–158

    Article  PubMed  CAS  Google Scholar 

  • Gradojević Z (1953) Ecology and development of Phytodecta fornicata Bruggm. (Col., Chrysomelidae). Newsletter of Serbian Academy of Science XXXI, no. 4, Institute for ecology and biogeography, Belgrade

  • Heath RL, McDonald G, Christeller JT, Lee M, Bateman K, West J, Van Heeswijck R, Anderson MA (1997) Proteinase inhibitors from Nicotiana alata enhance plant resistance to insect pests. J Insect Physiol 43:833–842

    Article  PubMed  CAS  Google Scholar 

  • Hilder VA, Gatehouse AMR, Sheerman SE, Barker RF, Boulter D (1987) A novel mechanism of insect resistance engineered into tobaco. Nature 330:160–163

    Article  CAS  Google Scholar 

  • Jouanin L, Bonadé-Bottino M, Girard C, Morrot G, Giband M (1998) Transgenic plants for insect resistance. Plant Sci 131:1–11

    Article  CAS  Google Scholar 

  • Kondo H, Abe K, Emori Y, Arai S (1990) Gene organization of oryzacystatin-II, a new cystatin superfamily member of plant origin, is closely related to that of oryzacystatin-I but different from those of animal cystatins. FEBS Lett 278:87–90

    Article  Google Scholar 

  • Laemmli UK (1970) Cleavege of structural proteins during the assembly of the head of bacteriophage T4. Nature 367:560–563

    Google Scholar 

  • Leple JC, Bonade-Bottino M, Augustin S, Pilate G, Le Tan VD, Delplanque A, Cornu D, Jouanin L (1995) Toxicity to Chrysomela tremulae (Coleoptera: Chrysomelidae) of transgenic poplars expressing a cysteine proteinase inhibitor. Mol Breed 1:319–328

    Article  CAS  Google Scholar 

  • Michaud D, Bernier-Vadnais N, Overney S, Yelle S (1995) Constitutive expression of digestive cysteine proteinase forms during development of the colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae). Insect Biochem Mol Biol 25:1041–1048

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Murdock LL, Brookhart G, Dunn PE, Foard DE, Kelly S, Kitch L, Shade RE, Shukle RH, Wofson JL (1987) Cysteine digestive proteinases in Coleoptera. Comp. Biochem Physiol 87B:783–787

    CAS  Google Scholar 

  • Narvaez-Vasquez J, Orozco-Cardenas ML, Ryan CA (1992) Differential expression of a chimeric CaMV-tomato proteinase inhibitor I gene in leaves of transformed nightshade, tobacco and alfalfa. Plant Mol Biol 20:1149–1157

    Article  PubMed  CAS  Google Scholar 

  • Ninković S, Miljuš-Đukić J, Nešković M (1995) Genetic transformation of alfalfa somatic embryos and their clonal propagation through repetitive somatic embryogenesis. Plant Cell Tissue Organ Cult 42:255–260

    Article  Google Scholar 

  • Ninković S, Miljuš-Djukić J, Nešković M (1998) Genetic engineering of herbicide-resistant alfalfa (Medicago sativa L. cv. Zaječarska 83). In: S Stamenković, I Mihaljev (eds) Proc 2nd Balkan Symp on Field Crops, no.1. Institute of Field and Vegetable Crops, Novi Sad, pp 457–460

  • Ninković S, Miljuš-Djukić J, Vinterhalter B, Nešković M (2004) Improved transformation of alfalfa somatic embryos using a superbinary vector. Acta Biol Crac Ser Bot 46:139–143

    Google Scholar 

  • Samac D, Smigocki A (2003) Expression of oryzacystatin I and II in alfalfa increases resistance to the root-lesion nematode. Phytopathol 93:799–806

    Article  CAS  Google Scholar 

  • Schuler TH, Poppy GM, Kerry BR, Denholm I (1998) Insect-resistant transgenic plants. Trends Biotechnol 16:168–175

    Article  CAS  Google Scholar 

  • Strizhov N, Keller M, Mathur J, Koncz-Kalman Z, Bosch D, Prudovsky E, Schell J, Sneh B, Koncz C, Zilberstein A (1996) A syntetic cryIC gene, encoding a Bacillus thuringiensis endotoxin, confers Spodoptera resistance in alfalfa and tobacco. Proc Natl Acad Sci USA 93:15012–15017

    Article  PubMed  CAS  Google Scholar 

  • Thomas J, Wasmann C, Echt C, Duun R, Bohnert H, McCoy T (1994) Introduction and expression of an insect proteinase inhibitor in alfalfa (Medicago sativa L.). Plant Cell Rep 14:31–36

    Article  CAS  Google Scholar 

  • Urwin P, Atkinson H, Waller D, McPherson M (1995) Engineered oryzacystatin-I expressed in transgenic hairy roots confers resistance to Globodera pallida. Plant J 8:121–131

    Article  PubMed  CAS  Google Scholar 

  • Uzelac B, Ninković S, Smigocki A, Budimir S (2007) Origin and development of secondary somatic embryos in transformed embryogenic cultures of Medicago sativa L. cv. Zaječarska 83. Biol Plant 51:1–6

    Article  Google Scholar 

  • Xu D, Xue Q, McElroy D, Mawal Y, Hilder VA, Wu R (1996) Constitutive expression of a cowpea trypsin inhibitor gene, CpTi, in transgenic rice plants confers resistance to two major rice insect pests. Mol Breed 2:167–173

    Article  CAS  Google Scholar 

  • Zhou X, Cao G, Lin R, Sun Y, Li W (1994) A rapid and efficient DNA extraction method of genus Fagopyrum for RAPD analysis. In: Javornik B, Bohanec B, Kreft I (eds) Proceedings of impact of plant biotechnology on agriculture. Biotechnical Faculty, Ljubljana, pp 171–175

    Google Scholar 

Download references

Acknowledgments

We would like to thank Dr. Lisa Jouanin from Laboratorie de Biologie celulaire, INRA, Versailles, France for kindly provided polyclonal OCI antibody and to Dr. Darka Šešlija from Evolutionary Department, IBISS, Belgrade, Serbia for the support provided for statistical analysis. This work was supported by the Ministry of Science and Environmental Protection, Republic of Serbia, grant No. 143026. We are very grateful to Dr Zoran Gradojević for helpful advice regarding the work with Phytodecta fornicata.

Author information

Authors and Affiliations

  1. Institute for Biological Research “S. Stanković”, University of Belgrade, Despota Stefana 142, Belgrade, 11060, Serbia

    Slavica Ninković, Jelica Lazarević, Branka Vinterhalter & Mirjana Nešković

  2. Institute for Genetics and Genetic Engineering, Bulevar Vojvode Stepe 444a, Belgrade, 11060, Serbia

    Jovanka Miljuš-Đukić, Svetlana Radović & Vesna Maksimović

  3. Molecular Plant Pathology Laboratory, USDA-ARS, Beltsville, MD, 20705, USA

    Ann Smigocki

Authors
  1. Slavica Ninković
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jovanka Miljuš-Đukić
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Svetlana Radović
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vesna Maksimović
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jelica Lazarević
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Branka Vinterhalter
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mirjana Nešković
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ann Smigocki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Slavica Ninković.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ninković, S., Miljuš-Đukić, J., Radović, S. et al. Phytodecta fornicata Brüggemann resistance mediated by oryzacystatin II proteinase inhibitor transgene . Plant Cell Tiss Organ Cult 91, 289–294 (2007). https://doi.org/10.1007/s11240-007-9296-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-007-9296-2

Keywords

Search

Navigation