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Plantibodies: versatile resistance genes against the polyphagous pathogens, tomato spotted wilt virus and root-knot nematode

Contract nr: FAIR-CT95-0905
Project nr: 905
Project type: SC
Starting date: 01/01/1996
Duration: 48 months
Total cost: 2813,000 EUR
EC Contribution: 2489,000 EUR
Scientific Officer: Marie-Françoise SCHARLL
Research topic: Plant health
Acronym: ANTIBODY MEDIATED RESISTANCE

Background:
This is the development of a project originally sponsored by the BIOTECH programme, which is oriented towards agriculture, the fundamental basis of which was established animals use antibodies to kill disease organisms. The idea is to give this mechanism to plants.

Objectives:
The objective was to apply the "plantibody technology", which was developed in the Biotechnology project "Plantibodies: a flexible approach to endow plants with new properties" (BIO2-CT92-0239), to obtain host-plant resistance against two economically important pathogens: tomato spotted wilt virus (TSWV) and the root knot nematode Meloidogyne incognita. The specific objectives were:

1) to obtain insight into the `molecular toolbox' used by root knot nematodes for establishing a successful relationship with their host plants;
2) to produce and characterise antibodies against antigens derived from both target pathogens and the cloning of genes encoding single chain antibody fragments in plant vectors; analysing resistance levels against both TSWV and M. incognita followed this step;
3) to safeguard the stability of in planta expressed single chain antibodies.

Description:
Antibodies to the tomato spotted wilt virus (TSWV) and to root knot nematodes (Meloidogyne incognita) will be developed and expressed in tomato.

Current situation/results:
Proof of concept was obtained, that stabilised single chain antibody constructs (addition of C-terminal KDEL sequence) directed against the nucleoprotein of TSWV results in a 100% protection against this virus. This is the first time that absolute, antibody mediated, transgenic resistance against a plant virus has been obtained. The highest level obtained thus far is 92% for the tobacco mosaic virus (TMV). This is an important result as TSWV is an economically-important pathogen.

Antibody-mediated resistance was also obtained against another economically important parasite, Meloidogyne incognita. The resistance level found was 40%. This can be considered as a breakthrough as, for the first time, antibody mediated transgenic resistance has been demonstrated for a complex pathogen/parasite. During this project, a leap forward was made regarding insight in the sedentary nematode - plant relationship. Several basis compatibility factors have been characterised, with the cell wall-degrading enzymes cellulase and pectate lyase as most prominent examples. Together, these results hold the promise of obtaining a resistance level that is, from an economic viewpoint, sufficient to be used in agricultural practice.

The stability of single chain antibodies expressed in plants has been safeguarded. High expression levels can now be obtained both intracellularly and extracellularly. Therewith, a good foundation was layed on the routine application of this promising technology.

Website: http://www.spg.wau.nl/nema/lma_bgr.htm


Coordinator
Arjen SCHOTS
Wageningen University
Lab. for Monoclonal Antibodies
Binnenhaven 10
P.O. BOX 8123

NL-6700 ES Wageningen
Tel.: +31 317 48 52 61
Fax: +31 317 48 52 67
E-mail: arjen.schots@lma.nema.wau.nl


Partners

  • Pierre ABAD
    I.N.R.A.
    Bld. Francis Meilland 123
    B.P. 2078
    F-06606 Antibes Cedex
    Tel.: +33-4-93.67.89.43
    Fax: +33-4-93.67.89.55

  • Ann DEPICKER
    Universiteit Gent
    K.L. Ledeganckstraat 35
    B- 9000 Gent
    Tel.: +32-9-264.51.74
    Fax: +32-9-264.53.49
    E-mail: anpic@gengenp.rug.ac.be

  • Rainer FISCHER
    Rheinnisch Westfalisch Technischen Hochschule Aachen
    Worringerweg 1
    D-52074 Aachen
    Tel.: +49-241-80.66.31
    Fax: +49-241-888.81.82
    E-mail: rainer.fischer@post.rwth-aachen.de

  • Rob GOLDBACH
    Wageningen University
    Laboratory of Virology
    Binnenhaven 11
    NL-6709 PD Wageningen
    Tel: +31-317 48 30 90
    Fax: +31-317-48 48 20
    E-mail: Rob.Goldbach@medew.viro.wau.nl

  • Eugenio BENVENUTO
    E.N.E.A. - Ente per le Nuove Tecnologie, l'energia e l'ambiente
    P.O. BOX 2400
    I-00060 Santa Maria di Galeria (Roma)
    Tel.: +39-6-30.48.63.47
    Fax: +39-6-30.48.65.45
    E-mail: benvenutoe@casaccia.enea.it

 
 
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