Grapevine is one of the major horticulture crops grown in temperate climates that is exposed to attacks from a variety of pathogens, including viruses. Viruses cause heavy crop losses, costing several million euros each year, so there is a necessity for developing a novel approach for disease control. This project will generate biologically safe, virus-resistant plants and recombinant antibodies will be generated by phage display to specifically recognise and inactivate key viral proteins. This will reduce the use of environment-unfriendly, undesirable chemical controls that are currently the only effective method for protecting vineyards from pathogen attack.
The objective of this proposal is to generate rootstocks and grapevine varieties that show sustainable resistance against viruses. This will be achieved by expression of recombinant antibodies, specifically binding to and inactivating viral proteins from selected Nepo- and Closteroviruses. Proof of principle demonstrating antibody-mediated pathogen resistance will be evaluated in tobacco followed by transgenic rootstocks and grapevine.
In Europe, grapevine plants are grafted onto a rootstock to assemble a whole plant. Nematodes, the vectors of Nepoviruses only infest the roots of a plant therefore Nepovirus resistance is only required for the production of rootstocks. Thus, Nepovirus-specific scFvs will be transformed only into rootstocks. Closteroviruses are transmitted by aerial vectors that do not attack rootstocks so Closterovirus-specific scFvs will be transformed only into the grapevine.
Progress to Date
All viral antigens were produced in bacteria and purified.Almost all anticipated immunisation has been completedand the first virus-specific scFvs are available.
Two plant expression vesctors were constructed to allow accumulation of virus-specific scFvs in the plant cell cytosol.
Optimised procedures for Agrobacterium-mediatedtransformation and regeneration of rootstocks have been established. Embryonic tissue has been produced for all grapevine and rootstock varieties and will be multiplied.
The first graevine plants have been transformed: regeneration of transgenic plants is in progress.
Difficulties in producing antigens with sufficient amounts and quality delayed immunisation for some of the viral antigens.
The project has generated various expression constructs and recombinant viral proteins.
ARABLE CROPS, HORTICULTURE, FRUIT
Scientist responsible for the project
||REKTOR DER RHEIN-WESTF. TECHNISCHEN HOCHSCHULE AACHEN
||01 January 2002
||1 312 860 €
|Total EC contribution
||1 249 985 €
- UNIVERSIT└ DEGLI STUDI DI BARI, Italy - IT
- CENTRUM GRUENE GENTECHNIK, STAATLICHE LEHR- UND FORSCHUNGSANSTALTFUER LANDWIRTSCHAFT, WEINBAU UND GARTENBAU, Germany - DE