The purpose of this project is to engineer plant oxylipin synthesis in vitro to assess their usefulness as 'green' antimicrobial and pesticidal chemoprotectants. Plant oxylipin biosynthesis will be manipulated in vivo to modify defence reactions for improved resistance to pests and pathogens. As a result of this work, an 'oxylipin tool kit' will be produced (made up of recombinant enzymes and oxylipins in analytical quantities). Extensive knowledge will have been generated on the in vivo role of oxylipins in plant-pest and -pathogen interactions by generating a series of transgenic lines of Arabidopsis, which will have altered oxylipin metabolism through over-expression and/or antisense suppression of the biosynthetic genes. The gained knowledge and tools will be applied to protect ornamental plants.
Plant oxylipins are a wide array of polyunsaturated fatty acid derivatives that participate in plant defence responses to pest and pathogen attack. Some of them have direct antimicrobial properties; others may act as regulators of plant defence gene expression. The low concentrations that the majority of these compounds attain in plants has so far precluded, for most of them, a direct evaluation of their involvement in plant defence responses, either as natural chemoprotectants or through regulation of gene expression.The aim of the project is to assess the potential of oxylipins as "green", environmental-friendly agrochemicals for plant protection against pests and pathogens, and to do a proof of concept using them in practice.The consortium will undertake the isolation and manipulation of plant enzymes and genes involved in oxylipin synthesis, in order to optimize and extend their production in vitro and in planta. The specific objectives of the project are:
1) to systematically evaluate the chemoprotective potential of a wide range of oxylipins as replacement sprays for existing agrochemical treatments which are to be discontinued in the near future.
2) to generate model transgenic Arabidopsis plants with enhanced resistance to pests and pathogens using oxylipin gene combinations to demonstrate the resistance to pests and pathogens.
3) to transfer the knowledge gained from 1) and 2) to the benefit of ornamental plant-pathogen/-pest control by:
3a) alternative "green" oxylipin protection, by giving the produced and characterised oxylipins to the industrial project partner, thus reducing the use of harmful chemical sprays in the horticulture industry, and
3b) genetically modifying Pelargonium for enhanced constitutive expression of oxylipins thus conferring improved pest and pathogen resistance. The eventual aim is to provide improved, commercially available cultivars of Pelargonium.
1) Selected oxylipins that either were not commercially available, or only at unaffordable costs, have been produced and used as chemoprotectants in test trials. The antimicrobial/insecticide effects of these oxylipins have been tested against a range of pests and pathogens. Enzymes of oxylipin biosynthesis have been expressed in vitro for their use in semi-enzymatic oxylipin synthesis procedures.
2) High-throughput semi-automated screens have been established for the identification of enzymes of oxylipin metabolism with new and improved properties generated through forced molecular evolution.
3) Bacterial vectors for expression of oxylipin enzymes in plants have been prepared, to be used in the generation of transgenic Arabidopsis thaliana plants with significantly altered oxylipin profiles.
4) A combined chromatographic method, allowing simultaneous analysis of more that 120 metabolites, has been established for the study of oxylipin profiles.
5) The effects of selected oxylipins on Arabidopis gene expression have been analysed.
6) Transformation protocols have been established for the generation of transgenic Pelargonium plants with altered oxylipin profile.
7) A detailed oxylipin profiling for arabidopsis plants challenged with fungal and bacterial pathogens has been established.
8) A detailed oxylipin profiling for Pelargonium plants challenged with fungal and bacterial pathogens has been established.
CROP PESTS AND DISEASES, GENOMICS
Scientist responsible for the project
Mr MARIANO ESTEBAN RODRIGUEZ
CAMPUS UNIVERSIDAD AUTONOMA
Spain - ES
Phone: +34 915854503
Fax: +34 915854506
||CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS
||01 January 2002
||4 271 392 €
|Total EC contribution
||2 799 952 €
|Web address of the project
- BIJVOET CENTER FOR BIOMOLECULAR RESEARCH, FACULTY OF CHEMISTRY,UTRECHT UNIVERSITY, Netherlands (The) - NL