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Commercial Success of ECLAIR Programme
AGRE-0005: Novel antifungal proteins applications in crop protection |
AGRE-0005: Novel antifungal proteins applications in crop protection
Science Background
Fungal infection is a major cause of loss of yield in crops worldwide. In addition, some fungi produce highly carcinogenic mycotoxins that can cause problems in animals fed with residues from agro-industrial processes, such as oilseed and grain processing. Despite the development of safer, more efficient, target-specific fungicides, there are environmental and other pressures (including economics) which encourage the search for novel methods of control.
Objectives
This project aimed to develop a novel method of fungal disease control based on the discovery of unique, broad spectrum antifungal proteins (AFPs) which occur in some plants but not in others. The genes encoding for these AFPs would be transgenically expressed in target crops. They would then be developed for commercial use in crop protection as part of an integrated approach to disease control.
Significant changes and results since end of ECLAIR
This project was continued under AIR in order to evaluate possible agro-industrial applications of highly potent antifungal proteins of plant seed origin. The AIR project was also coordinated by KU Leuven FA Janssens Laboratory of Genetics, Belgium and involved the industrial partner Zeneca Agrochemicals, UK. Both these organisations continue to collaborate on in-house research. Proteins and their corresponding genes are being developed, as well as synthetic peptides that mimic AFPs that could be used for other applications (food or medical). Material generated in ECLAIR is still in trials.
Results
At end of this ECLAIR project
A bioassay was used to screen some 800 wild and cultivated plant species for proteins with potent antifungal activity. Such proteins were found, and purified, in over 20 different plants. The proteins were classified on the basis of the amino acid sequence and found to have many properties in common (relatively small size, high isoelectric point and a high cysteine content). Genes for those proteins that inhibited fungal growth at around 1mg/ml and showed no toxic effects on animals or animal cells were isolated. The DNA fragments were linked to a promoter and used to transform tobacco plants. Expression of the protein was demonstrated, as was increased resistance to artificial inoculation with Alternaria longipes, the fungus causing "brown spot disease" of tobacco.
Current position
Zeneca, UK, the commercial partner in this project, continued the research in-house and participated in a continuation exploring the potential of this technology: AIR-CT94-1356: Agro-industrial applications of antifungal proteins from plant seeds. A substantial amount of basic research was conducted to establish how the antifungal proteins are folded, which peptide domains are important for antifungal activity, and by which mechanisms fungal growth inhibition is exerted. This information would allow the design and selection of novel AFPs with improved activity or altered selectivity. The major application was to express these proteins in economically important crops, as already had been done in tobacco, such that they acquire resistance to fungal diseases and require reduced chemical inputs. Special emphasis was given to the development of strategies aimed at minimizing the emergence of fungal resistance to antifungal proteins, such as conditional control of gene expression and combined expression of two antifungal proteins with different modes of action. In addition, the feasibility to produce antifungal proteins in substantial amounts, either by chemical synthesis or fermentation, was evaluated. This may open ways to novel applications as preservatives in e.g. degradable food and non-food coatings.
The AIR project increased understanding of the AFPs and generated AFP variants with improved antifungal potency. Material generated under AIR is still in trials, to establish if the proteins are expressed at a sufficiently high level or need to be combined with other proteins to give better activity. Proteins and their corresponding genes are being developed, as well as synthetic peptides that mimic AFPs that could be used for other applications (food or medical). KU Leuven intends to continue to fund research in-house in collaboration with Zeneca.
Impact
Commercial
Zeneca applied for 7 patents as a result of the ECLAIR project and a further 6-7 as a result of the AIR project.
Further Information
Broekaert, W.F., Cammue, B.P.A., De Bolle, M.F.C., Thevissen, K., De Samblanx, G.W., Osborn, R.W. (1997) Antimicrobial peptides in plants. Crit. Rev. Plant Sci. 16: 297-323
Antifungal agents: discovery and mode of action (1995) Eds: G.K. Dixon, L.G. Copping and D. Hollomon. BIOS Scientific Publishers, Oxford, UK. ISBN 1 85996 145 2
Contacts
Author
KU Leuven
Zeneca Agrichemicals
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