Plant husbandry – Process
Patent
1992-10-30
1995-06-06
Fox, David T.
Plant husbandry
Process
514 1, 530820, 530823, A01B 7900, A01N, C07K 1300
Patent
active
054218390
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
Antifungal polypeptides, e.g. lectins, have been isolated from monocotyledonous and dicotyledonous plants [Ramshaw, J. A. M. (1982) in Nucleic Acids and Proteins in Plants I. Encyclopedia of Plant Physiology, New Series, (Boulter, D. and Parthier, B. editors.) vol. 14 A, pp. 229-279, Springer, Berlin]. It has also been observed that the biosynthesis of thionine is initiated by infestation of the plant with pathogenic fungi. It is therefore assumed that this protein possesses antifungal activity [Bohlmann, H. et al. (1989) EMBO J. 7, 1559-1565; Broekaert, W. F. et al. (1989) Science 245, 1100-1102].
Antifungal polypeptides have also been isolated from microorganisms. For example, a similar protein has been detected in the fermentation broth of Aspergillus giganteus (Olson, B. M. & Goerner, G. L. (1965) Appl. Microbiol. 13, 314-321), but the amino acid sequence of said protein is two lysine residues shorter than that of the protein according to the invention.
We have now been able to isolate a polypeptide with antifungal action from Aspergillus giganteus and determine its sequence.
The nucleotide sequence of the peptide claimed has been published by Wnendt et al. (Nucl. Acids Res. 18, p. 3987, September 1990).
We have moreover found that expression, in plants, of the gene which codes for the polypeptide claimed in the main patent application cited above strongly inhibits the growth of phytopathogenic fungi on the plant.
SUMMARY OF THE INVENTION
A polypeptide with the amino acid sequence (SEQ ID NO: 1)
Ala-Thr-Tyr-Asn-Gly-Lys-Cys-Tyr-Lys-Lys-Asp-Asn-Ile-Cys-Lys-Tyr-Lys-Ala-Gln -Ser-Gly-Lys-Thr-Ala-Ile-Cys- Lys-Cys-Tyr-Val-Lys-Lys-Cys-Pro-Arg-Asp-Gly-Ala-Lys-Cys-Glu-Phe-Asp-Ser-Ty r-Lys-Gly-Lys-Cys-Tyr-Cys.
A process for the production of the polypeptide characterized above, which process is characterized in that Aspergillus giganteus is cultivated and the polypeptide is isolated.
A plant cell which expresses the gene of the antifungal peptide from Aspergillus giganteus, as well as a plant containing such cells, and plants, plant tissue or plant reproductive material grown from such a cell.
A process for the production of the plant cell, characterized in that the gene of the antifungal polypeptide from Aspergillus giganteus is expressed in the plant cell.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
The invention is described in detail below, especially in its preferred embodiments. The invention is further determined by the content of the Claims.
The polypeptide according to the invention is synthesized from Aspergillus giganteus with aerobic cultivation. Assimilable carbohydrates and sugar alcohols such as fructose, lactose or mannitol, and natural products containing carbohydrate, are suitable as preferred carbon sources. The following are possible as preferred nitrogenous nutrients: amino acids, peptides and proteins, as well as degradation products thereof, such as peptones or tryptones, and also meat extracts, ground seeds, for example of maize, wheat, beans, soya or the cotton plant, distillation residues from alcohol production, meat meals or yeast extracts, or else ammonium salts and nitrates. The nutrient solution can also contain for example chlorides, carbonates, sulfates or phosphates of the alkali metals or alkaline earth metals, iron, zinc and manganese, as additional inorganic salts.
The formation of the compound of the polypeptide according to the invention proceeds particularly well in a nutrient solution containing 0.5 to 2% of meat extracts, 0.5 to 2% of peptone, 0.5 to 4% of starch and 0.1 to 1% of sodium chloride, based in each case on the weight of the total nutrient solution.
The fermentation takes place aerobically, for example submersed with shaking or stirring in shake flasks or fermenters, with air or oxygen being introduced if appropriate. It can be carried out over a temperature range of about 18.degree. to 35.degree. C., preferably at about 25.degree. to 30.degree. C. The microorganism is cultivated under said conditions until the stationary phase i
REFERENCES:
patent: 3104204 (1963-09-01), Olson
patent: 4394443 (1983-07-01), Weissman et al.
Applied Microbiology, vol. 13, No. 3, May 1965, pp. 314-321; Olson, B. H., et al.: `Alpha sarcin, a new antitumor agent`.
Nucleic Acids Research, 18:13 (1990) Arlington, Va. US p. 3987, Wnendt et al.: `Cloning and nucleotide sequence of a cDNA Encoding the antifungal-protein of Aspergillus giganteus and preliminary characterization of the native gene`.
J. Cell. Biochem. Suppl, vol. 14E, 1990, & Symposium Apr. 16-22, 1990, p. 268, Broglie, R., et al.: Chitinase expression in transgenic plants: increased protection against a soil-borne fungal pathogen.
Dur. J. Biochem, vol. 193, No. 1, 1990, pp. 31-38; Nakaya, K., et al.: `Amino acid sequence and disulfide bridges of an antifungal-protein isolated from Aspergillus giganteus`.
Ar et al. 1986, Plant Physiol. 81:301-305.
An. 1985, Plant Physiol 79:568-570.
Potrykus, 1991, Annu. Rev. Plant Physiol. Plant Mol. Biol. 42:205-225.
Braun Peter
Eckes Peter
Hanel Heinz
Hilgenfeld Rolf
Logemann Jurgen
Fox David T.
Hoechst Aktiengesellschaft
Veitenheimer Erich E.
LandOfFree
Antifungal polypeptide and process for its production does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Antifungal polypeptide and process for its production, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Antifungal polypeptide and process for its production will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-983724