Mogen International Nv

Rijksuniversiteit Te Leiden

Syngenta Mogen B.v.

Zeneca Mogen B.v.

Leo Sjoerd Melchers

Bernardus Johannes Cornelissen

Marianne Beatrix Sela-Buurlage

Anne Silene Ponstein

Charles Peter Woloshuk

Elisabeth J Meulenhoff

John Ferdinand Bol

Hubertus J Linthorst

Petrus Josephus Van Den Elzen

Jeroen S Van Roekel

Alexandra Aleida Bres-Vloemans

Marion Apotheker-De Groot

Johanna Pieternella Els Van Deventer-Troost

Alexandra Aleida Vloemans

Maarten Hendrik Stuiver

Stephan Andreas Ohl

Saskia Kroon-Swart

Jerome Hubertus Henricus Victor Custers

J&uuml;rgen Logemann

Wessel Lageweg

The present invention provides an isolated protein obtainable from a plant source which has antifungal activity, specifically anti-Phytophthora activity and/or anti-Pythium activity and a molecular weight of about 55-65 kDa as judged by SDS PAGE-electrophoresis, an isolated DNA sequence comprising an open reading frame capable of encoding a protein according to the invention, preferably characterized in that it comprises an open reading frame which is capable of encoding a protein depicted in SEQ ID NO. 16, SEQ ID NO. 57, SEQ ID NO. 70, SEQ ID NO. 72 or SEQ ID NO. 74 or muteins thereof, and DNA capable of hybridizing therewith under stringent conditions. The invention further comprises plants incorporating chimeric DNA capable of encoding a protein according to the invention, and wherein the protein is expressed. Also shown is the carbohydrate and preferably hexose oxidating activity of said protein. Also methods are provided for combating fungi, especially Phytophthora and Pythium species, using a protein or a host cell capable of producing the protein.

Antifungal proteins, DNA coding therefore, and hosts incorporating same

The present invention provides an isolated protein obtainable from a plant source which has anti-Phytophthora activity and a molecular weight of about 60&plusmn;5 kDa as judged by SDS PAGE-electrophoresis, an isolated DNA sequence comprising an open reading frame capable of encoding a protein according to the invention, preferably characterized in that it comprises an open reading frame which is capable of encoding a protein as represented by amino acids 1 to 540 of SEQ ID NO: 6, or the precursor of said protein, and DNA capable of hybridising therewith under stringent conditions. The invention further comprises plants incorporating chimeric DNA capable of encoding a protein according to the invention, and wherein the protein is expressed. Also methods are provided for combatting fungi, especially,, using a protein or a host cell capable of producing the protein.

Antifungal proteins, DNA coding therefor, and hosts incorporating same

There is provided (a) a process to discharge into an extracellular space of a plant an intracellular osmotin protein that naturally has a vacuolar targeting sequence at its C-terminal end which naturally targets the protein into a plant vacuole; and (b) a recombinant polynucleotide for use in the process. The recombinant polynucleotide includes a promoter that is functional in plants; an open reading frame encoding an intracellular osmotin protein under control of the promoter, the open reading frame having been modified to target the intracellular osmotin protein to an apoplast by creating a translation stop codon in the open reading frame at the 3' end resulting in deletion of the C-terminal amino acids of the intracellular osmotin protein necessary for intracellular osmotin targeting; and a terminator operably linked to the modified open reading frame.

Polynucleotide encoding osmotin modified for apoplast targeting

Plants are provided with improved resistance against pathogenic fungi. They are genetically transformed with one or more polynucleotides which essentially comprise one or more genes encoding plant chitinases and .beta.-1,3-glucanases. Preferred are the intracellular forms of the said hydrolytic enzymes, especially preferred are those forms which are targeted to the apoplastic space of the plant by virtue of the modification of the genes encoding the said enzymes. Particularly preferred are plants exhibiting a relative overexpression of at least one gene encoding a chitinase and one gene encoding a .beta.-1,3-glucanase.

Transgenic fungal resistant plants expressing chitinase and glucanase,

Plants are provided with improved resistance against pathogenic fungi. They are genetically transformed with one or more polynucleotides which essentially comprise one or more genes encoding plant and .beta.-1,3-glucanases. Preferred are the intracellular forms of the said hydrolytic enzymes, especially preferred are those forms which are targeted to the apoplastic space of the plant by virtue of the modification of the genes encoding the said enzymes. Particularly preferred are plants exhibiting a relative overexpression of at least one gene encoding a .beta.-1,3-glucanase.

Process for obtaining fungal resistant plants with recombinant

Chimeric genes encoding antifungal chitin binding proteins (antifungal CBPs) with very low chitinase activity (10% or less than that of the class-I chitinases from tobacco). Also substantially pure DNA sequences encoding antifungal CBP are provided for the obtention of transgenic plants producing antifungal CBP. Plants expressing an antifungal CBP gene, optionally in combination with a plant expressible glucanase gene, show reduced susceptibility to fungi.

Antifungal chitin binding proteins and DNA coding therefor

Chitinase, DNA coding therefor and plants containing same

A method for inhibiting the growth and/or germination of a fungus that is susceptible to an osmotin protein by contacting the fungus, or causing the fungus to be contacted with, an effective amount of an osmotin protein. The fungus can be, for example, a plant pathogenic fungus, such as an Oomycete. In one embodiment of the invention, the effective amount of the osmotin protein is produced by a plant having incorporated into its genome a chimeric gene having a) an open reading frame encoding the osmotin protein, or a precursor of the osmotin protein, and having operably linked thereto b) a regulatory region which causes the chimeric gene to be expressed in the plant.

Antifungal preparations containing osmotin

Fungal resistant plants, process for obtaining fungal resistant plants

Antifungal preparations, and process for making such preparations

Growing community of inventors

Leiden, Netherlands

Leo Sjoerd Melchers