Cornell Research Foundation Inc.

Christopher K Hayes

Gary E Harman

Matteo Lorito

Antonio Di Pietro

Arne Tronsmo

Roxanne M Broadway

Christian P Kubicek

Sonja Klemsdahl

Antonio D Pietro

Felice Scala

A system for inhibiting the germination or growth of a fungus comprises (a) fungal cell wall degrading chitinolytic or glucanolytic enzyme and (b) antifungal cell membrane affecting: compound. Exemplified antifungal fungal cell membrane affecting compounds include flusilazole, miconazole, osmotin, gramicidin, valinomycin, phospholipase B, and trichorzianines. The system components (a) and (b) may be supplemented with polyene macrolide antibiotic, antifungal epithiodiketopiperazine antibiotic (e.g., gliotoxin), fungal cell wall biosynthesis inhibitor (e.g., L-sorbose) and/or detergent. Embodiments include method of contacting a plant which expresses cell wall degrading enzyme with antifungal fungal cell membrane affecting compound.

Combinations of fungal cell wall degrading enzyme and fungal cell membrane affecting compound

Two chitinases from,P1 (ATCC 74058) show chitin-containing-fungus-inhibiting activity. One is an endochitinase and the other is a chitobiase. Both have molecular weights of 40 kDa and isoelectric points of 3.9. A &lgr;gtll recombinant encoding for the endochitinase has been prepared (ATCC 55338) and the gene encoding for the endochitinase has been removed from the DNA of the recombinant by the restriction enzyme, Not I. Endochitinases and chitobiases including the two purified from,strain P1 demonstrate synergy with each other in anti-fungal effect.

Purified chitinases and use thereof

Two chitinases from Trichoderma harzianum P1 (ATCC 74058) show chitin-containing-fungus-inhibiting activity. One is an endochitinase and the other is a chitobiase. Both have molecular weights of 40 kDa and isoelectric points of 3.9. Endochitinases and chitobiases including the two purified from Trichoderma harzianum strain P1 demonstrate synergy with each other in antifungal effect. Isolated gene encoding for the endochitinase has the sequence set forth in the Sequence Listing as SEQ ID NO:1.

Gene encoding endochitinase

N-acetyl-.beta.-glucosaminidase isolated from Trichoderma harzianum strain P1 having accession No. ATCC 74058 has molecular mass of 72 kDa and isoelectric point of 4.6. Glucan 1,3-.beta.-glucosidase isolated from Trichoderma harzianum strain P1 has molecular mass of 78 kDa and isoelectric point of 6.2. Combinations of endochitinase, chitin 1,4-.beta.-chitobiosidase, N-acetyl-.beta.-glucosaminidase and glucan 1,3-.beta.-glucosidase have synergistic antifungal effect.

N-acetyl-.beta.-glucosaminidase isolated from Trichoderma harzianum

Antifungal composition comprising synergistic combination of fungal cell wall degrading enzyme selected from the , group consisting of chitinolytic enzymes, glucanolytic enzymes and cellulases and non-enzymatic fungicides selected from the group consisting of sterol synthesis inhibiting fungicides (e.g., flusilazole and miconazole) and thiol group inactivating fungicides which are not specific to fungal cell membranes (e.g., captan) are applied in an antifungal effective amount to fungus to be inhibited or to a locus to be protected from said fungus.

Antifungal synergistic combination of enzyme fungicide and non-enzymatic

Two chitinases from Trichoderma harzianum P1 (ATCC 74058) show chitin-containing-fungus-inhibiting activity. One is an endochitinase and the other is a chitobiase. Both have molecular weights of 40 kDa and isoelectric points of 3.9. Endochitinases and chitobiases including the two purified from Trichoderma harzianum strain P1 demonstrate synergy with each other in anti-fungal effect. Isolated gene encoding for the endochitinase has the sequence set forth in the Sequence Listing as SEQ ID NO:1.

Gene encoding for endochitinase

Combination of fungal cell wall degrading enzymes (e.g., chitinolytic enzymes) and antifungal bacteria (e.g., E. cloacae) which bind to fungal cell walls in the presence of said enzymes even when sugars are present, increases potency and range of use of the bacteria and fosters proliferation thereof.

Fungicidal compositions comprising chitinase and enterobacter cloacae,

Antifungal composition comprising synergistic combination of fungal cell wall degrading enzyme selected from the group consisting of chitinolytic enzymes, glucanolytic enzymes and cellulases and non-enzymatic fungicides selected from the group consisting of sterol synthesis inhibiting fungicides (e.g., flusilazole and miconazole) and thiol group inactivating fungicides which are not specific to fungal cell membranes (e.g., captan) are applied in an antifungal effective amount to fungus to be inhibited or to a locus to be protected from said fungus.

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Geneva, NY, United States of America

Christopher K Hayes