Methods for promoting plant health using free enzymes and microorganisms that overexpress enzymes

Abstract

Methods for stimulating plant growth and/or promoting plant health using free enzymes or recombinant microorganisms that overexpress enzymes are provided. Plant seeds coated with free enzymes or recombinant microorganisms that overexpress enzymes are also provided. Compositions comprising a fertilizer and an enzyme or a recombinant microorganism that overexpresses an enzyme are provided. Modified enzymes having ACC deaminase activity, recombinant microorganisms expressing the modified enzymes, plant seeds treated with the modified enzymes or recombinant microorganisms, and methods for stimulating plant growth and/or promoting plant health using the modified enzymes or recombinant microorganisms are also provided.

1 - 20 . (canceled) 
     
     
         21 . A method for protecting a plant from a pathogen comprising applying a free enzyme to a plant growth medium, a plant, a plant seed, or an area surrounding a plant or a plant seed, wherein the enzyme is selected from a phospholipase, a lipase a lactonase, a chitosanase, a protease, an acid phosphatase, a glucanase, an ACC deaminase, and combinations of any thereof. 
     
     
         22 . A method for protecting a plant from a pathogen comprising applying two or more free enzymes to a plant growth medium, a plant, a plant seed, or an area surrounding a plant or a plant seed, wherein the enzymes are independently selected from a phospholipase, a lipase, a lactonase, a chitosanase, a protease, a glucanase, and an ACC deaminase. 
     
     
         23 - 24 . (canceled) 
     
     
         25 . The method of  claim 21 , wherein the method comprises applying the enzyme to the plant growth medium. 
     
     
         26 - 29 . (canceled) 
     
     
         30 . The method of  claim 21 , wherein the method further comprises applying an agrochemical or agriculturally acceptable carrier to the plant growth medium, the plant, the plant seed, or the area surrounding a plant or a plant seed. 
     
     
         31 - 32 . (canceled) 
     
     
         33 . A method for protecting a plant from a pathogen, comprising applying a recombinant microorganism to a plant growth medium, a plant, a plant seed, or an area surrounding a plant or a plant seed, wherein:
 the recombinant microorganism expresses an enzyme, wherein expression of the enzyme is increased as compared to the expression level of the enzyme in a wild-type microorganism of the same kind under the same conditions;   the enzyme is selected from a phospholipase, a lipase, a lactonase, a chitosanase, a glucanase, a protease, and combinations of any thereof; and   the enzyme is expressed during vegetative growth of the recombinant microorganism.   
     
     
         34 - 36 . (canceled) 
     
     
         37 . A plant seed coated with a recombinant microorganism, wherein:
 the recombinant microorganism expresses an enzyme, wherein expression of the enzyme is increased as compared to the expression level of the enzyme in a wild-type microorganism of the same kind under the same conditions;   the enzyme is selected from a phospholipase, a lipase, a lactonase, a chitosanase, a glucanase, a protease, an ACC deaminase, and combinations of any thereof; and   the enzyme is expressed during vegetative growth of the recombinant microorganism.   
     
     
         38 - 59 . (canceled) 
     
     
         60 . The method of  claim 21 , wherein the method comprises applying the enzyme to an area surrounding a plant or plant seed. 
     
     
         61 - 63 . (canceled) 
     
     
         64 . The method of  claim 21 , wherein the method comprises applying the enzyme to roots of the plant. 
     
     
         65 - 66 . (canceled) 
     
     
         67 . The method of  claim 21 , wherein applying the enzyme to the plant seed comprises: (a) applying the enzyme to the plant seed at the time of planting; or (b) coating the plant seed with the enzyme. 
     
     
         68 . The method of  claim 67 , wherein the method comprises coating the plant seed with a seed coating formulation comprising:
 the enzyme; and   an agriculturally acceptable carrier; or   an agrochemical.   
     
     
         69 . A plant seed coated with a free enzyme, wherein the enzyme is selected from a phospholipase, a lipase, a xylanase, a xylosidase, a mannanase, a pectinase, a lactonase, a chitosanase, a protease, a phytase, an acid phosphatase, a non-cellulolytic glucanase, an ACC deaminase, and combinations of any thereof. 
     
     
         70 - 75 . (canceled) 
     
     
         76 . The plant seed of  claim 69 , wherein the plant seed is coated with a seed coating formulation comprising the enzyme; and
 an agriculturally acceptable carrier; or   an agrochemical.   
     
     
         77 - 172 . (canceled) 
     
     
         173 . The method of  claim 21 , wherein the enzyme comprises:
 a crude cell extract containing the enzyme;   a partially purified enzyme;   a substantially purified enzyme; or   an enzyme that is immobilized on a matrix or support.   
     
     
         174 - 227 . (canceled) 
     
     
         228 . The method of  claim 21 , wherein applying the enzyme comprises:
 an in-furrow treatment;   applying the enzyme to the soil; or   applying the enzyme foliarly.   
     
     
         229 . The plant seed of  claim 76 , wherein the agrochemical is not a fertilizer. 
     
     
         230 . The plant seed of  claim 76 , wherein the agrochemical comprises:
 an insecticide;   a nematicide;   an herbicide;   a fungicide;   a fungal inoculant; or   a bacterial inoculant.   
     
     
         231 . The plant seed of  claim 76 , wherein the agriculturally acceptable carrier comprises a dispersant, a surfactant, an additive, water, a thickener, an anti-caking agent, residue breakdown product, a composting formulation, a granular application, diatomaceous earth, an oil, a coloring agent, a stabilizer, a preservative, a polymer, a coating, or a combination thereof. 
     
     
         232 . The plant seed of  claim 37 , wherein the plant seed is coated with a seed coating formulation comprising the recombinant microorganism; and
 an agriculturally acceptable carrier; or   an agrochemical.   
     
     
         233 . The plant seed of  claim 232 , wherein the agrochemical is not a fertilizer. 
     
     
         234 . The plant seed of  claim 232 , wherein the agrochemical comprises:
 an insecticide;   a nematicide;   an herbicide;   a fungicide;   a fungal inoculant; or   a bacterial inoculant.   
     
     
         235 . The plant seed of  claim 232 , wherein the agriculturally acceptable carrier comprises a dispersant, a surfactant, an additive, water, a thickener, an anti-caking agent, residue breakdown product, a composting formulation, a granular application, diatomaceous earth, an oil, a coloring agent, a stabilizer, a preservative, a polymer, a coating, or a combination thereof. 
     
     
         236 . The method of  claim 33 , wherein the method comprises applying the recombinant microorganism to the plant growth medium. 
     
     
         237 . The method of  claim 33 , wherein the method further comprises applying an agrochemical or agriculturally acceptable carrier to the plant growth medium, the plant, the plant seed, or the area surrounding a plant or a plant seed. 
     
     
         238 . The method of  claim 33 , wherein the method comprises applying the recombinant microorganism to an area surrounding a plant or plant seed. 
     
     
         239 . The method of  claim 33 , wherein applying the recombinant microorganism to the plant seed comprises: (a) applying the recombinant organism to the plant seed at the time of planting; or (b) coating the plant seed with the recombinant organism. 
     
     
         240 . The method of  claim 239 , wherein the method comprises coating the plant seed with a seed coating formulation comprising:
 the recombinant microorganism; and   an agriculturally acceptable carrier; or   an agrochemical.   
     
     
         241 . The method of  claim 238 , wherein applying the recombinant microorganism to an area surrounding a plant comprises applying the recombinant microorganism to the soil. 
     
     
         242 . A formulation comprising a free enzyme, wherein the enzyme is selected from a phospholipase, a lipase, a lactonase, a chitosanase, a protease, a glucanase, an ACC deaminase, and combinations of any thereof; and an agriculturally acceptable carrier. 
     
     
         243 . A formulation comprising a recombinant microorganism and an agriculturally acceptable carrier, wherein:
 the recombinant microorganism expresses an enzyme, wherein expression of the enzyme is increased as compared to the expression level of the enzyme in a wild-type microorganism of the same kind under the same conditions;   the enzyme is selected from a phospholipase, a lipase, a lactonase, a chitosanase, a glucanase, a protease, and combinations of any thereof.