METHODS AND COMPOSITIONS FOR BIOPROTECTION OF TOMATOES FROM CLAVIBACTER MICHIGANENSIS SUBSP. MICHIGANENSIS
The present invention relates to compositions having antimicrobial activity against Clavibacter michiganensis subsp michiganensis (“Cmm”). The compositions comprises Bacillus pumilus strain deposited with ATCC® Patent Designation No. PTA-125304, culture medium or extracts comprising Micrococcin P1 produced by said Bacillus pumilus strain, and/or isolated Micrococcin P1. Further provided herein are methods of making and using the antimicrobial compositions to protect and treat tomatoes from Cmm infections.
1 . A method of controlling, suppressing and/or preventing an infection from Clavibacter michiganensis subsp. michiganensis (Cmm) in tomatoes, comprising:
providing an antimicrobial composition comprising at least one of (i) Bacillus pumilus strain deposited with ATCC® Patent Designation No. PTA-125304, (ii) a culture medium inoculated with said Bacillus pumilus strain and comprising Micrococcin P1 produced by said Bacillus pumilus strain, (ii) a cell-free extract of said Bacillus pumilus strain and comprising Micrococcin P1 produced by said Bacillus pumilus strain, and (iv) isolated Micrococcin P1 produced by said Bacillus pumilus strain; and
applying an effective amount of said antimicrobial composition to at least one of a tomato plant, a tomato root, a tomato leaf, a tomato fruit and a tomato stem;
wherein said applying provides for bioprotection of tomatoes against Cmm in a field.
2 . The method of claim 1 ,
wherein said bioprotection comprises at least one of enhancing resistance against Cmm, reducing damage caused by Cmm, enhancing plant antimicrobial response against Cmm, increasing plant antinematocidal activity, reducing pathological symptoms or lesions resulting from actions of Cmm, and increasing tomatoes yield; and
wherein said bioprotection is determined by comparing damages of tomatoes contacted or not with said antimicrobial composition.
3 . The method of claim 1 , wherein the antimicrobial composition further comprises a filtered fraction and/or a cell free supernatant of a microorganism mixture comprising Lactobacillus paracasei, Lactobacillus helveticus, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactococcus lactis, Bacillus amyloliquefaciens, Aspergillus oryzae, Saccharomyces cerevisiae, Candida utilis , and Rhodopseudomonas palustris , or a mixture thereof.
4 . The method of claim 1 , wherein said antimicrobial composition further comprises at least one strain of Bacillus subtilis , a culture medium inoculated with Bacillus subtilis , a cell-free extract of Bacillus subtilis or at least one metabolite of Bacillus subtilis.
5 . The method of claim 1 , wherein said antimicrobial composition further comprises at least one of an herbicide, an insecticide, a fungicide and a nutrient.
5 . The method of claim 1 , wherein the Micrococcin P1 has a molecular weight (MW) of about 1144 Dalton.
7 . The method of claim 1 , wherein the Micrococcin P1 has the following chemical structure:
8 . A composition for the bioprotection of tomatoes from, Clavibacter michiganensis subsp. michiganensis (Cmm), comprising:
an effective amount of isolated Micrococcin P1, wherein said Micrococcin P1 is produced by Bacillus pumilus strain deposited with ATCC® Patent Designation No. PTA-125304; and
an agriculturally acceptable carrier,
wherein said composition is formulated for application to at least one of a tomato plant, a tomato root, a tomato leaf, a tomato seed and a tomato stem; and
wherein said composition is effective in controlling, suppressing and/or preventing infection of tomatoes from Cmm when applied to tomatoes in a field.
9 . The composition of claim 8 , further comprising a cell-free supernatant and/or a filtered fraction of a microbial culture inoculated with one or more of Lactobacillus paracasei, Lactobacillus helveticus, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactococcus lactis, Bacillus amyloliquefaciens, Aspergillus oryzae, Saccharomyces cerevisiae, Candida utilis , and Rhodopseudomonas palustris.
11 . The composition of claim 8 , further comprising at least one strain of Bacillus subtilis , a culture medium inoculated with Bacillus subtilis , a cell-free extract of Bacillus subtilis or at least one metabolite of Bacillus subtilis.
12 . The composition of claim 8 , wherein the Micrococcin P1 has a molecular weight (MW) of about 1 1144 Dalton.
13 . The composition of claim 8 , wherein the Micrococcin P1 has the following chemical structure:
14 . A method of protecting tomatoes from Clavibacter michiganensis subsp. michiganensis (Cmm), comprising the step of:
applying an effective amount of the composition of claim 8 to a tomato plant, a tomato root, a tomato leaf, a tomato seed and/or a tomato stem,
wherein the effective amount is sufficient for bioprotection of the tomato plant from Cmm.