COMBINATION OF BACTERIAL BIOLOGICAL CONTROL AGENT AND FATTY ACIDS

Abstract

The present invention relates to a composition comprising a) at least one biological control agent which is a fungicidally active bacterium and/or a fungicidally active metabolite produced by the respective bacterium and b) one or more fatty acids or derivatives thereof selected from unsaturated and saturated C.sub.12-24 fatty acids, salts thereof, esters thereof or mixtures of any of the foregoing as well as methods of using this composition and related uses.

Claims

1. A composition comprising (a) at least one biological control agent which is a fungicidally active bacterium and/or a fungicidally active metabolite produced by the respective bacterium and (b) one or more fatty acids or derivatives thereof selected from unsaturated and saturated C.sub.12-24 fatty acids, salts thereof, esters thereof or mixtures of any of the foregoing.

2. The composition according to claim 1, wherein said biological control agent is a strain of a species selected from the group consisting of Bacillus subtilis, Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus mycoides, Bacillus licheniformis, and Paenibacillus sp.

3. The composition according to claim 1, wherein said biological control agent is selected from the group consisting of (B1.1) Bacillus subtilis strain QST713/AQ713, (B1.2) Bacillus pumilus strain QST2808, (B1.3) Bacillus pumilus strain GB34, (B1.4) Bacillus pumilus strain BU F-33, (B1.5) Bacillus amyloliquefaciens strain D747, (B1.6) Bacillus subtilis Y1336, (B1.7) Bacillus amyloliquefaciens strain MBI 600, (B1.8) Bacillus subtilis strain GB03, (B1.9) Bacillus subtilis var. amyloliquefaciens strain FZB24, (B1.10) Bacillus mycoides isolate J, (B1.11) Bacillus licheniformis strain SB3086, and (B1.12) a Paenibacillus sp. strain having Accession No. NRRL B-50972 or Accession No. NRRL B-67129 and/or a mutant of these strains having all the identifying characteristics of the respective strain, and/or a metabolite produced by the respective strain that exhibits activity against phytopathogens.

4. The composition according to claim 2, wherein said biological control agent is B. subtilis QST713.

5. The composition according to claim 1, wherein said one or more fatty acids or derivatives thereof are selected from unsaturated and saturated C.sub.14-20 fatty acids, salts thereof, esters thereof or mixtures of any of the foregoing.

6. The composition according to claim 1, wherein the one or more fatty acids or derivatives thereof comprises two or more fatty acids selected from C16:0 fatty acids, C16:1 fatty acids, C18:0 fatty acid, C18:1 fatty acids, C18:2 fatty acids, and C18:3 fatty acids, or salts thereof.

7. The composition according to claim 1, wherein the one or more fatty acids or derivatives thereof comprises oleic acid or a salt thereof.

8. The composition according to claim 1, wherein the one or more fatty acids or derivatives thereof comprise one or more metal salts of fatty acids, which one or more metal salts of fatty acids are obtainable by a process comprising (a) Providing a vegetable oil, preferably olive oil; (b) Hydrolyzing triglycerides in the vegetable oil; (c) Extracting fatty acids from the hydrolyzed vegetable oil; and (d) Forming the metal salts of the extracted fatty acids.

9. The composition according to claim 1, wherein the fatty acid derivatives are alkali metal salts of fatty acids, preferably potassium salts of fatty acids.

10. The composition according to claim 1, wherein said fatty acid derivatives salts of lithium, sodium, potassium, magnesium, calcium, aluminum, copper iron or zinc or a mixture thereof.

11. The composition according to claim 1, wherein component b) is one or more fatty acid.

12. The composition according to claim 1 which is a synergistic composition.

13. The composition according to claim 1 additionally comprising at least one auxiliary selected from the group consisting of extenders, solvents, spontaneity promoters, carriers, emulsifiers, dispersants, frost protectants, thickeners and adjuvants.

14. A seed coated with the composition according to claim 1.

15-17. (canceled)

18. A method for enhancing the fungicidal action of a biological control agent against plant pathogens comprising mixing a biological control agent as defined in claim 1 with one or more fatty acids or derivatives thereof as defined in claim 1 and applying said mixture to a plant or seed or a locus where said plant or seed is intended to be grown.

19. The method according to claim 18, wherein the plant pathogen is a strain of a genus selected from the group consisting of Venturia, Sclerotinia, Rhizoctonia, Phytium, Alternaria, Botrytis, Phytophthora, Puccinia, Blumeria, Fusarium and Septoria.

20. A method for reducing overall damage of plants and plant parts as well as losses in harvested fruits or vegetables caused by phytopathogens comprising the step of simultaneously or sequentially applying at least one biological control agent as defined claim 1 and one or more fatty acids or derivatives thereof as defined in claim 1 to a plant or seed or a locus where said plant or seed is intended to be grown.

21. A method for increasing crop yield and/or the quality of food commodities comprising the step of simultaneously or sequentially applying at least one biological control agent as defined in claim 1 and one or more fatty acids or derivatives thereof as defined in claim 1 to a plant or seed or a locus where said plant or seed is intended to be grown.

22. The method according to claim 18, wherein said applying is effected as a foliar or soil application or as a seed treatment.

23-24. (canceled)

25. A kit-of-parts comprising (a) at least one biological control agent which is a fungicidally active bacterium and/or a fungicidally active metabolite produced by the respective bacterium and (b) one or more fatty acids or derivatives thereof selected from unsaturated and saturated C.sub.12-24 fatty acids, salts thereof, esters thereof or mixtures of any of the foregoing, wherein (a) and (b) are spatially separated.

Description

EXAMPLE 1

Production of Fatty Acid Derivative

[0213] A salt derivative of fatty acids can be prepared by the following method. This fatty acid salt derivative is referred to as ABP-510 herein. ABP-510 is obtained by mixing in a reactor a selection of fatty acids (C14-C18) derived from olive oil following oil hydrolysis whereby the glycerine is separated from the fatty acids and the fatty acids are subjected to fractional distillation in order to achieve the required specification. The fatty acids are made to react in a temperature controlled blending vessel with such quantity of potassium hydroxide and water for the production of the liquid fatty acid salt until solubilization of the components occurs and a liquid is produced that will readily form a true solution when diluted in water at the recommended rates of application for use in the field.

EXAMPLE 2

Test of a B. subtilis Strain and Fatty Acids Against Scab

[0214] For this example, the Product Flipper® (AlphaBioControl) was used. To test efficacy of B. subtilis strain QST713 and/or a product comprising fatty acids in comparison to chemical fungicides, a field trial was set up in three replicates with 5 apple trees per plot as per the scheme according to table 1. On Mar. 30, 2018 the first spray was carried out in the presence of a natural infestation of Venturia inequalis ascospores (scab). The applications were carried out with an interval of 5-6 days until the second week of June.

TABLE-US-00001 TABLE 1 .sup.1Merpan: 80% captan, .sup.2Delan: 70% dithianon, .sup.3Serenade: Bacillus subtilis, strain QST 713 (1.0 × 10.sup.9 CFU/g), .sup.4Flipper: 479.8 g/l Carboxylic Acid Potassium Salt Treatment Number Treatment 1. UTC 2. 1.8 kg/ha Merpan.sup.1 in alternation with 0.425 kg/ha Delan DF.sup.2 3. 5.0 l/ha. Serenade.sup.3 4. 5.0 l/ha Serenade + 2.5 l/ha Flipper.sup.4 5. 5.0 l/ha. Serenade + 5 l/ha Flipper 6. 5 l/ha. Flipper

TABLE-US-00002 TABLE 2 Number of infested leaved per 80 leaves per plot Infested leaves per 80 leaves per plot Treatment Replicate Nov. 6, 2018 May 7, 2018 1 A 23 31 B 17 24 C 19 37 SUM 59 92 2 A 8 12 B 2 3 C 12 10 SUM 22 25 3 A 5 19 B 9 21 C 20 25 SUM 34 65 4 A 8 11 B 5 18 C 11 18 SUM 24 47 5 A 4 20 B 6 14 C 8 5 SUM 18 39 6 A 8 9 B 12 22 C 11 19 SUM 31 50

[0215] Evaluation:

[0216] 80 leaves per plot were evaluated at two dates for infestation with scab.

[0217] Results are depicted in table 2 and the resulting efficacy in table 3.

TABLE-US-00003 TABLE 3 Summary percentage efficacy control of Scab. Treatment % efficacy Number Treatment Nov. 6, 2018 May 7, 2018 1. UTC  0%  0% 2. 1.8 kg/ha Merpan in alternation 63% 73% with 0.425 kg/ha Delan DF 3. 5.0 l/ha. Serenade 42% 29% 4. 5.0 l/ha Serenade + 2.5 l/ha 59% 49% Flipper 5. 5.0 l/ha. Serenade + 5 l/ha 69% 58% Flipper 6. 5 l/ha. Flipper 47% 46%

EXAMPLE 3

Synergistic Action

[0218] A synergistic effect of fungicides is always present when the fungicidal activity of the active compound combinations exceeds the total of the activities of the active compounds when applied individually. The expected activity for a given combination of two active compounds can be calculated as follows (according to Colby's formula) (cf. Colby, S. R., “Calculating Synergistic and Antagonistic Responses of Herbicide Combinations”, Weeds 1967, 15, 20-22):

[0219] If

[0220] X is the efficacy when active compound A is applied at an application rate of m ppm (or g/ha),

[0221] Y is the efficacy when active compound B is applied at an application rate of n ppm (or g/ha),

[0222] E is the efficacy when the active compounds A and B are applied at application rates of m and n ppm (or g/ha), respectively, and

[0223] then

[00001]$E=X+Y-\frac{X.Math.Y}{100}$

[0224] The degree of efficacy expressed in % is denoted. 0% means an efficacy which corresponds to that of the control while an efficacy of 100% means that no disease is observed.

[0225] If the actual fungicidal activity exceeds the calculated value, then the activity of the combination is superadditive, i.e. a synergistic effect exists. In this case, the efficacy which was actually observed must be greater than the value for the expected efficacy (E) calculated from the abovementioned formula.

[0226] A further way of demonstrating a synergistic effect is the method of Tammes (cf. “Isoboles, a graphic representation of synergism in pesticides” in Neth. J. Plant Path., 1964, 70, 73-80).

[0227] The invention is illustrated by the following examples. However the invention is not limited to the examples.

EXAMPLE 4

Synergistic Action of a B. subtilis Strain and Fatty Acids; Late Blight Test on Tomato

[0228] Serenade ASO commercial product (1×10.sup.9 CFU/ml), FLiPPER commercial product (491.8 g/l potassium salt of fatty acids), or combinations thereof were diluted with deionized water to the desired concentration.

[0229] To test for preventive activity, young plants were sprayed with the preparation of products at the stated rates of application. After the spray coating had dried on, the plants are inoculated with an aqueous spore suspension of Phytophthora infestans. The plants were then placed in an incubation cabinet at approximately 20° C. and a relative atmospheric humidity of 100%.

TABLE-US-00004 TABLE 4 Late blight test (tomato)/preventive Application rate of Efficacy in % Active compounds product in % volume. found* calc.** Serenade ASO (QST713) 0.5 6 FLiPPER 0.5 24 Serenade + FLiPPER 1:1 0.5 + 0.5 42 29 *found = activity found **calc. = activity calculated using Colby's formula

[0230] The test was evaluated 5 days after inoculation. 0% means an efficacy which corresponds to that of the untreated control, while an efficacy of 100% means that no disease is observed.

[0231] Table 4 below clearly shows that the observed activity of the active compound combination according to the invention is greater than the calculated activity, i.e. a synergistic effect is present.

EXAMPLE 5

Synergistic action of a B. subtilis strain and fatty acids; Gray mold test on pepper

[0232] Serenade ASO commercial product, FLiPPER commercial product, or combinations thereof were diluted with deionized water to the desired concentration.

[0233] To test for preventive activity, young plants were sprayed with the preparations of products at the stated rates of application. After the spray coating had dried on, the plants were inoculated with a spore suspension of Botrytis cinerea. The plants were then placed in an incubation cabinet at approximately 20° C. and a relative atmospheric humidity of 100%.

[0234] The test was evaluated 3 days after the inoculation. 0% means an efficacy which corresponds to that of the untreated control, while an efficacy of 100% means that no disease is observed.

TABLE-US-00005 TABLE 5 Gray mold test (pepper)/preventive Application rate of Efficacy in % Active compounds product in % volume. found* calc.** Serenade ASO (QST713) 2 45 1 37 FLiPPER 2 56 1 14 Serenade + FLiPPER 1:1 2 + 2 82 76 1 + 1 62 46 *found = activity found **calc. = activity calculated using Colby's formula

[0235] Table 5 clearly shows that the observed activity of the active compound combination according to the invention is greater than the calculated activity, i.e. a synergistic effect is present.

Example 6

Synergistic Action of a B. subtilis Strain and Fatty Acids; Wheat Leaf Rust Test on Wheat

[0236] Serenade ASO commercial product, FLiPPER commercial product, or combinations thereof were diluted with deionized water to the desired concentration.

[0237] To test for preventive activity, young plants were sprayed with the preparation of products at the stated rates of application. After the spray coating had dried on, the plants were inoculated with an aqueous spore suspension of Puccinia tritici. The plants were then placed in an incubation cabinet at approximately 20° C. and a relative atmospheric humidity of 100%.

[0238] The test was evaluated 10 days after the inoculation. 0% means an efficacy which corresponds to that of the untreated control, while an efficacy of 100% means that no disease is observed.

[0239] Table 6 below clearly shows that the observed activity of the active compound combination according to the invention is greater than the calculated activity, i.e. a synergistic effect is present.

TABLE-US-00006 TABLE 6 Wheat leaf rust test (wheat)/preventive Application rate of Efficacy in % Active compounds product in % volume. found* calc.** Serenade ASO (QST713) 1 63 0.5 38 0.25 25 FLiPPER 1 71 0.5 25 0.25 0 Serenade + FLiPPER 1:1 1 + 1 100 89 0.5 + 0.5 95 54 0.25 + 0.25 79 25 *found = activity found **calc. = activity calculated using Colby's formula

EXAMPLE 7

Synergistic Action of a Paenibacillus spp. Strain and Fatty Acids; Cucurbit Powdery Mildew on Cucumbers

[0240] Paenibacillus spp. strain NRRL B-67615 in a suspension concentrate (SC) formulation (4×10.sup.6 CFU/ml), FLiPPER commercial product, or combinations thereof were diluted with deionized water to the desired concentration.

[0241] The application rate of Paenibacillus spp. refers to the amount of Paenibacillus spp. strain NRRL B-67615 formulated as a suspension concentrate.

[0242] To test for curative activity, young plants were inoculated with an aqueous spore suspension of Podosphaera xanthii in a greenhouse and 3 days later, the plants were sprayed with the preparation of test samples at the stated rate of application. After the spray coating had dried on, the plants were returned to the greenhouse.

[0243] The test was evaluated 4 days after the treatments were applied. 0% means an efficacy which corresponds to that of the untreated control, while an efficacy of 100% means that no disease is observed.

[0244] Table 7 below clearly shows that the observed activity of the active compound combination according to the invention is greater than the calculated activity, i.e. a synergistic effect is present.

TABLE-US-00007 TABLE 7 Cucurbit powdery mildew test (cucumber)/curative Application rate of material Efficacy in % Active compounds in % volume found* calc.** Paenibacillus spp. 0.5 0 NRRL B-67615 FLiPPER 2 53 Paenibacillus spp. 0.5 + 2 68 53 NRRL B-67615 + FLiPPER *found = activity found **calc. = activity calculated using Colby's formula

EXAMPLE 8

Synergistic Action of a Paenibacillus spp. Strain and Fatty Acids; Wheat Leaf Rust on Wheat

[0245] Paenibacillus spp. strain NRRL B-67615 in a suspension concentrate (SC) formulation, FLiPPER commercial product, or combinations thereof were diluted with deionized water to the desired concentration.

[0246] The application rate of Paenibacillus SC refers to the amount of Paenibacillus strain NRRL B-67615 formulated as a suspension concentrate.

[0247] To test for preventive activity, young plants were sprayed with the preparation of products at the stated rates of application. After the spray coating had dried on, the plants were inoculated with an aqueous spore suspension of Puccinia tritici. The plants were then placed in an incubation cabinet at approximately 20° C. and a relative atmospheric humidity of 100%.

[0248] The test was evaluated 10 days after the inoculation. 0% means an efficacy which corresponds to that of the untreated control, while an efficacy of 100% means that no disease is observed.

[0249] Table 8 below clearly shows that the observed activity of the active compound combination according to the invention is greater than the calculated activity, i.e. a synergistic effect is present.

TABLE-US-00008 TABLE 8 Wheat leaf rust test (wheat)/preventive Application rate of material Efficacy in % Active compounds in % volume found* calc.** Paenibacillus spp. 0.5 0 NRRL B-67615 FLiPPER 2 38 Paenibacillus NRRL 0.5 + 2 58 38 B-67615 + FLiPPER *found = activity found **calc. = activity calculated using Colby's formula