PHYTOSANITARY COMPOSITION COMPRISING ESSENTIAL OILS THAT POTENTIATE ANTIFUNGAL ACTIVITY

Abstract

The present invention relates to phytosanitary compositions with fungicidal properties that comprise one essential oil obtained from plants selected from oregano oil (Origanum vulgare) and thyme oil (Thymus vulgaris) or its active compounds carvacrol at a concentration between 0.1 and 530 ppm or thymol at a concentration between 0.31 and 530 ppm, or a combination thereof; and (2) potassium carbonate at a concentration between 3.5 and 25 mM, and an agent with known fungicidal properties for use, principally, in contact-protection against fungal infections in cultivated plants and post-harvest, and also in other antifungal applications. In said compositions, the effect of the agents that have known fungicidal properties is potentiated synergistically by the aforementioned essential oils.

Claims

1. A phytosanitary composition having antifungal activity comprising: (1) one essential oil obtained from plants selected from oregano oil (Origanum vulgare) and thyme oil (Thymus vulgaris) or its active compounds carvacrol at a concentration between 0.1 and 530 ppm or thymol at a concentration between 0.31 and 530 ppm, or a combination thereof; and (2) potassium carbonate at a concentration between 3.5 and 25 mM.

2. A phytosanitary composition according to claim 1, characterised in that the carvacrol is at a concentration between 22 and 310 ppm.

3. A phytosanitary composition according to claim 1, characterised in that the thymol is at a concentration between 22 and 350 ppm.

4. A phytosanitary composition according to claim 1, characterised in that the potassium carbonate is at a concentration between 10 and 25 mM.

5. A phytosanitary composition according to claim 1, characterised in that the composition is in liquid or solid form, such as a suspension, dispersion, emulsion, spray, microencapsulate or other type of mixture which remains stable over time or is incorporated in polymers, or waxes or any other similar support.

6. A phytosanitary composition according to claim 1, characterised in that the phytosanitary composition is formulated together with additives such as surfactants, polymers, alkalinising agents, pH-controlling agents, among others.

7. A phytosanitary composition according to claim 1, which further comprises a fertiliser selected from the group comprising compounds containing nitrogen and/or phosphorus, such as urea, melamine, hexamine, dicyanodiamide, ameline, cyanuric acid, melamine nitrate, triethyl phosphate and the like or mixtures thereof.

8. A phytosanitary composition according to claim 1, which further comprises a compound or product having chemical and/or biological activity used in agriculture, such as herbicides, insecticides, plant growth regulators and the like, or mixtures thereof.

Description

EXAMPLES

Example 1. Inhibition of Growth of the Fungus Botrytis cinerea by K.SUB.2.CO.SUB.3 .Alone

[0026] The fungus B. cinerea was cultured in PDB (potato dextrose broth) medium with different concentrations of K.sub.2CO.sub.3. The % inhibition, representing the extent to which growth was impeded in comparison with a control which did not have the compound(s) under test, was calculated in the following way:

[00001]$\%.Math..Math.\mathrm{Inhibition}=\frac{\mathrm{OD}.Math..Math.\left(\mathrm{control}\right)-\mathrm{OD}.Math..Math.\left(x\right)}{\mathrm{OD}.Math..Math.\left(\mathrm{control}\right)}100$

where OD(control) is the optical density of the control culture (without test compounds) and OD(x) is the optical density of the culture with the test substance(s). The optical density of the liquid culture was measured 24 hours after the start of culturing and the results are shown in Table I.

TABLE-US-00001 TABLE I Inhibition of the growth of B. cinerea by K.sub.2CO.sub.3 K.sub.2CO.sub.3 concentration 0 3.5 4.25 4.5 5 (mM) Inhibition SD (%) 0 48.2 53.3 67.2 71.0 2.3 2.4 1.9 1.6

[0027] As will be seen from the table above, with a K.sub.2CO.sub.3 concentration between 3.5 and 5 mM inhibition of the B. cinerea culture was observed.

Example 2. Inhibition of Growth of the Fungus Botrytis cinerea by Carvacrol

[0028] The fungus B. cinerea was cultured in PDB medium with different concentrations of carvacrol. The % inhibition was calculated as Example 1.

[0029] The optical density of the liquid culture was measured 24 hours after the start of culturing and the results are shown in Table II.

TABLE-US-00002 TABLE II Inhibition of the growth of B. cinerea by carvacrol Carvacrol concentration 0.1 0.31 1 3.1 10 31 100 (ppm) Inhibition 0 10.5 7.7 13.7 4.1 22.4 3.4 21.3 5.0 51.4 5.5 74.4 1.1 SD (%)

Example 3. Inhibition of the Fungus Botrytis cinerea by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0030] The fungus B. cinerea was cultured in a similar way to Example 1 with the difference that different concentrations of carvacrol were used in the medium and that a constant concentration of K.sub.2CO.sub.3 (3.5 mM) was used throughout. The 24 hour optical density of the culture was measured and the results are shown in Table III.

TABLE-US-00003 TABLE III Inhibition of B. cinerea by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) K.sub.2CO.sub.3 concentration 3.5 3.5 3.5 3.5 3.5 3.5 3.5 (mM) Carvacrol concentration 0.1 0.31 1 3.1 10 31 100 (ppm) Inhibition 50.2 3.0 58.8 2.2 65.7 2.7 82.4 1.9 88.0 1.5 100 0 100 0 SD (%)

[0031] It will be seen how the results are improved by adding carvacrol to K.sub.2CO.sub.3. With 10 ppm of carvacrol (see in Table II) only 21% inhibition is achieved, and with 3.5 mM of K.sub.2CO.sub.3 48% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus B. cinerea is increased up to 88%.

Example 4. Inhibition of Growth of the Fungus Botrytis cinerea by Thymol Alone

[0032] The fungus B. cinerea was cultured in a similar way to Example 1 with different concentrations of thymol. The 24 hour optical density of the culture was measured and the results are shown in Table IV.

TABLE-US-00004 TABLE IV Inhibition of the growth of B. cinerea by Thymol Thymol concentration 0.31 1 3.1 10 31 100 (ppm) Inhibition 0 0 2.1 2.9 10.7 2.3 12.6 2.4 32.0 1.7 88.2 0.6 SD (%)

Example 5. Inhibition of the Fungus Botrytis cinerea by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Thymol)

[0033] The fungus B. cinerea was cultured in a similar way to Example 1 with the difference that different concentrations of thymol were used in the medium and that a constant concentration of K.sub.2CO.sub.3 (3.5 mM) was used throughout. The 24 hour optical density of the culture was measured and the results are shown in Table V.

TABLE-US-00005 TABLE V Inhibition of B. cinerea by the composition according to this invention (K.sub.2CO.sub.3 + Thymol) K.sub.2CO.sub.3 concentration 3.5 3.5 3.5 3.5 3.5 3.5 (mM) Thymol concentration 0.31 1 3.1 10 31 100 (ppm) Inhibition 49.4 2.1 62.1 2.2 73.2 1.1 81.0 1.6 93.0 1.1 100 0 SD (%)

[0034] It will be seen how the results are improved by adding thymol to K.sub.2CO.sub.3. With 10 ppm of thymol only 13% inhibition is achieved, and with 3.5 mM of K.sub.2CO.sub.3 48% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus B. cinerea is increased up to some 81%.

Example 6. Inhibition of Growth of the Fungus Alternaria alternata by K.SUB.2.CO.SUB.3 .Alone

[0035] Alternaria alternata was cultured in a similar way to Example 1 with different concentrations of K.sub.2CO.sub.3. The 24 hour optical density of the culture was measured and the results are shown in Table VI.

TABLE-US-00006 TABLE VI Inhibition of the growth of A. alternata by K.sub.2CO.sub.3 K.sub.2CO.sub.3 concentration 0 3.5 4.25 4.5 5 (mM) Inhibition SD (%) 0 31.5 39.7 48.0 59.0 2.5 2.5 2.2 2.2

[0036] As will be seen from the table above, with a K.sub.2CO.sub.3 concentration between 3.5 and 5 mM inhibition of the A. alternata culture was observed.

Example 7. Inhibition of Growth of the Fungus Alternaria alternata by Carvacrol

[0037] The fungus A. alternata was cultured in PDB medium with different concentrations of carvacrol. The % inhibition was calculated as Example 1 and the results are shown in Table VII.

TABLE-US-00007 TABLE VII Inhibition of the growth of A. alternata by carvacrol Carvacrol concentration 0.1 0.31 1 3.1 10 31 100 (ppm) Inhibition 0 2.0 0.3 3.5 0.8 5.8 1.4 17.7 11.0 27.2 14.0 74.6 8.0 SD (%)

Example 8. Inhibition of the Fungus Alternaria alternata by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0038] The fungus A. alternata was cultured in a similar way to Example 1 with the difference that different concentrations of carvacrol were used in the medium and that a constant concentration of K.sub.2CO.sub.3 (3.5 mM) was used throughout. The 24 hour optical density of the culture was measured and the results are shown in Table VIII.

TABLE-US-00008 TABLE VIII Inhibition of A. alternata by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) K.sub.2CO.sub.3 concentration 3.5 3.5 3.5 3.5 3.5 3.5 3.5 (mM) Carvacrol concentration 0.1 0.31 1 3.1 10 31 100 (ppm) Inhibition 30.1 2.9 35.7 2.7 38.0 2.9 46.2 2.9 65.7 1.6 86.3 1.2 100 0 SD (%)

[0039] It will be seen how the results are improved by adding carvacrol to K.sub.2CO.sub.3. With 31 ppm of carvacrol (see Table VII) only 27% inhibition is achieved, and with 3.5 mM of K.sub.2CO.sub.3 32% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus A. alternata is increased up to some 86%.

Example 9. Inhibition of Growth of the Fungus Alternaria alternata by Thymol Alone

[0040] The fungus A. alternata was cultured in a similar way to Example 1 with different concentrations of thymol. The 24 hour optical density of the culture was measured and the results are shown in Table IX.

TABLE-US-00009 TABLE IX Inhibition of the growth of A. alternata by Thymol Thymol concentration 0.31 1 3.1 10 31 100 (ppm) Inhibition 0 0 4.1 2.9 10.4 2.6 16.0 2.0 29.0 1.5 69.4 0 SD (%)

Example 10. Inhibition of the Fungus Alternaria alternata by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Thymol)

[0041] The fungus A. alternata was cultured in a similar way to Example 1 with the difference that different concentrations of thymol were used in the medium and that a constant concentration of K.sub.2CO.sub.3 (3.5 mM) was used throughout. The 24 hour optical density of the culture was measured and the results are shown in Table X.

TABLE-US-00010 TABLE X Inhibition of A. alternata by the composition according to this invention (K.sub.2CO.sub.3 + Thymol) K.sub.2CO.sub.3 concentration (mM) 3.5 3.5 3.5 3.5 3.5 3.5 Thymol concentration 0.31 1 3.1 10 31 100 (ppm) Inhibition 33.7 2.2 38.7 2.7 45.3 2.4 55.5 1.2 71.8 1.3 100 0 SD (%)

[0042] It will be seen how the results are improved by adding thymol to K.sub.2CO.sub.3. With 31 ppm of thymol only 29% inhibition is achieved, and with 3.5 mM of K.sub.2CO.sub.3 32% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus A. alternata is increased up to some 72%.

Example 11. Inhibition of Growth of the Fungus Penicillium digitatum by K.SUB.2.CO.SUB.3 .Alone

[0043] Penicillium digitatum was cultured in a similar way to Example 1 with different concentrations of K.sub.2CO.sub.3. The 24 hour optical density of the culture was measured and the results are shown in Table XI.

TABLE-US-00011 TABLE XI Inhibition of the growth of P. digitatum by K.sub.2CO.sub.3 K.sub.2CO.sub.3 concentration 0 3.5 4.25 4.5 5 (mM) Inhibition SD (%) 0 29.1 33.0 36.9 39.5 2.1 1.7 1.6 1.8

[0044] As will be seen from the table above, with a K.sub.2CO.sub.3 concentration between 3.5 and 5 mM inhibition of the P. digitatum culture was observed.

Example 12. Inhibition of Growth of the Fungus Penicillium digitatum by Carvacrol Alone

[0045] The fungus P. digitatum was cultured in a similar way to Example 1 with different concentrations of carvacrol. The 24 hour optical density of the culture was measured and the results are shown in Table XII.

TABLE-US-00012 TABLE XII Inhibition of the growth of P. digitatum by Carvacrol Carvacrol 0.1 0.31 1 3.1 10 31 100 concentration (ppm) Inhibition 32.3 2.8 34.3 2.8 37.9 3.0 52.6 2.2 57.8 2.1 86.0 1.1 100 0 SD (%)

Example 13. Inhibition of the Fungus Penicillium digitatum by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0046] The fungus P. digitatum was cultured in a similar way to Example 1 with the difference that different concentrations of carvacrol were used in the medium and that a constant concentration of K.sub.2CO.sub.3 (3.5 mM) was used throughout. The 24 hour optical density of the culture was measured and the results are shown in Table XIII.

TABLE-US-00013 TABLE XIII Inhibition of P. digitatum by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) K.sub.2CO.sub.3 concentration 3.5 3.5 3.5 3.5 3.5 3.5 3.5 (mM) Carvacrol concentration 0.1 0.31 1 3.1 10 31 100 (ppm) Inhibition 65.9 2.4 66.2 2.9 72.4 2.2 88.6 1.9 96.5 1.4 100 0 100 0 SD (%)

[0047] It will be seen how the results are improved by adding carvacrol to K.sub.2CO.sub.3. With 10 ppm of carvacrol 58% inhibition is achieved, and with 3.5 mM of K.sub.2CO.sub.3 29% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus P. digitatum is increased up to 97%.

Example 14. Inhibition of Growth of the Fungus Penicillium digitatum by Thymol Alone

[0048] The fungus P. digitatum was cultured in a similar way to Example 1 with different concentrations of thymol. The 24 hour optical density of the culture was measured and the results are shown in Table XIV.

TABLE-US-00014 TABLE XIV Inhibition of the growth of P. digitatum by Thymol Thymol concentration 0.31 1 3.1 10 31 100 100 (ppm) Inhibition 28.2 3.9 24.2 6.0 36.3 2.3 36.2 2.0 50.7 2.0 78.3 2.2 95.6 0.5 SD (%)

Example 15. Inhibition of the Fungus Penicillium digitatum by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Thymol)

[0049] The fungus P. digitatum was cultured in a similar way to Example 1 with the difference that different concentrations of thymol were used in the medium and that a constant concentration of K.sub.2CO.sub.3 (3.5 mM) was used throughout. The 24 hour optical density of the culture was measured and the results are shown in Table XV.

TABLE-US-00015 TABLE XV Inhibition of P. digitatum by the composition according to this invention (K.sub.2CO.sub.3 + Thymol) K.sub.2CO.sub.3 concentration 3.5 3.5 3.5 3.5 3.5 3.5 (mM) Thymol concentration 0.31 1 3.1 10 31 100 (ppm) Inhibition 58.1 2.9 61.3 2.5 69.9 1.9 78.3 1.5 94.3 1.2 100 0 SD (%)

[0050] It will be seen how the results are improved by adding thymol to K.sub.2CO.sub.3. With 31 ppm of thymol (see Table XIV) only 51% inhibition is achieved, and with 3.5 mM of K.sub.2CO.sub.3 29% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus P. digitatum is increased up to some 94%.

Example 16. Inhibition of Growth of the Fungus Cercospora beticola by K.SUB.2.CO.SUB.3 .Alone

[0051] Cercospora beticola was cultured on PDA (potato dextrose agar) culture medium buffered to a pH value not exceeding 9.5. The inhibition degree, expressed as a percentage, was determined based on the growth relative to control that did not have the compound(s) to be tested. The % inhibition was calculated with the following formula:

[00002]$\%.Math..Math.\mathrm{Inhibition}=\frac{\mathrm{colony}.Math..Math.\mathrm{diameter}.Math..Math.\left(\mathrm{control}\right)-\mathrm{colony}.Math..Math.\mathrm{diameter}.Math..Math.\left(x\right)}{\mathrm{colony}.Math..Math.\mathrm{diameter}.Math..Math.\left(\mathrm{control}\right)}100$

wherein colony diameter (control) is the size of the control colony (without the compounds to be tested) and colony diameter (x) is the size of the colony with the substance(s) to be tested. A fixed K.sub.2CO.sub.3 concentration of 7.24 mM was tested in C. beticola. The results are shown in Table XVI.

TABLE-US-00016 TABLE XVI Inhibition of the growth of C. beticola by K.sub.2CO.sub.3 alone K.sub.2CO.sub.3 concentration (mM) 7.24 Inhibition SD (%) 25.0 1.2

Example 17. Inhibition of Growth of the Fungus Cercospora beticola by Carvacrol Alone

[0052] The fungus C. beticola was cultured in a similar way to Example 16 with 10 ppm of carvacrol. The % inhibition was calculated and the results are shown in Table XVII.

TABLE-US-00017 TABLE XVII Inhibition of the growth of C. beticola by carvacrol alone Carvacrol concentration (ppm) 10 Inhibition SD (%) 17.0 0.8

Example 18. Inhibition of Growth of the Fungus Cercospora beticola by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0053] The fungus C. beticola was cultured in a similar way to Example 16 with a fixed concentration of K.sub.2CO.sub.3 (7.24 mM) and carvacrol (10 ppm). The % inhibition was calculated and the results are shown in Table XVIII.

TABLE-US-00018 TABLE XVIII Inhibition of the growth of C. beticola by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) K.sub.2CO.sub.3 concentration (mM) 7.24 Carvacrol concentration (ppm) 10 Inhibition SD (%) 65.0 3.2

[0054] It will be seen how the results are improved by adding carvacrol to K.sub.2CO.sub.3. With 10 ppm of carvacrol (see Table XVII) only 17% inhibition is achieved, and with 7.24 mM of K.sub.2CO.sub.3 25% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus C. beticola is increased up to some 65%.

Example 19. Inhibition of the Fungus Botrytis cinerea by K.SUB.2.CO.SUB.3 .Alone

[0055] Leaves of tomato plants (var. Marmande) of 5 weeks old were treated with different concentrations of K.sub.2CO.sub.3, and 24 hours later, they were infected with the fungus Botrytis cinerea. Two weeks later, fungal infection was assessed in leaves. The % inhibition, representing the extent to which fungal growth was impeded in comparison with a control which did not have the compound(s) under test, was determined in the following way:

[00003]$\%.Math..Math.\mathrm{Inhibition}=\frac{\%.Math..Math.\mathrm{Infection}.Math..Math.\left(\mathrm{control}\right)-\%.Math..Math.\mathrm{Infection}.Math..Math.\left(x\right)}{\%.Math..Math.\mathrm{Infection}.Math..Math.\left(\mathrm{control}\right)}100$

where % Infection (control) is the percentage of fungal infection of the control leaves (without test compounds) and % Infection (x) is percentage of fungal infection of the treated leaves. The results are shown in Table XIX.

TABLE-US-00019 TABLE XIX Inhibition of the growth of B. cinerea by K.sub.2CO.sub.3 alone K.sub.2CO.sub.3 concentration 12 13 14 15 22 (mM) Inhibition SD (%) 13.4 13.5 13.8 13.8 15.3 2.5 2.1 2.3 3.0 2.4

Example 20. Inhibition of the Fungus Botrytis cinerea by Carvacrol Alone

[0056] Leaves of tomato plants treated with different concentrations of carvacrol were infected with the fungus B. cinerea in a similar way to Example 19. The % inhibition was calculated and the results are shown in Table XX.

TABLE-US-00020 TABLE XX Inhibition of the growth of B. cinerea by Carvacrol alone Carvacrol concentration 26 31 100 200 308 (ppm) Inhibition SD (%) 7.2 7.4 16.7 18.6 20.8 1.1 1.3 2.1 2.3 2.3

Example 21. Inhibition of Growth of the Fungus Botrytis cinerea by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0057] Leaves of tomato plants treated with different concentrations of carvacrol and carvacrol were infected with the fungus B. cinerea in a similar way to Example 19. The % inhibition was calculated and the results are shown in Table XXI.

TABLE-US-00021 TABLE XXI Inhibition of the growth of B. cinerea by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) K.sub.2CO.sub.3 concentration 14 22 15 15 13 (mM) Carvacrol concentration 26 31 100 200 308 (ppm) Inhibition SD (%) 24.3 28.1 48.3 60.0 72.8 2.1 2.8 2.8 2.6 2.4

[0058] It will be seen how the results are improved by adding carvacrol to K.sub.2CO.sub.3. With 308 ppm of carvacrol (see Table XX) only 21% inhibition is achieved, and with 13 mM of K.sub.2CO.sub.3 14% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus B. cinerea is increased up to some 73%.

Example 22. Inhibition of the Fungus Botrytis cinerea by Thymol Alone

[0059] Leaves of tomato plants treated with different concentrations of thymol were infected with the fungus B. cinerea in a similar way to Example 19. The % inhibition was calculated and the results are shown in Table XXII.

TABLE-US-00022 TABLE XXII Inhibition of the growth of B. cinerea by Thymol alone Thymol concentration (ppm) 35 100 200 350 Inhibition SD (%) 7.4 17.7 20.9 22.8 1.3 1.7 1.5 1.5

Example 23. Inhibition of Growth of the Fungus Botrytis cinerea by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Thymol)

[0060] Leaves of tomato plants treated with different concentrations of thymol and K.sub.2CO.sub.3 were infected with the fungus B. cinerea in a similar way to Example 19. The % inhibition was calculated and the results are shown in Table XXIII.

TABLE-US-00023 TABLE XXIII Inhibition of the growth of B. cinerea by the composition according to this invention (K.sub.2CO.sub.3 + Thymol) K.sub.2CO.sub.3 concentration (mM) 15 15 15 12 Thymol concentration (ppm) 35 100 200 350 Inhibition SD (%) 25.6 40.8 53.5 65.0 2.6 2.4 2.3 2.2

[0061] It will be seen how the results are improved by adding thymol to K.sub.2CO.sub.3. With 350 ppm of thymol (see Table XXII) only 23% inhibition is achieved, and with 12 mM of K.sub.2CO.sub.3 14% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus B. cinerea is increased up to some 65%.

Example 24. Inhibition of the Fungus Alternaria alternata by K.SUB.2.CO.SUB.3 .Alone

[0062] Leaves of tomato plants treated with different concentrations of K.sub.2CO.sub.3 were infected with the fungus A. alternata in a similar way to Example 19. The % inhibition was calculated and the results are shown in Table XXIV.

TABLE-US-00024 TABLE XXIV Inhibition of the growth of A. alternata by K.sub.2CO.sub.3 alone K.sub.2CO.sub.3 concentration 12 13 14 15 22 (mM) Inhibition SD (%) 6.5 7.0 7.2 7.8 8.6 2.2 1.1 1.1 1.4 1.5

Example 25. Inhibition of the Fungus Alternaria alternata by Carvacrol Alone

[0063] Leaves of tomato plants treated with different concentrations of carvacrol were infected with the fungus A. alternata in a similar way to Example 19. The % inhibition was calculated and the results are shown in Table XXV.

TABLE-US-00025 TABLE XXV Inhibition of the growth of A. alternata by Carvacrol alone Carvacrol concentration 26 31 100 200 308 (ppm) Inhibition SD (%) 4.3 5.8 11.4 16.1 18.4 0.7 0.9 1.3 1.2 1.2

Example 26. Inhibition of Growth of the Fungus Alternaria alternata by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0064] Leaves of tomato plants treated with different concentrations of carvacrol and K.sub.2CO.sub.3 were infected with the fungus B. cinerea in a similar way to Example 19. The % inhibition was calculated and the results are shown in Table XXVI.

TABLE-US-00026 TABLE XXVI Inhibition of the growth of A. alternata by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) K.sub.2CO.sub.3 concentration 14 22 15 15 13 (mM) Carvacrol concentration 26 31 100 200 308 (ppm) Inhibition SD (%) 18.4 17.3 43.9 60.9 70.8 2.1 2.4 3.2 2.8 3.5

[0065] It will be seen how the results are improved by adding carvacrol to K.sub.2CO.sub.3. With 308 ppm of carvacrol (see Table XXV) only 18% inhibition is achieved, and with 13 mM of K.sub.2CO.sub.3 7% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus A. alternata is increased up to some 71%.

Example 27. Inhibition of the Fungus Alternaria alternata by Thymol Alone

[0066] Leaves of tomato plants treated with different concentrations of thymol were infected with the fungus A. alternata in a similar way to Example 19. The % inhibition was calculated and the results are shown in Table XXVII.

TABLE-US-00027 TABLE XXVII Inhibition of the growth of A. alternata by Thymol alone Thymol concentration (ppm) 35 100 200 350 Inhibition SD (%) 10.0 15.2 19.3 21.3 1.4 2.0 2.3 2.6

Example 28. Inhibition of Growth of the Fungus Alternaria alternata by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Thymol)

[0067] Leaves of tomato plants treated with different concentrations of thymol and K.sub.2CO.sub.3 were infected with the fungus A. alternata in a similar way to Example 19. The % inhibition was calculated and the results are shown in Table XXVIII.

TABLE-US-00028 TABLE XXVIII Inhibition of the growth of A. alternata by the composition according to this invention (K.sub.2CO.sub.3 + Thymol) K.sub.2CO.sub.3 concentration (mM) 15 15 15 12 Thymol concentration (ppm) 35 100 200 350 Inhibition SD (%) 22.4 37.3 53.7 60.8 2.5 2.9 2.0 2.2

[0068] It will be seen how the results are improved by adding thymol to K.sub.2CO.sub.3. With 350 ppm of thymol (see Table XXVII) only 21% inhibition is achieved, and with 12 mM of K.sub.2CO.sub.3 7% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus A. alternata is increased up to some 61%.

Example 29. Inhibition of the Fungus Phytophthora infestans by K.SUB.2.CO.SUB.3 .Alone

[0069] Tomato plants (var. Marmande) of 5 weeks old were treated with 15 mM of K.sub.2CO.sub.3, and 24 hours later, they were infected with the fungus Phytophthora infestans. Two weeks later, fungal infection was assessed in leaves. The % inhibition, representing the extent to which growth was impeded in comparison with a control which did not have the compounds under test, was determined in the following way:

[00004]$\%.Math..Math.\mathrm{Inhibition}=\frac{\%.Math..Math.\mathrm{Infection}.Math..Math.\left(\mathrm{control}\right)-\%.Math..Math.\mathrm{Infection}.Math..Math.\left(x\right)}{\%.Math..Math.\mathrm{Infection}.Math..Math.\left(\mathrm{control}\right)}100$

where % Infection (control) is the percentage of fungal infection of the control plants (without test compound) and % Infection (x) is percentage of fungal infection of the treated plants. The results are shown in Table XXIX.

TABLE-US-00029 TABLE XXIX Inhibition of the growth of P. infestans by K.sub.2CO.sub.3 alone K.sub.2CO.sub.3 concentration (mM) 15 Inhibition SD (%) 18.5 3.6

Example 30. Inhibition of the Fungus Phytophthora infestans by Carvacrol Alone

[0070] Tomato plants were treated with different concentrations of carvacrol and subsequently infected with the fungus P. infestans in a similar way to Example 29. The % inhibition was calculated and the results are shown in Table XXX.

TABLE-US-00030 TABLE XXX Inhibition of the growth of P. infestans by Carvacrol alone Carvacrol concentration (ppm) 26 150 308 Inhibition SD (%) 3.2 0.8 17.1 2.3 35.0 4.8

Example 31. Inhibition of Growth of the Fungus Phytophthora infestans by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0071] Tomato plants were treated with different concentrations of carvacrol and a fixed concentration of K.sub.2CO.sub.3 and subsequently infected with the fungus P. infestans in a similar way to Example 29. The % inhibition was calculated and the results are shown in Table XXXI.

TABLE-US-00031 TABLE XXXI Inhibition of the growth of P. infestans by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) K.sub.2CO.sub.3 concentration (mM) 15 15 15 Carvacrol concentration (ppm) 26 150 308 Inhibition SD (%) 25.6 3.6 52.2 4.2 78.3 4.8

[0072] It will be seen how the results are improved by adding carvacrol to K.sub.2CO.sub.3. With 308 ppm of carvacrol (see Table XXX) only 35% inhibition is achieved, and with 15 mM of K.sub.2CO.sub.3 19% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus P. infestans is increased up to some 78%.

Example 32. Inhibition of the Fungus Phytophthora infestans by Thymol Alone

[0073] Tomato plants were treated with different concentrations of thymol and subsequently infected with the fungus P. infestans in a similar way to Example 29. The % inhibition was calculated and the results are shown in Table XXXII.

TABLE-US-00032 TABLE XXXII Inhibition of the growth of P. infestans by Thymol alone Thymol concentration (ppm) 35 150 350 Inhibition SD (%) 4.3 0.7 16.2 3.2 33.6 5.6

Example 33. Inhibition of Growth of the Fungus Phytophthora infestans by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Thymol)

[0074] Tomato plants were treated with different concentrations of thymol and a fixed concentration of K.sub.2CO.sub.3 and subsequently infected with the fungus P. infestans in a similar way to Example 29. The % inhibition was calculated and the results are shown in Table XXXIII.

TABLE-US-00033 TABLE XXXIII Inhibition of the growth of P. infestans by the composition according to this invention (K.sub.2CO.sub.3 + Thymol) K.sub.2CO.sub.3 concentration (mM) 15 15 15 Thymol concentration (ppm) 35 150 350 Inhibition SD (%) 26.3 2.2 49.3 3.6 74.2 5.2

[0075] It will be seen how the results are improved by adding thymol to K.sub.2CO.sub.3. With 350 ppm of thymol (see Table XXXII) only 34% inhibition is achieved, and with 15 mM of K.sub.2CO.sub.3 19% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus P. infestans is increased up to some 74%.

Example 34. Inhibition of the Fungus Leveillula Taurica by K.SUB.2.CO.SUB.3 .Alone

[0076] The fungus L. taurica was cultured in a similar way to Example 29 with different concentrations of K.sub.2CO.sub.3. The % inhibition was calculated and the results are shown in Table XXXIV.

TABLE-US-00034 TABLE XXXIV Inhibition of the growth of L. taurica by K.sub.2CO.sub.3 alone K.sub.2CO.sub.3 concentration (mM) 15 Inhibition SD (%) 13.9 0.5

Example 35. Inhibition of the Fungus Leveillula Taurica by Carvacrol Alone

[0077] Tomato plants were treated with different concentrations of carvacrol and subsequently infected with the fungus L. taurica in a similar way to Example 29. The % inhibition was calculated and the results are shown in Table XXXV.

TABLE-US-00035 TABLE XXXV Inhibition of the growth of L. taurica by Carvacrol alone. Carvacrol concentration (ppm) 26 150 308 Inhibition SD (%) 4.0 1.2 11.2 1.0 41.5 1.2

Example 36. Inhibition of Growth of the Fungus Leveillula Taurica by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0078] Tomato plants were treated with different concentrations of carvacrol and a fixed concentration of K.sub.2CO.sub.3 and subsequently infected with the fungus L. taurica in a similar way to Example 29. The % inhibition was calculated and the results are shown in Table XXXVI.

TABLE-US-00036 TABLE XXXVI Inhibition of the growth of L. taurica by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) K.sub.2CO.sub.3 concentration (mM) 15 15 15 Carvacrol concentration (ppm) 26 150 308 Inhibition SD (%) 18.6 3.3 78.3 0.6 92.0 0

[0079] It will be seen how the results are improved by adding carvacrol to K.sub.2CO.sub.3. With 308 ppm of carvacrol (see Table XXXV) only 42% inhibition is achieved, and with 15 mM of K.sub.2CO.sub.3 14% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus L. taurica is increased up to some 92%.

Example 37. Inhibition of the Fungus Leveillula Taurica by Thymol Alone

[0080] Tomato plants were treated with different concentrations of thymol and subsequently infected with the fungus L. taurica in a similar way to Example 29. The % inhibition was calculated and the results are shown in Table XXXVII.

TABLE-US-00037 TABLE XXXVII Inhibition of the growth of L. taurica by Thymol alone. Thymol concentration (ppm) 35 150 350 Inhibition SD (%) 10.5 5.4 38.2 5.5 51.2 1.0

Example 38. Inhibition of Growth of the Fungus Leveillula Taurica by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Thymol)

[0081] Tomato plants were treated with different concentrations of thymol and a fixed concentration of K.sub.2CO.sub.3 and subsequently infected with the fungus L. taurica in a similar way to Example 29. The % inhibition was calculated and the results are shown in Table XXX VIII.

TABLE-US-00038 TABLE XXXVIII Inhibition of the growth of L. taurica by the composition according to this invention (K.sub.2CO.sub.3 + Thymol) K.sub.2CO.sub.3 concentration (mM) 15 15 15 Thymol concentration (ppm) 35 150 350 Inhibition SD (%) 48.8 3.1 65.4 2.4 72.9 1.1

[0082] It will be seen how the results are improved by adding thymol to K.sub.2CO.sub.3. With 350 ppm of thymol (see Table XXXVII) only 51% inhibition is achieved, and with 15 mM of K.sub.2CO.sub.3 14% inhibition is achieved. However, when the two compounds are combined inhibition of growth of the fungus P. infestans is increased up to some 73%.

Example 39. Inhibition of the Fungus Pseudoperonospora cubensis by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0083] The efficacy of the composition of the present invention (K.sub.2CO.sub.3+carvacrol or thymol) was tested in field assays with cucumber, tomato, lettuce or potato plants to prevent Pseudoperonospora cubensis, Botrytis cinerea, Phytophthora infestans or Leveillula taurica growth. The efficacy of the fungicide was measured as follow:

[00005]$\%.Math..Math.\mathrm{Efficacy}=\frac{\%.Math..Math.\mathrm{Severity}.Math..Math.\left(\mathrm{control}\right)-\%.Math..Math.\mathrm{Severity}.Math..Math.\left(x\right)}{\%.Math..Math.\mathrm{Severity}.Math..Math.\left(\mathrm{control}\right)}100$

where % Severity (control) is the percentage of fungal severity of the control plants (without test compounds) and % Severity (x) is percentage of fungal severity of the treated plants. Cucumber plants were treated with different concentrations of K.sub.2CO.sub.3 and carvacrol and subsequently infected with Pseudoperonospora cubensis. The results are shown in Table XXXIX.

TABLE-US-00039 TABLE XXXIX Efficacy of Pseudoperonospora cubensis by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) in cucumber K.sub.2CO.sub.3 concentration 13.5 19 6.5 13.0 22.4 (mM) Carvacrol concentration 25.5 36.5 150 300 530 (ppm) Inhibition 16.7 26.7 35.5 45.3 64.2 SD (%) 3.3 4.2 3.9 5.3 7.3

[0084] The efficacy of the inhibition of growth of the fungus P. cubensis reached 64% by combining K.sub.2CO.sub.3 and carvacrol.

Example 40. Inhibition of the Fungus Pseudoperonospora cubensis by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Thymol)

[0085] The efficacy of the composition of the present invention (K.sub.2CO.sub.3+Thymol) was tested in cucumber to prevent P. cubensis growth. The efficacy of the antifungicide was measured and the results are shown in Table XL.

TABLE-US-00040 TABLE XL Efficacy of Pseudoperonospora cubensis by the composition according to this invention (K.sub.2CO.sub.3 + Thymol) in cucumber K.sub.2CO.sub.3 concentration (mM) 6.5 13.0 22.4 Thymol concentration (ppm) 150 300 530 Inhibition SD (%) 31.6 4.6 41.1 6.1 53.3 7.0

[0086] The efficacy of the inhibition of growth of the fungus P. cubensis reached 53% by combining K.sub.2CO.sub.3 and thymol.

Example 41. Inhibition of the Fungus Botrytis cinerea by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0087] The efficacy of the composition of the present invention (K.sub.2CO.sub.3+Carvacrol) was tested in tomato to prevent B. cinerea growth. The efficacy of the antifungicide was measured and the results are shown in Table XLI.

TABLE-US-00041 TABLE XLI Efficacy of Botrytis cinerea by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) in tomato K.sub.2CO.sub.3 concentration 13.5 19 6.5 13.0 22.4 (mM) Carvacrol concentration 25.5 36.5 150 300 530 (ppm) Inhibition SD (%) 21.7 27.8 38.5 69.8 78.3 4.4 4.1 5.6 8.3 6.5

[0088] The efficacy of the inhibition of growth of the fungus B. cinerea reached 78% by combining K.sub.2CO.sub.3 and carvacrol.

Example 42. Inhibition of the Fungus Botrytis cinerea by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Thymol)

[0089] The efficacy of the composition of the present invention (K.sub.2CO.sub.3+Thymol) was tested in tomato to prevent B. cinerea growth. The efficacy of the antifungicide was measured and the results are shown in Table XLII.

TABLE-US-00042 TABLE XLII Efficacy of Botrytis cinerea by the composition according to this invention (K.sub.2CO.sub.3 + Thymol) in tomato K.sub.2CO.sub.3 concentration (mM) 6.5 13.0 22.4 Thymol concentration (ppm) 150 300 530 Inhibition SD (%) 35.8 4.3 58.0 4.6 66.0 6.0

[0090] The efficacy of the inhibition of growth of the fungus B. cinerea reached 66% by combining K.sub.2CO.sub.3 and thymol.

Example 43. Inhibition of the Fungus Phytophthora infestans by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0091] The efficacy of the composition of the present invention (K.sub.2CO.sub.3+Carvacrol) was tested in lettuce to prevent P. infestans growth. The efficacy of the antifungicide was measured and the results are shown in Table XLIII.

TABLE-US-00043 TABLE XLIII Efficacy of P. infestans by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) in lettuce K.sub.2CO.sub.3 concentration (mM) 5.4 12.6 21.6 Carvacrol concentration (ppm) 132 309 529 Inhibition SD (%) 58.0 2.4 72.1 5.2 80.3 4.3

[0092] The efficacy of the inhibition of growth of the fungus P. infestans reached 80% by combining K.sub.2CO.sub.3 and carvacrol.

Example 44. Inhibition of the Fungus Phytophthora infestans by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0093] The efficacy of the composition of the present invention (K.sub.2CO.sub.3+Carvacrol) was tested in potato to prevent P. infestans growth. The efficacy of the antifungicide was measured and the results are shown in Table XLIV.

TABLE-US-00044 TABLE XLIV Efficacy of P. infestans by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) in potato K.sub.2CO.sub.3 concentration (mM) 5.4 12.6 21.6 Carvacrol concentration (ppm) 132 309 529 Inhibition SD (%) 68.6 2.9 71.7 3.4 86.7 4.1

[0094] The efficacy of the inhibition of growth of the fungus P. infestans reached 87% by combining K.sub.2CO.sub.3 and carvacrol.

Example 45. Inhibition of the Fungus Leveilulla Taurica by the Composition According to this Invention (K.SUB.2.CO.SUB.3.+Carvacrol)

[0095] The efficacy of the composition of the present invention (K.sub.2CO.sub.3+Carvacrol) was tested in tomato to prevent L. taurica growth. The efficacy of the antifungicide was measured and the results are shown in Table XLV.

TABLE-US-00045 TABLE XLV Efficacy of L. taurica by the composition according to this invention (K.sub.2CO.sub.3 + Carvacrol) in tomato K.sub.2CO.sub.3 concentration 5.7 13.2 24.5 5.4 12.6 21.6 (mM) Carvacrol concentration 11.0 25.6 47.5 132 309 529 (ppm) Inhibition 22.0 3.1 53.0 4.4 61.0 3.9 43.0 4.2 58.0 5.2 71.0 3.8 SD (%)

[0096] The efficacy of the inhibition of growth of the fungus L. taurica reached 71% by combining K.sub.2CO.sub.3 and carvacrol.