Antimicrobial and synergistic phytochemical composition, its preparation and applications

Abstract

The innovations provides an antimicrobial and synergistic phytochemical composition, consisting of two active ingredients (FZ and YC). This innovation also consists of formulation processes and its applications to fungi, bacteria and/or virus-caused plant diseases such as citrus Huanglongbing, grape downy mildew, strawberry gray mold etc. This innovation combines two ingredients (FZ and YC) with different modes of action, dramatically improving plant inducible acquired resistance. Therefore, this composition dramatically enhances the control efficacy of plant diseases caused by fungi, bacteria or virus, expands the antimicrobial spectrum and promotes the plant growth.

Claims

1. A phytochemical composition exhibiting antimicrobial effects comprising: a carrier; and a synergistic combination of active ingredients consisting essentially of a first active ingredient (FZ) and a second active ingredient (YC), wherein a) FZ includes: 1) rhubarb extract; 2) giant knotweed extract; 3) cassia seed extract; or 4) combinations thereof; and wherein b) YC includes: 1) canola pollen extract; 2) beewax extract; 3) synthesized brassinosteroid; or 4) combinations thereof.

2. The composition according to claim 1, wherein the FZ is selected from giant knotweed extract and cassia seed extract.

3. The composition according to claim 1, wherein the FZ is selected from giant knotweed extract.

4. The composition according to claim 1, wherein the YC is selected from Canola pollen extract, beewax extract, and synthesized brassinosteroid.

5. The composition according to claim 1, wherein the YC is selected from beewax extract.

6. The composition according to claim 1, wherein the percent content of FZ in the composition is 0.001-99.0% (w/w), and percent content of YC in the composition is 0.000001-60% (w/w).

7. The composition according to claim 6, wherein the percent content of FZ and YC in the composition is 0.01-60% (w/w) and 0.00001-40% (w/w), respectively.

8. The composition according to claim 1, wherein the weight ratio between FZ and YC (FZ:YC) is from 0.0001:1 to 1000000:1.

9. The composition according to claim 8, wherein the weight ratio between FZ and YC (FZ:YC) is from 0.001:1 to 100000:1.

10. The composition according to claim 9, wherein the weight ratio between FZ and YC (FZ:YC) is from 0.01:1 to 10000:1.

11. The composition according to claim 10, wherein the application concentration ratio between FZ and YC (FZ:YC) is from 1000 ppm:0.001 ppm to 1 ppm:500 ppm.

12. The composition according to claim 11 is from 500 ppm:0.01 ppm to 5 ppm:200 ppm.

13. The composition according to claim 1, where the composition is formulated as any one of the following formulation types: emulsifiable concentrate (EC), aqueous emulsion (EW), microemulison (ME), suspension concentrate (SC), oil dispersion (OD), capsule suspension (CS), wettable powder (WP), water dispersible granule (WDG). Soluble powder (SP), soluble granule (SG), suspension emulsion (SE), flowable concentrate for seed treatment (FS), aqueous solution (AS) and ready to use formulation.

14. The composition of claim 1, wherein the one or more rhubarb extracts include anthraquinones such as rhein, aloe-emodin, emodin, physcion, chrysophanol, physcion-glucoside, chrysophanol-glucoside, emodin-glucoside and rhein-glucoside; the one or more giant knotweed extracts, include anthraquinones such as emodin, physcion, chrysophenol, emodin-glucoside, physcion-glucosde, chrysophanol-glucoside and resveratrol; the one or more cassia seed extracts include anthraquinones such as emodin, chrysophanolphyscion, obtusin, obtusifolin and their corresponding glucosides; the one or more canola pollen extracts include brassinosteroids, such as brassinolide. 24-epibrassinolide, 28-epihomobrassinolidem, 28-homobrassinolide, dolicholide. typhasterol, 28-norcastasterone and 22, 23, 24-trisepibrassinolide; and the one or more beewax extracts, include brassinosteroids, such as brassinolide, 24-epibrassinolide, 28-epihomobrassinolidem, 28-homobrassinolide, dolicholide. typhasterol, 28-norcastasterone and 14-hydroxylated brassinosteroid; and the one or more synthesized brassinosteroids include one or more of brassinolide. 24-epibrassinolide, 28-epihomobrassinolidem, 28-homobrassinolide and typhasterol.

15. The composition of claim 1, wherein said combination has a synergistic antimicrobial effect.

16. A method for protecting a plant from diseases caused by fungi, bacteria or virus, comprising applying to the plant the composition of claim 1, thereby inducing plant resistance, promoting plant growth and/or enhancing plant yields.

17. The method of claim 16, wherein the plant is selected from the group consisting of: cucumber, pumpkin, squash, tomato, onion, shallots, beet, pepper, wheat, cereals, rice, sunflower, green bean, tobacco, flax, strawberry, grape, citrus, pears, apple, peony, chrysanthemum, rose and the plant diseases caused by fungi, bacteria, or virus is optionally: powdery mildew, downy mildew, late blight, pythium, damping-off disease, canker, black shank disease, Canker, early blight, rice blast, sheath blight, black scab, rust, glume spot disease, net blotch, leaf blight, botrytis disease, blast blight, bacterial wilt disease, bacterial stem rot disease and/or citrus huanglongbing.

18. The method of claim 17, wherein the plant is protected from cucumber and grape downy mildew and powdery mildew; cucumber and strawberry botrytis disease; citrus huanglongbing, rice blast; rice sheath blight and rice bacterial blight; peanut, pepper and tomato bacterial wilt; sunflower bacterial stem rot; and pear scab.

Description

DETAILED EXPERIMENTS

(1) By the following specific embodiments of the present invention is further illustrated, but the present invention is not limited only to the following examples. The content of the following examples is expressed by percent of weight.

(2) Some substances in the following examples were purchased from different companies: polyurea from Dongsheng Futian Jufu Company, organosiloxane from Mianyang Huili Huanyang Co. Ltd., sodium ligninsulfonate and calcium lignosulfonate from Henan Anyang Chemical Industry, polyoxyethylene styryl phenyl ether from Nanjing Taihua chemical industry, polypropylene alcohol from Beijing Baishun Chemical Tech Co. Ltd., sec-octyl phenol polyoxyethylene ether, fatty alcohol polyoxyethylene ether and polyoxyethylene fatty acid ester from Tianjin Hongmei Chemical Industry, kaolin from Anhui Tongling Guotai Non-metallic Materials Co., paraffin oil from Beijing Huaye Hongyu Chemical Co., Ltd, fatty alcohol polyglycol ether sulfate from Shanghai Youwen Chemical Co., urea (available from Annhui JinAo Chemical Co., Ltd., Alkyl phenol polyoxyethylene ether from Beijing Huayou Chemical Co., methylene bis-naphthalene sulfonate sodium, alkyl naphthalenesulfonic acid formaldehyde condensate and alkyl naphthalene sulfonic acid salt from Beijing Chemical Reagent Factory.

(3) Preparation methods of suspension spores used for spraying inoculation in the following examples is as follows: spray distilled water onto the diseased leaves, collect the spore solution and adjust it into a spore concentration of 10.sup.5/mL.

(4) Preparation of rhubarb extract: crush Rhubarb roots after drying, ferment, extract with ethanol-water mixture, concentrate, hydrolyze, neutralize, extract with ethyl acetate, concentrate to obtain rhubarb extracts.

(5) Preparation of giant knotweed extract: crush giant knotweed roots after drying, ferment, extract with ethanol-water mixture, concentrate, hydrolyze, neutralize, extract with ethyl acetate, concentrate to obtain giant knotweed extracts.

(6) Preparation of cassia seed extract: crush cassia seeds after drying, ferment, extract with ethanol-water mixture, concentrate, hydrolyze, neutralize, extract with ethyl acetate, concentrate to obtain cassia seed extracts.

(7) Preparation of canola pollen powder extract: crush the canola pollens, extract with ethanol-water, concentrate the extraction solution and then extract with ethyl acetate, concentrate the ethyl acetate layer to obtain canola pollen extract.

(8) Preparation of beeswax extract: emulsify the beewax with water and emulsifier to form a dispersion solution, add 2000-3000U/L degrading enzyme solution (e.g., enzyme solution from Pseudomonas fluorescens fermentation), adjust the pH and temperature, fully hydrolyze, extract the hydrolyzed solution with ethanol and n-hexane, concentrate to obtain the beewax extract.

(9) Preparation of synthetic brassinosteroids: usually from phytosteroid alcohols such as stigmasterol. Synthetic brassinosteroids can be obtained after a series of sequential reactions: sulfonylation, cyclization, oxidation, ring-opening, the catalytic oxidation and esterification.

(10) After purification, over 80% technical grade of active ingredients such as brassinolide, 80% typhasterol and 80% 14-hydroxylated brassinosteroid can be obtained from Canola pollen extract, beeswax extract, or synthetic brassinosteroids.

Example 1: Capsule Suspension: 0.5% of Active Ingredients

(11) Formula composition: rhubarb extract (4.995 g), 80% brassinolide technical grade (0.005 g), 150 solvent oil (solvent, 21 g), polyurea (capsule material, 3 g), polyvinyl alcohol (protecting agent, 1.0 g), organosiloxane (defoamers, 0.05 g), xanthan gum (thickener, 0.15 g), sodium lignin sulfonate (dispersing agent, 3 g), add water made up to 1 kg.

(12) Preparation method: The mixture of FZ, YC, No. 150 solvent oil and polyurea was added into another mixture consisting of polyvinyl alcohol, organosiloxane and water to emulsify, stirred until the polymerization reaction was completed; added xanthan gum and sodium lignin sulfonate to form a stable capsule suspension. The median capsule diameter was 5-8 microns. This product was coded as A1.

Example 2: Aqueous Emulsion: 5% of Active Ingredients

(13) Formula composition: giant knotweed extract (49.875 g), 80% typhasterol technical grade (0.125 g), No 150 solvent oil (50 g), polyoxyethylene styrene phenyl ether (emulsifier, 50 g), polypropylene alcohol (thickener, 80 g), diatomaceous earth (carrier, 30 g) add water made up to 1 kg.

(14) Preparation method: combined the FZ, YC, NO 150 solvent oil and polyoxyethylene styrene phenyl ether to make a homogenous oil phase; added diatomaceous earth into water to form a homogeneous aqueous phase; under high speed stirring, added the aqueous phase into oil phase to form 5% aqueous suspension with good dispersing property. This product was coded as A2.

Example 3: Microemulsion: 15% of Active Ingredients

(15) Formula composition: giant knotweed extract (75 g), canola pollen extract (75 g), N-methylpyrrolidone (co-solvent, 50 g), sec-octyl phenol ethoxylates (emulsifier, 100 g), ethoxylated castor oil (emulsifier, 200 g), ethylene glycol (antifreeze, 50 g), water made up to 1 kg.

(16) Preparation method: Made homogeneous oil phase by adding FZ, YC, N-methyl pyrrolidone, sec-octylphenol polyoxyethylene ether and ethoxylated castor oil: added glycol into water to form a uniform aqueous phase; under high speed stirring, the aqueous phase was added to the oil phase to form a microemulsion containing 15% of active ingredients. This product was coded as A3.

Example 4: Aqueous Suspension: 4.5% of Active Ingredients

(17) Formula composition: Cassia seed extract (44.8 g), beeswax extract (2 g), ethoxylated castor oil (emulsifier, 10 g), fatty alcohol polyethoxyethylene ether (emulsifier, 30 g), diatomaceous earth (thickener, 30 g), propylene glycol (antifreeze, 60 g), water made up to 1 kg.

(18) Preparation method: Water was used as a medium, the FZ, YC, ethoxylated castor oil, fatty alcohol polyoxyethylene ether, diatomaceous earth and glycol were added into sand mill to grind fine particle, forming aqueous suspensions containing 4.5% active ingredients. This product was coded as A4.

Example 5: Oil Dispersion, 50% of Active Ingredients

(19) Formula composition: giant knotweed extract (499.8 g), 80% 14-hydroxylated brassinosteroid technical grade (0.2 g), polyoxyethylene fatty alcohol ether (emulsifier, 100 g), polyoxyethylene fatty acid ester (emulsifier, 50 g), kaolin (thickener, 30 g), paraffin oil made up to 1 kg.

(20) Preparation method: Paraffin oil was used as the medium. The FZ, YC, polyoxyethylene fatty alcohol ether, polyoxyethylene fatty acid ester and kaolin were added into sand mill to grind fine particles, forming an oil suspension containing 50% active ingredient. This product was coded as A5.

Example 6: Soluble Granules, 20% of Active Ingredients

(21) Formula composition: Rhubarb extract (199.5 g), beeswax extract (0.5 g) calcium lignosulfonate (dispersants, 50 g), fatty alcohol polyglycol ether sulfate (wetting agent, 50 g), urea (carrier) to make up to 1 kg.

(22) Preparation method: The FZ, YC, calcium lignosulfonate, fatty alcohol polyglycol ether sulfate and urea were mixed and pulverized, added water, granulated it with a granulator with diameter of 1.0 mm sieve, dried, and sieved through 30 mesh sieve to prepare a water-dispersible granules containing 20% of active ingredients. This product was coded as A6.

Example 7: Emulsifier Concentrate, 70% of Active Ingredients

(23) Formula composition: Cassia extract (600 g), canola pollen extract (100 g), alkylphenol polyoxyethylene ether (emulsifier, 50 g), fatty alcohol polyoxyethylene ether (emulsifier, 50 g), No 150 solvent oil made up to 1 kg, mixed all and formed a homogeneous oil phase, i.e., emulsifier concentrate 70% of the active ingredient. This product was coded as A7.

Example 8: Wettable Powder, 80% of Active Ingredients

(24) Formula composition: Rhubarb extract (784 g), beeswax extract (16 g), sodium lignosulphonate (dispersant, 30 g), methylene-bis-naphthalene sulfonate (dispersing agent NNO, 20 g), soluble starch (carrier, 50 g), kaolin (carrier) made up to 1 kg.

(25) Preparation method: The components described above were thoroughly mixed, ground with a superfine grinding mill to obtain a wettable powder containing 80% of active ingredients. This product was coded as A8.

Example 9: Water Dispersible Granules, 90% of Active Ingredients

(26) Formula composition: giant knotweed extract (896 g), beeswax extract (4 g), calcium lignosulfonate (dispersants, 50 g), fatty alcohol polyglycol ether sulfate (wetting agent, 50 g), urea (vector) made up to 1 kg.

(27) Preparation method: The FZ, YC, calcium lignosulfonate, fatty alcohol polyglycol ether sulfate and urea were mixed and pulverized, added water, granulated with a granulator equipped with a diameter of 1.0 mm sieve, dried and then sieved through a 30 mesh sieve, forming water-dispersible granules containing 90% of active ingredients. Tis product was coded as A9

Example 10: Soluble Powder, 95% of Active Ingredients

(28) Formula composition: Giant knotweed extract (949.05 g), 80% brassinolide technical grade (0.95 g), alkylnaphthalene sulfonic acid formaldehyde condensate (30 g), alkylnaphthalene sulfonate (wetting agent, 20 g),

(29) Preparation method: The components described above were thoroughly mixed, ground with a superfine grinding mill to obtain soluble powder containing 95% of the active ingredient. This product was codes as A10.

Example 11: Aqueous Solution, 30% of Active Ingredients

(30) Formula composition: Giant knotweed extract (25 g), beeswax extract (275 g), ethanol (co-solvent, 50 g), ethylene glycol (antifreeze, 50 g), water made up to 1 kg.

(31) Preparation method: Mixed the giant knotweed extract, beewax extract and ethanol to form a homogeneous oil phase; added ethylene glycol into water to form a homogeneous aqueous phase; under high speed stirring, the oil phase was added to the aqueous phase to form a good aqueous solution containing 30% of active ingredients. This product was coded as A11.

(32) Control Formulation 1: Oil Dispersion, 3% Rhubarb Extract

(33) Formula composition: Rhubarb extract (30 g), polyoxyethylene fatty alcohol ethers (emulsifier, 30 g), polyoxyethylene fatty acid ester (emulsifier, 10 g), kaolin (thickener, 10 g), paraffin oil made up to 1 kg.

(34) Preparation method: Paraffin oil was used as the medium. The rhubarb extract, polyoxyethylene fatty alcohol ether, polyoxyethylene fatty acid ester and kaolin were added in sand grinding mill to obtain an oil dispersion containing 3% active ingredient. This product was coded as C1.

(35) Control Formulation 2: Microemulsion, 5% Giant Knotweed Extract

(36) Formula composition: giant knotweed extract (50 g), N-methylpyrrolidone (co-solvent, 20 g), sec-octylphenol polyoxylethylene ether (emulsifier, 40 g), ethoxylated castor oil (emulsifier, 80 g), ethylene glycol (antifreeze, 30 g), water made up to 1 kg.

(37) Preparation method: Made homogeneous oil phase by adding giant knotweed extract, N-methyl pyrrolidone, sec-octylphenol polyoxyethylene ether and ethoxylated castor oil: added glycol into water to form a uniform aqueous phase; under high speed stirring, the aqueous phase was added to the oil phase to form a microemulsion containing 5% of active ingredient. This product was coded as C2.

(38) Control Formulation 3: Soluble Powder, 10% Cassia Seed Extract

(39) Formula composition: Cassia extract (100 g), alkyl naphthalene sulfonic acid formaldehyde condensate (dispersant, 10 g), alkylnaphthalene sulfonate (wetting agent, 10 g), soluble starch made up to 1 kg

(40) Preparation method: The components described above were thoroughly mixed, ground with a superfine grinding mill to obtain soluble powder containing 10% of the active ingredients. This product was coded as C3.

(41) Control Formulation 4: Aqueous Solution, 0.1% Brassinolide

(42) Formula composition: 80% of brassinolide technical grade (1.25 g), ethanol (co-solvent, 10 g), ethylene glycol (antifreeze, 30 g), water made up to 1 kg.

(43) Preparation method: a homogeneous oil phase was made by mixing the brassinolide technical grade and alcohol. An aqueous phase was made by adding glycol into water; under high speed stirring, the oil phase was added to the aqueous phase to form a good aqueous solution containing 0.1% of the active ingredient. This product was coded as C4.

(44) Control Formulation 5: Aqueous Solution, 0.01% Typhasterol

(45) Formula composition: 80% of typhasterol technical grade (0.125 g), ethanol (co-solvent, 5 g), ethylene glycol (antifreeze, 30 g), water made up to 1 kg.

(46) Preparation method: a homogeneous oil phase was made by mixing the typhasterol technical grade and alcohol. An aqueous phase was made by adding glycol into water; under high speed stirring, the oil phase was added to the aqueous phase to form a good aqueous solution containing 0.01% of the active ingredient. This product was coded as C5.

(47) Control Formulation 6: Aqueous Solution, 0.1% Canola Pollen Extract

(48) Formula composition: Canola pollen extract (1 g), NO 150 solvent oil (solvent, 10 g), polyoxyethylene styryl phenyl ether (emulsifier, 40 g), polypropylene alcohol (thickener, 70 g), diatomaceous earth (carrier, 20 g) and water made up to 1 kg

(49) Preparation method: a homogeneous oil phase was made by mixing the canola pollen extract, No 150 solvent oil and polyoxyethylene styrene phenyl ether. An aqueous phase was made by adding polypropylene alcohol, diatomaceous earth and water; under high speed stirring, the oil phase was added to the aqueous phase to form a good aqueous solution containing 0.1% of the active ingredient. This product was coded as C6.

(50) Control Formulation 7: Soluble Powder, 1% Beewax Extract

(51) Formula composition: Beeswax extract (10 g), alkyl naphthalene sulfonic acid formaldehyde condensate (dispersant, 6 g), alkylnaphthalene sulfonate (wetting agent, 10 g), soluble starch made up to 1 kg.

(52) Preparation method: The components described above were thoroughly mixed, ground with a superfine grinding mill to obtain soluble powder containing 1% of the active ingredient. This product was coded as C7.

(53) Control Formulation 8: Aqueous Solution, 0.1% 14-Hydroxylated Brassinosteroids

(54) Formula composition: 80% of 14-hydroxylated brassinosteroids. (1.25 g), ethanol (co-solvent, 11 g), ethylene glycol (antifreeze, 25 g), water made up to 1 kg.

(55) Preparation method: a homogeneous oil phase was made by mixing 14-hydroxylated brassinosteroid technical grade and ethanol. An aqueous phase was made by adding ethylene glycol and water. Under high speed stirring, the oil phase was added to the aqueous phase to form a good aqueous solution containing 0.01% of the active ingredient. This product was coded as C8.

(56) Experimental examples 1-11 with compositions described above illustrated their applications against fungi-, bacteria- and/or virus-caused plant diseases.

(57) Colby equation was used to calculate the expected control effect of two active ingredients of FZ and YC,
Expected control effect=FZ+YC−(FZ*YC/100)

(58) FZ=observed control effect of FZ at the same concentration of FZ in the composition

(59) YC=observed control effect of YC at the same concentration of YC in the composition

(60) Partial experimental methods were as follows:

(61) Standard Test Method for Rice: pot assay was used. 12 pots were used for each group. Each group was divided into three treatments and a water blank control. Seven days prior to rice break period, whole rice plants were sprayed with the same volume of three treatment chemicals as that of water blank control until dripping. Twenty four hours after spraying, spore inoculation was conducted by spray inoculation. Six days after inoculation, spraying was repeated with water as a control, each treatment was repeated three times. Survey was conducted in rice maturity. The total number of rice plants and the number of diseased plants were recorded, and control was calculated.

(62) Cucumber experimental methods: pot assay was used. 12 pots were used for each group. Each group was divided into three treatments and a water blank control. At 2-leaf stage, whole cucumber plants were treated by foliar spraying, with the same volume of three treatment chemicals as that of water blank control until dripping. Twenty four hour later, all plants were inoculated with spores by foliar spraying. After the onset of investigated disease in the blank control treatment, disease progression in all other treatments was investigated and control effects were calculated.

(63) Orange Huanglongbing (Citrus HLB) Experimental Methods:

(64) In greenhouse condition, developing citrus plants (e.g., orange) with HLB symptoms were transferred into black bugs with an irrigation condition. Four groups of leaves (10 leaves/group) were selected for each plant. There were sprayed with diluted water solution of the FZ, YC and their combination, and water blank, respectively. Five replicates were set up. Four sprayings were performed with a 7-day interval. After 1 month from the last spraying, DNA was extracted from treated leaves, titers pf HLP was determined with PCR kits. Citrus plant leaves treated with water dilution of FZ, YC and their combinations were considered as treatment groups. Citrus plant leaves treated with water was considered as a control group.
control effect=100%*(control group-treatment group)/control group

Experimental Example 1: Antimicrobial and Synergistic Experiment of the Composition of Rhubarb Extract and Brassinolide Against Cucumber Downy Mildew

(65) TABLE-US-00001 TABLE 1 antimicrobial effect of individual components against cucumber downy mildew Product Percent of Control effect code Component Al Dilution (%) C1 Rhubarb 3.0% 600 60.5 extract C4 brassinolide 0.1% 25000 5.1

(66) TABLE-US-00002 TABLE 2 antimicrobial effect of the composition against cucumber downy mildew Expected Experimental Product Percent of control control code Component Al Dilution effect (%) effect (%) A1 Rhubarb 0.4995% 100 62.5 77.1 extract 80% 0.0005% brassinolide

Experimental Example 2: Antimicrobial and Synergistic Experiment of the Composition of Giant Knotweed Extract and Typhasterol Against Rice Blast

(67) TABLE-US-00003 TABLE 3 antimicrobial effect of individual components against rice blast Product Percent of Control effect code Component Al Dilution (%) C2 Giant knotweed 5.0% 1600 65.2 extract C5 typhasterol 0.01% 1600 20.3

(68) TABLE-US-00004 TABLE 4 antimicrobial effect of the composition against rice blast Expected Experimental Product Percent of control control code Component Al Dilution effect (%) effect (%) A2 Giant knotweed 0.49875% 1000 72.3 90.5 extract 80% 0.0125% typhasterol

Experimental Example 3: Antimicrobial and Synergistic Experiment of the Composition of Giant Knotweed Extract and Canola Pollen Extract Against Eggplant Phytophthora Fruitrot

(69) TABLE-US-00005 TABLE 5 antimicrobial effect of individual components against eggplant Phytophthora fruitrot Product Percent of Control effect code Component Al Dilution (%) C2 Giant knotweed 5.0% 1700 52.8 extract C6 Canola pollen 0.1% 32 7.9 extract

(70) TABLE-US-00006 TABLE 6 antimicrobial effect of the composition against eggplant Phytophthora fruitrot Expected Experimental Product Percent of control control code Component Al Dilution effect (%) effect (%) A3 Giant knotweed 7.5% 2500 56.5 71.3 extract Canola pollen 7.5% extract

Experimental Example 4: Antimicrobial and Synergistic Experiment of the Composition of Cassia Seed Extract and Beewax Extract Against Strawberry Root Rot

(71) TABLE-US-00007 TABLE 7 antimicrobial effect of individual components against strawberry root rot Product Percent of Control effect code Component Al Dilution (%) C3 Cassia seed 10% 3300 61.3 extract C7 beewax extract  1% 70000 25.4

(72) TABLE-US-00008 TABLE 8 antimicrobial effect of the composition against strawberry root rot Expected Experimental Product Percent of control control code Component Al Dilution effect (%) effect (%) A4 Cassia seed 4.48% 1500 71.1 87.9 extract beewax extract 0.02%

Experimental Example 5: Antimicrobial and Synergistic Experiment of the Composition of Giant Knotweed Extract and 14-Hydrolated Brassinosteroids Against Grape Downy Mildew

(73) TABLE-US-00009 TABLE 9 antimicrobial effect of individual components against grape downy mildew Product Percent of Control effect code Component Al Dilution (%) C2 Giant knotweed extract 5.0% 1600 66.8 C8 14-hydroxylated 0.1% 96000 15.7 brassinosteroids

(74) TABLE-US-00010 TABLE 10 antimicrobial effect of the composition against grape downy mildew Expected Experimental Product Percent of control control effect code Component Al Dilution effect (%) (%) A5 Giant knotweed 49.98% 16000 72.0 89.6 extract 80% 14- 0.02% hydroxylated brassino- steroids

Experimental Example 6: Antimicrobial and Synergistic Experiment of the Composition of Rhubarb Extract and Beewax Extract Against Tomato Leaf Curl Virus Disease

(75) TABLE-US-00011 TABLE 11 antimicrobial effect of individual components against tomato leaf curl virus disease Product Percent of Control effect code Component Al Dilution (%) C1 rhubarb 3.0% 300 36.3 extract C7 Beewax .sup. 1% 35000 28.0 extract

(76) TABLE-US-00012 TABLE 12 antimicrobial effect of the composition against tomato leaf curl virus disease Expected Experimental Product Percent of control control code Component Al Dilution effect (%) effect (%) A6 rhubarb 19.95% 2000 54.1 72.0 extract Beewax 0.05% extract

Experimental Example 7: Antimicrobial and Synergistic Experiment of the Composition of Cassia Seed Extract and Canola Pollen Extract Against Potato Early Blight

(77) TABLE-US-00013 TABLE 13 antimicrobial effect of individual components against potato early blight Product Percent of Control effect code Component Al Dilution (%) C3 Cassia seed  10% 2500 56.7 extract C6 Canola pollen 0.1% 150 10.2 extract

(78) TABLE-US-00014 TABLE 14 antimicrobial effect of the composition against potato early blight Expected Experimental Product Percent of control control code Component Al Dilution effect (%) effect (%) A7 Cassia seed 60% 15000 61.1 74.7 extract Canola pollen 10% extract

Experimental Example 8: Antimicrobial and Synergistic Experiment of the Composition of Rhubarb Extract and Beewax Extract Against Rose Powdery Mildew

(79) TABLE-US-00015 TABLE 15 antimicrobial effect of individual components against rose powdery mildew Product Percent of Control effect code Component Al Dilution (%) C1 rhubarb 3.0% 600 79.1 extract C7 beewax 1.0% 9700 3.1 extract

(80) TABLE-US-00016 TABLE 16 antimicrobial effect of the composition against rose powdery mildew Expected Experimental Product Percent of control control code Component Al Dilution effect (%) effect (%) A8 rhubarb 78.4% 15500 79.7 96.1 extract beewax 1.6% extract

Experimental Example 9: Antimicrobial and Synergistic Experiment of the Composition of Giant Knotweed Extract and Beewax Extract Against Soybean Rust

(81) TABLE-US-00017 TABLE 17 antimicrobial effect of individual components against soybean rust Product Percent of Control effect code Component Al Dilution (%) C2 Giant knotweed 5.0% 1550 67.4 extract C7 beewax extract 1.0% 70000 21.0

(82) TABLE-US-00018 TABLE 18 antimicrobial effect of the composition against soybean rust Expected Experimental Product Percent of control control code Component Al Dilution effect (%) effect (%) A9 Giant knotweed 89.6% 28000 74.2 91.6 extract beewax extract 0.4%

Experimental Example 10: Antimicrobial and Synergistic Experiment of the Composition of Giant Knotweed Extract and Brassinolide Against Pepper Bacterial Wilt

(83) TABLE-US-00019 TABLE 19 antimicrobial effect of individual components against pepper bacterial wilt Product Percent of Control effect code Component Al Dilution (%) C2 Giant knotweed 5.0% 26.3 43.9 extract C4 brassinolide 0.1% 658 11.7

(84) TABLE-US-00020 TABLE 20 antimicrobial effect of the composition against pepper bacterial wilt Expected Experimental Product Percent of control control code Component Al Dilution effect (%) effect (%) A10 Giant knotweed 94.905% 500 50.5 68.8 extract 80% 0.095% brassinolide

Experimental Example 11: Antimicrobial and Synergistic Experiment of the Composition of Giant Knotweed Extract and Beewax Extract Against Citrus Huanglongbing

(85) TABLE-US-00021 TABLE 21 antimicrobial effect of individual components against citrus Huanglongbing Product Percent of Control effect code Component Al Dilution (%) C2 Giant knotweed 5.0% 2000 6.7 extract C7 Beewax extract 1.0% 36.3 54.1

(86) TABLE-US-00022 TABLE 22 antimicrobial effect of the composition against citrus Huanglongbing Expected Experimental Product Percent of control control code Component Al Dilution effect (%) effect (%) A11 Giant knotweed 2.5% 1000 57.2 74.0 extract Beewax extract 27.5%

(87) Experimental examples (1-11) demonstrated that an obvious synergistic effect was observed for compositions of between FZ and YC within an appropriate ratio. When compared with individual components, compositions of FZ and YC significantly improved the control effect of plant diseases, therefore, reducing the amount of individual components, reducing farmers' expenses and also decreasing the impact on environment. In addition, the compositions of FZ and YC showed good efficacy against a variety of diseases on various crops, and their compositions displayed a broad antimicrobial spectrum.