KR101895387B1 - Method for producing eco-friendly composition for controlling plant diseases using microemulsion and microcapsule of plant extract - Google Patents

Abstract

The present invention relates to a microemulsion of a plant extract and a process for producing an eco-friendly plant disease prevention composition using microcapsules. More particularly, the present invention relates to a microemulsion prepared by emulsifying a plant extract and a vegetable oil having antimicrobial activity with an emulsifier, and microencapsulating the microemulsion using the biodegradable natural polysaccharide as a coating material, To a plant extract to disperse and stabilize the plant extract.
The composition for controlling environmental plant diseases according to the present invention can significantly improve antibacterial and bactericidal activity by mixing microemulsions and microcapsules at a certain ratio, and can not only maintain antibacterial and sterilizing power for a long period of time, And is capable of effectively inhibiting the generation and propagation of plant pathogenic bacteria such as various harmful bacteria and fungi.

Description

Technical Field [0001] The present invention relates to a microemulsion and a microcapsule of plant extract,

The present invention relates to a microemulsion prepared by emulsifying a plant extract and a vegetable oil, and a method for producing an eco-friendly plant disease prevention composition using microcapsules prepared by encapsulating the microemulsion prepared above.

Plant disease means that some or all of the plants are deteriorated physiologically due to the invasion of pathogenic organisms, and changes in morphology do not allow normal growth. In order to control the plant diseases by means of biological, chemical and poetic control measures against plant diseases caused by bacteria, fungi and virus pathogens, numerous chemical pesticides have been developed and used for controlling plant diseases, Various problems such as the resistance to plant pathogens and the accumulation of chemical pesticides have been caused by frequent use or overdosage of the allopathic remedy for synthetic drugs having the same action system having a pseudo-skeleton. Therefore, the demand for natural medicines is increasing. There is a growing demand for a composition for controlling plant diseases, which is eco-friendly and effective, and whose effect lasts for a long period of time. Accordingly, studies have been actively carried out to control the time of the active ingredient of the pesticide component so as to maintain the effect of the pesticide for a long period of time by applying sustained release to various active ingredients of the pesticide in a solid or liquid form and spraying the pesticide at an appropriate concentration at one time have.

As a part of this research, studies have been made on methods for producing sustained-release pesticides. For example, there have been proposed a method of putting active ingredients of pesticides into microcapsules, a method of encapsulating pesticidal active ingredients into cyclopentadienes, A method of preparing particles by mixing an active ingredient of the pesticide preparation alone or with an extender or the like and coating the prepared particle nuclei with wax or various resins have been reported. However, the above-described conventional methods have been pointed out as economical problems such as complicated manufacturing processes, expensive materials, and environmental problems that adversely affect the environment.

An example of a technique for extending the duration of the drug efficacy is disclosed in Korean Patent No. 1659332, and a Korean Patent Registration No. 1633290 discloses a preparation for environment-friendly crop disease prevention and control Discloses microcapsules containing a microbicidal active ingredient in Korean Patent Laid-Open Publication No. 2014-0115304. However, microemulsions prepared by emulsifying plant extracts of the present invention and vegetable oils, There has been no mention of a method for producing a composition for controlling environment-friendly plant disease using microcapsules prepared by encapsulating an emulsion.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide a method for producing a composition for controlling environment-friendly plant diseases, The present inventors have completed the present invention by confirming that it is possible to prevent and control the plant disease effectively.

In order to achieve the above object, the present invention provides a method for producing a W / O microemulsion, comprising: (1) mixing a first plant extract of antimicrobial activity and a vegetable oil,

(2) multiplying the W / O microemulsion with a chitosan solution containing chitosan of 30 to 54 kDa to form a W / O / W microemulsion;

(3) crosslinking the W / O / W microemulsion with sodium alginate solution to encapsulate the W / O / W microemulsion to prepare microcapsules; And

(4) The W / O / W microemulsion prepared in the step (2) and the microcapsules prepared in the step (3) are mixed together and the mixture is mixed with any one selected from the second plant extract, vinegar, fermented vinegar, A chitosan solution containing 30 to 54 kDa of chitosan is dispersed in a mixture to stabilize the mixture.

In addition, the present invention provides a composition for controlling environment-friendly plant diseases produced by the above-described method.

The present invention also provides a method of using the composition for controlling environmental plant disease according to the above production method as a disease controlling agent for crops, mushrooms, hydroponic crops, special crops, horticulture and crops.

The present invention relates to a microemulsion prepared by emulsifying a plant extract and a vegetable oil, and a method for producing an eco-friendly plant disease prevention composition using microcapsules prepared by encapsulating the microemulsion prepared above. More particularly, microemulsions obtained by emulsifying plant extracts and vegetable oils having antimicrobial activity with emulsifiers are prepared and microencapsulated using the biodegradable natural polysaccharides as a coating material. The present invention relates to a method for preparing a composition for controlling environment-friendly plant disease by mixing and dispersing the microemulsion and the microcapsule in a predetermined ratio to a plant extract. The composition for controlling environment-friendly plant disease using the microemulsion and microcapsule produced by the production method of the present invention is not only excellent in antibacterial and sterilizing power, but also has a long-lasting antibacterial and bactericidal effect as a sustained- .

FIG. 1 is a flowchart illustrating a method for preparing a plant-derived microemulsion and microcapsule-containing composition for controlling environment-friendly plant diseases according to the present invention.
Fig. 2 shows the average molecular weight of chitosan by the decomposition time decomposed by the photolysis method according to the present invention.
Fig. 3 shows the results of antimicrobial activity of chitosan by molecular weight.

(1) mixing an antimicrobial first plant extract and a vegetable oil to form a W / O microemulsion by high-speed stirring and emulsification;

(2) multiplying the W / O microemulsion with a chitosan solution containing chitosan of 30 to 54 kDa to form a W / O / W microemulsion;

(3) crosslinking the W / O / W microemulsion with sodium alginate solution to encapsulate the W / O / W microemulsion to prepare microcapsules; And

(4) The W / O / W microemulsion prepared in the step (2) and the microcapsules prepared in the step (3) are mixed together and the mixture is mixed with any one selected from the second plant extract, vinegar, fermented vinegar, And chitosan solution containing chitosan in an amount of 30 to 54 kDa is dispersed in a mixture to stabilize the mixture.

In the step (1), the first antimicrobial plant extract is preferably extracted from at least one selected from cinnamon, mustard, kelp, persimmon, saururus, But not limited to, canola oil, apricot seed oil, rose geranium oil, Thyme oil, Clove bud oil, , Castor oil, and nem oil, but is not limited thereto.

In the step (1), 5 to 30 parts by weight of the first plant extract and 5 to 30 parts by weight of the vegetable oil are mixed, more preferably 5 to 30 parts by weight of the apricot seeds and the bank extract are mixed with 5 to 30 Parts by weight of canola oil, and further 1 to 10 parts by weight of fermented alcohol or polyol may be mixed and used. 5 to 50% by weight of Tween-20 was added to 100 parts by weight of the mixed solution of the first plant extract and the vegetable oil, and emulsified at 5000 to 7000 rpm using a homogenizer , And emulsifying the mixture using an SPG membrane emulsifying apparatus to produce a W / O microemulsion.

The chitosan solution containing 30-54 kDa chitosan in the step (2) is prepared by (1) dissolving the polymer chitosan in a 0.5-1.0% (v / v) acetic acid solution to obtain a chitosan solution containing 1-5% (w / v) Preparing a solution; And

(2) a step of adding a photocatalyst of titanium dioxide (TiO ₂ ) to the chitosan acetic acid solution prepared in the step (1) and controlling the molecular weight of chitosan by irradiation with ultraviolet rays, , But the present invention is not limited thereto, and it is possible to manufacture it by adjusting it according to photolytic conditions.

The chitosan solution containing 30 to 54 kDa of chitosan in step (2) or (4) is preferably a chitosan solution of 30 to 54 kDa in an amount of 30 to 54 kDa dissolved in an organic acid, It does not.

In Step (2), 5 to 15 parts by weight of Tween-80, which is an emulsifier, is added to 100 parts by weight of the W / O microemulsion, and 1 to 3 wt. % Of a chitosan solution of 30 to 54 kDa is added thereto, and then the mixture is heated to 500 to 3,000 rpm It is preferable to stir for 10 to 30 minutes, or emulsify using an SPG membrane emulsifying apparatus to produce a W / O / W microemulsion, but the present invention is not limited thereto.

In step (3), 5 to 15 parts by weight of a 0.5 to 1.5% by weight sodium alginate solution is added to 100 parts by weight of the W / O / W microemulsion to prepare a microcapsule having a thin coating of soft capsules But is not limited thereto.

The second plant extract of step (4) is preferably a plant extract extracted from at least one selected from pine needles, gingko leaves, persimmon leaves, green tea leaves, ponytail, shrubs, But is not limited thereto.

The extraction solvent of the first plant extract or the second plant extract in the step (1) or (4) is water, C ₁ -C ₄ lower alcohol, acetone, acetic acid, citric acid, lactic acid or a mixture thereof. It does not. The extraction method in the present invention can be carried out by a person skilled in the art as much as necessary.

5 to 50 parts by weight of a W / C mixture of 100 parts by weight of a mixture of any one selected from the second plant extract, the vinegar solution, the fermented vinegar, and the microbial fermentation broth and the chitosan solution containing 30 to 54 kDa chitosan in the step (4) It is preferable that the mixed solution of the O / W microemulsion and the microcapsule is dispersed and stabilized. In the step (4), the mixing ratio of the W / O / W microemulsion and the microcapsule is 1/10 to 10/1 , More preferably in a volume ratio of 30 to 70: 70 to 30, and even more preferably in a volume ratio of 30:70, 50:50, or 70: 30, It is not.

The present invention also relates to a composition for controlling environmental plant diseases, which is produced by the above production method.

The plant diseases include S. aureus , E. coli , B. subtilis , P. aeruginosa , And plant disease caused by plant pathogens including Penicillium citrinum . The composition for inhibiting plant diseases according to the present invention is characterized by inhibiting the invasion of pathogens by destroying the cell walls and protoplasts of plant pathogens or by forming a physical coating on the surface of the crops and mixing microemulsions and microcapsules at a certain ratio , The drug release period can be prolonged and the effect can last for a long time.

The present invention also relates to a method for using the composition for controlling environment-friendly plant disease produced by the above-described method as a disease controlling agent for crops, mushrooms, hydroponic crops, special crops, horticultural crops and the like.

The method for controlling a disease caused by a plant disease control composition according to the present invention comprises diluting a stock solution of the plant disease control composition with water at an initial stage of growth of the crop to 500 to 1000 times the original amount of the composition for controlling the disease, It is preferable to dilute with 200 ~ 400 times of water and spray on the side at intervals of 1 ~ 2 weeks.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Example  1. Photolysis of chitosan

A 5% (w / v) chitosan solution was prepared by dissolving 1 kg of chitosan in 20 liters of a 0.5% (v / v) acetic acid solution. To the chitosan solution of 5% TiO ₂ ) was added and photolyzed by irradiation of ultraviolet ray for 72 hours while stirring.

As a result, as shown in FIG. 2, the decomposition rate of chitosan increased with ultraviolet irradiation time. When irradiated with ultraviolet rays for 72 hours, the average molecular weight decreased to 10,000 or less. The low molecular weight chitosan prepared by the photolysis method has a purity and a yield of almost 100%, has a uniform molecular weight, and is capable of arbitrarily controlling the molecular weight according to the ultraviolet irradiation period.

Example  2. Preparation of Apricot Seed Extract

100 g of the crude fine particle powder obtained by pulverizing dried apricot seeds was placed in 500 ml of 70% (v / v) methanol, heated and extracted with stirring at 40 to 65 ° C for 12 hours using a reflux condenser and then filtered. And concentrated under reduced pressure using a rotary evaporator.

Example  3. Manufacture of bank and pine leaf extract

 1 kg of the bank and pine needle were added to 5 liters of a 20% by weight acetic acid solution, respectively, and the mixture was immersed for 3 months while occasionally stirring at room temperature to extract and then filtered to prepare a bank extract. Sodium carbonate was added to the thus-prepared bank and pine leaf extract to neutralize it, and the mixture was heated and concentrated. The mixture was placed in a cellophane semipermeable tube and dialyzed for 3 days to remove impurities.

Example  4. Micro Emulsion  Produce

Step 1 : 30 parts by weight of the bank extract prepared in Example 2 and 3 and 10 parts by weight of the apricot extract were mixed to prepare a mixed plant extract.

15 parts by weight of canola oil; 5 parts by weight of apricot seed oil; And 10 parts by weight of polyethylene glycol (average molecular weight: 600) were mixed to prepare a vegetable oil. The mixture was added to the mixed plant extract to prepare a mixed solution.

Step 2 : 5 parts by weight of dihydroxy stearic acid PEG-30, which is a lipophilic emulsifier, was added to the mixed solution and emulsified by high-speed stirring at 5,000 to 7,000 rpm for 5 minutes to prepare a W / O micro To form an emulsion.

5 parts by weight of the addition of lauryl acrylate copolymer (Acrylic acid-Lauryl acrylate copolymer) - The W / O microemulsions of 50 was added 10 parts by weight of pH 6 buffered solution in parts by weight, and the hydrophilic polymer emulsions and acrylic acid: Step 3 (Average molecular weight: 30 kDa) was dissolved in a 0.5% (w / v) acetic acid solution and 30% by weight of a 3% (w / v) chitosan solution was added while stirring at 1,500 rpm. / W microemulsion was prepared.

Example  5. Manufacture of microcapsules

10 parts by weight of a buffer solution having a pH of 6 was added to 30 parts by weight of the W / O / W microemulsion prepared in Example 4, 15 parts by weight of 1% sodium alginate solution was added while stirring at 1,500 rpm, To prepare a microcapsule solution. The microcapsule solution was filtered, sufficiently washed with distilled water, and naturally dried to obtain microcapsule powder.

Example  6: For plant disease release agent  Preparation of composition

30 parts by weight of a microemulsion-microcapsule mixed solution obtained by mixing the microemulsion and microcapsules in a volume ratio of 50:50 was mixed with a pine needle extract and a 3% chitosan solution (average molecular weight: 30 kDa) at a volume ratio of 50:50 Was dispersed in 70 parts by weight of a mixed solution to prepare a plant disease control composition.

Experimental Example  1: Antimicrobial activity test 1

LB broth was diluted to 0, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.05, and 0.1% by diluting each sample solution of chitosan and chitosan with controlled molecular weights adjusted to pH 6, Staphylococcus aureus ; ATCC 25923), Escherichia coli ; ATCC 25922) (10 ⁴ to 10 ⁵ CFU / ml) were inoculated and cultured with shaking at 30 ° C for 24 hours.

The Staphylococcus aureus ; ATCC 25923) and Escherichia coli ( ATCC 25922) were diluted with physiological saline (0.85% (w / v) NaCl) and spread evenly on LB agar plate medium. After incubation at 30 ° C for 24 hours, Concentrations of chitosan solution and plant extracts with no change in bacterial counts were defined as the minimum inhibitory concentration (MIC,%).

As a result, the antimicrobial activity of the chitosan solution by the molecular weight was tested, and the antibacterial activity of the plant extract was shown in Table 1.

The chitosan solutions of 36 kDa and 54 kDa in the range of 30 to 54 kDa were obtained from S. aureus and Escherichia coli (E. coli) in the range of 30 to 54 kDa by measuring the minimum growth inhibitory concentration (MIC) The MIC value for E. coli was 0.04% to 0.06%, indicating that the antibacterial activity was increased by decreasing the MIC value. When the molecular weight of chitosan was over 54 kDa or less than 30 kDa, the antibacterial activity was lowered Respectively.

The results of measuring the minimum growth inhibitory concentration (MIC) of the plant extract according to the present invention as shown in Table 1 were as follows: Bank / apricot / pine leaf extract mixture (composition ratio 1: 3: 12 and 3: The antimicrobial activity of S. aureus and E. coli was 0.02 ~ 0.035%, which was higher than that of each extract alone.

Antimicrobial activity of plant extracts Name of sample
Minimum Growth Inhibitory Concentration (MIC,%) Escherichia coli (E. coli) Staphylococcus aureus (S. ureus a) Bank extract 0.075 0.060 Apricot Seed Extract 0.065 0.050 Pine leaf extract 0.100 0.080 Bank / apricot seeds / pine leaf extract mixed at a volume ratio of 1: 3: 12 0.035 0.025 Bank / apricot seeds / pine leaf extract mixed at a volume ratio of 3: 1: 12 0.030 0.020

Experimental Example  2. Antimicrobial activity test 2

(W / O / W) prepared in Example 4, microcapsules prepared in Example 5, and the like were mixed in a PDA (Potato dextrose agar) medium with different mixing ratios of the plant extracts adjusted to pH 6 The composition for controlling plant diseases prepared in Example 6 was diluted to 250 times and then sprayed with a solution of B. subtilis , P. aeruginosa , and Penicillium citrinum ( ATCC 6205) pathogenic fungus strain Number of test bacteria: 10 ⁴ to 10 ⁵ CFU / ml) was inoculated and cultured with shaking at 30 ° C for 24 hours.

The pathogenic bacteria and fungi were diluted with physiological saline (0.85% NaCl), spread evenly on agar plate medium, cultured at 30 ° C for 48 hours, counted the number of colonies (CFU) The results are shown in Table 2.

As a result of measuring the bacteria decrease rate (%) of the plant pathogens for the plant disease controlling composition of the present invention are shown in Table 2, and Bacillus subtilis (B. subtilis), Pseudomonas aeruginosa (P. aeruginosa) and Penny room Solarium sheet rinum ( For Penicillium citrinum) The bacterial reduction rate (%) was 250 to 500 times for the sample solution and 70 to 100% for the 500 times diluted solution.

Antimicrobial activity of multiple emulsions and compositions according to the invention
sample Bacterial Reduction Rate (%) B. subtilis P. aeruginosa P. citrimium 250 times 500 times 250 times 500 times 250 times 500 times The microemulsion of Example 4 90 85 85 80 85 70 The microcapsules of Example 5 85 80 80 75 80 75 The composition of Example 6 100 100 100 95 95 90

(%) = {(Number of bacteria in the control untreated group - number of the treated control group) / number of control untreated control group} x 100

Experimental Example  3. Effect of crop disease control

The control effect of the composition for controlling plant diseases prepared in Example 6 on blue mold (Penicilum spp.) Disease was tested in the mushroom cultivation test package.

In the pamphlet, the sample solution was diluted 500 times and treated 3 times at 7 days intervals before the onset of the mushroom cultivation test on the packaging medium, and 3 times at intervals of 7 days from the onset of onset or onset of onset, 10 days after the final treatment of the composition, the degree of disease was examined to evaluate the control effect. The pavement management was based on the general seedling method of the cultivator and the experiment was repeated 3 times by the nude method. After the final treatment, 1 m ² of mushroom culture area was used as a reference, and 10 to 20 mushrooms were included in one mushroom. The incidence rate (%) was defined as the percent area of the lesion occurrence area per mushroom medium in terms of percentage.

Table 3 shows the incidence (%) and control (%) of mushroom mushroom in Experimental Example 3.

Control effect against blue fungus disease of mushroom mushroom Test group
Outbreak rate (%) Control (%)
1 iteration 2 iterations 3 iterations Average The microemulsion of Example 4 Early onset 7.3 8.4 7.7 7.8 72.5 10 days before invention 6.8 5.7 6.4 6.3 77.8 The microcapsules of Example 5
Early onset 7.4 7.0 6.6 7.0 75.4 10 days before invention 4.7 6.3 5.8 5.6 80.3 The composition of Example 6 Early onset 4.5 3.9 5.0 4.5 84.2 10 days before invention 2.5 2.2 2.8 2.5 91.2 No treatment 27.3 25.6 32.4 28.4 -

The incidence (%) = (area of mushroom cultivated in diseased area in 1 / total area of mushroom in 1 mushroom) × 100

(%) = {(Incidence at the time of no treatment - incidence at the time of treatment) / incidence at the time of no treatment} × 100

As shown in Table 3, the microemulsion prepared by emulsifying the plant extract of the present invention and the vegetable oil, and the microcapsule prepared by encapsulating the microemulsion prepared above, were applied to the plant pest control composition 10 In the case of treatment before or at the onset of the onset, the control against blue fungus in the mushroom test package was 72.5%, 77.8%, 75.4%, 80.33%, 84.2% and 91.2%, respectively.

The microemulsion prepared by emulsifying the plant extract and vegetable oil or the microemulsion prepared by encapsulating the microemulsion prepared when the composition for controlling plant disease according to the present invention is treated at the early stage or 10 days before the onset of the disease, The effect of controlling the disease was remarkably increased as compared with the case where the microcapsules prepared by encapsulating the microemulsion were treated alone.

In addition, the plant disease prevention composition of the present invention was increased by about 7% as compared with the case where the composition was treated 10 days before the occurrence of the disease. Therefore, the composition for controlling a plant disease produced by the method of the present invention is diluted 250 to 500 times with the composition for controlling a plant disease of the present invention before the occurrence of a disease in the cultivation, .

Claims (12)

(1) Antimicrobial 1st plant extracts and canola oil, apricot seed oil, rose geranium oil extracted from at least one selected from cinnamon, mustard, gilt, persimmon, saururus, At least one vegetable oil selected from Rose geranium oil, Thyme oil, Clove bud oil, Castor oil and Neem oil is mixed and emulsified at a high speed to form a W / O microemulsion;
(2) multiplying the W / O microemulsion with a chitosan solution containing chitosan of 30 to 54 kDa to form a W / O / W microemulsion;
(3) crosslinking the W / O / W microemulsion with sodium alginate solution to encapsulate the W / O / W microemulsion to prepare microcapsules; And
(4) The W / O / W microemulsion prepared in the step (2) and the microcapsules prepared in the step (3) are mixed at a predetermined ratio, and the pine needles, ginkgo leaf, persimmon leaves, green tea leaves, A mixture of chitosan solution containing 30-54 kDa of chitosan and any one selected from the second plant extract, vinegar solution, fermentation vinegar, and microbial fermentation broth extracted from at least one selected from the at least one selected from the group consisting of tree saplings, And then stabilizing the plant-derived plant disease-preventing composition,
The plant diseases include S. aureus , E. coli , B. subtilis , P. aeruginosa , Or plant disease caused by plant pathogens including Penicillium citrinum or a blue fungal disease. delete delete The chitosan solution according to claim 1, wherein the chitosan solution contains 30 to 54 kDa of chitosan
(1) dissolving biodegradable chitosan in a 0.5 to 1.0% (v / v) acetic acid solution to prepare a 1 to 5% (w / v) chitosanic acid solution; And
(2) a step of adding a photocatalyst of titanium dioxide (TiO ₂ ) to the chitosan acetic acid solution prepared in the step (1) and controlling the molecular weight of chitosan by irradiation with ultraviolet light By weight of the composition. delete The method according to claim 1, wherein the extraction solvent of the first plant extract or the second plant extract in step (1) or step (4) is water, a C ₁ -C ₄ lower alcohol, acetone, acetic acid, citric acid, Wherein the composition is a mixture of a plant extract and a plant extract. The chitosan solution according to claim 1, wherein the chitosan solution containing 30-54 kDa chitosan in step (2) or step (4) is prepared by dissolving chitosan in 30-54 kDa in organic acid in 0.1-3.0 wt% Wherein the plant is a plant. 2. The method according to claim 1, wherein 100 parts by weight of a mixture of any one selected from the second plant extract, vinegar solution, fermentation vinegar, and microbial fermentation broth and the chitosan solution containing 30 to 54 kDa chitosan in step (4) To 50 parts by weight of a mixed solution of W / O / W microemulsion and microcapsules is dispersed and stabilized. The method according to claim 1, wherein the mixing ratio of the W / O / W microemulsion and the microcapsule in the step (4) is 1 to 10: . A composition for controlling environmental plant diseases according to any one of claims 1, 4, 6 to 9. delete A method of using the composition for controlling environmental plant disease according to claim 10 as a disease controlling agent for crops, mushrooms, hydroponic crops, special crops, horticultural crops or the like.

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