CN113519569B - Preparation method of medicament for early-stage defoliation of kiwi fruits - Google Patents

Abstract

A Chinese medicinal preparation for early stage of folium et cacumen Actinidiae chinensis is prepared from corn stalk, sugarcane peel, Chinese corktree branch, Chinese honeylocust fruit branch, Japanese pagodatree branch, purified water, and composite microbial inoculum by extracting, concentrating, drying, carbonizing, fermenting, mixing, and drying. The invention is environment-friendly, has solved the problem of environmental pollution of chemical agent, the bacteriostatic property test shows that the medicament of the invention has extremely strong killing effects to the leaf mold fungus causing the brown spot of kiwi fruit, the invention inhibits bacteria for 4 days, can effectively kill the leaf mold fungus, can reasonably symbiotic between the microorganisms in the fermentation process, will not have the microorganism antagonism problem, and the extract after extracting mainly contains bacteriostatic components such as phellodendrine, saponin of Chinese honeylocust, rutin, etc., can enhance the bacteriostatic effect of the microorganism fermentation broth, play a role of synergy, the storage process is long, the product property will not change color after being placed for 12 months, and the bacteriostatic effect will not decline, worth the market popularizing and applying.

Description

Preparation method of medicament for early-stage defoliation of kiwi fruits

Technical Field

The invention mainly relates to the technical field of disease control of kiwifruits, and particularly relates to a preparation method of a medicament for early defoliation of kiwifruits.

Background

When the kiwi fruits are planted, a serious leaf drop phenomenon often occurs, the leaf drop in successive years seriously affects the tree vigor and the yield, and at present, the leaf drop phenomenon of the kiwi fruits is caused mainly in two aspects, namely physiological leaf drop and pathological brown spot. Aiming at the physiological defoliation, the corresponding problems can be solved mainly by means of reasonable pruning, scientific irrigation, reasonable loading and the like, and the pathological brown spot is difficult to treat.

The cercospora brown spot of the kiwi fruit belongs to a fungal disease, pathogenic bacteria of the cercospora brown spot are phyllosticta, the cercospora brown spot mainly damages leaves and branches and sometimes damages the surfaces of fruits, the scab mainly originates from the leaf margin and also has leaf surfaces, the scab is originally water-stain-shaped and green, and then expands along the leaf margin or inwards to form irregular brown scab, the disease condition is rapidly diffused under the rainy and high-humidity conditions, the scab is mildewed and rotted due to brown stain, under the normal climate, the periphery of the scab is dark brown, the center of the scab is brown to light brown, and a plurality of black speck particles, namely a pathogenic conidiophore, are scattered or densely grown on the scab. The damaged leaves curl towards the leaf surface at high temperature, are easy to crack, dry and shed in the later period, the disease spots in the middle of the leaf surface are obviously smaller than those in the leaf, and the disease spots penetrate through the leaf back and are brownish yellow.

At present, the main effective means for preventing and treating the brown spot of the kiwi fruit is chemical prevention and treatment, the specific method is carried out at the initial infection stage of pathogenic bacteria, the bactericide selects and uses pesticides for preventing and treating fungal diseases, and the chemical pesticide prevents and treats the soil, surface water, underground water and agricultural products to cause pollution and further enters a biologic chain, so that the disease has serious, long-term and potential harmfulness to all environmental organisms and human health, and therefore, the problem that the kiwi fruit brown spot can be prevented and treated by replacing the chemical pesticide is urgently needed to be solved in the kiwi fruit planting at present.

Disclosure of Invention

The invention aims to provide a preparation method of a medicament for preventing and treating early defoliation of kiwi fruits caused by phyllosticta.

The invention is realized by the following steps:

a preparation method for a medicament for early-stage alternaria leaf of Chinese gooseberry is characterized in that the medicament is prepared by taking corn straws, sugarcane peels, phellodendron amurense branches, Chinese honeylocust branches, pagodatree branches, purified water and a composite microbial inoculum as raw materials and respectively extracting, concentrating and drying, carbonizing, fermenting, mixing and drying; wherein the extraction is to extract phellodendron amurense branches, Chinese honeylocust fruit branches and pagodatree branches by adding a solvent; the compound microbial inoculum is composed of bacillus licheniformis, trichoderma viride, lactococcus lactis, streptomyces termitorum and the like.

The Bacillus licheniformis (Bacillus licheniformis) is GDMCC1.182, the Trichoderma viride (Trichoderma viride) is GDMCC3.602, the Lactococcus lactis (Lactococcus lactis) is CGMCC1.12794, and the Streptomyces termiticus (Streptomyces termitum) is GDMCC 802609; the strains are all commercial products.

Further, the extraction method comprises the steps of putting branches of phellodendron amurense, branches of Chinese honeylocust fruit and branches of sophora japonica into a universal pulverizer, pulverizing and sieving the branches with a sieve of 10-20 meshes to obtain mixed coarse powder, adding purified water which is 10-15 times of the total mass of the mixed coarse powder, setting the temperature to be 80-90 ℃, extracting for 2-3 hours, filtering with filter cloth after the extraction is finished, collecting filtrate and filter cakes, and storing the filtrate and the filter cakes respectively for later use; the mass ratio of the phellodendron amurense branches to the Chinese honeylocust tree branches is 1: 1-3: 2-5.

Further, the concentration and drying are carried out by putting the filtrate into a vacuum reduced pressure concentration tank, setting the vacuum degree at-0.05 to-0.08 MPa and the temperature at 60 to 70 ℃, carrying out reduced pressure concentration to obtain thick paste with the relative density of 1.25 to 1.30(60 ℃), putting the thick paste into a reduced pressure vacuum drying box, setting the vacuum degree at-0.05 to-0.08 MPa and the temperature at 60 to 70 ℃, carrying out vacuum drying for 24 to 36 hours to obtain dry paste, putting the dry paste into a universal pulverizer for pulverizing, and sieving by a 100-mesh sieve to obtain extract powder for later use.

Further, the carbonization is to mix and crush the corn straws and the sugarcane peels, to obtain mixed powder of the corn straws and the sugarcane peels after sieving the mixed powder by a sieve of 10-20 meshes, to add the filter cakes collected in the extraction step, to mix the mixed powder evenly, to place the mixed powder in a vacuum carbonization furnace, to set the vacuum degree to-30 to-50 pa, to set the carbonization temperature to 300-400 ℃, to preserve heat for 2-3 hours, to naturally cool the mixed powder, and to collect carbonized materials for later use; the mass ratio of the corn straws to the sugarcane peels is 2: 3-5, the mass ratio of mixed powder of the corn straws and the sugarcane peels to a filter cake is 10: 2-4, the corn straws and the sugarcane peels are crushed and mixed and then are mixed with the filter cake, the mixed powder of the corn straws and the sugarcane peels with specific particle sizes is coated on the surface of the filter cake to form complete coating, the anaerobic carbonization process is matched, anaerobic partial carbonization is realized through the carbonization, the mixed powder of the corn straws and the sugarcane peels coated on the outer layer is carbonized from outside to inside, the carbonization amount of the mixed powder is 20% -50% of the total mixed powder, and the filter cake inside is not carbonized; in addition, also carry out dry heat sterilization to mixed powder when carbonizing, provide aseptic matrix for follow-up fermentation, and mixed powder surface carbonization, form the hole, play certain supporting role to the fermentation heap, this hole structure can provide better respiration for the microorganism of follow-up fermentation simultaneously, and wrap up in the filter cake of inside and not carbonized mixed powder, the hole on accessible surface lasts for follow-up fermentation microorganism provides nutrient substance, guarantee smooth fermentation, can not be because of the participation of materials such as filter cake, lead to the fermentation environment to worsen.

In order to ensure more thorough fermentation, the specific partially carbonized fermentation substrate is matched with a specific two-stage fermentation means to improve the fermentation efficiency, specifically, the prepared carbonized mixed powder is placed in a fermentation tank, purified water is added, the mixture is uniformly stirred, lactococcus lactis and bacillus licheniformis are added, the fermentation temperature is set to be 28-32 ℃, the fermentation is carried out for 8-10 days, the first stage of fermentation is finished, then trichoderma viride and streptomyces termitomycetes are added, the fermentation temperature is kept to be 28-32 ℃, the fermentation is continued for 30-35 days, the fermentation is finished, and the fermentation liquid is filtered and collected for later use; the mass ratio of the carbonized mixed powder to the purified water is 1: 2-5, and the mass ratio of the viable bacteria of the carbonized mixed powder, lactococcus lactis, bacillus licheniformis, trichoderma viride and streptomyces termiticus is 100: 0.15-0.25: 0.25-0.35.

Lactococcus lactis is an anaerobic bacterium, a large amount of oxygen is needed in the growth process of the bacillus licheniformis, the lactobacillus lactis has a biological oxygen deprivation function, oxygen around the lactococcus lactis can be absorbed in the fermentation process, the fermentation of the lactococcus lactis is promoted, on the other hand, various nutrient substances including vitamins, amino acids, organic acids and growth promotion factors can be generated in the fermentation process of the bacillus licheniformis, the vitamins generated by the metabolism of the bacillus licheniformis can be consumed in the metabolism process of the lactococcus lactis, B vitamins such as folic acid and the like are synthesized at the same time, more comprehensive nutrient substances are provided for fermentation, the metabolites of the lactococcus lactis can rapidly reduce the surrounding pH in the fermentation process, the mixed powder obtained by the carbonization process of a specific part is weakly alkaline after purified water is added, the pH of the whole fermentation environment can be neutralized to be weakly acidic, and a better fermentation environment is provided for microbial fermentation, the optimum pH value of the trichoderma viride fermentation is 6.0-6.5, the optimum pH value of the streptomyces termitorum fermentation is 6.0-7.5, and due to the existence of the mixed powder obtained by the partial carbonization process, the pH value cannot be reduced due to the fermentation product of lactococcus lactis, so that the fermentation of the trichoderma viride and the streptomyces termitorum is inhibited; on the other hand, the part of carbonized mixed powder with holes formed on the surface can play a supporting role for a fermentation pile to ensure the respiration effect in the fermentation process of the strains, and meanwhile, the mixed powder with holes can also convey nutrient substances for the microbial fermentation process, and the special two-stage fermentation process is matched, so that the microbial fermentation efficiency of the invention is greatly improved; in the aspect of bacteriostasis, bacillus licheniformis has an oxygen deprivation function and can compete for oxygen around the trichoderma virens so as to play a role in bacteriostasis, and trichoderma viride is metabolized to generate toxic protein, so that amino acid absorption and protein synthesis obstacles of the trichoderma virens can be inhibited, and the formation of polysome is reduced, so that the growth of the trichoderma virens is weakened, glucanase generated by the trichoderma viride can also enable fungal cell walls to release an inducer of glucan source, the defense reaction of kiwi fruits is started, and the generation and accumulation of phenolic compounds and lignin related to disease resistance of the kiwi fruits are promoted. Meanwhile, the protease generated by the trichoderma viride can degrade pathogenic bacteria for digesting plant cell walls, directly inhibit the germination of the pathogenic bacteria, passivate the enzyme of the pathogenic bacteria and prevent the pathogenic bacteria from immersing into plant cells; the streptomyces termitomyces phyllosticta has strong antagonistic action, and the extracted extract powder is finally matched, so that the pesticide disclosed by the invention has a strong killing effect on phyllosticta, and meanwhile, the defense reaction of kiwi fruits can be started, and the effects of treating early-stage alternaria leaf drop of kiwi fruits caused by phyllosticta are achieved through matching.

If the mixed drying is not well controlled, the bottle spraying condition can occur in the freeze drying process, so that the dried product has irregular properties, on the other hand, if the mixed drying is not well controlled, the properties of the product in the storage process can be changed, and the bacteriostatic ability is reduced, in order to solve the problem, the prepared fermentation liquor is added with sorbitol and stirred for dissolution, then the prepared extract powder is added, the mixture is continuously stirred until the mixture is uniformly dispersed, the mixture is placed in a freeze drying box, the temperature is rapidly frozen to-38 to-35 ℃, the mixture is kept for 120 to 150 minutes, then the mixture is vacuumized and dried, the temperature is increased to-15 to-10 ℃ at the temperature increasing rate of 5 to 8 ℃/hour, and the constant temperature is kept for 90 to 120 minutes; heating to 0 ℃ at the heating rate of 5-8 ℃/hour, and keeping the temperature at 0 ℃ for 400-450 minutes; heating to 10-15 ℃ at the heating rate of 5-8 ℃/h, keeping the temperature for 360-420 minutes at constant temperature, heating to 25-30 ℃ at 8-10 ℃/h, keeping the temperature for 60-120 minutes, taking out the product after drying, wherein the water content of the product is lower than 6%, and thus obtaining the product; the mass ratio of the fermentation liquor to the sorbitol to the extract powder is 100: 150-180: 10-20. Sorbitol is a polyhydroxy compound, has hygroscopic property and is very easy to dissolve in water, can be used as an excipient of fermentation liquor and extract powder, is matched with a freeze-drying technical means, can prevent the phenomenon of bottle spraying in the freeze-drying process, ensures that the product can be smoothly freeze-dried and is convenient to store, has good chemical stability, can penetrate through or cover the periphery of the fermentation liquor and the extract powder by using larger amount of sorbitol, is matched with a specific freeze-drying mode, can ensure that the product is not damaged by factors such as illumination, oxidation and the like, ensures that the product cannot be discolored in the storage process, and cannot reduce the bacteriostatic ability.

The invention has the following beneficial effects:

the medicament prepared by the preparation method for the early-stage alternaria leaf of the kiwi fruit is environment-friendly, and solves the problem of environmental pollution caused by chemical agents, and the bacteriostatic performance test shows that the medicament has extremely strong killing effect on the phyllosticta causing the brown spot of the kiwi fruit, the invention can effectively kill the phyllosticta after bacteriostasis for 4 days, the microorganisms can reasonably symbiotic in the fermentation process, the microbial antagonism problem can not occur, and the extracted extract contains the bacteriostatic components such as phellodendrine, saponin of Chinese honeylocust fruit, rutin and the like, so that the bacteriostatic effect of the microbial fermentation liquor can be enhanced, the synergistic effect can be achieved, the storage process is long, the product is placed for 12 months, the product property cannot change color, the bacteriostatic effect cannot be reduced, and the medicament is worthy of market popularization and application.

Detailed Description

The present invention is described in detail below by way of examples, it being necessary to note that the following examples are provided only for illustrating the present invention and are not to be construed as limiting the scope of the present invention, and modifications or substitutions of the method, steps or conditions of the present invention may be made without departing from the spirit and spirit of the present invention.

Example 1

A preparation method of a medicament for early-stage alternaria leaf of kiwi fruit comprises the following steps:

1. extraction of

Placing the branches of phellodendron amurense, the branches of Chinese honeylocust fruit and the branches of sophora japonica in a universal pulverizer, pulverizing and sieving with a 20-mesh sieve to obtain mixed coarse powder, adding purified water 15 times of the total mass of the mixed coarse powder, setting the temperature at 90 ℃, extracting for 2 hours, filtering with filter cloth after extraction is finished, collecting filtrate and filter cakes, and storing the filtrate and the filter cakes respectively for later use; the mass ratio of the phellodendron amurense branches to the Chinese honey locust tree branches to the sophora japonica branches is 1:3: 5.

2. Concentrating and drying

Putting the filtrate prepared in the step 1 into a vacuum reduced pressure concentration tank, setting the vacuum degree to be-0.05-0.08 MPa and the temperature to be 70 ℃, carrying out reduced pressure concentration to obtain thick paste with the relative density of 1.29(60 ℃), putting the thick paste into a reduced pressure vacuum drying box, setting the vacuum degree to be-0.05-0.08 MPa and the temperature to be 70 ℃, carrying out vacuum drying for 24 hours to obtain dry paste, putting the dry paste into a universal pulverizer for pulverization, and sieving by a 100-mesh sieve to obtain extract powder for later use.

3. Carbonizing

Mixing and crushing corn straws and sugarcane peels, sieving the mixture by a 20-mesh sieve to obtain mixed powder of the corn straws and the sugarcane peels, adding the filter cakes collected in the extraction step, uniformly mixing, placing the mixture in a vacuum carbonization furnace, setting the vacuum degree to be-30 to-50 pa, setting the carbonization temperature to be 400 ℃, preserving the heat for 2 hours, naturally cooling, and collecting carbonized materials for later use; the mass ratio of the corn straws to the sugarcane peels is 2:5, and the mass ratio of the mixed powder of the corn straws and the sugarcane peels to the filter cake is 10: 4.

4. Fermentation of

Firstly, placing the prepared carbonized mixed powder in a fermentation tank, adding purified water, uniformly stirring, adding lactococcus lactis and bacillus licheniformis, setting the fermentation temperature to be 32 ℃, fermenting for 8 days, ending the first stage of fermentation, then adding trichoderma viride and streptomyces termitomyces, keeping the fermentation temperature to be 32 ℃, continuing to ferment for 30 days, ending the fermentation, filtering, and collecting fermentation liquor for later use; the mass ratio of the carbonized mixed powder to the purified water is 1:5, and the mass ratio of the viable bacteria of the carbonized mixed powder, the lactococcus lactis, the bacillus licheniformis, the trichoderma viride and the streptomyces termitorum is 100: 0.25:0.25:0.35:0.35.

5. Mixing and drying

Adding sorbitol into the prepared fermentation liquor, stirring for dissolving, adding the prepared extract powder, continuously stirring until the mixture is uniformly dispersed, subpackaging the mixture into 100ml glass bottles, then placing the glass bottles into a freeze drying box, quickly freezing the mixture to-35 ℃, keeping the temperature for 120 minutes, then vacuumizing and drying, heating to-10 ℃ at the heating rate of 8 ℃/hour, and keeping the temperature for 120 minutes at constant temperature; then heating to 0 ℃ at the heating rate of 8 ℃/hour, and keeping the temperature at 0 ℃ for 400 minutes; heating to 15 ℃ at the heating rate of 8 ℃/hour, keeping the constant temperature for 420 minutes, heating to 30 ℃ at the heating rate of 10 ℃/hour, keeping the constant temperature for 120 minutes, taking out the product after drying, and detecting that the water content of the product is 4.3 percent to obtain the product; the mass ratio of the fermentation liquor to the sorbitol to the extract powder is 100:180: 20.

Example 2

A preparation method of a medicament for early-stage leaf cast of kiwi fruits comprises the following steps:

1. extraction of

Placing the branches of phellodendron amurense, the branches of Chinese honeylocust fruit and the branches of sophora japonica in a universal pulverizer, pulverizing and sieving with a 16-mesh sieve to obtain mixed coarse powder, adding purified water with the total mass being 12 times of that of the mixed coarse powder, setting the temperature at 85 ℃, extracting for 2.5 hours, filtering with filter cloth after the extraction is finished, collecting filtrate and filter cakes, and storing the filtrate and the filter cakes respectively for later use; the mass ratio of the phellodendron amurense branches to the Chinese honey locust branches is 1:2: 4.

2. Concentrating and drying

Putting the filtrate prepared in the step 1 into a vacuum reduced pressure concentration tank, setting the vacuum degree to be-0.05-0.08 MPa and the temperature to be 65 ℃, carrying out reduced pressure concentration to obtain thick paste with the relative density of 1.28(60 ℃), putting the thick paste into a reduced pressure vacuum drying box, setting the vacuum degree to be-0.05-0.08 MPa and the temperature to be 65 ℃, carrying out vacuum drying for 32 hours to obtain dry paste, putting the dry paste into a universal pulverizer for pulverization, and sieving by a 100-mesh sieve to obtain extract powder for later use.

3. Carbonizing

Mixing and crushing corn straws and sugarcane peels, sieving the mixture by a 16-mesh sieve to obtain mixed powder of the corn straws and the sugarcane peels, adding the filter cakes collected in the extraction step, uniformly mixing, placing the mixture in a vacuum carbonization furnace, setting the vacuum degree to be-30 to-50 pa, setting the carbonization temperature to be 350 ℃, preserving the heat for 2.5 hours, naturally cooling, and collecting carbonized materials for later use; the mass ratio of the corn straws to the sugarcane peels is 2:4, and the mass ratio of the mixed powder of the corn straws and the sugarcane peels to the filter cake is 10: 3.

4. Fermentation of

Firstly, placing the prepared carbonized mixed powder in a fermentation tank, adding purified water, uniformly stirring, adding lactococcus lactis and bacillus licheniformis, setting the fermentation temperature to be 30 ℃, fermenting for 9 days, finishing the first stage of fermentation, then adding trichoderma viride and streptomyces termitomyces, keeping the fermentation temperature to be 30 ℃, continuing to ferment for 32 days, finishing the fermentation, filtering, and collecting fermentation liquor for later use; the mass ratio of the carbonized mixed powder to the purified water is 1:4, and the mass ratio of the viable count of the carbonized mixed powder, the lactococcus lactis, the bacillus licheniformis, the trichoderma viride and the streptomyces termitorum is 100: 0.20:0.20:0.30:0.30.

5. Mixing and drying

Adding sorbitol into the prepared fermentation liquor, stirring for dissolving, adding the prepared extract powder, continuously stirring until the mixture is uniformly dispersed, subpackaging the mixture into 100ml glass bottles, then placing the glass bottles into a freeze drying box, quickly freezing the mixture to-38 ℃, keeping the temperature for 150 minutes, then vacuumizing and drying, heating to-15 ℃ at the heating rate of 5 ℃/hour, and keeping the constant temperature for 120 minutes; then heating to 0 ℃ at the heating rate of 5 ℃/hour, and keeping the temperature at 0 ℃ for 400 minutes; heating to 10 ℃ at the heating rate of 5 ℃/hour, keeping the constant temperature for 360 minutes, heating to 25 ℃ at the heating rate of 8 ℃/hour, keeping the constant temperature for 120 minutes, taking out the product after drying, and detecting that the water content of the product is 4.7 percent to obtain the product; the mass ratio of the fermentation liquor to the sorbitol to the extract powder is 100:180: 20.

Example 3

A preparation method of a medicament for early-stage alternaria leaf of kiwi fruit comprises the following steps:

1. extraction of

Placing the branches of phellodendron amurense, the branches of Chinese honeylocust fruit and the branches of sophora japonica in a universal pulverizer, pulverizing and sieving with a 10-mesh sieve to obtain mixed coarse powder, adding purified water with the total mass 10 times of that of the mixed coarse powder, setting the temperature at 80 ℃, extracting for 3 hours, filtering with filter cloth after extraction is finished, collecting filtrate and filter cakes, and storing the filtrate and the filter cakes respectively for later use; the mass ratio of the phellodendron amurense branches to the Chinese honey locust branches is 1:1: 2.

2. Concentrating and drying

Putting the filtrate prepared in the step 1 into a vacuum reduced pressure concentration tank, setting the vacuum degree to be-0.05-0.08 MPa and the temperature to be 60 ℃, carrying out reduced pressure concentration to obtain thick paste with the relative density of 1.30(60 ℃), putting the thick paste into a reduced pressure vacuum drying box, setting the vacuum degree to be-0.05-0.08 MPa and the temperature to be 70 ℃, carrying out vacuum drying for 36 hours to obtain dry paste, putting the dry paste into a universal pulverizer for pulverization, and sieving by a 100-mesh sieve to obtain extract powder for later use.

3. Carbonizing

Mixing and crushing corn straws and sugarcane peels, sieving the mixture by a sieve of 10 meshes to obtain mixed powder of the corn straws and the sugarcane peels, adding the filter cakes collected in the extraction step, uniformly mixing, placing the mixture into a vacuum carbonization furnace, setting the vacuum degree to be minus 30 to minus 50pa, setting the carbonization temperature to be 300 ℃, preserving the heat for 3 hours, naturally cooling, and collecting carbonized materials for later use; the mass ratio of the corn straws to the sugarcane peels is 2:3, and the mass ratio of the mixed powder of the corn straws and the sugarcane peels to the filter cake is 10: 2.

4. Fermentation of

Firstly, placing the prepared carbonized mixed powder in a fermentation tank, adding purified water, uniformly stirring, adding lactococcus lactis and bacillus licheniformis, setting the fermentation temperature to be 28 ℃, fermenting for 10 days, finishing the first stage of fermentation, then adding trichoderma viride and streptomyces termitorum, keeping the fermentation temperature at 28 ℃, continuing to ferment for 35 days, finishing the fermentation, filtering, and collecting fermentation liquor for later use; the mass ratio of the carbonized mixed powder to the purified water is 1:2, and the mass ratio of the viable bacteria of the carbonized mixed powder, lactococcus lactis, bacillus licheniformis, trichoderma viride and streptomyces termitorum is 100: 0.15:0.15:0.25:0.25.

5. Mixing and drying

Adding sorbitol into the prepared fermentation liquor, stirring for dissolving, adding the prepared extract powder, continuously stirring until the mixture is uniformly dispersed, subpackaging the mixture into 100ml glass bottles, then placing the glass bottles into a freeze drying box, quickly freezing the mixture to-35 ℃, keeping the temperature for 150 minutes, then vacuumizing and drying, heating to-10 ℃ at the heating rate of 8 ℃/hour, and keeping the constant temperature for 120 minutes; then heating to 0 ℃ at the heating rate of 8 ℃/hour, and keeping the temperature at 0 ℃ for 450 minutes; heating to 10 ℃ at the heating rate of 8 ℃/hour, keeping the constant temperature for 420 minutes, heating to 30 ℃ at the heating rate of 8 ℃/hour, keeping the constant temperature for 120 minutes, taking out the product after drying is finished, and detecting that the water content of the product is 5.2 percent to obtain the product; the mass ratio of the fermentation liquor to the sorbitol to the extract powder is 100:150: 10.

Example 4:

the invention relates to a comparison test of bacteriostatic efficacy of phyllosticta:

comparative sample 1: the product obtained after drying the fermentation broth obtained in the fermentation process of example 1, i.e. the sample obtained without adding the extracted extract powder with respect to example 1.

Comparative sample 2: the extract powder obtained in the extraction method of example 1 was obtained, i.e. a sample obtained without addition of fermentation broth, relative to example 1.

Comparative sample 3: the concrete method is that the extract powder prepared by the extraction method of the embodiment 1 only cancels the carbonization step, and specifically comprises the steps of mixing the powdered activated carbon purchased from Jinhui filter material Co., Ltd of Gongyi city with the filter cake, wherein the mass ratio of the powdered activated carbon to the filter cake is 10:4, and then carrying out the subsequent fermentation step and the mixing and drying step according to the method of the embodiment 1 to obtain the sample.

Culture medium: 50g of glucose, 30g of peptone, 40g of agar and 2000ml of distilled water were mixed uniformly to prepare a medium.

The method comprises the following steps: the products prepared in example 1, example 2, example 3, comparative sample 1, comparative sample 2 and comparative sample 3 were diluted with purified water at a mass ratio of 1:100, added to the above culture medium, respectively, stirred uniformly, added with 1ml of a phyllosticta solution (about 300 fungi), stirred sufficiently, left to stand at room temperature for 5 days, observed the growth day by day, and subjected to phyllosticta counting, the results of which are shown in the following table:

time (sky)	1 day	2 days	3 days	4 days	5 days
						Example 1	287	169	72	——	——
Example 2	306	172	69	——	——
						Example 3	298	157	41	——	——
Comparative sample 1	293	227	183	176	127
						Comparative sample 2	283	261	256	239	203
Comparative sample 3	301	207	162	127	62

Remarking: in the table, "-" indicates no detection.

From the above results, the bacteriostatic ability of the products prepared in examples 1, 2 and 3 of the present invention against phyllosticta fungi is significantly stronger than that of the control sample 1 and 2, and the phyllosticta fungi cannot be detected after the products are left to be cultured at room temperature for 4 days in examples 1, 2 and 3, which indicates that the agent prepared by the present invention can kill phyllosticta fungi for 4 days, while the phyllosticta fungi can still be detected after the products are cultured at room temperature for 5 days in comparative samples 1 and 2, which indicates that the bacteriostatic ability of the fermentation liquid and the extract of the present invention is synergistic, and the killing performance is provided, while the performance of inhibiting phyllosticta fungi of comparative sample 3 is stronger than that of comparative sample 1 and 2, but the bacteriostatic ability of inhibiting phyllosticta fungi is weaker than that of examples 1, 2 and 3 of the present invention, which indicates that the carbonation process of the present invention has a positive effect on the bacteriostatic effect of the final product, therefore, the invention is effective to early defoliation caused by kiwifruit infected by phyllosticta.

Example 5:

the invention is used for the inspection test of the medicament storage stability of early defoliation of kiwi fruit:

the placing conditions are as follows: at a temperature of 25 ℃ and a relative humidity of 60%.

Test samples:

sample 1: the sample prepared in example 1.

Sample 2: example 2 sample prepared.

Sample 3: example 3 sample prepared.

Sample 4 (control): according to the preparation method of the embodiment 1, sorbitol is not added in the mixing process, and the product is obtained by freeze drying after the sorbitol is directly and uniformly mixed.

The test method comprises the following steps: after 12 months of storage, samples were taken to observe the properties and bacteriostatic properties of the phyllosticta (bacteriostatic test was performed as in example 4), and the results were compared with those of the freshly prepared product, as shown in the following table.

The test results show that the product prepared without adding sorbitol is easy to discolor in the placing process, and the bacteriostatic performance is obviously reduced, but the product can be placed for 12 months and still keeps the original bacteriostatic ability, so that the product still has strong bacteriostatic ability within at least 12 months.

Claims (1)

1. A preparation method of a medicament for early-stage alternaria leaf of kiwi fruits is characterized by taking corn straws, sugarcane peels, phellodendron amurense branches, Chinese honeylocust branches, pagodatree branches, purified water and a composite microbial inoculum as raw materials and preparing the medicament according to the following steps:

(1) extraction: placing the phellodendron amurense branches, the Chinese honeylocust fruit branches and the sophora japonica branches in a universal pulverizer, pulverizing and sieving by a sieve with 10-20 meshes to obtain mixed coarse powder, adding purified water with the total mass being 10-15 times of that of the mixed coarse powder, setting the temperature to be 80-90 ℃, extracting for 2-3 hours, filtering by using filter cloth after extraction is finished, collecting filtrate and filter cakes, and storing the filtrate and the filter cakes respectively for later use; the mass ratio of the phellodendron amurense branches to the Chinese honeylocust tree branches is 1: 1-3: 2-5;

(2) concentrating and drying: putting the filtrate obtained in the step (1) into a vacuum reduced pressure concentration tank, setting the vacuum degree to be-0.05 to-0.08 MPa and the temperature to be 60 to 70 ℃, carrying out reduced pressure concentration to obtain thick paste with the relative density of 1.25 to 1.30, putting the thick paste into a reduced pressure vacuum drying box, setting the vacuum degree to be-0.05 to-0.08 MPa and the temperature to be 60 to 70 ℃, carrying out vacuum drying for 24 to 36 hours to obtain dry paste, putting the dry paste into a universal pulverizer for pulverization, and sieving by a 100-mesh sieve to obtain extract powder for later use;

(3) carbonizing: mixing and crushing corn straws and sugarcane peels, sieving the mixture by a sieve with 10-20 meshes to obtain mixed powder of the corn straws and the sugarcane peels, adding the filter cakes collected in the step (1), uniformly mixing the mixture, placing the mixture in a vacuum carbonization furnace, setting the vacuum degree to be minus 30 to minus 50pa, setting the carbonization temperature to be 300-400 ℃, preserving the heat for 2-3 hours, naturally cooling the mixture, and collecting carbonized materials for later use; the mass ratio of the corn straws to the sugarcane peels is 2: 3-5, and the mass ratio of the mixed powder of the corn straws and the sugarcane peels to the filter cake is 10: 2-4;

(4) fermentation: the fermentation is realized by two-stage fermentation, firstly, the prepared carbonized mixed powder is placed in a fermentation tank, purified water is added, the mixture is uniformly stirred, lactococcus lactis and bacillus licheniformis are added, the fermentation temperature is set to be 28-32 ℃, the fermentation is carried out for 8-10 days, the first stage fermentation is finished, then trichoderma viride and streptomyces termitomyces are added, the fermentation temperature is kept to be 28-32 ℃, the fermentation is continued for 30-35 days, the fermentation is finished, the filtration is carried out, and fermentation liquor is collected for later use; the mass ratio of the carbonized mixed powder to the purified water is 1: 2-5, and the mass ratio of the viable bacteria of the carbonized mixed powder, lactococcus lactis, bacillus licheniformis, trichoderma viride and streptomyces termiticus is 100: 0.15-0.25: 0.25-0.35;

(5) mixing and drying: adding sorbitol into the fermentation liquor prepared in the step (4), stirring and dissolving, adding the extract powder prepared in the step (2), continuously stirring until the mixture is uniformly dispersed, placing the mixture in a freeze drying box, quickly freezing the mixture to-38 to-35 ℃, keeping the temperature for 120-150 minutes, then vacuumizing and drying, heating the mixture to-15 to-10 ℃ at the heating rate of 5-8 ℃/hour, and keeping the temperature for 90-120 minutes at constant temperature; heating to 0 ℃ at the heating rate of 5-8 ℃/hour, and keeping the temperature at 0 ℃ for 400-450 minutes; heating to 10-15 ℃ at the heating rate of 5-8 ℃/h, keeping the temperature for 360-420 minutes at constant temperature, heating to 25-30 ℃ at 8-10 ℃/h, keeping the temperature for 60-120 minutes, taking out the product after drying, wherein the water content of the product is lower than 6%, and thus obtaining the product; the mass ratio of the fermentation liquor to the sorbitol to the extract powder is 100: 150-180: 10-20.