Preparation method of medicament for preventing and treating kiwi fruit diseases

Abstract

The invention relates to a composite medicament for preventing and treating kiwi fruit diseases, which is prepared by the following steps: preparing mixed powder from sugarcane, coptis chinensis, purslane, dandelion, isatis root and scutellaria baicalensis as raw materials, fermenting, collecting filtrate, adding mannitol, auxin IAA and Tween 20 into the filtrate, and finally freeze-drying. According to the present invention, the fermentation is performed by using Streptomyces albidoflavus, Streptomyces mononomycin, Bacillus spinizenii, Bacillus thuringiensis and Trichoderma viride as the fermentation bacterial agent, and the fermentation method comprises the following steps: using Streptomyces albidoflavus, Streptomyces mononomycin, Bacillus spinizenii, Bacillus thuringiensis and Trichoderma viride as the fermentation bacterial agent, by spraying the composite agent, the prevention efficiency is high, the infection rate of the cultivation method for the kiwi fruits suffering from the gray mold after the kiwi fruits are cultivated for 3 years is 0, the survival rate of the kiwi fruits is up to 100%, the fruit bearing and falling rate is lower than 2% after the kiwi fruits are cultivated for 3 years, the yield can be up to about 1500 kg / mu, and the gray mold infection rate is as low as 1% or below after the mature kiwi fruits are stored for 7 days at normal temperature.

Description

[0001] The present invention is a divisional application of patent application number 202311673852.2, and the invention name is “A cultivation method for preventing and controlling kiwifruit gray mold disease”. Technical Field

[0002] The invention relates to the technical field of kiwifruit disease prevention and treatment, and in particular to a method for preparing a medicament for preventing and treating kiwifruit diseases. Background Art

[0003] Kiwi fruit is rich in nutrients. In addition to containing organic substances such as actinine, proteolytic enzymes, and sugars, as well as trace elements such as calcium, potassium, zinc, and selenium, and 17 amino acids required by the human body, it is also rich in vitamin C, gluconic acid, citric acid, etc. It has a sweet and sour taste and is deeply loved by the public. However, kiwi fruit likes warmth and humidity, and is very susceptible to gray mold. Gray mold mainly occurs during the flowering period, young fruit period, and storage period. Gray mold is caused by infection by Botrytis cinerea, and flowers, fruits, leaves, and stems can all be infected. After the flowers are infected, they turn brown and rot and fall off. The infected petals fall on the leaves and fruits to further infect the young fruits and leaves. The infected leaves cause leaf spots. The hairs of the fruits turn brown in the early stage of infection, followed by local rot, causing fruit drop. When the humidity is high, a gray-black mold layer will form on the fruit surface. If the prevention and control are not in place during the growth process, the incidence of gray mold during the subsequent fruit storage period will also increase. In the prior art, chemical fungicides such as pyrimethanil, procymidone, and isoprodinil are often used to spray kiwifruit orchards to prevent and control gray mold. However, chemical agents cannot be used frequently, and the control effect is limited. Therefore, it is necessary to seek an efficient and environmentally friendly agent for preventing and controlling gray mold, as well as a kiwifruit cultivation method based on the agent, to achieve efficient prevention and control of gray mold, thereby improving the yield and quality of kiwifruit. Summary of the invention

[0004] The present invention aims to provide a composite agent for preventing and controlling kiwifruit diseases, which can effectively improve the prevention and control effect of kiwifruit gray mold, significantly improve the survival rate of kiwifruit cultivation, reduce the kiwifruit fruit drop rate, and reduce the infection rate of kiwifruit to gray mold during storage.

[0005] Another object of the present invention is to provide a method for preparing the composite agent for preventing and controlling gray mold of kiwifruit.

[0006] The purpose of the present invention is achieved through the following technical solutions: A composite agent for preventing and controlling diseases of kiwifruit, specifically for preventing and controlling gray mold, characterized in that: sugar cane, coptis root, purslane, dandelion, isatis root, and scutellaria are used as raw materials to prepare a mixed powder, the filtrate is collected after fermentation, mannitol, auxin IAA and Tween 20 are added to the filtrate, and finally freeze-dried to obtain the compound agent, wherein the fermentation is carried out by using Streptomyces albicans ( Streptomyces albidoflavus )、Streptomyces monomycinicola( Streptomyces monomycini)、Bacillus stutzeri( Bacillus spizizenii ), Bacillus thuringiensis ( Bacillus thuringiensis ) and Trichoderma viride ( Trichoderma green ) is a fermentation agent.

[0007] When fermentation agents were prepared using sugarcane, coptis root, purslane, dandelion, isatis root, and scutellaria as raw materials for the prevention and treatment of gray mold, it was found that during long-term use, the pathogenic bacteria would develop drug resistance, the prevention and treatment effect of the agent would gradually weaken, and the efficacy would decrease. In the continuous exploration, it was found that by adding the growth hormone IAA to the fermentation broth, IAA played an induced antagonistic role and also inhibited the resistance of pathogenic bacteria to the fermentation agent. At the same time, by freeze-drying the mixture of fermentation broth and IAA, it was found that the induced antagonistic effect of IAA on pathogenic bacteria was further enhanced. In addition, as a growth hormone, IAA not only prevents and treats gray mold, but also helps the growth and reproduction of kiwifruit to a certain extent and improves the quality of the fruit.

[0008] Furthermore, the mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomyciniensis, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride is 100:0.15:0.15:0.20:0.20:0.25, and the mass ratio of sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum officinale, Radix Isatidis and Scutellaria baicalensis in the mixed powder is 10:2~3:3~5:1~2:1~2:4~6.

[0009] Furthermore, the mass ratio of the fermentation broth, mannitol and Tween 20 is 100:50-60:1-2, and the mass proportion of the auxin IAA in the prepared composite agent is 0.3-0.5%.

[0010] A method for preparing a composite agent for preventing and controlling kiwifruit diseases, characterized in that it comprises the following steps: (1) Sugarcane, coptis root, purslane, dandelion, isatis root and scutellaria baicalensis are mixed and crushed to prepare a mixed powder; (2) After the mixed powder is boiled and cooled, a fermentation agent is added to perform fermentation treatment, and the residue and filtrate are collected after filtering, where the filtrate is the fermentation liquid; (3) Add mannitol, auxin IAA and Tween 20 to the fermentation broth and freeze-dry to obtain a composite agent.

[0011] Furthermore, in the step (1), the mass ratio of sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria baicalensis is 10:2-3:3-5:1-2:1-2:4-6.

[0012] Furthermore, the steaming in step (2) is steaming in water for 2 to 3 hours, which sterilizes the product while killing enzymes and preserving glycosides, thereby inhibiting the effective ingredients from being decomposed by enzymes.

[0013] Furthermore, the fermentation agent used in step (2) includes Streptomyces albicans ( Streptomyces whitish-yellow )、Streptomyces monomycinicola( Streptomyces monomycini )、Bacillus stutzeri( Bacillus spizizen ), Bacillus thuringiensis ( Bacillus thuringiensis ) and Trichoderma viride ( Trichoderma green ).

[0014] Furthermore, the fermentation in step (2) is specifically to add Streptomyces albus and Streptomyces monomycinicus in the cooled mixed powder in sequence, ferment at 28-30°C for 3 days, then add Bacillus stutzeri, Bacillus thuringiensis and Trichoderma viride, maintain the temperature at 30-32°C and continue fermenting for 10-15 days, filter after the fermentation is completed, collect the filter residue and the filtrate, and the filtrate is the fermentation liquid.

[0015] Furthermore, in the step (2), the mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomyciniensis, Bacillus stutzeri, Bacillus thuringiensis and Trichoderma viride is 100:0.15:0.15:0.20:0.20:0.25.

[0016] Furthermore, in step (3), the mass ratio of the fermentation broth, mannitol and Tween 20 is 100:50-60:1-2, and the mass proportion of the auxin IAA in the prepared composite agent is 0.3-0.5%.

[0017] Furthermore, in the freeze-drying step (3), the temperature is first lowered to -30~-35°C, maintained for 150~180 minutes, and then vacuum dried, heated to -10~-15°C at a heating rate of 5~8°C / h, and maintained at a constant temperature for 150~180 minutes; then heated to 0°C at a heating rate of 5~8°C / h, and maintained at 0°C for 200~250 minutes; then heated to 15°C at a heating rate of 5~8°C / h, and maintained at a constant temperature for 300~350 minutes; finally heated to 30~32°C at 8~10°C / h, and maintained at a constant temperature for 120~150 minutes, and drying is completed.

[0018] Most specifically, a method for preparing a composite agent for preventing and controlling gray mold, characterized in that it comprises the following steps: (1) Preparation of mixed powder Mix sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria, put them in a universal grinder, grind them through a 10-mesh sieve to obtain a mixed powder, wherein the mass ratio of sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria is 10:2-3:3-5:1-2:1-2:4-6; (2) Preparation of fermentation broth The mixed powder is placed in a steamer and steamed for 2-3 hours. The mixed powder is taken out, cooled, and a fermentation agent is added for fermentation. Specifically, Streptomyces albus and Streptomyces monomycinis are first added, and fermentation is carried out at 28-30° C. for 3 days. Then, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride are added, and the temperature is maintained at 30-32° C. and fermentation is continued for 10-15 days. After the fermentation is completed, the mixture is filtered to collect the filter residue and the filtrate, and the filtrate is the fermentation liquid. The mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomycinis, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride is 100: 0.15: 0.15: 0.20: 0.20: 0.25; (3) Preparation of compound medicines Mannitol, auxin IAA and Tween 20 are added to the fermentation broth prepared in step (2) in sequence, stirred and dissolved, and then freeze-dried, specifically at -30 ~ -35 ° C, maintained for 150 ~ 180 minutes, and then vacuum dried, heated to -10 ~ -15 ° C at a heating rate of 5 ~ 8 ° C / h, and kept at a constant temperature for 150 ~ 180 minutes; then heated to 0 ° C at a heating rate of 5 ~ 8 ° C / h, and kept at 0 ° C for 200 ~ 250 minutes; then heated to 15 ° C at a heating rate of 5 ~ 8 ° C / h, and kept at a constant temperature for 300 ~ 350 minutes; finally heated to 30 ~ 32 ° C at 8 ~ 10 ° C / h, and kept at a constant temperature for 120 ~ 150 minutes to prepare a composite agent, wherein the mass ratio of the fermentation broth, mannitol and Tween 20 is 100:50 ~ 60:1 ~ 2, and the mass proportion of auxin IAA in the prepared composite agent is 0.3 ~ 0.5%.

[0019] A cultivation method for preventing and controlling gray mold of kiwifruit, characterized in that: sugarcane, coptis root, purslane, dandelion, isatis root and scutellaria are fermented to prepare filter residue and fermentation liquid, the fermentation liquid and auxin IAA are prepared into a composite agent, the filter residue is buried at the root of the kiwifruit seedlings 2-3 months after planting, and the composite agent solution is sprayed on the seedlings, and then the composite agent solution is sprayed every 2-3 months until the kiwifruit is ripe and picked, the composite agent solution is obtained by diluting the composite agent with water 200-300 times, and spraying 150-200 mL per kiwifruit plant.

[0020] Furthermore, the fermentation specifically comprises the following steps: using sugarcane, coptis root, purslane, dandelion, isatis root and scutellaria to prepare a mixed powder, steaming and cooling the mixture, then sequentially adding Streptomyces albus and Streptomyces monomycinum, fermenting the mixture at 28-30°C for 3 days, then adding Bacillus stutzeri, Bacillus thuringiensis and Trichoderma viride, maintaining the temperature at 30-32°C and continuing the fermentation for 10-15 days, filtering the mixture after the fermentation is completed, collecting the filter residue and the filtrate, and the filtrate is the fermentation liquid.

[0021] Furthermore, the mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomyciniensis, Bacillus stutzeri, Bacillus thuringiensis and Trichoderma viride is 100: 0.15: 0.15: 0.20: 0.20: 0.25.

[0022] Furthermore, the composite agent is prepared by sequentially adding mannitol, auxin IAA and Tween 20 into the fermentation broth, stirring and dissolving the mixture, and then freeze-drying the mixture.

[0023] Furthermore, the mass ratio of the fermentation broth, mannitol and Tween 20 is 100:50-60:1-2, and the mass proportion of the auxin IAA in the prepared composite agent is 0.3-0.5%.

[0024] Furthermore, the freeze-drying process is to first lower the temperature to -30~-35°C, maintain it for 150~180min, then vacuum dry, heat it to -10~-15°C at a heating rate of 5~8°C / h, and maintain it at a constant temperature for 150~180min; then heat it to 0°C at a heating rate of 5~8°C / h, and maintain it at a constant temperature for 200~250min; then heat it to 15°C at a heating rate of 5~8°C / h, and maintain it at a constant temperature for 300~350min; finally, heat it to 30~32°C at 8~10°C / h, and maintain it at a constant temperature for 120~150min, and the drying is completed.

[0025] Most specifically, a cultivation method for preventing and controlling gray mold of kiwifruit is characterized by comprising the following steps: 1. Select seedlings and set up scaffolds Select kiwifruit seedlings with a height of 30-40 cm and a root diameter of 1.0-1.3 cm for planting. The male-female ratio of kiwifruit seedlings is 8-10:1. The planting time is March to April each year. The row spacing between two kiwifruit seedlings is 4-6m. After planting, water each seedling at a watering rate of 500-600ml / plant. Install vertical support columns 40-60 cm away from the kiwi seedlings. The height of the support columns is 2.5-3.0 m. Connect each vertical support column with a horizontal support column on the top of the support column and fix it with a small wire. Build the frame based on each ridged part. 2. Fertilization Within six months after planting, apply compound fertilizer once a month, with each application amount of 60-80g / plant. After six months of planting, apply compound fertilizer once every two months, with each application amount of 280-300g / plant. 3. Preparation of compound medicine (1) Preparation of mixed powder Mix sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria, put them in a universal grinder, grind them through a 10-mesh sieve to obtain a mixed powder, wherein the mass ratio of sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria is 10:2-3:3-5:1-2:1-2:4-6; (2) Preparation of fermentation broth The mixed powder is placed in a steamer and steamed for 2-3 hours. The mixed powder is taken out, cooled, and a fermentation agent is added for fermentation. Specifically, Streptomyces albus and Streptomyces monomycinis are first added, and fermentation is carried out at 28-30° C. for 3 days. Then, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride are added, and the temperature is maintained at 30-32° C. and fermentation is continued for 10-15 days. After the fermentation is completed, the mixture is filtered to collect the filter residue and the filtrate, and the filtrate is the fermentation liquid. The mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomycinis, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride is 100: 0.15: 0.15: 0.20: 0.20: 0.25; (3) Preparation of compound medicines Mannitol, auxin IAA and Tween 20 are sequentially added to the fermentation broth prepared in step (2), stirred and dissolved, and then freeze-dried, specifically at -30~-35°C, maintained for 150~180min, then vacuum dried, heated to -10~-15°C at a heating rate of 5~8°C / h, and maintained at a constant temperature for 150~180min; then heated to 0°C at a heating rate of 5~8°C / h, and maintained at 0°C for 200~250min; then heated to 15°C at a heating rate of 5~8°C / h, and maintained at a constant temperature for 300~350min; finally heated to 30~32°C at 8~10°C / h, and maintained at a constant temperature for 120~150min to obtain a composite agent, wherein the mass ratio of the fermentation broth, mannitol and Tween 20 is 100:50~60:1~2, and the mass proportion of auxin IAA in the prepared composite agent is 0.3~0.5%; 4. Prevention and control of gray mold 2 to 3 months after the seedlings are planted, the fermented filter residue is buried at the roots of the seedlings, and the seedlings are sprayed with a compound agent solution. Thereafter, the compound agent solution is sprayed every 2 to 3 months until the kiwifruit is ripe and ready to be picked. The compound agent solution is prepared by adding 200 to 300 times the mass of the compound agent to water, and is sprayed at a rate of 150 to 200 mL per kiwifruit plant.

[0026] The present invention has the following technical effects: By spraying the composite agent prepared by the present invention for preventing and controlling gray mold of kiwi fruit, the prevention efficiency is high, the infection rate of the cultivation method of kiwi fruit suffering from gray mold after cultivation for 3 years is 0, the survival rate of kiwi fruit is as high as 100%, the fruit drop rate is less than 2% after three years of cultivation, the yield is high, and the yield can be as high as about 1500kg / mu. After the mature kiwi fruit is stored at room temperature for 7 days, the gray mold infection rate is as low as less than 1%. The composite agent of the present invention has a very strong killing effect on gray mold, and can effectively kill gray mold after 3 days of bacteriostasis.

[0027] In the composite agent of the present invention, the fermentation products of Chinese medicinal materials such as coptis root, dandelion, purslane, isatis root, scutellaria baicalensis, etc. are combined with the auxin IAA to enhance the induced antagonistic effect on gray mold, play a synergistic effect, and at the same time be environmentally friendly, solving the problem of environmental pollution caused by chemical agents. DETAILED DESCRIPTION

[0028] The present invention is described in detail below through examples. It is necessary to point out that the following examples are only used to further illustrate the present invention and cannot be understood as limiting the scope of protection of the present invention. Those skilled in the art can make some non-essential improvements and adjustments to the present invention based on the above-mentioned contents of the present invention.

[0029] The fermentation microorganisms in the fermentation agent used in the present invention are as follows: Streptomyces alboflavinus was purchased from China General Microbiological Culture Collection (CGMCC) with the collection number CGMCC4.252.

[0030] Streptomyces monomycinensis, Bacillus stutzeri, Bacillus thuringiensis and Trichoderma viride were purchased from Guangdong Microbiological Culture Collection Center (GDMCC), among which the collection number of Streptomyces monomycinensis is GDMCC 4.201; the collection number of Bacillus stutzeri is GDMCC 1.136; the collection number of Bacillus thuringiensis is GDMCC 1.147; and the collection number of Trichoderma viride is GDMCC3.602.

[0031] Example 1 A method for preparing a composite agent for preventing and controlling gray mold comprises the following steps: (1) Preparation of mixed powder Mix sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria, put them in a universal grinder, grind them through a 10-mesh sieve to obtain a mixed powder, wherein the mass ratio of sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria is 10:2.5:4:1.5:1.5:5; (2) Preparation of fermentation broth The mixed powder is placed in a steamer and steamed for 2.5 hours. The mixed powder is taken out, cooled, and a fermentation agent is added for fermentation. Specifically, Streptomyces albus and Streptomyces monomycinis are first added, and fermentation is carried out at 28-30° C. for 3 days. Then, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride are added, and the temperature is maintained at 30-32° C. and continued to ferment for 12 days. After the fermentation is completed, the mixture is filtered to collect the filter residue and the filtrate, and the filtrate is the fermentation liquid. The mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomycinis, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride is 100: 0.15: 0.15: 0.20: 0.20: 0.25; (3) Preparation of compound medicines Mannitol, auxin IAA and Tween 20 are added to the fermentation broth prepared in step (2) in sequence, stirred and dissolved, and then freeze-dried, specifically at -30°C, maintained for 180 minutes, then vacuum dried, heated to -10°C at a heating rate of 6°C / h, and maintained at a constant temperature for 180 minutes; then heated to 0°C at a heating rate of 6°C / h, and maintained at 0°C for 240 minutes; then heated to 15°C at a heating rate of 6°C / h, and maintained at a constant temperature for 320 minutes; finally heated to 30°C at 10°C / h, and maintained at a constant temperature for 150 minutes to obtain a composite agent, wherein the mass ratio of the fermentation broth, mannitol and Tween 20 is 100:55:1.5, and the mass proportion of auxin IAA in the prepared composite agent is 0.4%.

[0032] Example 2 A cultivation method for preventing and controlling gray mold of kiwi fruit comprises the following steps: 1. Select seedlings and set up scaffolds Before planting kiwifruit, choose sandy soil, with a row spacing of 2.5 meters, turn the soil to form ridges, dig trenches between ridges, and the trench depth is 20-30 cm to facilitate drainage. Select kiwifruit seedlings with a height of 30-40 cm and a root stem diameter of 1.0-1.3 cm for planting. The male-female ratio of kiwifruit seedlings is 10:1. The planting time is March to April each year. The row spacing of two kiwifruit seedlings is 4-6 meters. After planting, water each seedling with a watering amount of 500-600 ml / plant; Install vertical support columns 40-60 cm away from the kiwi seedlings. The height of the support columns is 2.5-3.0 m. Connect each vertical support column with a horizontal support column on the top of the support column and fix it with a small wire. Build the frame based on each ridged part. 2. Fertilization Within six months after planting, apply compound fertilizer once a month, with each application amount of 60-80g / plant. After six months of planting, apply compound fertilizer once every two months, with each application amount of 280-300g / plant. 3. Prevention and control of gray mold Two months after the seedlings are planted, the fermented filter residue is buried at the roots of the seedlings, and the seedlings are sprayed with the solution prepared by the composite agent prepared in Example 1. Thereafter, the composite agent solution is sprayed once every two months until the kiwifruit is ripe and ready to be picked. The composite agent solution is prepared by adding 250 times the mass of the composite agent to water, and is sprayed at a rate of 150-200 mL per kiwifruit plant.

[0033] Comparative Example 1 According to the method of Example 2, kiwi fruit was cultivated. The agent used in the process of preventing and controlling gray mold was prepared by directly concentrating and evaporating the fermentation liquid prepared in Example 1, adding mannitol and Tween 20, and then freeze-drying it into powder. That is, the auxin IAA was not added to the agent, and the rest was consistent with Example 1.

[0034] Comparative Example 2 According to the method of Example 2, kiwifruit was cultivated, and the agent used in the process of preventing and controlling gray mold was auxin IAA, instead of fermentation liquid compound. And through multiple tests, it was found that when auxin IAA was used alone, when the IAA aqueous solution of the same concentration as in Example 2 was sprayed, the concentration of auxin IAA in the sprayed composite agent solution was only 16 μg / mL, and the control effect on gray mold was not obvious. When the concentration of the sprayed IAA aqueous solution was 60-80 μg / mL, the resistance of kiwifruit to gray mold was most significantly improved. Therefore, during the cultivation process, an aqueous solution with an auxin IAA concentration of 60 μg / mL was used to replace the composite agent solution used in Example 2 for gray mold control, and the remaining steps were consistent with Example 1.

[0035] Comparative Example 3 Kiwi fruit was cultivated according to the method of Example 2, and the agents used in the process of preventing and controlling gray mold were as follows: When preparing the compound medicine, the fermentation liquid is directly concentrated and evaporated, then freeze-dried into powder, and then directly mixed with IAA to form the compound medicine.

[0036] 50 mL of the pharmaceutical solution used in Example 1, Comparative Example 1, Comparative Example 2 and Comparative Example 3, and 50 mL of the prepared low-concentration IAA (the same concentration as the IAA in the composite pharmaceutical solution of Example 2) were taken as a group and added into 50 g of beef extract peptone liquid culture medium respectively, and stirred evenly. 1 mL of gray mold (Botrytis cinerea) (the number of viable bacteria was about 2000) was added into the culture medium, and cultured in a sterile environment at a temperature of 25°C for 5 days. The growth was observed daily, and the number of gray mold was counted. The results are shown in Table 1.

[0037] Table 1: Resistance of different pesticides to Botrytis cinerea

[0038] Note: “——” in the table means not detected.

[0039] It can be seen that the number of live Botrytis cinerea bacteria decreased sharply when the composite agent prepared by the present invention was used for the antibacterial test of Botrytis cinerea. On the 4th day of the test, no live Botrytis cinerea bacteria were detected in the culture medium. When a single Chinese medicine agent (the agent in Example 1) was used, it can be seen that the number of live Botrytis cinerea bacteria was also continuously decreasing, but the rate of decrease was slow. When a single auxin IAA was used, it could only have a significant inhibitory effect on Botrytis cinerea at a certain concentration. When the concentration was low, the inhibitory effect of IAA on Botrytis cinerea was extremely poor. When the Chinese medicine agent and IAA were directly mixed and used, the effect of inhibiting Botrytis cinerea was significantly improved compared with the single Chinese medicine agent and the single auxin IAA, but the effect was still slightly worse than that of the composite agent prepared by the present invention.

[0040] Example 3 A cultivation method for preventing and controlling gray mold of kiwi fruit comprises the following steps: 1. Select seedlings and set up scaffolds Select kiwifruit seedlings with a height of 30-40 cm and a root diameter of 1.0-1.3 cm for planting. The male-female ratio of kiwifruit seedlings is 8:1. The planting time is March to April each year. The row spacing of two kiwifruit seedlings is 4-6m. After planting, water each seedling at a watering rate of 500-600ml / plant. Install vertical support columns 40-60 cm away from the kiwi seedlings. The height of the support columns is 2.5-3.0 m. Connect each vertical support column with a horizontal support column on the top of the support column and fix it with a small wire. Build the frame based on each ridged part. 2. Fertilization Within six months after planting, apply compound fertilizer once a month, with each application amount of 60-80g / plant. After six months of planting, apply compound fertilizer once every two months, with each application amount of 280-300g / plant. 3. Preparation of compound medicine (1) Preparation of mixed powder Mix sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria, put them in a universal grinder, grind them through a 10-mesh sieve to obtain a mixed powder, wherein the mass ratio of sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria is 10:2:3:1:1:4; (2) Preparation of fermentation broth The mixed powder is placed in a steamer and steamed for 3 hours. The mixed powder is taken out, cooled, and a fermentation agent is added for fermentation. Specifically, Streptomyces albus and Streptomyces monomycinis are first added, and fermentation is carried out at 28-30° C. for 3 days. Then, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride are added, and the temperature is maintained at 30-32° C. and fermentation is continued for 15 days. After the fermentation is completed, the mixture is filtered to collect the filter residue and the filtrate, and the filtrate is the fermentation liquid. The mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomycinis, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride is 100: 0.15: 0.15: 0.20: 0.20: 0.25; (3) Preparation of compound medicines Mannitol, auxin IAA and Tween 20 are sequentially added to the fermentation broth prepared in step (2), stirred and dissolved, and then freeze-dried, specifically at -35°C, maintained for 150 minutes, then vacuum dried, heated to -15°C at a heating rate of 5°C / h, and maintained at a constant temperature for 150 minutes; then heated to 0°C at a heating rate of 5°C / h, and maintained at 0°C for 200 minutes; then heated to 15°C at a heating rate of 5°C / h, and maintained at a constant temperature for 300 minutes; finally heated to 32°C at 8°C / h, and maintained at a constant temperature for 120 minutes to obtain a composite agent, wherein the mass ratio of the fermentation broth, mannitol and Tween 20 is 100:50:1, and the mass proportion of auxin IAA in the prepared composite agent is 0.3%; 4. Prevention and control of gray mold Two months after the seedlings are planted, the fermented filter residue is buried at the roots of the seedlings, and the seedlings are sprayed with a solution of the compound agent. Thereafter, the compound agent solution is sprayed once every three months until the kiwifruit is ripe and ready to be picked. The solution of the compound agent is prepared by adding 200 times the mass of the compound agent to water, and is sprayed at a rate of 150 to 200 mL per kiwifruit plant.

[0041] Example 4 A cultivation method for preventing and controlling gray mold of kiwi fruit comprises the following steps: 1. Select seedlings and set up scaffolds Select kiwifruit seedlings with a height of 30-40 cm and a root diameter of 1.0-1.3 cm for planting. The male-female ratio of kiwifruit seedlings is 9:1. The planting time is March to April each year. The row spacing of two kiwifruit seedlings is 4-6m. After planting, water each seedling at a watering amount of 500-600ml / plant. Install vertical support columns 40-60 cm away from the kiwi seedlings. The height of the support columns is 2.5-3.0 m. Connect each vertical support column with a horizontal support column on the top of the support column and fix it with a small wire. Build the frame based on each ridged part. 2. Fertilization Within six months after planting, apply compound fertilizer once a month, with each application amount of 60-80g / plant. After six months of planting, apply compound fertilizer once every two months, with each application amount of 280-300g / plant. 3. Preparation of compound medicine (1) Preparation of mixed powder Mix sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria, put them in a universal grinder, grind them through a 10-mesh sieve to obtain a mixed powder, wherein the mass ratio of sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria is 10: 3: 5: 2: 2: 6; (2) Preparation of fermentation broth The mixed powder is placed in a steamer and steamed for 2 hours. The mixed powder is taken out, cooled, and a fermentation agent is added for fermentation. Specifically, Streptomyces albus and Streptomyces monomycinii are first added, and fermentation is carried out at 28-30° C. for 3 days. Then, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride are added, and the temperature is maintained at 30-32° C. and continued to ferment for 10 days. After the fermentation is completed, the mixture is filtered to collect the filter residue and the filtrate, and the filtrate is the fermentation liquid. The mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomycinii, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride is 100: 0.15: 0.15: 0.20: 0.20: 0.25; (3) Preparation of compound medicines Mannitol, auxin IAA and Tween 20 are sequentially added to the fermentation broth prepared in step (2), stirred and dissolved, and then freeze-dried, specifically at -30°C, maintained for 160 minutes, then vacuum dried, heated to -12°C at a heating rate of 8°C / h, and maintained at a constant temperature for 160 minutes; then heated to 0°C at a heating rate of 8°C / h, and maintained at 0°C for 250 minutes; then heated to 15°C at a heating rate of 8°C / h, and maintained at a constant temperature for 350 minutes; finally heated to 30°C at 10°C / h, and maintained at a constant temperature for 140 minutes to obtain a composite agent, wherein the mass ratio of the fermentation broth, mannitol and Tween 20 is 100:60:2, and the mass proportion of auxin IAA in the prepared composite agent is 0.5%; 4. Prevention and control of gray mold Two months after the seedlings are planted, the fermented filter residue is buried at the roots of the seedlings, and the seedlings are sprayed with a solution of the compound agent. Thereafter, the compound agent solution is sprayed every 2.5 months until the kiwifruit is ripe and ready to be picked. The solution of the compound agent is prepared by adding 300 times the mass of the compound agent to water, and is sprayed at a rate of 150 to 200 mL per kiwifruit plant.

[0042] 7 mu of land was selected in the same planting area as the experimental planting site, and the 7 mu of land was divided into 7 groups, each with 1 mu. The blank group was planted according to the method of Example 1, but without gray mold control treatment, and the kiwifruit of the same seedlings was planted according to the methods of Examples 2, 3, 4, Comparative Example 1, Comparative Example 2, and Comparative Example 3. The kiwifruit was planted for 3 years, and the survival rate after 6 months of planting, the average fruit drop rate of young fruits after the third year of planting, the yield (kg / mu) in the third year of planting, and the gray mold infection rate of kiwifruit fruits harvested in the third year of planting after storage for 1 week at room temperature (temperature of 25-30°C, humidity of 60-70%) were recorded respectively. The results are shown in Table 2.

[0043] Table 2:

[0044] During the kiwifruit planting process, no prevention and control measures were taken for the plants to prevent gray mold. The survival rate of kiwifruit seedlings was only 86% after 6 months of planting. In the third year of planting, the average fruit drop rate of kiwifruit young fruits was 13.4%, and the final yield was 1061kg / mu. After the harvested mature kiwifruit was stored at room temperature for 7 days, it was found that the infection rate of gray mold in kiwifruit fruits reached 31.3%. The use of a single agent prepared with Chinese herbal ingredients and a single auxin IAA for gray mold prevention and control both had different degrees of prevention and control effects on gray mold. However, in addition to enhancing the resistance of kiwifruit to gray mold, the auxin IAA can also promote the growth of kiwifruit at a certain concentration. However, low concentrations of IAA cannot achieve a good prevention and control effect, and its use concentration can only be increased. In addition, the high-frequency use of higher concentrations of auxin IAA has an inhibitory effect on the growth of kiwifruit. Although the fruit drop rate is reduced, the final kiwifruit yield is slightly lower than that of the blank group. Compared with the use of a single Chinese medicine and a single growth hormone IAA, the direct mixing of the medicine prepared by Chinese medicine fermentation and the growth hormone IAA has a certain improvement in the prevention and control of kiwifruit gray mold, but the effect is not significant. By preparing a composite medicine made of Chinese medicine fermentation and IAA by freeze-drying and using it to prevent and control gray mold, the survival rate of the planted kiwifruit plants reached 100%, the gray mold infection rate was 0, and the young fruit drop rate was less than 2% after three years. The yield reached about 1500kg / mu. After the harvested kiwifruit was stored at room temperature for 7 days, the gray mold infection rate was only less than 1%.

Claims

1. A composite agent for preventing and controlling kiwifruit diseases, specifically for preventing and controlling gray mold, characterized in that: The invention is to prepare a mixed powder from sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria baicalensis as raw materials, collect the filtrate after fermentation, add mannitol, auxin IAA and Tween 20 to the filtrate, and finally freeze-dry the mixture. The fermentation is carried out by using Streptomyces albicans ( Streptomyces albidoflavus )、Streptomyces monomycinicola( Streptomyces monomycini )、Bacillus stutzeri( Bacillus spizizenii ), Bacillus thuringiensis ( Bacillus thuringiensis ) and Trichoderma viride ( Trichoderma viride ) is a fermentation agent.

2. A composite agent for preventing and controlling kiwifruit diseases as claimed in claim 1, characterized in that: The mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomycinus, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride is 100:0.15:0.15:0.20:0.20:0.25, and the mass ratio of sugarcane, coptis root, purslane, dandelion, isatis root and scutellaria baicalensis in the mixed powder is 10:2~3:3~5:1~2:1~2:4~6.

3. A composite agent for preventing and controlling kiwifruit diseases as claimed in claim 1 or 2, characterized in that: The mass ratio of the fermentation broth, mannitol and Tween 20 is 100:50-60:1-2, and the mass proportion of the auxin IAA in the prepared composite agent is 0.3-0.5%.

4. A method for preparing a composite agent for preventing and controlling kiwifruit diseases, characterized in that: The steps include: (1) Sugarcane, coptis root, purslane, dandelion, isatis root and scutellaria baicalensis are mixed and crushed to prepare a mixed powder; (2) After the mixed powder is boiled and cooled, a fermentation agent is added to perform fermentation treatment, and the residue and filtrate are collected after filtering, where the filtrate is the fermentation liquid; (3) Add mannitol, auxin IAA and Tween 20 to the fermentation broth and freeze-dry to obtain a composite agent.

5. The method for preparing a composite agent for preventing and controlling kiwifruit diseases according to claim 4, characterized in that: In the step (1), the mass ratio of sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria baicalensis is 10:2-3:3-5:1-2:1-2:4-6.

6. A method for preparing a composite agent for preventing and controlling kiwifruit diseases according to claim 4 or 5, characterized in that: The fermentation agent used in step (2) includes Streptomyces albicans ( Streptomyces albidoflavus )、Streptomyces monomycinicola( Streptomyces monomycini )、Bacillus stutzeri( Bacillus spizizenii ), Bacillus thuringiensis ( Bacillus thuringiensis ) and Trichoderma viride ( Trichoderma viride ).

7. A method for preparing a composite agent for preventing and controlling kiwifruit diseases according to any one of claims 4 to 6, characterized in that: The fermentation in step (2) is specifically to add Streptomyces albus and Streptomyces monomycinicus in turn to the cooled mixed powder, ferment at 28-30°C for 3 days, then add Bacillus stutzeri, Bacillus thuringiensis and Trichoderma viride, keep the temperature at 30-32°C and continue fermenting for 10-15 days, filter after the fermentation is completed, collect the filter residue and filtrate, and the filtrate is the fermentation liquid.

8. A method for preparing a composite agent for preventing and controlling kiwifruit diseases according to any one of claims 4 to 7, characterized in that: In the step (2), the mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomycinii, Bacillus stutzeri, Bacillus thuringiensis and Trichoderma viride is 100:0.15:0.15:0.20:0.20:0.

25.

9. A method for preparing a composite agent for preventing and controlling kiwifruit diseases according to any one of claims 4 to 8, characterized in that: In the step (3), the mass ratio of the fermentation broth, mannitol and Tween 20 is 100:50-60:1-2, and the mass proportion of the auxin IAA in the prepared composite agent is 0.3-0.5%.

10. A method for preparing a composite agent for preventing and treating gray mold of kiwi fruit, characterized in that: The steps include: (1) Preparation of mixed powder Mix sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria, put them in a universal grinder, grind them through a 10-mesh sieve to obtain a mixed powder, wherein the mass ratio of sugar cane, coptis root, purslane, dandelion, isatis root and scutellaria is 10:2-3:3-5:1-2:1-2:4-6; (2) Preparation of fermentation broth The mixed powder is placed in a steamer and steamed for 2-3 hours. The mixed powder is taken out, cooled, and a fermentation agent is added for fermentation. Specifically, Streptomyces albus and Streptomyces monomycinis are first added, and fermentation is carried out at 28-30° C. for 3 days. Then, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride are added, and the temperature is maintained at 30-32° C. and fermentation is continued for 10-15 days. After the fermentation is completed, the mixture is filtered to collect the filter residue and the filtrate, and the filtrate is the fermentation liquid. The mass ratio of the mixed powder, Streptomyces albus, Streptomyces monomycinis, Bacillus stuartii, Bacillus thuringiensis and Trichoderma viride is 100: 0.15: 0.15: 0.20: 0.20: 0.25; (3) Preparation of compound medicines Mannitol, auxin IAA and Tween 20 are added to the fermentation broth prepared in step (2) in sequence, stirred and dissolved, and then freeze-dried, specifically at -30 ~ -35 ° C, maintained for 150 ~ 180 minutes, and then vacuum dried, heated to -10 ~ -15 ° C at a heating rate of 5 ~ 8 ° C / h, and kept at a constant temperature for 150 ~ 180 minutes; then heated to 0 ° C at a heating rate of 5 ~ 8 ° C / h, and kept at 0 ° C for 200 ~ 250 minutes; then heated to 15 ° C at a heating rate of 5 ~ 8 ° C / h, and kept at a constant temperature for 300 ~ 350 minutes; finally heated to 30 ~ 32 ° C at 8 ~ 10 ° C / h, and kept at a constant temperature for 120 ~ 150 minutes to prepare a composite agent, wherein the mass ratio of the fermentation broth, mannitol and Tween 20 is 100:50 ~ 60:1 ~ 2, and the mass proportion of auxin IAA in the prepared composite agent is 0.3 ~ 0.5%.

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