Preparation method of a medicament for preventing and controlling kiwifruit diseases

Abstract

A compound agent for preventing and controlling kiwifruit diseases is prepared by taking sugarcane, Coptis chinensis, purslane, dandelion, Isatis indigotica, and Scutellaria baicalensis as raw materials to make a mixed powder, fermenting it, collecting the filtrate, adding mannitol, auxin IAA, and Tween 20 to the filtrate, and finally freeze-drying. The fermentation is carried out using Streptomyces albidoflavus ( Streptomyces albidoflavus ), Streptomyces monomycini ( Streptomyces monomycini ), Bacillus sphaericus ( Bacillus spizizenii ), Bacillus thuringiensis ( Bacillus thuringiensis ), and Trichoderma viride ( Trichoderma viride ) as fermentation inoculants. By spraying the above compound agent, the present invention has a high prevention efficiency. After 3 years of cultivation, the infection rate of the kiwifruit cultivation method with gray mold is 0, the survival rate of kiwifruit is as high as 100%, the fruits are produced after three years of cultivation, the fruit drop rate is less than 2%, the yield can be as high as about 1500 kg / mu, and the gray mold infection rate of the mature kiwifruit is as low as within 1% after being stored at room temperature for 7 days.

Description

[0001] This invention is a divisional application of the patent application with the application number 202311673852.2 and the invention title of "A Cultivation Method for Controlling Gray Mold of Kiwifruit". Technical Field

[0002] This invention relates to the technical field of kiwifruit disease control, and specifically relates to a preparation method of a medicament for controlling kiwifruit diseases. Background Art

[0003] Kiwifruit contains rich nutritional components. In addition to containing organic substances such as actinidine, proteolytic enzymes, and sugars, as well as trace elements such as calcium, potassium, zinc, selenium, and 17 amino acids required by the human body, it also contains rich vitamin C, gluconic acid, citric acid, etc. Its taste is sour and sweet, and it is deeply loved by the public. However, kiwifruit likes warmth and humidity and is extremely susceptible to gray mold. Gray mold mainly occurs during the flowering period, young fruit period, and storage period. Gray mold is caused by the infection of Botrytis cinerea, and flowers, fruits, leaves, and stems can all be diseased. After the flowers are infected, they turn brown and rot and fall off. The infected petals fall on the leaves and fruits, further infecting the young fruits and leaves. The infection of the leaves causes leaf spots. At the initial stage of fruit damage, the hairs turn brown, and then local rot occurs, resulting in fruit drop. When the humidity is high, a grayish-black mold layer appears on the fruit surface. If the control is not in place during the growth process, the incidence rate of gray mold during the subsequent fruit storage period will also increase. In the prior art, for spraying medicaments in kiwifruit orchards to control gray mold, chemical fungicides such as pyrimethanil, procymidone, and iprodione are often used for spraying. However, chemical medicaments cannot be used frequently, and the control effect is limited. Therefore, it is necessary to seek an efficient and environmentally friendly medicament for controlling gray mold, as well as a kiwifruit cultivation method based on this medicament, to achieve efficient control of gray mold, thereby improving the yield and quality of kiwifruit. Summary of the Invention

[0004] The purpose of this invention is to provide a compound medicament for controlling kiwifruit diseases, which can effectively improve the control effect on kiwifruit gray mold, significantly improve the survival rate of kiwifruit cultivation, reduce the kiwifruit fruit drop rate, and at the same time reduce the infection rate of gray mold during the kiwifruit storage period.

[0005] Another purpose of this invention is to provide a preparation method of the above compound medicament for controlling kiwifruit gray mold.

[0006] The purpose of this invention is achieved through the following technical solutions:

[0007] A compound medicament for controlling kiwifruit diseases, specifically for controlling gray mold, is characterized in that it is made from sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis to form a mixed powder, the filtrate is collected after fermentation, mannitol, auxin IAA, and Tween 20 are added to the filtrate, and finally freeze-drying is carried out. The fermentation is carried out using Streptomyces albidoflavus ( Streptomyces albidoflavus ), Streptomyces monomycini ( Streptomyces monomycini ), Bacillus sphaericus ( Bacillus spizizenii ), Bacillus thuringiensis ( Bacillus thuringiensis ), and Trichoderma viride ( Trichoderma green ) are used as fermentation inoculants.

[0008] When using sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis as raw materials to ferment and prepare a fermentation inoculant for controlling Botrytis cinerea, it is found that during long-term use, the pathogenic bacteria will develop drug resistance, and the control effect of the inoculant gradually weakens, and the drug efficacy decreases. During continuous exploration, it is found that adding auxin IAA to the fermentation broth, IAA plays an induced antagonistic role, and at the same time inhibits the drug resistance of the pathogenic bacteria to the fermentation inoculant. At the same time, by freeze-drying the mixture of the fermentation broth and IAA, it is found that the induced antagonistic effect of IAA on the pathogenic bacteria is further enhanced. In addition, as a growth hormone, IAA helps the growth and reproduction of kiwifruit to a certain extent while controlling Botrytis cinerea, and improves the fruit quality.

[0009] Further, the mass ratio of the mixed powder, Streptomyces albidoflavus, Streptomyces monomycini, Bacillus sphaericus, 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 mongolicum, Isatis indigotica, and Scutellaria baicalensis in the mixed powder is 10:2 - 3:3 - 5:1 - 2:1 - 2:4 - 6.

[0010] Further, the mass ratio of the fermentation broth, mannitol, and Tween 20 is 100:50 - 60:1 - 2, and the mass fraction of auxin IAA in the prepared compound medicament is 0.3 - 0.5%.

[0011] A preparation method of a compound medicament for controlling kiwifruit diseases, which is characterized by comprising the following steps:

[0012] (1) Mix and pulverize sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis to make a mixed powder;

[0013] (2) After steaming and cooling the mixed powder, add a fermentation inoculant for fermentation treatment, filter and collect the filter residue and filtrate, and the filtrate is the fermentation broth;

[0014] (3) Add mannitol, auxin IAA, and Tween 20 to the fermentation broth, and freeze-dry to obtain the compound medicament.

[0015] Further, in the step (1), the mass ratio of sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis is 10:2 - 3:3 - 5:1 - 2:1 - 2:4 - 6.

[0016] Further, in the step (2), the steaming is carried out by steaming in water, and the steaming time is 2 to 3 hours. While sterilizing, it inactivates enzymes to preserve glycosides and inhibits the decomposition of active ingredients by enzymes.

[0017] Further, the fermentation inoculant used in the step (2) includes Streptomyces albidoflavus ( Streptomyces whitish-yellow ), Streptomyces monomycini ( Streptomyces monomycini ), Bacillus subtilis ( Bacillus spizizen ), Bacillus thuringiensis ( Bacillus thuringiensis ), and Trichoderma viride ( Trichoderma green ).

[0018] Further, in the step (2), the fermentation is specifically carried out by sequentially adding Streptomyces albidoflavus and Streptomyces monomycini to the cooled mixed powder, fermenting at 28 to 30 °C for 3 days, and then adding Bacillus subtilis, Bacillus thuringiensis, and Trichoderma viride, and continuing to ferment at 30 to 32 °C for 10 to 15 days. After the fermentation is completed, filtration is carried out to collect the filter residue and the filtrate, and the filtrate is the fermentation broth.

[0019] Further, the mass ratio of the mixed powder, Streptomyces albidoflavus, Streptomyces monomycini, Bacillus subtilis, Bacillus thuringiensis, and Trichoderma viride in the step (2) is 100: 0.15: 0.15: 0.20: 0.20: 0.25.

[0020] Further, the mass ratio of the fermentation broth, mannitol, and Tween 20 in the step (3) is 100: 50 - 60: 1 - 2, and the mass proportion of indole - 3 - acetic acid (IAA) in the prepared compound medicament is 0.3 - 0.5%.

[0021] Further, in the step (3), the freeze - drying is carried out by first reducing the temperature to - 30 to - 35 °C, maintaining for 150 to 180 minutes, then carrying out vacuum drying, heating at a heating rate of 5 - 8 °C / h to - 10 to - 15 °C, and maintaining at a constant temperature for 150 to 180 minutes; then heating at a heating rate of 5 - 8 °C / h to 0 °C, maintaining at 0 °C for 200 - 250 minutes; then heating at a heating rate of 5 - 8 °C / h to 15 °C, maintaining at a constant temperature for 300 - 350 minutes, and finally heating at 8 - 10 °C / h to 30 - 32 °C, maintaining at a constant temperature for 120 - 15 minutes to complete the drying.

[0022] Most specifically, a preparation method of a compound medicament for controlling Botrytis cinerea is characterized by including the following steps:

[0023] (1) Prepare the mixed powder

[0024] Mix sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis, place them in a universal grinder, pulverize them through a 10-mesh sieve to obtain a mixed powder. The mass ratio of sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis is 10:2 - 3:3 - 5:1 - 2:1 - 2:4 - 6;

[0025] (2)Prepare the fermentation broth

[0026] Put the mixed powder in a steamer and steam it over water for 2 - 3 hours. Take out the mixed powder and let it cool. Then add a fermentation inoculant for fermentation. Specifically, first add Streptomyces flavovirens and Streptomyces monomycini, and ferment at 28 - 30°C for 3 days. Then add Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride, and continue to ferment at 30 - 32°C for 10 - 15 days. After fermentation, filter to collect the filter residue and the filtrate. The filtrate is the fermentation broth. The mass ratio of the mixed powder, Streptomyces flavovirens, Streptomyces monomycini, Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride is 100:0.15: 0.15: 0.20: 0.20:0.25;

[0027] (3)Prepare the compound medicament

[0028] Add mannitol, auxin IAA, and Tween 20 to the fermentation broth prepared in step (2) in sequence, stir to dissolve, and then perform freeze-drying. Specifically, at -30 ~ -35°C, keep it for 150 - 180 minutes, then vacuum dry. Heat it at a heating rate of 5 - 8°C / h to -10 ~ -15°C and keep it at a constant temperature for 150 - 180 minutes; then heat it at a heating rate of 5 - 8°C / h to 0°C and keep it at 0°C for 200 - 250 minutes; then heat it at a heating rate of 5 - 8°C / h to 15°C and keep it at a constant temperature for 300 - 350 minutes. Finally, heat it at a heating rate of 8 - 10°C / h to 30 - 32°C and keep it at a constant temperature for 120 - 150 minutes to obtain the compound medicament. The mass ratio of the fermentation broth, mannitol, and Tween 20 is 100:50 - 60:1 - 2. The mass proportion of auxin IAA in the prepared compound medicament is 0.3 - 0.5%.

[0029] A cultivation method for preventing and controlling gray mold of kiwifruit, characterized in that: it uses sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis to ferment to prepare filter residue and fermentation broth. The fermentation broth and auxin IAA are prepared into a compound medicament. After the kiwifruit seedlings are planted for 2 - 3 months, bury the filter residue at the roots of the seedlings, and spray the compound medicament solution on the seedlings. Then spray the compound medicament solution every 2 - 3 months until the kiwifruit is ripe and picked. The compound medicament solution is obtained by diluting the compound medicament with water by 200 - 300 times, and is sprayed at 150 - 200 mL per kiwifruit plant.

[0030] Further, the fermentation is specifically carried out by making a mixed powder from sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica and Scutellaria baicalensis. After steaming and cooling it in a water bath, Streptomyces microflavus and Streptomyces monomycini are added successively, and it is fermented at 28-30 °C for 3 days. Then, Bacillus sphaericus, Bacillus thuringiensis and Trichoderma viride are added, and the temperature is maintained at 30-32 °C for continuous fermentation for 10-15 days. After the fermentation is completed, it is filtered to collect the filter residue and the filtrate, and the filtrate is the fermentation broth.

[0031] Further, the mass ratio of the mixed powder, Streptomyces microflavus, Streptomyces monomycini, Bacillus sphaericus, Bacillus thuringiensis and Trichoderma viride is 100: 0.15: 0.15: 0.20: 0.20: 0.25.

[0032] Further, the composite agent is prepared by adding mannitol, auxin IAA and Tween 20 to the fermentation broth in sequence, stirring and dissolving them, and then performing freeze-drying.

[0033] Further, 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%.

[0034] Further, for the freeze-drying, the temperature is first lowered to -30 to -35 °C and maintained for 150-180 min, then vacuum drying is carried out, and the temperature is raised at a heating rate of 5-8 °C / h to -10 to -15 °C and kept constant for 150-180 min; then the temperature is raised at a heating rate of 5-8 °C / h to 0 °C, and kept constant at 0 °C for 200-250 min; then the temperature is raised at a heating rate of 5-8 °C / h to 15 °C and kept constant for 300-350 min, and finally the temperature is raised to 30-32 °C at a heating rate of 8-10 °C / h and kept constant for 120-150 min to complete the drying.

[0035] Most specifically, a cultivation method for preventing and controlling Botrytis cinerea of kiwifruit is characterized by including the following steps:

[0036] I. Selecting seedlings and building a trellis

[0037] 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 from March to April every year. The row spacing between two kiwifruit seedlings is 4-6 m. After planting, water each seedling, and the watering amount is 500-600 ml / plant;

[0038] Install vertical support columns at a distance of 40-60 cm from the kiwifruit seedlings. The support columns are 2.5-3.0 m high. At the top of the support columns, connect the vertical support columns with horizontal support columns and fix them with small iron wires, and build a trellis with each ridged part as a unit;

[0039] II. Fertilization

[0040] Within half a year after planting, apply compound fertilizer once a month, with the amount of each application being 60 - 80 g / plant. After half a year of planting, apply compound fertilizer once every two months, with the amount of each application being 280 - 300 g / plant;

[0041] III. Preparation of Compound Agent

[0042] (1) Preparation of mixed powder

[0043] Take sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis, mix them, place them in a universal grinder, crush them through a 10 - mesh sieve to obtain mixed powder. The mass ratio of sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis is 10:2 - 3:3 - 5:1 - 2:1 - 2:4 - 6;

[0044] (2) Preparation of fermentation broth

[0045] Put the mixed powder in a steamer and steam it for 2 - 3 h over water. Take out the mixed powder, let it cool, and add a fermentation inoculant for fermentation. Specifically, first add Streptomyces albidoflavus and Streptomyces monomycini, and ferment at 28 - 30 °C for 3 days. Then add Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride, and continue to ferment at 30 - 32 °C for 10 - 15 days. After fermentation, filter to collect the filter residue and filtrate. The filtrate is the fermentation broth. The mass ratio of the mixed powder, Streptomyces albidoflavus, Streptomyces monomycini, Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride is 100:0.15:0.15:0.20:0.20:0.25;

[0046] (3) Preparation of compound agent [[ID=2 cuatro]]<end>

[0047] Add mannitol, auxin IAA, and Tween 20 to the fermentation broth prepared in step (2) in sequence, stir to dissolve, and then perform freeze - drying. Specifically, at - 30 - 35 °C, keep for 150 - 180 min, then vacuum - dry. Heat at a heating rate of 5 - 8 °C / h to - 10 - 15 °C, and keep at a constant temperature for 150 - 180 min; then heat at a heating rate of 5 - 8 °C / h to 0 °C, keep at 0 °C for 200 - 250 min; then heat at a heating rate of 5 - 8 °C / h to 15 °C, keep at a constant temperature for 300 - 350 min, and finally heat at 8 - 10 °C / h to 30 - 32 °C, keep at a constant temperature for 120 - 150 min to obtain the compound agent. 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 obtained compound agent is 0.3 - 0.5%;

[0048] IV. Prevention and control of Botrytis cinerea

[0049] After 2 to 3 months of seedling planting, fermented filter residue is buried at the roots of the seedlings, and a compound medicament solution is sprayed on the seedlings. After that, the compound medicament solution is sprayed once every 2 to 3 months until the kiwifruit is ripe and picked. The compound medicament solution is prepared by adding compound medicament to 200 to 300 times its mass of water, and is sprayed in an amount of 150 to 200 mL per kiwifruit plant.

[0050] The present invention has the following technical effects:

[0051] By spraying the compound medicament for preventing gray mold of kiwifruit prepared by the present invention, the prevention efficiency is high. After 3 years of cultivation, the infection rate of the cultivation method of kiwifruit suffering from gray mold is 0, the survival rate of kiwifruit is as high as 100%, the fruit drops after 3 years of cultivation, and the fruit drop rate is less than 2%. The yield is high, and the yield can be as high as about 1500 kg / mu. After 7 days of storage at room temperature, the gray mold infection rate of the mature kiwifruit is as low as within 1%. The compound medicament of the present invention has a strong killing effect on Botrytis cinerea, and can effectively kill Botrytis cinerea after 3 days of antibacterial.

[0052] In the compound medicament of the present invention, through the fermentation products of traditional Chinese medicines such as Coptis chinensis, Taraxacum mongolicum, Portulaca oleracea, Isatis indigotica, and Scutellaria baicalensis, in combination with auxin IAA, the induced antagonistic effect on Botrytis cinerea can be enhanced, playing a synergistic effect, and at the same time being environmentally friendly, solving the problem of environmental pollution caused by chemical medicaments. Specific embodiments

[0053] The present invention will be specifically described below through examples. It is necessary to point out here that the following examples are only used to further illustrate the present invention and cannot be understood as limiting the protection scope of the present invention. Those skilled in the art can make some non-essential improvements and adjustments to the present invention according to the above content of the present invention.

[0054] The fermentation microorganisms in the fermentation inoculant used in the present invention are as follows:

[0055] Streptomyces albidoflavus is purchased from the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC 4.252.

[0056] Streptomyces monomycini, Bacillus cereus, Bacillus thuringiensis, and Trichoderma viride are purchased from the Guangdong Provincial Microbiological Culture Collection Center (GDMCC). Among them, the preservation number of Streptomyces monomycini is GDMCC 4.201; the preservation number of Bacillus cereus is GDMCC 1.136; the preservation number of Bacillus thuringiensis is GDMCC 1.147; the preservation number of Trichoderma viride is GDMCC 3.602.

[0057] Example 1

[0058] A preparation method of a composite agent for preventing and controlling gray mold, comprising the following steps:

[0059] (1) Prepare a mixed powder

[0060] Take sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis, mix them, place them in a universal crusher, crush them through a 10-mesh sieve to obtain a mixed powder, and the mass ratio of sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis is 10:2.5:4:1.5:1.5:5;

[0061] (2) Prepare a fermentation broth

[0062] Put the mixed powder in a steamer and steam it for 2.5 h over water. Take out the mixed powder, let it cool, and add a fermentation inoculant for fermentation. Specifically, first add Streptomyces microflavus and Streptomyces monomycini, and ferment at 28-30 °C for 3 days. Then add Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride, and continue to ferment at 30-32 °C for 12 days. After fermentation, filter to collect the filter residue and filtrate. The filtrate is the fermentation broth, and the mass ratio of the mixed powder, Streptomyces microflavus, Streptomyces monomycini, Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride is 100:0.15:0.15:0.20:0.20:0.25;

[0063] (3) Prepare the composite agent

[0064] Add mannitol, auxin IAA, and Tween 20 to the fermentation broth prepared in step (2) in sequence, stir and dissolve them, and then perform freeze-drying. Specifically, at -30 °C, keep it for 180 min, then vacuum dry it, heat it at a heating rate of 6 °C / h to -10 °C, and keep it at a constant temperature for 180 min; then heat it at a heating rate of 6 °C / h to 0 °C, keep it at 0 °C for 240 min; then heat it at a heating rate of 6 °C / h to 15 °C, keep it at a constant temperature for 320 min, and finally heat it to 30 °C at a heating rate of 10 °C / h and keep it at a constant temperature for 150 min to obtain the composite agent. 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%.

[0065] Example 2

[0066] A cultivation method for preventing and controlling gray mold of kiwifruit, comprising the following steps:

[0067] I. Select seedlings and set up a trellis

[0068] Before planting kiwifruit, select sandy soil. With a row spacing of 2.5 meters, turn over the soil to form ridges, dig trenches between the ridges, with the trench depth being 20 - 30 cm for convenient drainage. 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 10:1. The planting time is from March to April every year. The row spacing between two kiwifruit seedlings is 4 - 6 m. After planting, water each seedling, with the watering amount being 500 - 600 ml / plant;

[0069] Install vertical support columns at a distance of 40 - 60 cm from the kiwifruit seedlings. The support columns are 2.5 - 3.0 m high. At the top of the support columns, connect each vertical support column with a horizontal support column and fix them with small iron wires, and set up the trellis in units of each ridged part;

[0070] II. Fertilization

[0071] Within half a year after planting, apply compound fertilizer once a month, with the amount of each fertilization being 60 - 80 g / plant. After planting for half a year, apply compound fertilizer once every two months, with the amount of each fertilization being 280 - 300 g / plant;

[0072] III. Prevention and control of gray mold

[0073] Two months after planting the seedlings, bury the fermented filter residue at the roots of the seedlings and spray the medicament solution prepared by compound medicament in Example 1 on the seedlings. After that, spray the compound medicament solution once every two months until the kiwifruit is ripe and picked. The medicament solution prepared by compound medicament is prepared by adding 250 times the mass of water to the compound medicament, and is sprayed in an amount of 150 - 200 mL per kiwifruit plant.

[0074] Comparative Example 1

[0075] Cultivate kiwifruit according to the method of Example 2. The medicament used in the process of preventing and controlling gray mold is obtained by directly concentrating and evaporating the fermentation broth prepared in Example 1, adding mannitol and Tween 20, and then freeze - drying into powder, that is, auxin IAA is not added to the medicament, and the rest is the same as in Example 1.

[0076] Comparative Example 2

[0077] Cultivate kiwifruit according to the method of Example 2. The medicament used in the process of preventing and controlling Botrytis cinerea is auxin IAA, and the fermentation broth compound is not adopted. And through multiple experiments, it is found that when auxin IAA is used alone and the same concentration of IAA aqueous solution as in Example 2 is sprayed, the concentration of auxin IAA in the sprayed compound medicament solution is only 16 μg / mL, and the control effect on Botrytis cinerea is not obvious. When the concentration of the sprayed IAA aqueous solution is 60 - 80 μg / mL, the resistance of kiwifruit to Botrytis cinerea is improved most significantly. Therefore, during cultivation, an aqueous solution with an auxin IAA concentration of 60 μg / mL is used to replace the compound medicament solution used in Example 2 for the prevention and control of Botrytis cinerea, and the remaining steps are the same as those in Example 1.

[0078] Comparative Example 3

[0079] Cultivate kiwifruit according to the method of Example 2. The medicaments used in the process of preventing and controlling Botrytis cinerea are as follows:

[0080] When preparing the compound medicament, the fermentation broth is directly concentrated and evaporated, then freeze-dried into powder, and then directly mixed with IAA to form the compound medicament.

[0081] Take 50 mL of the medicament solutions 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 of IAA as in the compound medicament solution of Example 2) separately as a group, and add them to 50 g of beef extract peptone liquid medium respectively, stir evenly, add 1 mL of Botrytis cinerea (Botrytis cinerea) (the viable bacteria count is about 2000) to the medium, place and culture at 25 °C in a sterile environment for 5 days, observe the growth situation daily, and conduct the statistics of the number of Botrytis cinerea. The results are shown in Table 1.

[0082] Table 1: Resistance of different medicaments to Botrytis cinerea

[0083]

[0084] Note: “——” in the table indicates not detected.

[0085] It can be seen that when the compound medicament prepared by the present invention is used for the antibacterial test of Botrytis cinerea, the viable count of Botrytis cinerea decreases sharply. On the 4th day of the test, the viable count of Botrytis cinerea could no longer be detected in the culture medium. When using a single traditional Chinese medicine medicament (the medicament in Example 1), it can be seen that the viable count of Botrytis cinerea also decreases continuously, but the reduction rate is slow. When using a single auxin IAA, it can only have an obvious inhibitory effect on Botrytis cinerea at a certain concentration. When the concentration is low, the antibacterial effect of IAA on Botrytis cinerea is extremely poor. When the traditional Chinese medicine medicament and IAA are directly mixed and used, the inhibitory effect on Botrytis cinerea is significantly improved compared with that of the single traditional Chinese medicine medicament and the single auxin IAA, but the effect is still slightly worse than that of the compound medicament prepared in the present invention.

[0086] Example 3

[0087] A cultivation method for preventing and controlling kiwifruit gray mold disease includes the following steps:

[0088] I. Seedling selection and trellis erection

[0089] Select kiwifruit seedlings with a height of 30 - 40 cm and a rootstock diameter of 1.0 - 1.3 cm for planting. The male - female ratio of kiwifruit seedlings is 8:1. The planting time is from March to April every year. The row spacing between two kiwifruit seedlings is 4 - 6 m. After planting, water each seedling, and the watering amount is 500 - 600 ml / plant;

[0090] Install vertical support columns at a distance of 40 - 60 cm from the kiwifruit seedlings. The support columns are 2.5 - 3.0 m high. At the top of the support columns, connect each vertical support column with a horizontal support column and fix it with small iron wires, and erect the trellis in units of each ridged part;

[0091] II. Fertilization

[0092] Within half a year after planting, apply compound fertilizer once a month, and the amount of each fertilization is 60 - 80 g / plant. After planting for half a year, apply compound fertilizer once every two months, and the amount of each fertilization is 280 - 300 g / plant;

[0093] III. Preparation of compound medicament

[0094] (1) Preparation of mixed powder

[0095] Take sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis, mix them, place them in a universal pulverizer, pulverize and pass through a 10 - mesh sieve to obtain a mixed powder. The mass ratio of sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis is 10:2:3:1:1:4;

[0096] (2) Preparation of fermentation broth

[0097] The mixed powder is placed in a steamer and steamed over water for 3 hours. Then, the mixed powder is taken out, cooled, and fermented by adding a fermentation inoculant. Specifically, Streptomyces albidoflavus and Streptomyces monomycini are added first and fermented at 28 - 30 °C for 3 days. Then, Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride are added, and fermentation continues at 30 - 32 °C for 15 days. After fermentation, filtration is carried out to collect the filter residue and filtrate. The filtrate is the fermentation broth. The mass ratio of the mixed powder, Streptomyces albidoflavus, Streptomyces monomycini, Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride is 100:0.15:0.15:0.20:0.20:0.25;

[0098] (3)Preparation of the composite medicament

[0099] 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, it is carried out at -35 °C for 150 min, then vacuum dried. The temperature is raised to -15 °C at a rate of 5 °C / h and kept constant for 150 min; then the temperature is raised to 0 °C at a rate of 5 °C / h, and kept constant at 0 °C for 200 min; then the temperature is raised to 15 °C at a rate of 5 °C / h and kept constant for 300 min. Finally, the temperature is raised to 32 °C at a rate of 8 °C / h and kept constant for 120 min to obtain the composite medicament. The mass ratio of the fermentation broth, mannitol, and Tween 20 is 100:50:1. The mass proportion of auxin IAA in the prepared composite medicament is 0.3%;

[0100] IV. Prevention and control of gray mold

[0101] Two months after the seedlings are planted, the fermented filter residue is buried at the roots of the seedlings, and the composite medicament solution is sprayed on the seedlings. After that, the composite medicament solution is sprayed once every 3 months until the kiwifruit is ripe and picked. The solution prepared from the composite medicament is prepared by adding water 200 times the mass of the composite medicament, and it is sprayed in an amount of 150 - 200 mL per kiwifruit plant.

[0102] Example 4

[0103] A cultivation method for preventing and controlling gray mold of kiwifruit includes the following steps:

[0104] I. Seedling selection and trellis building

[0105] Kiwi seedlings with a height of 30 - 40 cm and a root diameter of 1.0 - 1.3 cm are selected for planting. The male-female ratio of the kiwi seedlings is 9:1. The planting time is from March to April every year. The row spacing between two kiwi seedlings is 4 - 6 m. After planting, each seedling is watered with a watering amount of 500 - 600 ml / seedling;

[0106] Install vertical support columns at a distance of 40 - 60 cm from the kiwifruit seedlings. The support columns are 2.5 - 3.0 m high. Connect the vertical support columns at the top with horizontal support columns and fix them with small iron wires. Set up the trellis for each ridged part.

[0107] II. Fertilization

[0108] Within half a year after planting, apply compound fertilizer once a month, with a dosage of 60 - 80 g / plant each time. After half a year of planting, apply compound fertilizer once every two months, with a dosage of 280 - 300 g / plant each time.

[0109] III. Preparation of compound medicament

[0110] (1) Preparation of mixed powder

[0111] Take sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis, mix them, place them in a universal grinder, pulverize and pass through a 10 - mesh sieve to obtain mixed powder. The mass ratio of sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis is 10:3:5:2:2:6.

[0112] (2) Preparation of fermentation broth

[0113] Put the mixed powder in a steamer and steam it for 2 h over water. Take out the mixed powder, let it cool, and add a fermentation inoculant for fermentation. Specifically, first add Streptomyces albidoflavus and Streptomyces monomycini, and ferment at 28 - 30 °C for 3 days. Then add Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride, and continue to ferment at 30 - 32 °C for 10 days. After fermentation, filter to collect the filter residue and filtrate. The filtrate is the fermentation broth. The mass ratio of the mixed powder, Streptomyces albidoflavus, Streptomyces monomycini, Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride is 100:0.15:0.15:0.20:0.20:0.25.

[0114] (3) Preparation of compound medicament

[0115] Add mannitol, auxin IAA, and Tween 20 to the fermentation broth prepared in step (2) in sequence, stir and dissolve them, and then perform freeze - drying. Specifically, at - 30 °C, keep it for 160 min, then vacuum - dry. Heat it at a heating rate of 8 °C / h to - 12 °C and keep it at a constant temperature for 160 min; then heat it at a heating rate of 8 °C / h to 0 °C and keep it at 0 °C for 250 min; then heat it at a heating rate of 8 °C / h to 15 °C and keep it at a constant temperature for 350 min; finally, heat it at a heating rate of 10 °C / h to 30 °C and keep it at a constant temperature for 140 min to obtain the compound medicament. The mass ratio of the fermentation broth, mannitol, and Tween 20 is 100:60:2. The mass proportion of auxin IAA in the prepared compound medicament is 0.5%.

[0116] IV. Control of Botrytis cinerea

[0117] Two months after the seedlings are planted, fermented filter residues are buried at the roots of the seedlings, and a compound medicament solution is sprayed on the seedlings. After that, the compound medicament solution is sprayed once every 2.5 months until the kiwifruits are ripe and picked. The solution prepared from the compound medicament is obtained by mixing the compound medicament with 300 times its mass of water, and is sprayed in an amount of 150 - 200 mL per kiwifruit plant.

[0118] Seven mu of land is selected as the experimental planting area in the same planting region. The seven mu of land is divided into seven groups, with each group being 1 mu. They are planted respectively according to the method of Example 1, but without the gray mold control treatment method as the blank group, and kiwifruits of the same seedlings are planted according to the methods of Example 2, Example 3, Example 4, Comparative Example 1, Comparative Example 2, and Comparative Example 3. After three years of planting, the survival rate after six months of planting, the average fruit drop rate of young fruits in the third year of planting, the yield (kg / mu) in the third year of planting, and the gray mold infection rate of the kiwifruits harvested in the third year of planting after being stored for one week at room temperature (temperature is 25 - 30 °C, humidity is 60 - 70%) are recorded respectively. The results are shown in Table 2.

[0119] Table 2:

[0120]

[0121] During the process of kiwifruit cultivation, no control measures against Botrytis cinerea were adopted for the plants. After 6 months of planting, the survival rate of kiwifruit seedlings was only 86%. By the third year of planting, the average fruit drop rate of young kiwifruit was 13.4%, and the final yield was 1061 kg / mu. After storing the harvested mature kiwifruit at room temperature for 7 days, it was found that the infection rate of Botrytis cinerea on the kiwifruit reached 31.3%. Using medicaments prepared with traditional Chinese medicine components alone and the single auxin IAA for the prevention and control of Botrytis cinerea both achieved different degrees of prevention and control effects. However, in addition to enhancing the resistance of kiwifruit to Botrytis cinerea, auxin IAA can also promote the growth of kiwifruit at a certain concentration. But low-concentration IAA cannot achieve good prevention and control effects, and only the application concentration can be increased. Moreover, frequent use of a higher concentration of auxin IAA inhibits the growth of kiwifruit. Although the fruit drop rate decreased, the final yield of kiwifruit was slightly lower than that of the blank group. Directly mixing the medicament prepared by fermenting traditional Chinese medicine and auxin IAA improved the prevention and control effect on Botrytis cinerea of kiwifruit to a certain extent compared with using a single traditional Chinese medicine medicament and a single auxin IAA, but the effect was not significant. By preparing a composite medicament from the medicament fermented from traditional Chinese medicine and IAA through freeze-drying and using it to prevent and control Botrytis cinerea, the survival rate of the planted kiwifruit plants reached 100%, the infection rate of Botrytis cinerea was 0, the fruit drop rate of young fruits after three years was less than 2%, and the yield reached about 1500 kg / mu. After storing the harvested kiwifruit at room temperature for 7 days, the infection rate of Botrytis cinerea was only within 1%.

Claims

1. A compound agent for preventing and controlling kiwifruit diseases, specifically for preventing and controlling gray mold, characterized in that: It is prepared by making a mixed powder from sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica, and Scutellaria baicalensis, collecting the filtrate after fermentation, adding mannitol, auxin IAA, and Tween 20 to the filtrate, and finally performing freeze-drying. The fermentation is carried out using Streptomyces albidoflavus ( Streptomyces albidoflavus ), Streptomyces monomycini ( Streptomyces monomycini ), Bacillus subtilis ( Bacillus spizizenii ), Bacillus thuringiensis ( Bacillus thuringiensis ), and Trichoderma viride ( Trichoderma viride ) as fermentation inoculants. The fermentation is carried out by sequentially adding Streptomyces albidoflavus and Streptomyces monomycini to the mixed powder, fermenting at 28 - 30 °C for 3 days, then adding Bacillus subtilis, Bacillus thuringiensis, and Trichoderma viride, and maintaining the temperature at 30 - 32 °C to continue fermentation for 10 - 15 days. After the fermentation is completed, filtration is carried out to collect the filter residue and the filtrate, and the filtrate is the fermentation broth.

2. The compound medicament for preventing and controlling kiwifruit diseases according to claim 1, wherein: The mass ratio of the mixed powder, Streptomyces albidoflavus, Streptomyces monomycini, Bacillus spizizenii, 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 mongolicum, Isatis indigotica 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 described 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 fraction of auxin IAA in the prepared compound medicament is 0.3-0.5%.

4. A preparation method of a compound medicament for preventing and controlling kiwifruit diseases, characterized in that, Comprising the following steps: (1) Mix and pulverize sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica and Scutellaria baicalensis to prepare a mixed powder; (2)After steaming and cooling the mixed powder, add a fermentation inoculum for fermentation treatment to obtain a fermentation broth. The fermentation inoculum includes Streptomyces albidoflavus ( Streptomyces albidoflavus ), Streptomyces monomycini ( Streptomyces monomycini ), Bacillus subtilis ( Bacillus spizizenii ), Bacillus thuringiensis ( Bacillus thuringiensis ), and Trichoderma viride ( Trichoderma viride ). The fermentation treatment is to sequentially add Streptomyces albidoflavus and Streptomyces monomycini to the cooled mixed powder, ferment at 28 - 30 °C for 3 days, then add Bacillus subtilis, Bacillus thuringiensis, and Trichoderma viride, and continue to ferment at 30 - 32 °C for 10 - 15 days. After fermentation, filter to collect the filter residue and filtrate. The filtrate is the fermentation broth; (3) Add mannitol, auxin IAA and Tween 20 to the fermentation broth, and freeze-dry to obtain the compound medicament.

5. The preparation method of a composite medicament for preventing and controlling kiwifruit diseases according to claim 4, characterized in that:

6. The preparation method of a composite medicament for preventing and controlling kiwifruit diseases according to claim 5, characterized in that: In the step (1), the mass ratio of sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica and Scutellaria baicalensis is 10: 2-3: 3-5: 1-2: 1-2: 4-6.

7. The preparation method of a compound medicament for preventing and controlling kiwifruit diseases according to claim 6, characterized in that: In the step (2), the mass ratio of the mixed powder, Streptomyces albidoflavus, Streptomyces monomycini, Bacillus spizizenii, Bacillus thuringiensis and Trichoderma viride is 100: 0.15: 0.15: 0.20: 0.20: 0.

25.

8. A preparation method of a composite medicament for preventing and controlling gray mold of kiwifruit, 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 fraction of auxin IAA in the prepared compound medicament is 0.3-0.5%. Comprising the following steps: (1) Prepare a mixed powder Take sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica and Scutellaria baicalensis, mix them, place them in a universal pulverizer, pulverize them through a 10-mesh sieve to obtain a mixed powder, and the mass ratio of sugarcane, Coptis chinensis, Portulaca oleracea, Taraxacum mongolicum, Isatis indigotica and Scutellaria baicalensis is 10: 2-3: 3-5: 1-2: 1-2: 4-6; (2) Prepare a fermentation broth The mixed powder is placed in a steamer and steamed over water for 2 - 3 hours. Then, take out the mixed powder, let it cool, and add a fermentation inoculum for fermentation. Specifically, first add Streptomyces albidoflavus and Streptomyces monomycini, and ferment at 28 - 30 °C for 3 days. Then add Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride, and continue to ferment at 30 - 32 °C for 10 - 15 days. After fermentation, filter to collect the filter residue and filtrate. The filtrate is the fermentation broth. The mass ratio of the mixed powder, Streptomyces albidoflavus, Streptomyces monomycini, Bacillus sphaericus, Bacillus thuringiensis, and Trichoderma viride is 100:0.15:0.15:0.20:0.20:0.25; (3)Preparation of the composite agent In the fermentation broth prepared in step (2), add mannitol, auxin IAA, and Tween 20 in sequence and stir to dissolve, then perform freeze-drying. Specifically, at -30 to -35 °C, hold for 150 - 180 minutes, then perform vacuum drying, heat up at a heating rate of 5 - 8 °C / h to -10 to -15 °C, and keep it constant for 150 - 180 minutes; then heat up at a heating rate of 5 - 8 °C / h to 0 °C, keep it constant at 0 °C for 200 - 250 minutes; then heat up at a heating rate of 5 - 8 °C / h to 15 °C, keep it constant for 300 - 350 minutes, and finally heat up at 8 - 10 °C / h to 30 - 32 °C, keep it constant for 120 - 150 minutes to obtain the composite agent. 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 obtained composite agent is 0.3 - 0.5%.

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