A fermented compound medicinal fertilizer and its preparation method and application

Abstract

The invention discloses a fermentation-type composite medicinal fertilizer and a preparation method and application thereof, and belongs to the technical field of microorganisms and bio-organic fertilizers. The medicinal fertilizer of the invention comprises the following raw materials: 50-80 parts of fermented Chinese medicine residues, 5-10 parts of microbial agents, 10-20 parts of humic acid, and 5-10 parts of composite organic and inorganic nutrients. The invention realizes the recycling and reuse of waste resources such as Chinese medicine residues through two-step efficient fermentation, converts them into nutrients that can be absorbed by plants, and the active substances produced by fermentation have a good inhibitory effect on tobacco bacterial wilt pathogens; after application, the dual use effect of fertilizers and pesticides is achieved, the amount and frequency of pesticide use are reduced, and the economic and social benefits are remarkable, which is worthy of application and promotion.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and bio-organic fertilizers, and specifically relates to a fermentation-type composite medicinal fertilizer and a preparation method and application thereof. Background Art

[0002] Pesticide fertilizer usually refers to a compound with the integrated functions of pesticide and fertilizer, which is produced by mixing pesticides and fertilizers in a certain proportion and through a specific process. The pesticide can be a herbicide, insecticide or fungicide, and the fertilizer can be either a single substance or a multi-nutrient compound fertilizer, which has one or more functions such as killing / inhibiting insects, killing / inhibiting bacteria, killing / inhibiting weeds, and regulating crop growth, while also providing nutrition for crops or improving fertilizer utilization. Since pesticide fertilizer is a new type of compound (or material) that integrates the dual functions of pesticides and fertilizers, it not only has the basic function of ensuring agricultural production and harvest, but also has the advantages of being easy to use and reducing environmental pollution.

[0003] Traditional fertilizers can be classified into four categories according to their composition and preparation methods: ① Chemical pesticides and fertilizers are mixed into preparations with the dual functions of pesticides and fertilizers according to a certain process. ② Biological fertilizers. Microorganisms with insecticidal or antibacterial properties (such as Bacillus thuringiensis) and plant hormones (such as Brassinolide) are used to form green fertilizers (such as patent CN102775230A). Due to the survival characteristics of microorganisms, the types and scope of use of such fertilizers are very limited. ③ Organic fertilizers. After harmless treatment of industrial and agricultural waste or plant residues, they are combined with plant materials with insecticidal or antibacterial properties (such as humic acid, etc.) or chemical pesticides and inorganic fertilizers to form organic fertilizers with dual functions of nutrition and plant protection. Organic fertilizers have the advantages of being naturally degradable, safe for humans and animals, and not easy to develop drug resistance, but such plant-based fertilizers have the disadvantages of complicated production process and high labor. ④ Inorganic compounds or single substances that have the dual functions of medicine and fertilizer, such as potassium permanganate, lime sulfur and elemental sulfur. Potassium permanganate has strong oxidizing properties and is a widely used inorganic disinfectant. Moreover, since potassium and manganese are both essential elements for plant growth, potassium permanganate can be used for plant seed dressing to sterilize and kill insects. However, due to the strong oxidizing properties of potassium permanganate, its drug concentration and application range as a medicine fertilizer are greatly limited.

[0004] Among these fertilizers, biological fertilizers can significantly increase crop yield and quality, improve soil health, and are green and environmentally friendly. They can reduce environmental pollution, enhance plant disease resistance, and have significant economic benefits, so they have become a research hotspot. However, the current biological fertilizers have complex ingredients and unstable efficacy and fertilizer effects. In actual planting, it is difficult to promote effective utilization. Therefore, how to develop an efficient, stable, and targeted biological fertilizer is a technical problem that needs to be solved urgently. Summary of the invention

[0005] The present invention aims at the problems existing in the prior art of biological medicine fertilizer. By screening specific microorganisms to effectively ferment the traditional Chinese medicine residues, the nutrients in the residues are efficiently decomposed, and at the same time, an efficient antibacterial effect is given to the residues, thereby achieving multiple goals at one stroke.

[0006] In order to achieve the above technical objectives, the technical solution adopted by the present invention is:

[0007] A fermented compound medicinal fertilizer comprises the following raw materials: 50-80 parts of fermented Chinese medicinal residues, 5-10 parts of microbial agents, 10-20 parts of humic acid, and 5-10 parts of compound organic and inorganic nutrients.

[0008] Furthermore, the preparation method of the fermented Chinese medicine residue is:

[0009] (1) Collecting Chinese medicine residue, mushroom residue and straw, crushing them into particles with a particle size of 0.5-1 cm, adding zeolite powder, sucrose and urea, stirring and mixing them evenly, adjusting the water content to 60%-70%, and sterilizing to obtain a premix;

[0010] (2) adjusting the pH of the premix to 5.8-6.5, adding anaerobic fermentation liquid, and performing anaerobically fermentation at 30-37° C. for 15-20 days to obtain pre-fermented medicinal residue;

[0011] (3) Turn the pre-fermented medicinal residues over, adjust the pH to 6.9-7.5, spray aerobic fermentation liquid, mix well, and then pile and ferment at room temperature. Turn the pile once a day to ventilate and cool the compost. The composting is fermented for 5-7 days. When the temperature of the compost pile no longer rises and begins to drop, the composting is finished. The compost is then air-dried or dried, and crushed to obtain the fermented Chinese medicinal residues.

[0012] Furthermore, the Chinese medicine residue is selected from the waste materials after processing one or a combination of several of honeysuckle, astragalus, codonopsis, American ginseng, salvia miltiorrhiza, dandelion and isatis indigotica.

[0013] Furthermore, the mushroom residue is one or more of the group consisting of enoki mushroom residue, oyster mushroom residue, and king oyster mushroom residue; the mass ratio of the Chinese medicine residue, mushroom residue, and straw is 10:1.5-2.5:3-5, the added amount of zeolite powder is 1% of the mass of the Chinese medicine residue, the added amount of sucrose is 1-3% of the mass of the Chinese medicine residue, and the added amount of urea is 0.5-1.5% of the mass of the Chinese medicine residue.

[0014] Furthermore, the anaerobic fermentation bacterial liquid is a mixture of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus, and the volume ratio of the two is 1:1; the amount of the anaerobic fermentation bacterial liquid added is 0.1-0.5% of the mass of the premix; the preparation method of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus bacterial liquid is: the seed liquid of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus are inoculated into the fermentation medium at an inoculation amount of 5% for fermentation, and the fermentation conditions are as follows: the initial pH of the fermentation tank is 7.0, the fermentation temperature is 30-37°C, and after natural fermentation to pH4.5-4.9, alkali solution is added to maintain the constant pH of the fermentation liquid at 5.5-5.9 until the effective viable bacteria count of the fermentation liquid is 1×10 8 -1×10 9 CFU / mL is the fermentation end point, and the fermentation broths of the two bacteria are mixed in a volume ratio of 1:1 to obtain an anaerobic fermentation broth; the strain number of the Lactococcus lactis is CGMCC No.1.15072, and the original preservation time is January 25, 2015; the strain number of the Lactobacillus delbrueckii subspecies bulgaricus is CGMCC No.1.16075, and the original preservation time is February 15, 2017. Both Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus can be purchased through the China General Microorganism Collection Center without the need for repeated biological preservation.

[0015] Furthermore, the preparation method of Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus seed solution is: inoculate the activated target strains into MRS liquid culture medium, culture at 30-35°C for 2-3 days, and obtain the seed solution of Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus.

[0016] Furthermore, the composition and preparation method of the fermentation medium are as follows: 10.0 g of peptone, 10.0 g of beef extract, 5.0 g of yeast extract, 20.0 g of glucose, 5.0 g of sodium acetate, 2.0 g of diamine citrate, 1.0 g of Tween-80, 0.4 g of dilithium hydrogen phosphate, 0.58 g of magnesium sulfate, 0.29 g of manganese sulfate, 20.0 g of calcium carbonate, and 15.0 g of agar are added to distilled water in sequence and the distilled water is supplemented to 1000 mL, the pH is adjusted to 6.3, heated after stirring, boiled for 2 minutes, and sterilized at 121°C and 0.1 MPa for 30 minutes.

[0017] Furthermore, the solid-liquid ratio of the aerobic fermentation bacterial liquid and the pre-fermentation medicinal residue is 100mL:10kg; the preparation method of the aerobic bacterial liquid is:

[0018] (1) inoculating a Bacillus megaferium strain with a preservation number of CGMCC No. 27648 stored at low temperature onto a beef extract peptone agar medium and culturing it at 26-32° C. for 15-20 hours, then picking a single colony and inoculating it onto an LB slant medium and culturing it at 26-32° C. for 10-20 hours, then washing the surface of the medium with sterile water, and using the eluate as an inoculum;

[0019] (2) adding 1% of the volume of the inoculum obtained in step (1) to the LB liquid, and culturing with shaking at 26-32° C. for 15-20 hours to obtain a seed liquid;

[0020] (3) The seed solution of step (2) was inoculated into corn starch medium at an inoculation ratio of 0.05-1%, and then cultured at 26-32°C and a shaking speed of 150-200 rpm for 30-40 hours to obtain an effective viable count of 1×10 8 -1×10 9 CFU / mL of aerobic fermentation broth.

[0021] Furthermore, the Bacillus megaterium has a deposit number of CGMCC No. 27648, a deposit date of June 16, 2023, and is deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration, and the deposit address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. This strain was donated by others.

[0022] Furthermore, the composition of the corn starch culture medium is: 3g beef extract, 25g corn starch, 3g CaCl2, 7g yeast extract, 1000mL deionized water, and the pH value of the culture medium is natural.

[0023] Furthermore, the microbial agent is obtained by mixing EM bacteria and Bacillus subtilis in a mass ratio of 1:5.

[0024] Furthermore, the composite organic-inorganic nutrient is obtained by mixing urea, potassium dihydrogen phosphite and brown algae oligosaccharide in a mass ratio of 5:0.5:0.1-0.3.

[0025] A method for preparing a fermentation-type compound medicinal fertilizer comprises the following steps:

[0026] A. Preparation of fermented Chinese medicine residues;

[0027] B. Mix and crush the fermented Chinese medicine residue, microbial agent, humic acid and composite organic and inorganic nutrients by weight, add the mixture into a disc granulator, start the disc granulator, and evenly spray water or starch solution on the powder mixture during the rotation of the disc until the mixed powder is rolled into particles; screen the particles, add the particles into a screening machine, and screen to obtain fertilizer particles with a particle size range of 3.5-4.5 mm; and dry to obtain finished medicinal fertilizer particles.

[0028] The invention discloses an application of a fermented compound pesticide fertilizer in preventing and controlling tobacco bacterial wilt.

[0029] The organic fertilizer has an effective live bacteria count of ≥500 million / g and an application amount of 5-10g / plant.

[0030] Traditional Chinese medicine residues contain many high-purity Chinese medicine nutrients, which are excellent resources for preparing bio-organic fertilizers. After the traditional Chinese medicine residues are fully mixed with organic matrix or other organic wastes, stable and nutritious bio-organic fertilizers are formed by fermentation, which can effectively improve crop quality and increase crop yield. Not only does it reduce the demand for chemical fertilizers for crops, but it also makes good use of the traditional Chinese medicine residues. However, the traditional Chinese medicine residues contain a large number of substances that are difficult to degrade and utilize, such as cellulose, lignin and other components. Therefore, screening can degrade cellulose, lignin and other substances, and can also efficiently decompose the microorganisms of the traditional Chinese medicine residues is a major problem currently studied. Therefore, the present invention designs a two-step fermentation process, the first step of anaerobic pre-fermentation + the second step of aerobic fermentation.

[0031] In the first step of anaerobic fermentation, the selected Lactococcus lactis liquid and Lactobacillus delbrueckii subspecies bulgaricus bacteria work together to decompose the toxic components in traditional Chinese medicine, thereby reducing the toxic side effects of the residue. In addition, fermentation can also degrade anti-nutritional factors such as phytic acid, oxalate, and tannin in traditional Chinese medicine into small molecules that are more easily absorbed by plants. At the same time, a variety of organic acids (such as lactic acid), B vitamins and other growth-promoting factors will be produced during the fermentation process. These substances can not only improve the soil microecological environment, but also enhance the immunity and stress resistance of plants.

[0032] The second step is aerobic fermentation. The selected Bacillus megaterium converts cellulose, lignin and other components in the traditional Chinese medicine residue into small molecules through efficient fermentation, which are more easily absorbed and utilized by crops. Bacillus megaterium can produce antimicrobial peptides or other active substances, which have a high inhibitory effect on the pathogen of tobacco wilt.

[0033] Beneficial effects:

[0034] (1) The present invention designs a two-step fermentation process to achieve efficient fermentation of Chinese medicine residues. The active ingredients of the Chinese medicine residues are effectively decomposed and utilized, which not only provides nutrients for crops, but also plays a role in disease resistance and antibacterial, reduces the use of pesticides, and achieves multiple goals at one stroke;

[0035] (2) Add auxiliary microorganisms such as EM bacteria and Bacillus subtilis. Both EM bacteria and Bacillus subtilis can secrete plant growth hormones, such as indoleacetic acid and gibberellins, which can promote the root development of crops, enhance photosynthesis efficiency, and improve the stress resistance of crops. At the same time, Bacillus subtilis can produce a variety of antibacterial substances, such as subtilisin and polymyxin, which have a significant inhibitory effect on a variety of soil-borne pathogens;

[0036] (3) Add humus, urea, etc. to provide supplementary nutrients for crops, add potassium dihydrogen phosphite and brown algae oligosaccharides. Potassium dihydrogen phosphite can promote the rooting of crops, and phosphite has a certain inhibitory effect on pathogens while efficiently supplying P and K. The addition of brown algae oligosaccharides can induce the synthesis of endogenous plant growth hormones (such as IAA, GA, etc.), thereby promoting the rooting, growth and overall growth of crops. Secondly, it can improve the utilization rate of fertilizers (especially nitrogen fertilizers);

[0037] (4) The biopesticide fertilizer composed of the above raw materials can realize the recycling and reuse of waste resources such as Chinese medicine residues, converting them into nutrients that can be absorbed by plants. At the same time, the active substances produced by fermentation have a good inhibitory effect on tobacco wilt pathogens. After application, it can achieve the dual use effects of fertilizer and pesticide, reduce the amount and frequency of pesticide use, and has significant economic and social benefits. It is worthy of application and promotion. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 This is a diagram showing the antibacterial effect of Bacillus megaterium of the present invention;

[0039] Figure 2 This is a diagram of the field planting experiment results of the present invention. DETAILED DESCRIPTION

[0040] The technical solution of the present invention is further described below in conjunction with specific embodiments, but is not limited thereto.

[0041] Example 1

[0042] A fermented compound medicinal fertilizer comprises the following raw materials: 50 parts of fermented Chinese medicinal residues, 10 parts of microbial agents, 10 parts of humic acid, and 10 parts of compound organic and inorganic nutrients.

[0043] The preparation method of the fermented Chinese medicine residue is as follows:

[0044] (1) Collecting Chinese medicine residue, mushroom residue and straw, crushing them into particles with a particle size of 0.5-1 cm, adding zeolite powder, sucrose and urea, stirring and mixing them evenly, adjusting the water content to 60%-70%, and sterilizing to obtain a premix;

[0045] (2) adjusting the pH of the premix to 5.8-6.5, adding anaerobic fermentation liquid, and performing anaerobically fermentation at 30-37° C. for 15 days to obtain pre-fermented medicinal residue;

[0046] (3) Turn the pre-fermented medicinal residues over, adjust the pH to 6.9-7.5, spray aerobic fermentation liquid, mix well, and then pile and ferment at room temperature. Turn the pile once a day to ventilate and cool the compost. The composting is fermented for 5-7 days. When the temperature of the compost pile no longer rises and begins to drop, the composting is finished. The compost is then air-dried or dried, and crushed to obtain the fermented Chinese medicinal residues.

[0047] The Chinese medicine residue is selected from waste materials after processing of honeysuckle, astragalus and codonopsis.

[0048] The mushroom residue is Flammulina velutipes residue; the mass ratio of Chinese medicine residue, mushroom residue and straw is 10:1.5.5:3, the added amount of zeolite powder is 1% of the mass of the Chinese medicine residue, the added amount of sucrose is 1% of the mass of the Chinese medicine residue, and the added amount of urea is 0.5% of the mass of the Chinese medicine residue.

[0049] The anaerobic fermentation bacterial liquid is a mixture of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus, and the volume ratio of the two is 1:1; the amount of the anaerobic fermentation bacterial liquid added is 0.1% of the mass of the premix; the preparation method of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus bacterial liquid is as follows: the seed liquid of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus are inoculated into the fermentation medium at an inoculation amount of 5% for fermentation, and the fermentation conditions are as follows: the initial pH of the fermentation tank is 7.0, the fermentation temperature is 30-37°C, and after the natural fermentation reaches pH4.5-4.9, alkali solution is added to maintain the constant pH of the fermentation liquid at 5.5-5.9 until the effective viable bacteria count of the fermentation liquid is 1×10 8 -1×10 9 CFU / mL is the fermentation end point, and the fermentation broths of the two bacteria are mixed in a volume ratio of 1:1 to obtain an anaerobic fermentation broth; the strain number of the Lactococcus lactis is CGMCC No.1.15072, and the original preservation time is January 25, 2015; the strain number of the Lactobacillus delbrueckii subspecies bulgaricus is CGMCC No.1.16075, and the original preservation time is February 15, 2017. Both Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus can be purchased through the China General Microorganism Collection Center without the need for repeated biological preservation.

[0050] The preparation method of Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus seed solution is as follows: the activated target strains are inoculated into MRS liquid culture medium, and cultured at 30-35°C for 2 days to obtain the seed solution of Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus.

[0051] The composition and preparation method of the fermentation medium are as follows: 10.0 g of peptone, 10.0 g of beef extract, 5.0 g of yeast extract, 20.0 g of glucose, 5.0 g of sodium acetate, 2.0 g of diamine citrate, 1.0 g of Tween-80, 0.4 g of dilithium hydrogen phosphate, 0.58 g of magnesium sulfate, 0.29 g of manganese sulfate, 20.0 g of calcium carbonate, and 15.0 g of agar are sequentially added into distilled water and the distilled water is supplemented to 1000 mL, the pH value is adjusted to 6.3, the mixture is heated after stirring, boiled for 2 min, and sterilized at 121° C. and 0.1 MPa for 30 min.

[0052] The solid-liquid ratio of the aerobic fermentation bacterial liquid and the pre-fermentation medicinal residue is 100 mL: 10 kg; the preparation method of the aerobic bacterial liquid is:

[0053] (1) inoculating a Bacillus megaferium strain with a preservation number of CGMCC No. 27648 stored at low temperature onto a beef extract peptone agar medium and culturing it at 26-32° C. for 15 hours, then picking a single colony and inoculating it onto an LB slant medium and culturing it at 26-32° C. for 10 hours, then washing the surface of the medium with sterile water, and using the eluate as an inoculum;

[0054] (2) adding 1% of the volume of the inoculum obtained in step (1) to the LB liquid, and culturing with shaking at 26-32° C. for 15 hours to obtain a seed liquid;

[0055] (3) The seed solution of step (2) was inoculated into corn starch medium at an inoculation ratio of 0.05%, and then cultured at 26-32°C and a shaking speed of 150-200 rpm for 30-40 hours to obtain an effective viable count of 1×10 8 -1×10 9 CFU / mL of aerobic fermentation broth.

[0056] The Bacillus megaterium has a deposit number of CGMCC No. 27648 and a deposit date of June 16, 2023. It is deposited in the General Microbiology Center of the China Culture Collection Administration, and the deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. This strain was donated by others.

[0057] The corn starch culture medium is composed of: 3 g beef extract, 25 g corn starch, 3 g CaCl2, 7 g yeast extract, and 1000 mL deionized water. The pH value of the culture medium is natural.

[0058] The microbial agent is obtained by mixing EM bacteria and Bacillus subtilis in a mass ratio of 1:5.

[0059] The composite organic-inorganic nutrient is obtained by mixing urea, potassium dihydrogen phosphite and brown algae oligosaccharide in a mass ratio of 5:0.5:0.1.

[0060] A method for preparing a fermentation-type compound medicinal fertilizer comprises the following steps:

[0061] A. Preparation of fermented Chinese medicine residues;

[0062] B. Mix and crush the fermented Chinese medicine residue, microbial agent, humic acid and composite organic and inorganic nutrients by weight, add the mixture into a disc granulator, start the disc granulator, and evenly spray water or starch solution on the powder mixture during the rotation of the disc until the mixed powder is rolled into particles; screen the particles, add the particles into a screening machine, and screen to obtain fertilizer particles with a particle size range of 3.5-4.5 mm; and dry to obtain finished medicinal fertilizer particles.

[0063] Example 2

[0064] A fermented compound medicinal fertilizer comprises the following raw materials: 65 parts of fermented Chinese medicinal residues, 7 parts of microbial agents, 20 parts of humic acid, and 5 parts of compound organic and inorganic nutrients.

[0065] The preparation method of the fermented Chinese medicine residue is as follows:

[0066] (1) Collecting Chinese medicine residue, mushroom residue and straw, crushing them into particles with a particle size of 0.5-1 cm, adding zeolite powder, sucrose and urea, stirring and mixing them evenly, adjusting the water content to 60%-70%, and sterilizing to obtain a premix;

[0067] (2) adjusting the pH of the premix to 5.8-6.5, adding anaerobic fermentation liquid, and performing anaerobically fermentation at 30-37° C. for 17 days to obtain pre-fermented medicinal residue;

[0068] (3) Turn the pre-fermented medicinal residues over, adjust the pH to 6.9-7.5, spray aerobic fermentation liquid, mix well, and then pile and ferment at room temperature. Turn the pile once a day to ventilate and cool the compost. The composting is fermented for 5-7 days. When the temperature of the compost pile no longer rises and begins to drop, the composting is finished. The compost is then air-dried or dried, and crushed to obtain the fermented Chinese medicinal residues.

[0069] The Chinese medicine residue is selected from the waste materials after processing of honeysuckle, astragalus, codonopsis, American ginseng, salvia miltiorrhiza, dandelion and isatis indigotica.

[0070] The mushroom residue is Pleurotus ostreatus residue; the mass ratio of Chinese medicine residue, mushroom residue and straw is 10:1.5:5, the added amount of zeolite powder is 1% of the mass of the Chinese medicine residue, the added amount of sucrose is 3% of the mass of the Chinese medicine residue, and the added amount of urea is 0.5% of the mass of the Chinese medicine residue.

[0071] The anaerobic fermentation bacterial liquid is a mixture of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus, and the volume ratio of the two is 1:1; the amount of the anaerobic fermentation bacterial liquid added is 0.1-0.5% of the mass of the premix; the preparation method of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus bacterial liquid is as follows: the seed liquid of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus are inoculated into the fermentation medium at an inoculation amount of 5% for fermentation, and the fermentation conditions are as follows: the initial pH of the fermentation tank is 7.0, the fermentation temperature is 30-37°C, and after the natural fermentation reaches pH4.5-4.9, alkali solution is added to maintain the constant pH of the fermentation liquid at 5.5-5.9 until the effective live bacteria count of the fermentation liquid reaches 1×10 8 -1×10 9 CFU / mL is the fermentation end point, and the fermentation broths of the two bacteria are mixed in a volume ratio of 1:1 to obtain an anaerobic fermentation broth; the strain number of the Lactococcus lactis is CGMCC No.1.15072, and the original preservation time is January 25, 2015; the strain number of the Lactobacillus delbrueckii subspecies bulgaricus is CGMCC No.1.16075, and the original preservation time is February 15, 2017. Both Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus can be purchased through the China General Microorganism Collection Center without the need for repeated biological preservation.

[0072] The preparation method of Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus seed solution is the same as that in Example 1.

[0073] The composition and preparation method of the fermentation medium are the same as those in Example 1.

[0074] The solid-liquid ratio of the aerobic fermentation bacterial liquid and the pre-fermentation medicinal residue is 100 mL: 10 kg; the preparation method of the aerobic bacterial liquid is:

[0075] (1) inoculating a Bacillus megaferium strain with a preservation number of CGMCC No. 27648 stored at low temperature onto a beef extract peptone agar medium and culturing it at 26-32° C. for 15 hours, then picking a single colony and inoculating it onto an LB slant medium and culturing it at 26-32° C. for 15 hours, then washing the surface of the medium with sterile water, and using the eluate as an inoculum;

[0076] (2) adding 1% of the volume of the inoculum obtained in step (1) to the LB liquid, and culturing with shaking at 26-32° C. for 20 hours to obtain a seed liquid;

[0077] (3) The seed solution of step (2) was inoculated into corn starch medium at a 1% inoculation ratio, and then cultured at 26-32°C and a shaking speed of 150-200 rpm for 30-40 hours to obtain an effective viable count of 1×10 8 -1×10 9 CFU / mL of aerobic fermentation broth.

[0078] The Bacillus megaterium is the same as in Example 1.

[0079] The composition of the corn starch culture medium is the same as that of Example 1.

[0080] The microbial agent is obtained by mixing EM bacteria and Bacillus subtilis in a mass ratio of 1:5.

[0081] The composite organic-inorganic nutrient is obtained by mixing urea, potassium dihydrogen phosphite and brown algae oligosaccharide in a mass ratio of 5:0.5:0.3.

[0082] A method for preparing a fermentation-type compound medicinal fertilizer comprises the following steps:

[0083] A. Preparation of fermented Chinese medicine residues;

[0084] B. Mix and crush the fermented Chinese medicine residue, microbial agent, humic acid and composite organic and inorganic nutrients by weight, add the mixture into a disc granulator, start the disc granulator, and evenly spray water or starch solution on the powder mixture during the rotation of the disc until the mixed powder is rolled into particles; screen the particles, add the particles into a screening machine, and screen to obtain fertilizer particles with a particle size range of 3.5-4.5 mm; and dry to obtain finished medicinal fertilizer particles.

[0085] Example 3

[0086] A fermented compound medicinal fertilizer comprises the following raw materials: 80 parts of fermented Chinese medicinal residues, 5 parts of microbial agents, 15 parts of humic acid, and 8 parts of compound organic and inorganic nutrients.

[0087] The preparation method of the fermented Chinese medicine residue is as follows:

[0088] (1) Collecting Chinese medicine residue, mushroom residue and straw, crushing them into particles with a particle size of 0.5-1 cm, adding zeolite powder, sucrose and urea, stirring and mixing them evenly, adjusting the water content to 60%-70%, and sterilizing to obtain a premix;

[0089] (2) adjusting the pH of the premix to 5.8-6.5, adding anaerobic fermentation liquid, and performing anaerobically fermentation at 30-37° C. for 20 days to obtain pre-fermented medicinal residue;

[0090] (3) Turn the pre-fermented medicinal residues over, adjust the pH to 6.9-7.5, spray aerobic fermentation liquid, mix well, and then pile and ferment at room temperature. Turn the pile once a day to ventilate and cool the compost. The composting is fermented for 5-7 days. When the temperature of the compost pile no longer rises and begins to drop, the composting is finished. The compost is then air-dried or dried, and crushed to obtain the fermented Chinese medicinal residues.

[0091] The Chinese medicine residue is selected from the waste materials after processing of honeysuckle, astragalus, codonopsis, American ginseng, salvia miltiorrhiza, dandelion and isatis indigotica.

[0092] The mushroom residue is a mixture of enoki mushroom residue, oyster mushroom residue and king oyster mushroom residue; the mass ratio of traditional Chinese medicine residue, mushroom residue and straw is 10:2.5:5, the added amount of zeolite powder is 1% of the mass of the traditional Chinese medicine residue, the added amount of sucrose is 3% of the mass of the traditional Chinese medicine residue, and the added amount of urea is 1.5% of the mass of the traditional Chinese medicine residue.

[0093] The anaerobic fermentation bacterial liquid is a mixture of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus, and the volume ratio of the two is 1:1; the amount of the anaerobic fermentation bacterial liquid added is 0.1-0.5% of the mass of the premix; the preparation method of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus bacterial liquid is as follows: the seed liquid of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus are inoculated into the fermentation medium at an inoculation amount of 5% for fermentation, and the fermentation conditions are as follows: the initial pH of the fermentation tank is 7.0, the fermentation temperature is 30-37°C, and after the natural fermentation reaches pH4.5-4.9, alkali solution is added to maintain the constant pH of the fermentation liquid at 5.5-5.9 until the effective live bacteria count of the fermentation liquid reaches 1×10 8 -1×10 9 CFU / mL is the fermentation end point, and the fermentation broths of the two bacteria are mixed in a volume ratio of 1:1 to obtain an anaerobic fermentation broth; the strain number of the Lactococcus lactis is CGMCC No.1.15072, and the original preservation time is January 25, 2015; the strain number of the Lactobacillus delbrueckii subspecies bulgaricus is CGMCC No.1.16075, and the original preservation time is February 15, 2017. Both Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus can be purchased through the China General Microorganism Collection Center without the need for repeated biological preservation.

[0094] The preparation method of Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus seed solution is the same as that in Example 1.

[0095] The composition and preparation method of the fermentation medium are the same as those in Example 1.

[0096] The solid-liquid ratio of the aerobic fermentation bacterial liquid and the pre-fermentation medicinal residue is 100 mL: 10 kg; the preparation method of the aerobic bacterial liquid is:

[0097] (1) inoculating a Bacillus megaferium strain with a preservation number of CGMCC No. 27648 stored at low temperature onto a beef extract peptone agar medium and culturing it at 26-32° C. for 20 hours, then picking a single colony and inoculating it onto an LB slant medium and culturing it at 26-32° C. for 20 hours, then washing the surface of the medium with sterile water, and using the eluate as an inoculum;

[0098] (2) adding 1% of the volume of the inoculum obtained in step (1) to the LB liquid, and culturing with shaking at 26-32° C. for 20 hours to obtain a seed liquid;

[0099] (3) The seed solution of step (2) was inoculated into corn starch medium at an inoculation ratio of 0.05-1%, and then cultured at 26-32°C and a shaking speed of 150-200 rpm for 30-40 hours to obtain an effective viable count of 1×10 8 -1×10 9 CFU / mL of aerobic fermentation broth.

[0100] The Bacillus megaterium is the same as in Example 1.

[0101] The corn starch culture medium is the same as that in Example 1.

[0102] The microbial agent is obtained by mixing EM bacteria and Bacillus subtilis in a mass ratio of 1:5.

[0103] The composite organic-inorganic nutrient is obtained by mixing urea, potassium dihydrogen phosphite and brown algae oligosaccharide in a mass ratio of 5:0.5:0.3.

[0104] A method for preparing a fermentation-type compound medicinal fertilizer comprises the following steps:

[0105] A. Preparation of fermented Chinese medicine residues;

[0106] B. Mix and crush the fermented Chinese medicine residue, microbial agent, humic acid and composite organic and inorganic nutrients by weight, add the mixture into a disc granulator, start the disc granulator, and evenly spray water or starch solution on the powder mixture during the rotation of the disc until the mixed powder is rolled into particles; screen the particles, add the particles into a screening machine, and screen to obtain fertilizer particles with a particle size range of 3.5-4.5 mm; and dry to obtain finished medicinal fertilizer particles.

[0107] Comparative Example 1

[0108] A fermented compound medicinal fertilizer comprises the following raw materials: 50 parts of fermented Chinese medicinal residues, 10 parts of microbial agents, 10 parts of humic acid, and 10 parts of compound organic and inorganic nutrients.

[0109] The preparation method of the fermented Chinese medicine residue is as follows:

[0110] (1) Collecting Chinese medicine residue, mushroom residue and straw, crushing them into particles with a particle size of 0.5-1 cm, adding zeolite powder, sucrose and urea, stirring and mixing them evenly, adjusting the water content to 60%-70%, and sterilizing to obtain a premix;

[0111] (2) The premix is ​​adjusted to a pH of 6.9-7.5, sprayed with aerobic fermentation liquid, mixed evenly, and then piled and fermented at room temperature, the pile is turned over once a day, and the compost is ventilated and cooled. The compost is fermented for 20-22 days. When the temperature no longer changes, the composting is completed, air-dried or oven-dried, and crushed to obtain fermented Chinese medicine residue.

[0112] In this comparative example, except that only one aerobic fermentation step is provided, the other raw materials and preparation process are the same as those of Example 1.

[0113] Comparative Example 2

[0114] In this comparative example, except that only Lactococcus lactis was used in the anaerobic fermentation process, the other raw materials and preparation process were the same as those in Example 1. That is:

[0115] A fermented compound medicinal fertilizer comprises the following raw materials: 50 parts of fermented Chinese medicinal residues, 10 parts of microbial agents, 10 parts of humic acid, and 10 parts of compound organic and inorganic nutrients.

[0116] The preparation method of the fermented Chinese medicine residue is as follows:

[0117] (1) Collecting Chinese medicine residue, mushroom residue and straw, crushing them into particles with a particle size of 0.5-1 cm, adding zeolite powder, sucrose and urea, stirring and mixing them evenly, adjusting the water content to 60%-70%, and sterilizing to obtain a premix;

[0118] (2) adjusting the pH of the premix to 5.8-6.5, adding anaerobic fermentation liquid, and performing anaerobically fermentation at 30-37° C. for 15 days to obtain pre-fermented medicinal residue;

[0119] (3) Turn the pre-fermented medicinal residues over, adjust the pH to 6.9-7.5, spray aerobic fermentation liquid, mix well, and then pile and ferment at room temperature. Turn the pile once a day to ventilate and cool the compost. The composting is fermented for 5-7 days. When the temperature of the compost pile no longer rises and begins to drop, the composting is finished. The compost is then air-dried or dried, and crushed to obtain the fermented Chinese medicinal residues.

[0120] The anaerobic fermentation bacterial liquid is Lactococcus lactis; the amount of the anaerobic fermentation bacterial liquid added is 0.1% of the mass of the premix; the preparation method of the Lactococcus lactis liquid is as follows: Lactococcus lactis is inoculated into a fermentation medium at an inoculation amount of 5% for fermentation, and the fermentation conditions are as follows: the initial pH of the fermentation tank is 7.0, the fermentation temperature is 30-37°C, and after natural fermentation to pH 4.5-4.9, alkali solution is added to maintain the constant pH of the fermentation liquid at 5.5-5.9 until the effective viable bacteria count of the fermentation liquid is 1×10 8 -1×10 9 CFU / mL is the fermentation end point, and the anaerobic fermentation bacterial liquid is obtained; the strain number of Lactococcus lactis is CGMCC No.1.15072, and the original preservation time is January 25, 2015. Lactococcus lactis can be purchased through the China General Microorganism Collection Center without repeated biological preservation.

[0121] Comparative Example 3

[0122] In this comparative example, except that only Lactobacillus delbrueckii subspecies bulgaricus was used in the anaerobic fermentation process, the remaining raw materials and preparation process were the same as those in Example 1. That is:

[0123] The anaerobic fermentation bacterial liquid is Lactobacillus delbrueckii subspecies bulgaricus bacterial liquid; the amount of the anaerobic fermentation bacterial liquid added is 0.1% of the mass of the premix; the preparation method of the Lactobacillus delbrueckii subspecies bulgaricus bacterial liquid is as follows: the seed liquid of Lactobacillus delbrueckii subspecies bulgaricus is inoculated into the fermentation medium at an inoculation amount of 5% for fermentation, and the fermentation conditions are as follows: the initial pH of the fermentation tank is 7.0, the fermentation temperature is 30-37°C, and after natural fermentation to pH4.5-4.9, alkali solution is added to maintain the constant pH of the fermentation liquid at 5.5-5.9 until the effective viable bacteria count of the fermentation liquid is 1×10 8 -1×10 9 CFU / mL is the fermentation end point, and the anaerobic fermentation bacterial liquid is obtained; the strain number of the Lactobacillus delbrueckii subspecies bulgaricus is CGMCC No.1.16075, and the original preservation time is February 15, 2017. Lactobacillus delbrueckii subspecies bulgaricus can be purchased through the China General Microorganism Collection Center without the need for repeated biological preservation.

[0124] Comparative Example 4

[0125] In this comparative example, except that the ratio of the lactic acid Lactococcus liquid to the Lactobacillus delbrueckii subspecies bulgaricus was changed to 2:1 during the anaerobic fermentation process, the other raw materials and preparation process were the same as those in Example 1. That is:

[0126] The anaerobic fermentation bacterial liquid is a mixture of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus, and the volume ratio of the two is 2:1; the amount of the anaerobic fermentation bacterial liquid added is 0.1% of the mass of the premix; the preparation method of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus bacterial liquid is as follows: the seed liquid of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus are inoculated into the fermentation medium at an inoculation amount of 5% for fermentation, and the fermentation conditions are as follows: the initial pH of the fermentation tank is 7.0, the fermentation temperature is 30-37°C, and after the natural fermentation reaches pH4.5-4.9, alkali solution is added to maintain the constant pH of the fermentation liquid at 5.5-5.9 until the effective viable bacteria count of the fermentation liquid is 1×10 8 -1×10 9 CFU / mL is the fermentation end point, and the fermentation broths of the two bacteria are mixed in a volume ratio of 2:1 to obtain an anaerobic fermentation broth; the strain number of the Lactococcus lactis is CGMCC No.1.15072, and the original preservation time is January 25, 2015; the strain number of the Lactobacillus delbrueckii subspecies bulgaricus is CGMCC No.1.16075, and the original preservation time is February 15, 2017. Both Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus can be purchased through the China General Microorganism Collection Center without the need for repeated biological preservation.

[0127] Comparative Example 5

[0128] In this comparative example, except that the ratio of the lactic acid Lactococcus liquid to the Lactobacillus delbrueckii subspecies bulgaricus was changed to 1:2 during the anaerobic fermentation process, the other raw materials and preparation process were the same as those in Example 1. That is:

[0129] The anaerobic fermentation bacterial liquid is a mixture of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus, and the volume ratio of the two is 1:2; the amount of the anaerobic fermentation bacterial liquid added is 0.1% of the mass of the premix; the preparation method of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus bacterial liquid is as follows: the seed liquid of Lactococcus lactis and Lactobacillus delbruechii subsp.bulgaricus are inoculated into the fermentation medium at an inoculation amount of 5% for fermentation, and the fermentation conditions are as follows: the initial pH of the fermentation tank is 7.0, the fermentation temperature is 30-37°C, and after the natural fermentation reaches pH4.5-4.9, alkali solution is added to maintain the constant pH of the fermentation liquid at 5.5-5.9 until the effective viable bacteria count of the fermentation liquid is 1×10 8 -1×10 9CFU / mL is the fermentation end point, and the fermentation broths of the two bacteria are mixed in a volume ratio of 1:2 to obtain an anaerobic fermentation broth; the strain number of the Lactococcus lactis is CGMCC No.1.15072, and the original preservation time is January 25, 2015; the strain number of the Lactobacillus delbrueckii subspecies bulgaricus is CGMCC No.1.16075, and the original preservation time is February 15, 2017. Both Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus can be purchased through the China General Microorganism Collection Center without the need for repeated biological preservation.

[0130] Comparative Example 6

[0131] In this comparative example, except that the commercially available Bacillus megaterium strain was used in the aerobic fermentation bacterial liquid, the other raw materials and preparation process were the same as those in Example 1. That is:

[0132] The preparation method of the aerobic bacterial liquid is as follows:

[0133] (1) inoculating a low-temperature stored Bacillus megaterium strain on beef extract peptone agar medium and culturing it at 26-32° C. for 15 hours, then picking a single colony and inoculating it on LB slant medium and culturing it at 26-32° C. for 10 hours, then washing the surface of the medium with sterile water, and using the eluate as the inoculum;

[0134] (2) adding 1% of the volume of the inoculum obtained in step (1) to the LB liquid, and culturing with shaking at 26-32° C. for 15 hours to obtain a seed liquid;

[0135] (3) The seed solution of step (2) was inoculated into corn starch medium at an inoculation ratio of 0.05%, and then cultured at 26-32°C and a shaking speed of 150-200 rpm for 30-40 hours to obtain an effective viable count of 1×10 8 -1×10 9 CFU / mL of aerobic fermentation broth.

[0136] The Bacillus megaterium strain was purchased from Shanghai Collection Biotechnology Center.

[0137] Performance Testing

[0138] Verification of antibacterial performance of strains: Co-culture method: Inoculate the Bacillus megaterium strain with the preservation number of CGMCC No. 27648 on beef extract peptone agar medium and culture it at 26-32°C for 15-20 hours to obtain a bacterial cake. Mix 1mL of tobacco Ralstonia solanacearum liquid with 99mL BGT medium and pour it on the plate, place the bacterial cake in the center of the plate, use blank Gao's No. 1 solid medium as a control, culture at 28°C for 2d, measure the size of the inhibition zone by the cross method, and set up 3 replicates for each group of experiments. Use the Oxford cup method to retest the antibacterial effect: Use a sterile puncher (6mm) to punch the Bacillus megaterium strain and inoculate it in Gao's No. 1 liquid medium, culture it at 28°C and 160r / min for 7d, draw 2mL of fermentation liquid and centrifuge it at 10000r / min for 10min, take the supernatant, filter and sterilize it with a 0.22μm microporous filter membrane to obtain the fermentation filtrate. Mix 1 mL of tobacco Ralstonia solanacearum bacterial liquid with 99 mL of BGT medium and pour on the plate. Place a sterilized Oxford cup in the center of the plate. Add 200 μL of fermentation filtrate to the Oxford cup. Use blank Gao's No. 1 liquid medium as a control. Culture at 28°C for 2 days. Measure the diameter of the inhibition zone by the cross method. Set up 3 replicates for each group of experiments.

[0139] After testing, it was found that the diameter of the antibacterial zone around the Bacillus megaterium cake produced by the co-culture method was 45.5±0.1mm, and the diameter of the antibacterial zone produced by the Oxford cup method was 47.9±0.1mm. The strain has a good inhibitory effect on Ralstonia solanacearum. Figure 1 shown.

[0140] Verification of enzyme production capacity of two strains in anaerobic fermentation broth:

[0141] The method for preparing seed solution of Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus is as follows: inoculate the activated target bacteria into MRS liquid culture medium, culture at 30-35°C for 2-3 days, and obtain the seed solution of Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus.

[0142] The seed liquid of different experimental groups was added to the fermentation enzyme production medium and cultured statically at (30-35)°C. Samples were taken after 5 days and centrifuged at 4°C and 6000r / min for 10min. The supernatant was taken for the determination of cellulase and ligninase activities.

[0143] Set up four experimental groups, each experimental group is repeated 5 times, and the results are averaged. The experimental groups are:

[0144] S1: Lactococcus lactis seed solution;

[0145] S2: Lactobacillus delbrueckii subsp. bulgaricus seed solution;

[0146] S3: a mixture of Lactococcus lactis seed solution and Lactobacillus delbrueckii subsp. bulgaricus seed solution in a volume ratio of 2:1;

[0147] S4: a mixture of Lactococcus lactis seed solution and Lactobacillus delbrueckii subsp. bulgaricus seed solution in a volume ratio of 1:2;

[0148] S5: a mixture of Lactococcus lactis seed solution and Lactobacillus delbrueckii subspecies bulgaricus seed solution in a volume ratio of 1:1;

[0149] The composition of the fermentation medium for enzyme production is as follows: liquid fermentation medium (1L): 200g peeled potatoes, 3g KH2PO4, 1g MgSO4·7H2O, 5g ammonium tartrate, 20g corn straw powder, adjusted to pH 5.0 with H2SO4. 50mL of medium was placed in each 250mL Erlenmeyer flask. The medium was heated at 1×10 5 Sterilize at 400 Pa for 30 min.

[0150] Determination method:

[0151] Method for determining CMC enzyme activity. Add 4 mL of 0.625% CMC-Na solution prepared with pH 6.00.2 mol / L sodium phosphate buffer to a 25 mL stoppered test tube and preheat in a 60°C water bath for 5 min. Add 1.0 mL of the original enzyme solution, shake well, react accurately at 60°C for 30 min, then add 2 mL of DNS reagent, put it in a boiling water bath for 5 min to develop color, immediately place it in ice water to cool to room temperature, make up to 20.0 mL, and shake well. Use the enzyme solution inactivated in a boiling water bath for 5 min under the same conditions as the blank, and measure the absorbance at a wavelength of 490 nm. Then calculate the amount of glucose generated on the corresponding glucose standard curve. Definition of enzyme activity unit: Under the above reaction conditions, the amount of enzyme required to produce 1 μg of reducing sugar per minute is 1 cellulase activity unit (U).

[0152] Determination of ligninase activity. Laccase: 10mL reaction system contains 50mmol / L sodium succinate buffer solution (pH4.5), 0.4mmol / L guaiacol and 0.5mL enzyme solution. After reacting at 30℃ for 30min, the OD value is measured at 465nm. Enzyme activity is defined as: with the inactivated enzyme solution reaction mixture as the control, the optical density increase of 0.01 per minute is 1 enzyme activity unit.

[0153] Lignin peroxidase (resveratrol method): 2mmol / L resveratrol, 50mmol / L tartaric acid buffer (pH2.5), 0.4mmol / L H2O2, measured at room temperature, OD value measured at 310nm UV light. Enzyme activity is defined as: 1 enzyme activity unit is defined as an increase of 0.01 in optical density per minute.

[0154] Manganese-dependent peroxidase: The reaction system contains 3.4 mL of 50 mmol / L sodium lactate buffer (pH 4.5), 0.1 mL of 1.6 mmol / L manganese sulfate solution, and 0.1 mL of 1.6 mmol H2O2 to start the reaction. The OD change value is measured at room temperature under 240 nm ultraviolet light for 4 minutes. Enzyme activity is defined as: an increase of 0.01 in optical density per minute is 1 enzyme activity unit.

[0155] Table 1 Enzyme activity determination results

[0156]

[0157] Product index determination:

[0158] Experimental group: The medicated fertilizer was prepared according to the method of Examples 1-3 and Comparative Examples 1-6 of the present invention.

[0159] Seed germination test and product physical and chemical index determination:

[0160] Sterile distilled water was added to the sample at a volume ratio of 1:10. 5 mL of compost extract was taken with a pipette and added to a sterile culture dish lined with filter paper. 10 plump pakchoy seeds were evenly placed on the surface. Sterile water was used as a control. Three parallel tests were conducted for each treatment. The culture dish was placed in a constant temperature incubator at 25°C for 48 hours to determine the seed germination rate GI. It is generally believed that a seed germination rate of more than 50% indicates that the organic fertilizer is basically mature. Other relevant indicators are tested for compost product performance according to the corresponding methods in the national quality standard for organic fertilizers NY525-2021.

[0161] Table 2 Performance indexes of pesticide fertilizer

[0162]

[0163] Table 3 Performance index of pesticide fertilizer

[0164]

[0165] Planting experiment:

[0166] The tobacco variety is Yunyan 87, provided by the Jinggu County Branch of Pu'er City Company of Yunnan Tobacco Company.

[0167] Test location:

[0168] A diseased tobacco field in Jinggu Dai and Yi Autonomous County, Pu'er City, Yunnan Province. Tobacco bacterial wilt has occurred in this experimental field for many years, and the incidence is serious.

[0169] A randomized block design was used with 10 treatments in total. The control group (CK) was treated with local conventional fertilization (nitrogen application rate of 105 kg / hm2).2 , the fertilizer ratio is m(N):m(P2O5):m(K2O)=1:1.2:3);

[0170] The treatment groups were:

[0171] A1: Example 1+conventional fertilization;

[0172] A2: Example 2+conventional fertilization;

[0173] A3: Example 3+conventional fertilization;

[0174] A4: Comparative Example 1 + conventional fertilization;

[0175] A5: Comparative Example 2 + conventional fertilization;

[0176] A6: Comparative Example 3 + conventional fertilization;

[0177] A7: Comparative Example 4 + conventional fertilization;

[0178] A8: Comparative Example 5 + conventional fertilization;

[0179] A9: Comparative Example 6 + conventional fertilization;

[0180] CK: conventional fertilization;

[0181] Each treatment group was repeated 3 times, with 5 rows in each plot, 12 tobacco plants in each row, 1.20m row spacing, and 0.55m plant spacing. Transplanting was carried out on May 1, 2023, and the treatment group was fertilized with fungicide (5g / plant) 25 days after the tobacco seedlings were transplanted. Other cultivation and management measures were carried out according to local conventional methods. Field survey:

[0182] The occurrence of tobacco bacterial wilt at maturity was investigated according to GB / T 23222-2008 "Grading and Investigation Methods for Tobacco Pests and Diseases", and the incidence rate, disease index and control effect were calculated. Figure 2 shown.

[0183] From the data in the figure, we can see that the fertilizer of the present invention achieves effective inhibition of tobacco bacterial wilt, and changes the fermentation process and the comparative documents 1-6 of the composition of aerobic fermentation strains. Due to the decline in fermentation effect and the weakening of the synergistic effect between strains, the active substances produced by fermentation are reduced, thus leading to a decline in the prevention effect.

[0184] It should be noted that the above embodiments are only partial embodiments of the preferred methods of implementing the present invention, rather than all embodiments. Obviously, based on the above embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

Claims

1. A fermentation-type compound fertilizer, characterized in that: The invention comprises the following raw materials: 50-80 parts of fermented Chinese medicine residues, 5-10 parts of microbial agents, 10-20 parts of humic acid and 5-10 parts of composite organic and inorganic nutrients.

2. The fermentation-type compound fertilizer according to claim 1, characterized in that: The preparation method of the fermented Chinese medicine residue is as follows: (1) Collecting Chinese medicine residue, mushroom residue and straw, crushing them into particles with a particle size of 0.5-1 cm, adding zeolite powder, sucrose and urea, stirring and mixing them evenly, adjusting the water content to 60%-70%, and sterilizing to obtain a premix; (2) adjusting the pH of the premix to 5.8-6.5, adding anaerobic fermentation liquid, and performing anaerobically fermentation at 30-37° C. for 15-20 days to obtain pre-fermented medicinal residue; (3) Turn the pre-fermented medicinal residues over, adjust the pH to 6.9-7.5, spray aerobic fermentation liquid, mix well, and then pile and ferment at room temperature. Turn the pile once a day to ventilate and cool the compost. The composting is fermented for 5-7 days. When the temperature of the compost pile no longer rises and begins to drop, the composting is finished. The compost is then air-dried or dried, and crushed to obtain the fermented Chinese medicinal residues.

3. The fermentation-type compound fertilizer according to claim 2, characterized in that: The Chinese medicine residue is selected from the waste materials after processing one or a combination of several of honeysuckle, astragalus, codonopsis, American ginseng, salvia miltiorrhiza, dandelion and isatis indigotica.

4. The fermentation-type compound fertilizer according to claim 2, characterized in that: The mushroom residue is one or more of the group consisting of enoki mushroom residue, oyster mushroom residue, and king oyster mushroom residue; the mass ratio of the Chinese medicine residue, mushroom residue, and straw is 10:1.5-2.5:3-5; the added amount of zeolite powder is 1% of the mass of the Chinese medicine residue; the added amount of sucrose is 1-3% of the mass of the Chinese medicine residue; and the added amount of urea is 0.5-1.5% of the mass of the Chinese medicine residue.

5. The fermentation-type compound fertilizer according to claim 2, characterized in that: The anaerobic fermentation bacterial liquid is a Lactococcus lactis liquid ( Lactococcus lactis ) and Lactobacillus delbrueckii subsp. bulgaricus ( Lactobacillus delbruechii subsp. bulgaricus ) in a volume ratio of 1:1; the amount of anaerobic fermentation bacterial solution added is 0.1-0.5% of the mass of the premix; the preparation method of Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus bacterial solution is as follows: the seed solution prepared after activation of Lactococcus lactis and Lactobacillus delbrueckii subspecies bulgaricus is inoculated into the fermentation medium at an inoculation rate of 5% for fermentation, and the fermentation conditions are as follows: the initial pH of the fermentation tank is 7.0, the fermentation temperature is 30-37°C, and after natural fermentation to pH4.5-4.9, alkali solution is added to maintain the constant pH of the fermentation liquid at 5.5-5.9 until the effective viable bacteria count of the fermentation liquid is 1×10 8 -1×10 9 CFU / mL is the fermentation endpoint. The fermentation broths of the two bacteria are mixed in a volume ratio of 1:1 to obtain an anaerobic fermentation broth; the strain number of the Lactococcus lactis is CGMCC No.1.15072, and the strain number of the Lactobacillus delbrueckii subspecies bulgaricus is CGMCC No.1.16075.

6. The fermentation-type compound fertilizer according to claim 2, characterized in that: The solid-liquid ratio of the aerobic fermentation bacterial liquid and the pre-fermented medicinal residue is 100 mL: 10 kg; the preparation method of the aerobic bacterial liquid is: (1) The Bacillus megaterium strain with the deposit number of CGMCC No. 27648 stored at low temperature ( Bacillus megaferium ) inoculated on beef extract peptone agar medium and cultured at 26-32°C for 15-20 hours, then picked a single colony and inoculated on LB slant medium and cultured at 26-32°C for 10-20 hours, then washed the surface of the medium with sterile water, and the eluate was used as the inoculation solution; (2) adding 1% of the volume of the inoculum obtained in step (1) to the LB liquid, and culturing with shaking at 26-32° C. for 15-20 hours to obtain a seed liquid; (3) The seed solution of step (2) was inoculated into corn starch medium at an inoculation ratio of 0.05-1%, and then cultured at 26-32°C and a shaking speed of 150-200 rpm for 30-40 hours to obtain an effective viable count of 1×10 8 -1×10 9 CFU / mL of aerobic fermentation broth.

7. The fermentation-type compound fertilizer according to claim 1, characterized in that: The microbial agent is obtained by mixing EM bacteria and Bacillus subtilis in a mass ratio of 1:

5.

8. The fermentation-type compound fertilizer according to claim 1, characterized in that: The composite organic-inorganic nutrient is obtained by mixing urea, potassium dihydrogen phosphite and brown algae oligosaccharide in a mass ratio of 5:0.5:0.1-0.

3.

9. A method for preparing the fermentation-type compound medicinal fertilizer according to any one of claims 1 to 8, characterized in that: The following steps are involved: A. Preparation of fermented Chinese medicine residues; B. Mix and crush the fermented Chinese medicine residue, microbial agent, humic acid and composite organic and inorganic nutrients by weight, add the mixture into a disc granulator, start the disc granulator, and evenly spray water or starch solution on the powder mixture during the rotation of the disc until the mixed powder is rolled into particles; screen the particles, add the particles into a screening machine, and screen to obtain fertilizer particles with a particle size range of 3.5-4.5 mm; and dry to obtain finished medicinal fertilizer particles.

10. Use of the fermented compound fertilizer according to any one of claims 1 to 8 in preventing and treating tobacco bacterial wilt.

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