Biological material combination and method for anaerobic disinfection of soil

Abstract

The invention belongs to the field of soil treatment, and particularly relates to a biological material combination and method for anaerobic disinfection of soil. According to the invention, anaerobic bacteria, enzyme and small molecular biochar are combined and applied to soil anaerobic disinfection, so that the efficiency of soil anaerobic disinfection is improved, and the soil film mulching time is shortened; the soil activity can be improved, the soil acidity and alkalinity are adjusted, salinization is reduced, and the utilization rate of the fertilizer is increased; the soil-borne disease occurrence degree can be reduced in the plant growth process, root growth is promoted, premature senility is prevented, and the yield and quality of crops are improved. By applying the biological material composition, the problem of continuous cropping soil obstacles can be effectively prevented and treated, the disinfection effect is better than that of traditional anaerobic disinfection treatment, the method is more environmentally friendly than chemical disinfection treatment, use of pesticide and chemical fertilizer can be reduced, and the method is a novel soil disinfection technology worthy of popularization and application.

Description

Technical Field

[0001] The invention belongs to the field of soil treatment, and in particular relates to a biological material combination and method that can be used for anaerobic disinfection of soil. Background Art

[0002] With the popularity of greenhouse planting, the soil in the greenhouse has been planted for many years and has suffered from continuous cropping. Continuous cropping refers to the phenomenon that after the same kind or family of closely related crops are planted continuously in the same greenhouse soil, according to daily planting management, it will cause poor crop growth, reduced yield, reduced fruit quality, and aggravated soil-borne pests and diseases. Continuous cropping is also called repeated cropping. The soil of continuous cropping usually accumulates a large number of pathogens. After long-term continuous cropping of crops, the pests and diseases in the greenhouse gradually increase, causing serious yield reduction. People realize that the soil needs to be disinfected. Early soil disinfection generally applies soil fumigants (such as methyl bromide, dazomethane, and methacryloyl) to the soil to prevent and control soil-borne diseases, and the application of chemical agents and the environmental safety problems caused by pesticide residues in the later period. In order to solve this problem, compared with the disinfection of chemical agents, biological and environmentally friendly disinfection technologies have gradually received attention, such as anaerobic disinfection technology, film sealing and sun drying method, and biological improvement method.

[0003] As an environmentally friendly soil disinfection technology, anaerobic disinfection technology can effectively prevent and treat most pathogenic microorganisms such as pathogens and nematodes in the soil. This technology creates an anaerobic environment, reduces the proportion of aerobic pathogenic microorganisms, and reduces the redox potential of the soil through the volatile organic acids and alcohols produced by anaerobic fermentation metabolism, thereby inhibiting soil pathogens in continuously cropped soil.

[0004] The disinfection effect of anaerobic disinfection technology is related to factors such as soil texture, carbon source type, soil temperature, humidity, film type and sealing time during implementation. Anaerobic disinfection technology is suitable for soil disinfection in high temperature seasons. The disinfection time generally lasts 30 to 45 days, and the disinfection duration is relatively long; the original anaerobic microbial structure and number of the soil limit the disinfection effect. In view of the above factors, the disinfection effect of anaerobic disinfection technology on soil pathogens is unstable, and the market response is general, which seriously affects the application and promotion of this technology in production. Summary of the invention

[0005] The purpose of the present invention is to provide a biological material combination and method that can be used for anaerobic disinfection of soil to make up for the shortcomings of conventional anaerobic disinfection technology.

[0006] Specifically, the present invention first provides a biomaterial combination, which includes: anaerobic bacteria, enzymes and small molecule biocarbon; wherein the anaerobic bacteria are a mixture of yeast, lactobacillus and methanobacterium, and the yeast is one or more of Saccharomyces cerevisiae, Pichia pastoris, and Pichia kluyveri; the lactobacillus is one or more of Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus bulgaricus; the methanobacterium is Mathanobacteriumthermoautrophicum, Methanobucterium bryantii, Methanobacterium formicum, and Methanobacterium tumefaciens. ormicicum); the enzyme is selected from one or more of protease, chitinase, lipase, endo-β-1,3 glucanase, endo-β-1,4 glucanase, and amylase; the small molecule biocarbon is selected from one or more of glucose, sucrose, fructose, humic acid, and chitosan oligosaccharide.

[0007] The invention also provides a soil anaerobic disinfection method, which comprises: applying the biological material combination into the soil.

[0008] The present invention combines anaerobic bacteria, enzymes and small molecule biochar for anaerobic soil disinfection, and inhibits the growth of pathogens by producing volatile organic acids and alcohols, thereby improving the efficiency of anaerobic soil disinfection and shortening the soil film covering time; it can also improve the vitality of the soil, adjust the soil pH, reduce salinization, and improve the utilization rate of fertilizers; and it is beneficial to reduce the occurrence of soil-borne diseases during plant growth, promote root growth, prevent premature aging, and improve the yield and quality of crops. It has been verified by the examples that the application of the biomaterial combination of the present invention can effectively prevent and control the problem of continuous cropping soil obstacles, and the disinfection effect is better than that of traditional anaerobic disinfection treatment, and it is more environmentally friendly than chemical disinfection, and can reduce the use of pesticides and fertilizers. It is a new soil disinfection technology worthy of promotion and application. DETAILED DESCRIPTION

[0009] The specific embodiments of the present invention are described in detail below. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention. Those skilled in the art may make various modifications and changes to the present invention without departing from the scope or spirit of the present invention. For example, a feature described or illustrated as part of one embodiment may be used in another embodiment to produce a further embodiment.

[0010] Unless otherwise indicated, the meaning of all terms (including technical and scientific terms) used to disclose the present invention is the same as that commonly understood by those of ordinary skill in the art to which the present invention belongs. By way of further guidance, the following definitions are used to better understand the teachings of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0011] The terms "and / or", "or / and", and "and / or" used in this article include any one of two or more related listed items, and also include any and all combinations of related listed items, and the arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions that are all connected by "logical and", and undoubtedly includes technical solutions that are all connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution that is all connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the combination of four of A, B, C, and D (that is, the technical solution that is all connected by "logical AND").

[0012] As used herein, the terms "comprising", "including" and "comprising" are synonymous and are inclusive or open-ended and do not exclude additional, unrecited members, elements or method steps.

[0013] Numerical ranges expressed as endpoints herein include all numbers and fractions subsumed within the range, as well as the recited endpoints.

[0014] The present invention relates to concentration values, and its meaning includes fluctuations within a certain range. For example, it can fluctuate within the corresponding accuracy range. For example, 2% can allow fluctuations within the range of ±0.1%. For values ​​that are large or do not require too fine control, its meaning is also allowed to include greater fluctuations. For example, 100mM can allow fluctuations within the range of ±1%, ±2%, ±5%, etc. Involving molecular weight, its meaning is allowed to include fluctuations of ±10%.

[0015] In the present invention, descriptions such as "plurality" and "multiple" refer to quantities greater than or equal to 2 unless otherwise specified.

[0016] In the present invention, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0017] In the present invention, “preferred”, “better”, “more preferred” and “suitable” are only used to describe implementation methods or examples with better effects. It should be understood that they do not constitute limitations on the scope of protection of the present invention.

[0018] In the present invention, "optionally", "optional", "optionally", "optionally", "optional", "optional", "optional" means optional, that is, it means to be selected from any one of the two parallel schemes of "yes" or "no". If multiple "optional" or "optional" appear in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" or "optional" is independent.

[0019] The invention relates to a biomaterial combination, which comprises: anaerobic bacteria, enzymes and small molecule biocarbon; the anaerobic bacteria are a mixed bacteria of yeast, lactobacillus and methanobacterium, the yeast is one or more of Saccharomyces cerevisiae, Pichia pastoris and Pichia kluyveri; the lactobacillus is one or more of Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactobacillus bulgaricus; the methanobacterium is Mathanobacterium thermoautrophicum, Methanobucterium bryantii, Methanobacterium formicum and Methanobacterium truncatum. ormicicum); the enzyme is selected from one or more of protease, chitinase, lipase, endo-β-1,3 glucanase, endo-β-1,4 glucanase, and amylase; the small molecule biocarbon is selected from one or more of glucose, sucrose, fructose, humic acid, and chitosan oligosaccharide.

[0020] The present invention finds that the above-mentioned anaerobic bacteria can increase the number of anaerobic bacteria in the soil and produce a large amount of volatile organic acids and alcohol substances, which is more conducive to soil disinfection and improves the efficiency of soil disinfection; the above-mentioned enzymes can accelerate the degradation of organic materials and accelerate the reaction process; the above-mentioned small molecule biochar can increase the activity of soil anaerobic bacteria, and further provide energy for the rapid reproduction of anaerobic bacteria, thereby ensuring the smooth progress of the reaction. By comprehensively using the above-mentioned anaerobic bacteria, enzymes and small molecule biochar, the efficiency of anaerobic disinfection of the soil can be significantly improved, and the vitality of the soil can be significantly improved. At the same time, its application scenarios are more than traditional soil disinfection technology, providing farmers with a more reliable bio-enhanced disinfection technology, which can more effectively solve the problem of soil-borne diseases in various scenarios.

[0021] In some embodiments, the anaerobic bacteria include Saccharomyces cerevisiae, Lactobacillus acidophilus and thermoautotrophic methanobacterium in a mass ratio of 1:0.8-1.2:1.5-2.5, wherein the effective bacterial content of the yeast is 150-250 million CFU / g, the effective bacterial content of the lactobacillus is 800-1.2 billion CFU / g, and the effective bacterial content of the methanobacterium is 800-1200 million CFU / g. The present invention further found that the above anaerobic bacteria combination has a better anaerobic disinfection effect in soil.

[0022] As an example, the mass ratio of the saccharomyces cerevisiae, Lactobacillus acidophilus and thermoautotrophic methanobacterium can be any value within the above numerical range, such as 1:0.8:2.5, 1:1.2:1.5, 1:0.9:1.8, 1:1.1:2.2, 1:1:2, etc. In a preferred embodiment, the mass ratio of the saccharomyces cerevisiae, Lactobacillus acidophilus and thermoautotrophic methanobacterium is 1:1:2.

[0023] In some specific embodiments, the effective bacterial content of the yeast is 150 million CFU / g, 180 million CFU / g, 190 million CFU / g, 200 million CFU / g, 210 million CFU / g, 220 million CFU / g or 250 million CFU / g.

[0024] In some specific embodiments, the effective bacterial content of the lactobacillus is 800 million CFU / g, 850 million CFU / g, 900 million CFU / g, 950 million CFU / g, 980 million CFU / g, 1 billion CFU / g, 1.02 billion CFU / g, 1.05 billion CFU / g, 1.1 billion CFU / g, 1.15 billion CFU / g or 1.20 billion CFU / g.

[0025] In some specific embodiments, the effective bacterial content of the Methanobacterium is 8 billion CFU / g, 8.5 billion CFU / g, 9 billion CFU / g, 9.5 billion CFU / g, 9.8 billion CFU / g, 10 billion CFU / g, 10.2 billion CFU / g, 10.5 billion CFU / g, 11 billion CFU / g, 11.5 billion CFU / g or 12 billion CFU / g.

[0026] In some specific embodiments, those skilled in the art can adjust the effective bacteria content and quality of the strains, but the final effective bacteria dosage in the anaerobic bacteria is still within the scope of the present invention, and such technical solutions also belong to the protection scope of the present invention.

[0027] In some embodiments, the enzyme comprises a protease and an amylase in a mass ratio of 1:0.8-1.2, wherein the enzyme activity of the protease is 80,000-120,000 U / g, and the enzyme activity of the amylase is 1500-2500 U / g. The present invention further finds that the above-mentioned combined enzyme has a higher degradation efficiency for organic materials and is more conducive to accelerating the anaerobic disinfection reaction process.

[0028] As an example, the mass ratio of the protease to the amylase can be any value within the above numerical range, such as 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, etc. In a preferred embodiment, the mass ratio of the protease to the amylase is 1:1.

[0029] In some specific embodiments, the enzymatic activity of the protease is 80,000 U / g, 85,000 U / g, 90,000 U / g, 95,000 U / g, 98,000 U / g, 100,000 U / g, 102,000 U / g, 105,000 U / g, 110,000 U / g, 115,000 U / g or 120,000 U / g.

[0030] In some specific embodiments, the enzymatic activity of the amylase is 1500 U / g, 1800 U / g, 1900 U / g, 1950 U / g, 2000 U / g, 2050 U / g, 2100 U / g, 2200 U / g or 2500 U / g.

[0031] In some specific embodiments, those skilled in the art can adjust the enzyme activity and enzyme quality of the enzyme, but the final amount of enzyme activity unit (U) in the enzyme is still within the scope of the present invention, and such technical solutions also belong to the protection scope of the present invention.

[0032] In some embodiments, the small molecule biochar includes humic acid and glucose in a mass ratio of 1:0.8-1.2. The present invention further finds that the small molecule biochar is more conducive to improving the effect of anaerobic reaction, thereby improving the soil disinfection effect.

[0033] As an example, the mass ratio of humic acid to glucose can be any value within the above numerical range, such as 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, etc. In a preferred embodiment, the mass ratio of humic acid to glucose is 1:1.

[0034] In some embodiments, the mass ratio of the anaerobic bacteria, enzymes and small molecule biochar is 5-10: 1-3: 40-60. The present invention also finds that by combining and applying in the above ratio, it will further help ensure the disinfection effect of the biological material combination on the soil.

[0035] As an example, the mass ratio of the anaerobic bacteria, enzyme and small molecule biochar can be any value within the above numerical range, such as 1:1:45, 2:2:40; 3:5:50, 5:3:46, 5:6:45, 7:8:58, 10:10:60, etc.

[0036] In some specific embodiments, the mass ratio of the anaerobic bacteria, enzyme and small molecule biochar is 5-6:1-2:40-60.

[0037] In specific implementation, the anaerobic bacteria, enzymes and small molecule bio-carbon can be packaged separately and then combined to form the bio-material combination; some of the materials can also be mixed and then combined, such as mixing the anaerobic bacteria and enzymes and packaging them separately, and packaging the small molecule bio-carbon separately, and then combining them to form the bio-material combination. Those skilled in the art can adjust the packaging methods of the components mentioned in the bio-material combination based on common sense, which all fall within the scope of protection of the present invention.

[0038] In specific implementation, the above-mentioned components can be obtained through commercial channels.

[0039] The invention also relates to a method for anaerobic disinfection of soil, which comprises: applying the biological material combination into the soil.

[0040] In some embodiments, the application amount of the anaerobic bacteria is 5-10 kg / mu. As an example, the application amount of the anaerobic bacteria can be 5 kg / mu, 6 kg / mu, 7 kg / mu, 8 kg / mu, 9 kg / mu, 10 kg / mu, or any value within the above numerical range.

[0041] In some embodiments, the application amount of the enzyme is 1-3 kg / mu. As an example, the application amount of the enzyme can be 1 kg / mu, 2 kg / mu, 3 kg / mu or any value within the above numerical range.

[0042] In some embodiments, the application amount of the small molecule biochar is 40-60 kg / mu. As an example, the application amount of the small molecule biochar can be 40 kg / mu, 42 kg / mu, 45 kg / mu, 48 kg / mu, 50 kg / mu, 52 kg / mu, 55 kg / mu, 58 kg / mu, 60 kg / mu, or any value within the above numerical range.

[0043] In some embodiments, the soil is soil in a protected facility (including a solar greenhouse, spring and autumn greenhouse, plant factory, etc.).

[0044] In some embodiments, the soil anaerobic disinfection method includes: before turning the soil, mixing the anaerobic bacteria and enzymes with organic materials and applying them to the soil; during the turning and film covering period, irrigating the soil to create an anaerobic environment, and applying the small molecule biochar at the same time. The above application method will further help improve the soil disinfection effect of the biological material combination.

[0045] In some specific embodiments, when the mixture of anaerobic bacteria, enzymes and organic materials is applied to the soil, the application method is broadcasting.

[0046] In some embodiments, the organic material is one or more of straw, sheep manure, and cow manure, and the carbon-nitrogen ratio is 20 to 30:1, and the application amount is 1 to 3 tons / mu.

[0047] As an example, in a specific implementation, the carbon-nitrogen ratio of the organic material can be 20:1, 22:1, 25:1, 27:1, 30:1, or any value within the above range.

[0048] As an example, in a specific implementation, the amount of the organic material used can be 1 ton / mu, 1.2 tons / mu, 1.3 tons / mu, 1.5 tons / mu, 1.7 tons / mu, 1.8 tons / mu, 2 tons / mu, or any value within the above numerical range.

[0049] In some embodiments, the irrigation is performed using a drip tape and / or a micro-spray tape; when irrigating the soil, water is first poured until a small amount of water accumulates on the ground surface without overflowing, and then a diluted solution of the small molecule biochar (such as a 50-100 times diluted solution) is applied to the soil through the drip tape and / or the micro-spray tape. After the application, the drip tape and / or the micro-spray tape is rinsed with water (the time may be 5-10 minutes).

[0050] In some embodiments, the soil anaerobic disinfection method comprises the following steps:

[0051] (1) When the temperature is stably above 25°C, clear the weeds and debris (such as diseased crop residues and mulch) from the soil;

[0052] (2) Applying organic materials, anaerobic bacteria and enzymes to the soil;

[0053] (3) Till the soil more than 20 cm, deep loosen to 40-50 cm, and till until the soil is loose, flat and compacted;

[0054] (4) Laying drip irrigation tape and / or micro-sprinkler tape, and laying ground film;

[0055] (5) irrigating the soil with a drip irrigation tape and / or a micro-sprinkler tape, and applying the small molecule biochar to the soil through the drip irrigation tape and / or the micro-sprinkler tape;

[0056] (6) Start to seal the container and continue to seal the container for more than 15 days. Remove the film and let it air-dry for 3 to 7 days.

[0057] In some embodiments, in step (4), the density of laying the drip irrigation tape is one tape at an interval of 1 to 1.5 m, and the density of laying the micro-sprinkler tape is one tape at an interval of 2 to 4 m; the ground film is covered by the reverse buckle method.

[0058] In some embodiments, in step (6), during the sealing period, when the temperature is stably above 25°C, the sealing is carried out for 15 to 17 days, and the film is removed and the film is exposed to the sun for 3 to 5 days; when it is rainy, the sealing time is extended and the film is removed and the film is exposed to the sun for 3 to 7 days.

[0059] In the specific implementation, the person skilled in the art can confirm other detailed operations in combination with common sense to be more conducive to the soil disinfection effect. For example, when applying organic materials, anaerobic bacteria and enzymes to the soil, first mix the three evenly, and pay special attention to the edges and corners of the plot when applying; when laying the ground film, it is necessary to prevent the residual uncovered soil on the ground film; when sealing, if there are gaps or holes in the shed, it is necessary to glue it; and so on.

[0060] Embodiments of the present invention will be described in detail below in conjunction with examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples that do not specify specific conditions are preferably referred to the guidance provided in the present invention, and can also be based on the experimental manual or normal conditions in this area, and can also be based on other experimental methods known in the art, or according to the conditions recommended by the manufacturer.

[0061] In the following specific embodiments, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument test accuracy or operation accuracy are allowed.

[0062] Example 1

[0063] In order to effectively judge the effect of the biomaterial combination of the present invention, soils planted with vegetables for many consecutive years were selected to carry out plot tests. The disinfection evaluation indicators included changes in soil physical and chemical properties, temperature, pH, EC and other indicators, as well as pathogenic microorganism isolation and detection (Fusarium, Phytophthora) and the like.

[0064] This embodiment first provides a biomaterial combination, wherein the anaerobic bacteria is a mixture of saccharomyces cerevisiae, lactobacillus acidophilus and thermoautotrophic methanobacterium in a mass ratio of 1:1:2; the enzyme is a mixture of protease and amylase in a mass ratio of 1:1, and the small molecule biocarbon is a mixture of humic acid and glucose in a mass ratio of 1:1. Among them, the effective bacterial content of saccharomyces cerevisiae is 200 million CFU / g, the effective bacterial content of lactobacillus acidophilus is 1 billion CFU / g, and the effective bacterial content of thermoautotrophic methanobacterium is 10 billion CFU / g. The enzyme activity of protease is 100,000 U / g, and the enzyme activity of amylase is 2000 U / g.

[0065] Experimental treatment: The mass ratios of biological material combination, anaerobic bacteria dosage, enzyme dosage and small molecule bio-carbon were 5:1:40, 5:1:60, 5:2:40, 5:2:60, 6:1:40, 6:1:60, 6:2:40 and 6:2:60, respectively, totaling 8 combinations. The treatments were carried out according to the following implementation steps. Conventional anaerobic treatment was a treatment method without adding a biological material combination (only adding organic materials). The sealing time of the experimental treatment group was 15 days, and the conventional anaerobic treatment was 30 days.

[0066] 1. Implementation steps

[0067] The implementation area is the soil for growing vegetables for many years, and the area of ​​the area is 1 square meter.

[0068] This embodiment further provides a method for anaerobic disinfection of soil, which comprises the following steps:

[0069] (1) Clean the soil of diseased and crop residues, weeds, and mulch film;

[0070] (2) Spreading organic materials, anaerobic bacteria and enzymes; the organic material was a mixture of straw and sheep manure, with a carbon-nitrogen ratio of 25:1 and an application rate of 2 tons per mu; the application rate of anaerobic bacteria was 5 kg / mu or 6 kg / mu, and the application rate of enzymes was 1 kg / mu or 2 kg / mu; the specific application rates of anaerobic bacteria and enzymes per mu in each test group were set according to the different groups of the test treatment;

[0071] (3) The soil is tilled to 20 cm, deep tilled to 40-50 cm, and rotary tilled until the soil is loose, flat, and compacted;

[0072] (4) Laying drip irrigation tape and ground film;

[0073] (5) First, water is irrigated until a small amount of water accumulates on the ground surface without overflowing, and then the small molecule biochar is diluted and applied to the ground in the form of drip irrigation. After the application is completed, the drip irrigation belt is rinsed with water; wherein, the per mu dosage of the small molecule biochar is 40 kg or 60 kg; the specific per mu dosage of the small molecule biochar in each test group is set for different groups;

[0074] (6) Keep the container sealed for 15 days, then remove the film and air dry for 5 days.

[0075] 2. Soil disinfection effect evaluation method

[0076] a. Monitor soil temperature changes, pH changes, EC changes, and soil physical and chemical properties (nitrogen, phosphorus, and potassium) during the test

[0077] b. After soil disinfection, mix the test soil thoroughly, and weigh 5g of fresh soil sample to send to the laboratory for isolation and detection of pathogenic microorganisms. The pathogenic microorganisms isolated and detected in the test are Fusarium spp. and Phytophthora spp. Among them, the soil-borne pathogen Fusarium spp. was isolated using Komada's method (Komada et al. 1975). Phytophthora spp. and Pythium spp. were isolated using Masago's method (Masago et al. 1977).

[0078] c. Weigh 10 g of the sterilized and mixed soil, add 30 mL of sterilized deionized water, shake at room temperature for 1 hour, aspirate 5 mL of the suspension, add it to a sterilized culture dish, cover it with filter paper, and place 20 Shanghai green seeds on it. After 2-3 days, investigate the germination rate, root length and stem length of the seedlings.

[0079] Through the above data, the control effect of the biological material combination and the soil anaerobic disinfection method of the present invention on soil-borne diseases can be evaluated more comprehensively and objectively.

[0080] 3. Soil disinfection effect

[0081] After disinfection, the above indicators (soil temperature, pH, EC, soil physical and chemical properties) after disinfection by the biomaterial combination of the present invention are better than those of other treatments. In terms of temperature, the soil temperature of the biomaterial combination of 5:1:40 is slightly higher than that of other combinations, the soil pH and EC are reduced, and the nitrogen, phosphorus and potassium contents are increased (Table 1).

[0082] Table 1 Physical and chemical data of soils in each test treatment

[0083]

[0084] After the disinfection with different biological material combinations, the soil pathogenic microorganisms were separated. The results in Table 2 showed that compared with other treatments, the disinfection with the 5:1:40 biological material combination had a disinfecting effect of 96.84% and 96.43% on Phytophthora and Fusarium, respectively. Compared with other treatments, the 5:1:40 biological material combination had the best disinfecting effect.

[0085] Table 2 The disinfecting effect of pathogens

[0086]

[0087] After disinfection, the germination test results of each treatment group are shown in Table 3. Compared with other treatments, the 5:1:40 biological material combination disinfection had a germination rate, root length, and stem length of 95%, 9.15 cm, and 3.02 cm, respectively. Compared with other treatments, all indicators were better.

[0088] Table 3 Seed germination safety test

[0089] Experimental treatment Germination rate Root length (cm) Stem length (cm) 5：1：40 95％ 9.15 3.02 5：2：40 95％ 8.75 2.81 5：1：60 95％ 8.62 2.65 5：2：60 95％ 8.71 2.58 6：1：40 95％ 8.80 2.58 6：1：60 95％ 8.36 2.57 6：2：40 95％ 8.18 2.65 6：2：60 95％ 8.11 2.49 Conventional anaerobic treatment 90％ 7.78 2.45 Soil before disinfection 85％ 7.23 2.45

[0090] Example 2

[0091] In order to effectively judge the effect of the anaerobic soil disinfection technology of the present invention, soils planted with vegetables for many consecutive years were selected to carry out a plot test of the new disinfection technology. The disinfection evaluation indicators included changes in soil physical and chemical properties, temperature, pH, EC and other indicators, as well as pathogenic microorganism isolation and detection (Fusarium, Phytophthora).

[0092] 1. Implementation steps

[0093] First, the soil to be treated (10 square meters) is cleaned of diseased and damaged plants, and then the soil is disinfected according to the soil anaerobic disinfection method in Example 1. The test treatments are respectively the biological material combination of the present invention at a ratio of 5:1:40, biological shed bacterial agent treatment, and conventional anaerobic treatment. Among them, the biological material combination of the present invention at a ratio of 5:1:40 and conventional anaerobic treatment are the same as in Example 1. In the biological shed bacterial agent disinfection, the biological shed bacterial agent is a common shed product on the market (the effective strains are Bacillus subtilis and Bacillus licheniformis), which is applied together with the organic material in the implementation step (2), and the other treatments remain unchanged. The blank treatment is unsterilized soil. The treatment time of the biological shed bacterial agent and the blank treatment is 30 days.

[0094] 2. Soil effect evaluation

[0095] The evaluation method is similar to that in Example 1.

[0096] 3. Soil disinfection effect

[0097] After the disinfection, the above indicators (soil temperature, pH, EC, soil physical and chemical properties) after disinfection by the biomaterial combination of the present invention are better than those of other treatments in terms of temperature, pH, EC, soil physical and chemical properties, etc. In terms of temperature, the soil temperature treated by the biomaterial combination of the present invention is slightly higher than that of other treatments, the soil pH and EC are reduced, and the nitrogen, phosphorus and potassium contents are increased.

[0098] Table 4 Soil physical and chemical properties test

[0099]

[0100] After the disinfection, the soil pathogenic microorganisms were separated. The results in Table 5 showed that the disinfection effect of the biological material combination of the present invention on Phytophthora and Fusarium was 95.2% and 95.24% respectively compared with other treatments. Compared with conventional anaerobic disinfection and biological shed sterilizing agents, the disinfection effect was better.

[0101] Table 5 Pathogen disinfecting effect of each treatment

[0102]

[0103] After the disinfection, the germination test results of each treatment group are shown in Table 6. Compared with the blank control, the germination rate, root length and stem length of the Shanghai green were 95%, 9.3 cm and 3.4 cm respectively. Compared with other treatments, all indicators were better.

[0104] Table 6 Seed germination safety test

[0105] Experimental treatment Germination rate Root length (cm) Stem length (cm) Biomaterial combination of the present invention 95％ 9.30 3.40 Biological greenhouse bacteria agent 90％ 8.80 3.24 Conventional anaerobic treatment 85％ 7.67 3.09 Blank control 80％ 7.03 2.60

[0106] Example 3

[0107] In order to verify the soil disinfection effect of this technology in preventing and controlling soil-borne diseases, a greenhouse with continuous strawberry planting was selected to carry out field effect verification tests of the biological material combination and biological greenhouse sterilizing agent of the present invention. After soil disinfection, soil samples were collected, and then germination tests and pathogen detection tests were carried out to fully understand the specific effects of the technology in field application.

[0108] 1. Implementation steps

[0109] Referring to Example 2, the biological material combination (5:1:40) and the biological shed bacteria agent of the present invention were added in proportion to disinfect the soil. The implementation area was one treatment for half an acre of land.

[0110] 2. Soil disinfection effect evaluation method

[0111] The soil before and after disinfection was sampled using a 5-point sampling method, and 500 g of fresh soil was taken for soil physical and chemical property testing and pathogen isolation testing. The pathogens isolated and tested in the experiment mainly included Fusarium, Phytophthora, etc., and the survival rate of crops in the seedling stage and the growth index during the growth period were tracked and investigated, so as to more comprehensively evaluate the control effect of the biomaterial combination and the bio-stifling microbial agent of the present invention on soil-borne crop diseases.

[0112] Field surveys are conducted in the early stages of planting, the middle and late stages of crop growth. The survey content specifically includes testing of soil physical and chemical properties before and after disinfection, isolation and testing of soil-borne pathogenic microorganisms (Fusarium, Phytophthora), seedling survival rate, crop plant height, stem thickness and other indicators.

[0113] 3. Soil disinfection effect

[0114] After soil disinfection was completed, the disinfected soil was tested and the results are shown in Table 7.

[0115] It can be seen from the results that after soil disinfection, the contents of alkaline nitrogen, available phosphorus and quick-acting potassium increased, and pH and EC decreased in the disinfection of the biological material combination of the present invention compared with the disinfection of biological greenhouse bacteria.

[0116] Table 7 Soil physical and chemical properties after soil disinfection

[0117]

[0118] After the disinfection, the soil pathogenic microorganisms were separated. The results in Table 8 showed that compared with the blank control, the disinfecting effects of the biological material combination disinfection of the present invention on Phytophthora and Fusarium were 96.25% and 98%, respectively, which were better than the disinfecting effect of the biological sterilizing agent disinfection, indicating that this method has a strong killing effect on soil-borne pathogenic microorganisms.

[0119] Table 8 Detection of soil pathogens

[0120]

[0121] According to the five-point sampling method, the number of live and dead seedlings of the test crops was investigated and the survival rate of the crops was calculated. The calculation formula is as follows:

[0122] Survival rate = (number of live seedlings) / (total number surveyed)*100%.

[0123] As can be seen from Table 9, the survival rate of the biological material combination of the present invention is better than that of the biological shed sterilizing agent and the blank control at the seedling stage. The highest survival rate of strawberry seedlings is obtained by the biological material combination disinfection, and the survival rate of the corresponding treatment area is 98.67%.

[0124] Table 9 Seedling survival rate of different treatments

[0125]

[0126] As shown in Table 10, compared with the blank control treatment and the biological shed sterilization treatment, the strawberry plant height and stem thickness were significantly better after the biological material combination disinfection treatment of the present invention.

[0127] Table 10 Effects of different treatment conditions on plant height and stem diameter of strawberry

[0128] Experimental treatment Plant height(cm) Stem diameter (mm) Biomaterial combination of the present invention 23.19 16.38 Biological greenhouse bacteria agent 22.25 15.27 Blank control 21.92 14.58

[0129] In general, the disinfection of the biological material combination of the present invention has a good effect in the treatment and prevention of pathogens. It can reduce the residual chemical components in the soil, improve the soil's own resistance to external pressure, and gradually restore soil health by sealing the soil in a biological way.

[0130] It can be seen from the above that the biomaterial combination of the present invention and the soil anaerobic disinfection method using the same can effectively shorten the disinfection time and improve the disinfection efficiency; reduce the salinity of the soil, improve the soil vitality, and improve the arable land quality of the soil; reduce the content of Fusarium and Phytophthora in the soil, reduce the occurrence of soil-borne diseases, ensure the survival rate of crops, and promote the normal growth of crops (stem length and stem thickness), and maintain the homeostasis of the soil ecosystem.

[0131] Therefore, the biomaterial combination of the present invention and the soil anaerobic disinfection method using the biomaterial combination have at least the following application effects:

[0132] 1) Shortened soil processing time;

[0133] 2) It not only adjusts the soil pH, but also reduces salinization, which is beneficial to improve the utilization rate of fertilizers;

[0134] 3) Reduce the occurrence of soil-borne diseases during plant growth, promote root growth, prevent premature aging, and improve crop yield and quality.

[0135] The above-mentioned embodiments only express several implementation methods of the present invention, and the description is relatively specific and detailed, but it cannot be understood as limiting the scope of the invention patent. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. A biomaterial combination comprising: Anaerobic bacteria, enzymes and small molecule biochar; The anaerobic bacteria are a mixture of yeast, lactobacillus and methanobacterium, wherein the yeast is one or more of Saccharomyces cerevisiae, Pichia pastoris and Pichia kluyveri; the lactobacillus is one or more of Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactobacillus bulgaricus; the methanobacterium is one or more of Mathanobacterium thermoautrophicum, Methanobucterium bryantii and Methanobacterium ormicicum; The enzyme is selected from one or more of protease, chitinase, lipase, endo-β-1,3 glucanase, endo-β-1,4 glucanase, and amylase; The small molecule biocarbon is selected from one or more of glucose, sucrose, fructose, humic acid and chitosan oligosaccharide.

2. The biomaterial combination according to claim 1, wherein: The anaerobic bacteria include yeast, lactobacillus and methanobacterium in a mass ratio of 1:0.8-1.2:1.5-2.5, wherein the effective bacterial content of the yeast is 150 million to 250 million CFU / g, the effective bacterial content of the lactobacillus is 800 million to 1.2 billion CFU / g, and the effective bacterial content of the methanobacterium is 8 billion to 12 billion CFU / g.

3. The biomaterial combination according to claim 1 or 2, wherein: The enzyme comprises a protease and an amylase in a mass ratio of 1:0.8-1.2, wherein the enzyme activity of the protease is 80,000-120,000 U / g, and the enzyme activity of the amylase is 1500-2500 U / g.

4. The biomaterial combination according to any one of claims 1 to 3, wherein: The small molecule biochar comprises humic acid and glucose in a mass ratio of 1:0.8-1.

2.

5. The biomaterial combination according to any one of claims 1 to 4, wherein: The mass ratio of the anaerobic bacteria, the enzyme and the small molecule biocarbon is 5-10:1-3:40-60.

6. A method for anaerobic disinfection of soil, comprising: The biomaterial combination according to any one of claims 1 to 5 is applied to soil.

7. The method for anaerobic soil disinfection according to claim 6, wherein: The application amount of the anaerobic bacteria is 5-10 kg / mu, the application amount of the enzyme is 1-3 kg / mu, and the application amount of the small molecule biochar is 40-60 kg / mu.

8. The method for anaerobic disinfection of soil according to claim 6 or 7, comprising: Before tilling the soil, the anaerobic bacteria and enzymes are mixed with organic materials and then applied to the soil; After the soil is turned over and covered with film, the soil is irrigated under the film and the small molecule biochar is applied at the same time.

9. The method for anaerobic soil disinfection according to claim 8, wherein: The organic material is one or more of straw, cow dung, and sheep dung, and the carbon-nitrogen ratio is 20-30:1, and the application amount is 1-3 tons / mu.

10. The method for anaerobic soil disinfection according to claim 8 or 9, wherein: The irrigation is carried out using a drip irrigation tape and / or a micro-sprinkler tape; When irrigating the soil, water is first poured until a small amount of water accumulates on the ground surface without overflowing, and then the diluted solution of the small molecule biochar is applied to the soil through the drip irrigation tape and / or the micro-spraying tape. After the application is completed, the drip irrigation tape and / or the micro-spraying tape is rinsed with water.

11. The method for anaerobic disinfection of soil according to any one of claims 6 to 10, comprising the following steps: (1) When the temperature is stable above 25°C, clear the weeds and debris on the soil surface; (2) Applying organic materials, anaerobic bacteria and enzymes to the soil; (3) Till the soil more than 20 cm, deep loosen to 40-50 cm, and till until the soil is loose, flat and compacted; (4) Laying drip irrigation tape and / or micro-sprinkler tape, and laying ground film; (5) irrigating the soil with a drip irrigation tape and / or a micro-sprinkler tape, and applying the small molecule biochar to the soil through the drip irrigation tape and / or the micro-sprinkler tape; (6) Start to seal the container and continue to seal the container for more than 15 days. Remove the film and let it air-dry for 3 to 7 days.

12. The method for anaerobic soil disinfection according to claim 11, wherein: In step (4), the density of laying the drip irrigation tape is one tape at an interval of 1 to 1.5 m, and the density of laying the micro-sprinkler tape is one tape at an interval of 2 to 4 m; the ground film is covered by the reverse buckle method.

13. The method for anaerobic soil disinfection according to claim 11 or 12, wherein: In step (6), during the sealing period, when the temperature is stably above 25°C, the sealing is carried out for 15 to 17 days, and the film is removed and the leaves are exposed to the sun for 3 to 5 days; when it is rainy, the sealing time is extended and the film is removed and the leaves are exposed to the sun for 3 to 7 days.

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