CN117820040A - Rice full-element biological organic fertilizer and preparation method and application thereof - Google Patents
Rice full-element biological organic fertilizer and preparation method and application thereof Download PDFInfo
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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- Fertilizers (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a rice full-element biological organic fertilizer and a preparation method and application thereof. The rice whole-element biological organic fertilizer comprises an active functional bacterial component, an organic fertilizer component, an amino acid component and an inorganic compound fertilizer component, wherein the active functional bacterial component contains bacillus bailii for promoting growth and reducing cadmium, bacillus amyloliquefaciens for resisting diseases and promoting growth and streptomyces for resisting diseases and promoting growth of rice; the organic fertilizer component contains decomposed sawn timber ash, mushroom residues, bran, chicken manure and pig manure. The invention also discloses a preparation method of the rice full-element biological organic fertilizer. The fertilizer has the advantages of organic fertilizer, inorganic fertilizer, microbial fertilizer and amino acid fertilizer, can improve the utilization rate of the fertilizer, improve the physicochemical property, nutrition state and biological activity of soil, promote the growth and propagation of rhizosphere microorganisms of rice, enhance the disease resistance level of the rice, provide comprehensive and balanced nutrition for the growth of the rice, and obviously improve the yield and quality of the rice.
Description
Technical Field
The invention belongs to the technical field of microbial fertilizers and preparation thereof, and relates to a rice full-element biological organic fertilizer, a preparation method and application thereof.
Background
The agricultural land is 8% of the cultivated land in the world, 21% of the grain in the world is produced, but the consumption of chemical fertilizer accounts for 35% of the world, and the usage of chemical fertilizer in unit area is 3 times of the average world level. The continuous and large-scale application of chemical fertilizer for a long time causes the continuous reduction of organic matters in soil, the soil is reduced in fertility due to salinization, acidification and hardening, water source is polluted, ecological environment is destroyed, and human and animal health is endangered. The original high-yield field has a trend of developing to a barren thin low-yield field.
Aiming at the problems of reduced basic soil productivity, resource exhaustion, environmental pollution and the like existing in the soil in China, the green fertilizer industry is preferentially developed.
The method is a main way for applying organic matters and improving the diversity of soil microbial population structures. The biological organic fertilizer is widely focused on the characteristics of regulating plant growth, increasing crop yield, improving crop quality, reducing fertilizer use, improving soil quality, protecting ecological environment and the like, and becomes an irreplaceable agricultural product in the development of green ecological agriculture. However, although a single organic fertilizer can fertilize soil, the nutrient content is low, the release is slow, the yield increasing effect on crops in the season is not obvious, the microbial function in the existing bio-organic fertilizer is single, the effect is low, and the effect on crops in the season is unstable.
The full-element biological organic fertilizer is a novel compound fertilizer integrating compost, fertilizer, biological fertilizer and amino acid fertilizer, and has the functions of promoting the growth of crops in the season, remarkably inhibiting the acidification of soil, effectively regulating and controlling the microbial system of the soil and continuously improving the soil foundation soil fertility. The application of the full-element biological organic fertilizer can simultaneously improve the content of organic matters in soil and the biological activity of the soil, so that soil mineralization microorganisms, soil nitrifying bacteria, soil nitrogen-fixing bacteria, soil phosphate and potassium dissolving bacteria and the like are in a good balance relationship, and the soil fertility is continuously and stably output.
Disclosure of Invention
The invention aims at providing a preparation method of a rice full-element biological organic fertilizer. The rice whole-element biological organic fertilizer prepared by the method has the advantages of organic fertilizer, inorganic fertilizer, microbial fertilizer and amino acid fertilizer, can improve the fertilizer utilization rate, the physicochemical property, the nutrition state and the biological activity of soil, promote the growth and propagation of rhizosphere microorganisms of rice, enhance the disease resistance level of the rice, provide comprehensive and balanced nutrition for the growth of the rice, and obviously improve the yield and the quality of the rice.
A preparation method of a rice full-element biological organic fertilizer comprises the following steps:
(1) Firstly, carrying out liquid fermentation on bacillus bailii 20-10 and bacillus amyloliquefaciens CWJ2, and then adding the fermentation liquor into a decomposed organic fertilizer for solid state fermentation;
(2) Carrying out solid fermentation on streptomyces Ahn 65;
(3) Mixing the fermentation products obtained in the step (1) and the step (2) with amino acid and an inorganic compound fertilizer to obtain a rice full-element biological organic fertilizer;
the preservation numbers of bacillus belicus 20-10, bacillus amyloliquefaciens CWJ2 and streptomycete Ahn65 are CCTCC NO: M20221980,CCTCC NO:M20221983,CCTCC NO:M20221981 respectively.
The step (1) specifically comprises:
a. activating strains: b, bacillus belicus 20-10 and bacillus amyloliquefaciens CWJ2 are separately inoculated on a solid LB culture medium to activate strains, and then are separately inoculated on an LB liquid culture medium to be cultured, so as to prepare seed liquid;
b. fermentation culture: inoculating seed liquid into a fermentation tank for culture according to the inoculum size of 0.5-2% by volume, and introducing sterile air for fermentation culture;
the fermentation liquid of the two bacteria contains more than 50 hundred million viable spores per milliliter, and when the spore proportion reaches more than 90%, the fermentation liquid is placed in a tank, and the pH value is 7.0-8.0, so that no mixed bacteria pollution is caused;
c. the carbon-nitrogen ratio of the decomposed organic fertilizer is adjusted to be 20:1-30:1, the weight ratio of water is adjusted to be 40-60%, and fermentation liquid of bacillus bailii and bacillus amyloliquefaciens is inoculated according to the proportion of 0.5-2% of inoculum size, and the fermentation liquid is uniformly mixed for natural composting fermentation.
Further, the step (1) more specifically includes:
a. activating strains: inoculating bacillus belicus 20-10 and bacillus amyloliquefaciens CWJ2 on a solid LB culture medium, culturing for 16-30 hours at 30 ℃ and then activating strains, inoculating the strains into 100mL LB liquid culture medium by using a sterile ring, and culturing for 16-24 hours at 30 ℃ and 180rpm to prepare seed liquid;
b. fermentation culture: sterilizing a fermentation tank, filling a culture medium, sterilizing, maintaining the pressure, cooling to 30 ℃, inoculating fermentation liquor of a seed tank into the fermentation tank for culture according to the inoculation amount of 0.5-2%, and introducing sterile air; culturing for 60-72 h at 30 ℃ with aeration rate of 1.5 Vols/vol.min and oxygen content of 35% by volume;
the formula of the fermentation medium comprises: 2% of maltose, 1% of starch, 1.25% of yeast extract, 0.1% of NaCl, 0.3% of ammonium sulfate, 0.1% of calcium carbonate and the balance of water, wherein the initial pH value is 7.2-7.5;
c. the carbon-nitrogen ratio of the decomposed organic fertilizer is adjusted to be respectively 20:1-30:1, the weight ratio of water is adjusted to be 40-60%, fermentation liquid of bacillus bailii and bacillus amyloliquefaciens is inoculated according to the proportion of 0.5-2% of inoculum size, and natural composting fermentation is carried out after uniform mixing; when the temperature reaches above 50 ℃, turning the pile for 2-3 times per day; completing the compost fermentation until the germination index of the organic fertilizer is more than or equal to 90%;
the decomposed organic fertilizer is prepared by decomposing saw dust, mushroom residue, bran, chicken manure and pig manure according to the weight ratio of 0.5:0.5:2-3:2-3:4.
Preferably, in the step c, the carbon-nitrogen ratio of the decomposed organic fertilizer is adjusted to be 25:1, the weight ratio of water is adjusted to be 40%, fermentation liquid of bacillus belicus and bacillus amyloliquefaciens is inoculated according to the proportion of 0.5% of inoculum size, and natural composting fermentation is carried out after uniform mixing.
The step (2) specifically comprises:
a. strain activation
The streptomycete Ahn65 is accessed into an ISP2 solid culture medium for activation, and then transferred into an ISP2 liquid culture medium for culture to prepare seed liquid;
b. solid fermentation
Inoculating the seed liquid of streptomyces Ahn65 into sterile bran according to the mass ratio of 1-5%, standing for culturing, and stopping culturing when the spore content of the streptomyces reaches more than 50 hundred million/g;
c. and drying the solid fermentation product of the streptomyces Ahn65 to prepare the streptomyces bacterial powder.
Further, the step (2) more specifically includes:
a. strain activation
Activating Streptomyces Ahn65 in ISP2 solid culture medium, transferring to 50mL ISP2 culture solution, and shake culturing at 28deg.C and 180rpm for 1-2d to obtain seed solution;
b. solid fermentation
Inoculating the culture solution of streptomyces Ahn65 into the sterile bran at a mass ratio of 1-5%, supplementing water to a mass ratio of 40-60%, standing at 28 ℃ for culturing for 10-20d, and stopping culturing when the spore content of the streptomyces reaches more than 50 hundred million/g;
c. and (3) drying the solid fermentation product of the streptomycete Ahn65 in a 40 ℃ incubator for 24 hours to prepare the streptomycete bacterial powder.
Further, adding streptomycete powder into the product of the step (1) according to the mass ratio of 1-2%, uniformly mixing, and drying at low temperature until the water content is less than or equal to 30%, thereby obtaining the bio-organic fertilizer rich in three functional microorganisms of bacillus belicus, bacillus amyloliquefaciens and streptomycete, wherein the effective viable count of each microorganism is not less than 2 hundred million/g.
Adding 5-8% of amino acid raw powder and inorganic compound fertilizer into the prepared bio-organic fertilizer, and uniformly mixing to obtain the full-element bio-organic fertilizer; the total biological organic fertilizer has effective viable count content of 6-80 hundred million/g, organic matter content of 30-55%, total nutrient content of NPK of 8-11%, moisture content of less than or equal to 30%, total amino acid of 5-10%, free amino acid of 0.5-5%, chloride ion content of less than or equal to 2.0% and pH of 6.0-7.0.
The invention also provides a rice full-element biological organic fertilizer prepared by the method.
The invention also provides application of the rice full-element biological organic fertilizer, which can promote rice growth, improve soil nutrition and enhance the disease resistance level of rice.
Compared with the prior art of a patent of a compound microbial agent and application thereof in rice growth promotion, disease resistance and cadmium reduction (application number CN 2023110559849), the invention has the following beneficial effects:
(1) The full-element biological organic fertilizer has more remarkable yield increasing effect on rice, and the yield of the rice can be increased by 41.59% -42.71% by using 150 kg/mu, which is far higher than the yield increasing effect (19.35%) of the composite microbial agent in patent CN 2023110559849.
(2) The highest prevention effect of the full-element biological organic fertilizer on the rice leaf blast is 66.08 percent, which is higher than the prevention effect (56.74 percent) of the composite microbial inoculum in CN2023110559849 on the rice leaf blast.
(3) The full-element biological organic fertilizer disclosed by the invention not only promotes the growth of rice and improves the resistance of rice to rice blast, but also can effectively improve soil fertility and microbial diversity, and the contents of soil organic matters, total nitrogen, effective phosphorus and quick-acting potassium are respectively increased by 8.82%, 13.46%, 7.80%, 197.87% and 39.23% compared with the blank, so that the total number of soil microorganisms is more and the microbial abundance is greater.
The Streptomyces Ahn65 adopted by the invention is classified and named as Streptomyces sp, the preservation number is CCTCC NO: M20221981, the preservation date is 2022, 12 months and 19 days, the preservation unit is China Center for Type Culture Collection (CCTCC), and the preservation unit address is in the Wuchang district eight-path 299 number Wuhan university in Wuhan, hubei province.
The Bacillus belicus 20-10 is classified and named Bacillus velezensis, and has a preservation number of CCTCC NO: M20221980, a preservation date of 2022, 12 months and 19 days, a preservation unit of China Center for Type Culture Collection (CCTCC), and a preservation unit address of eight-path 299 universities in Wuchang district of Wuhan, hubei province.
The bacillus amyloliquefaciens CWJ2 is classified and named Bacillus amyloliquefaciens, the preservation number is CCTCCNO: M20221983, the preservation date is 2022, 12 months and 19 days, the preservation unit is China Center for Type Culture Collection (CCTCC), and the preservation unit address is in the eight-path 299-number university of Wuchang district of Wuhan, hubei province.
Detailed Description
The present invention will be described in further detail with reference to examples.
Example 1: preparation of full-element biological organic fertilizer
(1) Preparation of active functional bacteria:
firstly, bacillus bailii and bacillus amyloliquefaciens are respectively and independently fermented by a fermentation tank, and the operation steps and the process conditions are as follows:
a. strain activation
Inoculating bacillus belicus 20-10 and bacillus amyloliquefaciens CWJ2 on a solid LB culture medium, culturing for 24 hours at 30 ℃ and then activating strains, then digging a spoon of thalli by a sterile inoculating loop, inoculating the thalli into 100mL LB liquid culture medium (500 mL triangular flask) sterilized for 30min at 121 ℃, and culturing for 24 hours at 30 ℃ and 180rpm to prepare seed liquid.
b. Fermentation tank culture
Firstly, cleaning and sterilizing a fermentation tank, filling a fermentation culture medium, then sterilizing, maintaining the pressure and cooling to 30 ℃, inoculating seed liquid into the fermentation tank for culture according to the inoculation amount of 1% by volume, and introducing sterile air;
the sterilization adopts high pressure steam sterilization, namely at 121 ℃ and the pressure of 0.4kg/cm 2 Sterilizing for 30min under the condition; after inoculation of the fermentation tank, culturing for 72 hours at 30 ℃, wherein the ventilation rate is 1.5 Vols/vol.min, and the oxygen content is 35% by volume;
LB medium formula: 1% of tryptone, 5% of yeast powder, 0.5% of sodium chloride, water with the pH value of 7.2 and 1.8-2.0% of agar into a solid culture medium;
the formula of the fermentation medium comprises: 2% of maltose, 1% of starch, 1.25% of yeast extract, 0.1% of NaCl, 0.3% of ammonium sulfate, 0.1% of calcium carbonate and the balance of water, wherein the initial pH value is 7.2-7.5;
the percentage content of the formula components of the culture medium is weight percentage;
fermentation broth requirements: the fermentation liquid contains more than 50 hundred million viable spores per milliliter, and is placed in a tank when the spore proportion reaches more than 90%, and the pH value is 7.0-8.0, so that the mixed bacteria pollution is avoided;
c. the carbon-nitrogen ratio of the decomposed organic fertilizer is adjusted to be 25:1 respectively, the weight ratio of water is adjusted to be 40%, fermentation liquid of bacillus belicus and bacillus amyloliquefaciens is inoculated according to the proportion of 0.5% of inoculum size, and natural composting fermentation is carried out after uniform mixing. When the temperature reaches above 50 ℃, turning the pile for 2-3 times per day; completing the compost fermentation until the germination index of the organic fertilizer is more than or equal to 90%;
the decomposed organic fertilizer is prepared by decomposing saw dust, mushroom residue, bran, chicken manure and pig manure according to the weight ratio of 0.5:0.5:2:3:4.
(2) Preparation of active functional streptomycete preparation:
a. strain activation
Streptomyces Ahn65 was inoculated into ISP2 solid medium for activation, and then transferred to 50mL ISP2 medium, followed by shaking culture at 28℃and 180rpm for 1-2d, to prepare seed solution.
b. Solid fermentation
Inoculating the culture solution of streptomyces Ahn65 into sterile bran at a volume-mass ratio of 1%, supplementing water to a volume-mass ratio of 40%, standing at 28 ℃ for 20d, and stopping culturing when the spore content of the streptomyces reaches more than 50 hundred million/g.
c. Streptomycete preparation
And (3) drying the solid fermentation product of the streptomyces Ahn65 in a 40 ℃ incubator for about 24 hours to prepare streptomyces powder, and bagging, sealing and preserving. The effective viable count reaches 200-500 hundred million/g by adopting a flat plate dilution plating counting method.
(3) Preparing a biological organic fertilizer:
adding the streptomycete preparation into the bio-organic fertilizer which is a product obtained by secondary fermentation of the functional bacteria prepared in the step (1) according to the mass ratio of 1%, uniformly mixing, transferring into a low-temperature dryer for drying until the water content is less than or equal to 30%, and obtaining the bio-organic fertilizer which is rich in three functional microorganisms of bacillus belicus, bacillus amyloliquefaciens and streptomycete, wherein the effective viable count of each microorganism is not less than 2 hundred million/g.
(4) Preparing a full-element biological organic fertilizer:
the amino acid raw powder is purchased from Shandong Weifang Lloyd Biotechnology Co., ltdContains 80% of plant source amino acids. The compound fertilizer was purchased from Shandong Shindanli fertilizer Co., ltd., total nutrient (N+P) 2 O 5 +K 2 The O) content was 50%. And (3) feeding the prepared bio-organic fertilizer into a storage bin, adding amino acid raw powder and compound fertilizer with the mass percent of 6.9% respectively, uniformly mixing, and packaging to obtain the full-element bio-organic fertilizer.
The total biological organic fertilizer has effective viable count content of 6-80 hundred million/g, organic matter content of 30-55%, total nutrient content of NPK of 8-11%, moisture content of less than or equal to 30%, total amino acid of 5-10%, free amino acid of 0.5-5%, chloride ion content of less than or equal to 2.0% and pH of 6.0-7.0.
Example 2: exploring experiment of organic fertilizer component:
1. preparing an organic fertilizer component:
preparing organic fertilizers 1 with different decomposition degrees: uniformly mixing chicken manure and pig manure which are treated at a high temperature of 85 ℃ for 2 days with crushed sawn wood ash, mushroom residues and bran in a weight ratio of 3:4:0.5:0.5:2, transferring into a large solid composting tank, adjusting the carbon-nitrogen ratio to 30:1, adjusting the water content to 50%, adding an organic material decomposition agent for composting, turning and ventilating when the composting temperature reaches above 60 ℃, turning and composting for 3 times per day, fermenting for 15 days, taking out part of the organic fertilizer as incompletely decomposed organic fertilizer 1, continuously composting for about 20 days, and properly adding a small amount of water until the germination index of the organic fertilizer is more than or equal to 90%, decomposing completely, and stopping composting to obtain the decomposed organic fertilizer 1;
preparation of the organic fertilizer 2 decomposed to different degrees: uniformly mixing pig manure and bran which are subjected to high-temperature treatment at 85 ℃ for 2 days after sieving and crushing according to the weight ratio of 8:2, transferring into a large solid composting tank, adjusting the carbon-nitrogen ratio to 30:1, adjusting the water content to 50%, adding an organic material decomposing agent for composting, turning and ventilating when the composting temperature reaches above 60 ℃, turning and ventilating for 3 times per day, composting and fermenting for 15 days, and taking out part of the organic material decomposed agent as incompletely decomposed organic fertilizer 2; and composting is continued until the germination index of the organic fertilizer is more than or equal to 90% until the organic fertilizer is completely decomposed, and composting is stopped, so that the decomposed organic fertilizer 2 is obtained.
Organic fertilizer 3: some brands of organic fertilizers purchased in the farmer market of marjoram in the Changsha city.
2. Secondary fermentation of functional bacteria:
1) And (3) fumbling an organic fertilizer formula for secondary fermentation:
different kinds of functional microorganisms including bacillus bailii, bacillus amyloliquefaciens and streptomycete are developed, and conditions and levels of secondary fermentation are performed in organic fertilizers with different raw material ratios, different decomposition degrees and different sources.
Spore suspensions of bacillus belicus, bacillus amyloliquefaciens fermentation broth and streptomycete are respectively inoculated into the 5 different organic fertilizers according to the inoculation amount of 1%, water is supplemented to 40% of the volume weight ratio of water, the carbon-nitrogen ratio is regulated to 25:1, and secondary composting fermentation is carried out after full stirring and uniform mixing, and meanwhile, the treatment without inoculating microorganisms is set as a control. When the temperature reaches above 50 ℃, turning the pile for 2-3 times per day. Sampling on day 3, diluting the sample by adopting a flat plate dilution coating method, and then respectively carrying out dilution gradient 10 -5 -10 -7 The number of colonies in different dilution gradients was counted and the average number of target microorganisms in the samples was counted according to dilution factors.
As shown in Table 1, when Bacillus bailii and Bacillus amyloliquefaciens both use decomposed organic fertilizer 1 as the secondary fermentation raw material, the proliferation level is highest, then decomposed organic fertilizer 2 is subjected to secondary fermentation in non-decomposed organic fertilizer 1, non-decomposed organic fertilizer 2 and commercial organic fertilizer, the proliferation level is not high, which indicates that the decomposition degree of the organic fertilizer is a key index for secondary fermentation of functional bacteria, and in addition, the material proportion and nutritional composition of the organic fertilizer also affect the secondary fermentation of the functional bacteria. In this experiment, the secondary fermentation of Streptomyces in all organic fertilizers did not show that this strain did not proliferate effectively, and it is possible that Streptomyces was unsuitable for the secondary fermentation in organic fertilizers.
Table 1 horizontal units of secondary fermentation of microorganisms in different organic fertilizers: *10 8 CFU/g
2) Fumbling secondary fermentation conditions
According to the conclusion, the decomposed organic fertilizer 1 is selected as the organic fertilizer for secondary fermentation, and the condition of secondary fermentation is searched.
The organic fertilizer water volume and weight ratio are respectively adjusted to be 40%, 50% and 60%, functional bacteria bacillus bailii and bacillus amyloliquefaciens are inoculated according to the inoculum size of 1%, and the organic fertilizer water volume and weight ratio are uniformly stirred to carry out indoor normal-temperature solid fermentation. When the temperature reaches above 50 ℃, turning the pile for 2-3 times per day. After natural composting fermentation for 3d, determining the bacterial count of bacillus bailii and bacillus amyloliquefaciens for secondary fermentation by a plate counting method. The results are shown in Table 2, and the number of secondary fermentation bacteria of Bacillus bailii and Bacillus amyloliquefaciens decreases with increasing water content, preferably 40%.
Table 2 influence units of different water contents on the number of functional bacteria secondary fermentation bacteria: *10 8 CFU/g
The carbon-nitrogen ratio of the decomposed organic fertilizer 1 is adjusted to 25-30: 1, the volume and weight ratio of water is 40%, bacillus beliae and bacillus amyloliquefaciens are respectively inoculated according to the inoculum sizes of 0.5%, 1% and 2%, when the temperature reaches more than 50 ℃, the stack turning is started, and the stack turning is carried out for 2-3 times per day. After natural composting fermentation for 3d, determining the bacterial count of bacillus bailii and bacillus amyloliquefaciens for secondary fermentation by a plate counting method. As a result, table 3 shows that the number of Bacillus bailii and Bacillus amyloliquefaciens each had a maximum inoculum size of 2%, but in view of cost and convenience, it is preferable to select an inoculum size of 0.5%.
Table 3 effect unit of different inoculum size on number of functional bacteria secondary fermentation bacteria: *10 8 CFU/g
The carbon-nitrogen ratio of the decomposed organic fertilizer 1 is adjusted to be 20: 1. 25: 1. 30:1, adjusting the volume weight ratio of water to 40%, inoculating bacillus belicus and bacillus amyloliquefaciens according to the proportion of 0.5% of inoculum size, uniformly mixing, carrying out natural composting fermentation, and turning the pile for 2-3 times per day when the temperature reaches above 50 ℃. After natural composting fermentation for 3d, determining the bacterial count of bacillus bailii and bacillus amyloliquefaciens for secondary fermentation by a plate counting method. The results are shown in Table 4, where the secondary fermentation carbon to nitrogen ratio of Bacillus belicus and Bacillus amyloliquefaciens is 25:1 is preferable. At this time, the highest fermentation level of Bacillus bailii was 59.32 hundred million/g, whereas the highest fermentation level of Bacillus amyloliquefaciens was 15.85 hundred million/g. Saving a lot of cost for the mass proliferation and application of functional microorganisms.
Table 4 influence unit of different carbon nitrogen ratios on the number of functional bacteria secondary fermentation bacteria: *10 8 CFU/g
Example 3: full-element bio-organic fertilizer rice field test:
(1) And (3) test design:
the field test is carried out in Yiyang city of Hunan province in 7-11 months of 2023, the field area is 5 mu, the soil texture is sticky soil, the soil type is red yellow mud, rape is sowed on previous crops, and then the milk vetch green manure is sowed. Basic physical and chemical properties of soil: pH 5.24, organic matter 38.93g/kg, total nitrogen 2.22g/kg, available phosphorus 7.97mg/kg, quick-acting potassium 176.17mg/kg and Cd 0.32mg/kg.
The test field is provided with 4 treatments, each treatment comprises 3 cells, each cell comprises 12 cells, and the area of each cell is 20m 2 And 3m wide protection rows are arranged around the test field. The test cell ridges are all covered by agricultural films from above the plow bottom layer, so that fertilizer and water strings are prevented from being drained and irrigated. The following 4 treatments are set, and the field management and the pest control are completely consistent.
Treatment 1: the full-element biological organic fertilizer and the fertilizer are applied in a decrement way, namely 80 percent of the conventional fertilizer is applied before the rice is transplanted, namely 16kg/6 of the compound fertilizer is applied67m 2 As a base fertilizer, 150kg/667m of full-element biological organic fertilizer is applied one week after rice transplanting 2 。
Treatment 2: the inactivated full-element biological organic fertilizer and the fertilizer are subjected to decrement fertilization, namely 80 percent of conventional fertilization is applied before rice transplanting, namely 16kg/667m of compound fertilizer is applied 2 As a base fertilizer, 150kg/667m of inactivated whole-element biological organic fertilizer is applied one week after rice transplanting 2 。
Treatment 3: conventional fertilization, wherein 20kg/667m of compound fertilizer is applied according to the amount of locally recommended fertilizer application 2 。
Treatment 4: the blank (CK) was not fertilized.
(2) Test results:
the application of the full-element bio-organic fertilizer can obviously promote the growth and tillering of rice, the plant height of the rice is respectively increased by 25.10 percent and 14.96 percent compared with a blank control and conventional fertilization, and the tillering number is respectively increased by 75.78 percent and 57.39 percent compared with the blank control and conventional fertilization. The wet weight and dry weight of the plot are increased by 41.59% and 42.71% respectively compared with the blank control, and 18.83% and 17.74% compared with the conventional fertilization, which is the first creation of the full-element bio-organic fertilizer for paddy rice, and the yield increase level is far higher than that of the bio-organic fertilizer and the organic fertilizer sold in the market at present, and is shown in tables 5 and 6.
TABLE 5 influence of different treatments on economic Properties of Rice
TABLE 6 Effect of different treatments on Rice yield
Meanwhile, the full-element biological organic fertilizer can also improve the disease resistance of rice, has a disease index prevention effect on rice leaf blast as high as 66.08% (table 7), and can effectively improve the nutrition indexes such as nitrogen, phosphorus, potassium and the like of soil and the content of organic matters in the soil (table 8). The treatment of the full-element bio-organic fertilizer remarkably improves the abundance and diversity of soil microorganisms, and the indexes respectively representing the number of species (OTU number), the total number of species index (ace index, chao index) and the biological diversity (shannon) of the species of the soil microorganisms are remarkably higher than those of a control (Table 9).
TABLE 7 Effect of different treatments on late rice leaf blast
TABLE 8 Effect of different treatments on late rice rhizosphere soil nutrient content
TABLE 9 Effect of different treatments on rhizosphere microbial diversity
Claims (10)
1. The preparation method of the rice full-element biological organic fertilizer is characterized by comprising the following steps of:
(1) Firstly, carrying out liquid fermentation on bacillus bailii 20-10 and bacillus amyloliquefaciens CWJ2, and then adding the fermentation liquor into a decomposed organic fertilizer for solid state fermentation;
(2) Carrying out solid fermentation on streptomyces Ahn 65;
(3) Mixing the fermentation products obtained in the step (1) and the step (2) with amino acid and an inorganic compound fertilizer to obtain a rice full-element biological organic fertilizer;
the preservation numbers of bacillus belicus 20-10, bacillus amyloliquefaciens CWJ2 and streptomycete Ahn65 are CCTCC NO: M20221980,CCTCC NO:M20221983,CCTCC NO:M20221981 respectively.
2. The method according to claim 1, wherein step (1) specifically comprises:
a. activating strains: b, bacillus belicus 20-10 and bacillus amyloliquefaciens CWJ2 are separately inoculated on a solid LB culture medium to activate strains, and then are separately inoculated on an LB liquid culture medium to be cultured, so as to prepare seed liquid;
b. fermentation culture: inoculating seed liquid into a fermentation tank for culture according to the inoculum size of 0.5-2% by volume, and introducing sterile air for fermentation culture;
the fermentation liquid of the two bacteria contains more than 50 hundred million viable spores per milliliter, and when the spore proportion reaches more than 90%, the fermentation liquid is placed in a tank, and the pH value is 7.0-8.0, so that no mixed bacteria pollution is caused;
c. the carbon-nitrogen ratio of the decomposed organic fertilizer is adjusted to be 20:1-30:1, the weight ratio of water is adjusted to be 40-60%, and fermentation liquid of bacillus bailii and bacillus amyloliquefaciens is inoculated according to the proportion of 0.5-2% of inoculum size, and the fermentation liquid is uniformly mixed for natural composting fermentation.
3. The method according to claim 2, wherein,
a. activating strains: inoculating bacillus belicus 20-10 and bacillus amyloliquefaciens CWJ2 on a solid LB culture medium, culturing for 16-30 hours at 30 ℃ and then activating strains, inoculating the strains into 100mL LB liquid culture medium by using a sterile ring, and culturing for 16-24 hours at 30 ℃ and 180rpm to prepare seed liquid;
b. fermentation culture: sterilizing a fermentation tank, filling a culture medium, sterilizing, maintaining the pressure, cooling to 30 ℃, inoculating fermentation liquor of a seed tank into the fermentation tank for culture according to the inoculation amount of 0.5-2%, and introducing sterile air; culturing for 60-72 h at 30 ℃ with aeration rate of 1.5 Vols/vol.min and oxygen content of 35% by volume;
the formula of the fermentation medium comprises: 2% of maltose, 1% of starch, 1.25% of yeast extract, 0.1% of NaCl, 0.3% of ammonium sulfate, 0.1% of calcium carbonate and the balance of water, wherein the initial pH value is 7.2-7.5;
c. the carbon-nitrogen ratio of the decomposed organic fertilizer is adjusted to be respectively 20:1-30:1, the weight ratio of water is adjusted to be 40-60%, fermentation liquid of bacillus bailii and bacillus amyloliquefaciens is inoculated according to the proportion of 0.5-2% of inoculum size, and natural composting fermentation is carried out after uniform mixing; when the temperature reaches above 50 ℃, turning the pile for 2-3 times per day; completing the compost fermentation until the germination index of the organic fertilizer is more than or equal to 90%;
the decomposed organic fertilizer is prepared by decomposing saw dust, mushroom residue, bran, chicken manure and pig manure according to the weight ratio of 0.5:0.5:2-3:2-3:4.
4. The preparation method of claim 3, wherein in step c, the carbon nitrogen ratio of the decomposed organic fertilizer is adjusted to be 25:1, the weight ratio of water is adjusted to be 40%, and fermentation liquid of bacillus belicus and bacillus amyloliquefaciens is inoculated according to the proportion of 0.5% of inoculation amount, and natural composting fermentation is carried out after uniform mixing.
5. The preparation method according to claim 1, wherein the step (2) specifically comprises:
a. strain activation
The streptomycete Ahn65 is accessed into an ISP2 solid culture medium for activation, and then transferred into an ISP2 liquid culture medium for culture to prepare seed liquid;
b. solid fermentation
Inoculating the seed liquid of streptomyces Ahn65 into sterile bran according to the mass ratio of 1-5%, standing for culturing, and stopping culturing when the spore content of the streptomyces reaches more than 50 hundred million/g;
c. and drying the solid fermentation product of the streptomyces Ahn65 to prepare the streptomyces bacterial powder.
6. The method according to claim 5, wherein,
a. strain activation
Activating Streptomyces Ahn65 in ISP2 solid culture medium, transferring to 50mL ISP2 culture solution, and shake culturing at 28deg.C and 180rpm for 1-2d to obtain seed solution;
b. solid fermentation
Inoculating the culture solution of streptomyces Ahn65 into the sterile bran at a mass ratio of 1-5%, supplementing water to a mass ratio of 40-60%, standing at 28 ℃ for culturing for 10-20d, and stopping culturing when the spore content of the streptomyces reaches more than 50 hundred million/g;
c. and (3) drying the solid fermentation product of the streptomycete Ahn65 in a 40 ℃ incubator for 24 hours to prepare the streptomycete bacterial powder.
7. The process according to any one of claim 1 to 6, wherein,
adding streptomycete powder into the product of the step (1) according to the mass ratio of 1-2%, uniformly mixing, and drying at low temperature until the water content is less than or equal to 30%, thereby obtaining the bio-organic fertilizer rich in three functional microorganisms of bacillus bailii, bacillus amyloliquefaciens and streptomycete, wherein the effective viable count of each microorganism is not less than 2 hundred million/g.
8. The method according to claim 7, wherein,
adding 5-8% of amino acid raw powder and inorganic compound fertilizer into the prepared bio-organic fertilizer, and uniformly mixing to obtain the full-element bio-organic fertilizer; the total biological organic fertilizer has effective viable count of 6-80 hundred million/g, organic matter content of 30-55%, total nutrient of NPK of 8-11%, moisture content of less than or equal to 30%, total amino acid of 5-10%, free amino acid of 0.5-5%, chloride ion content of less than or equal to 2.0% and pH of 6.0-7.0.
9. A rice whole-cell bio-organic fertilizer prepared by the method of any one of claims 1-8.
10. The use of the rice whole-element biological organic fertilizer as claimed in claim 9, wherein the rice growth is promoted, the soil nutrition is improved, and the disease resistance level of the rice is enhanced.
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