CN115057731A - Special bio-organic fertilizer for Chinese chestnut mycorrhiza and preparation method thereof - Google Patents
Special bio-organic fertilizer for Chinese chestnut mycorrhiza and preparation method thereof Download PDFInfo
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Abstract
The invention provides a special biological organic fertilizer for Chinese chestnut mycorrhiza and a preparation method thereof, relating to the technical field of biological organic fertilizers, wherein the special biological organic fertilizer is prepared from the following raw materials: 60-80 parts of chestnut mushroom dregs, 3-7 parts of bran, 1-5 parts of biochar, 8-12 parts of soybean meal and 0.1-1 part of composite mycorrhizal fungi agent; the compound mycorrhizal fungi agent comprises cellulose degrading bacillus, phosphate solubilizing aspergillus niger, potassium bacteria bacillus licheniformis, mycorrhizal fusarium, bolete fusca and bolete edulis; the raw materials are naturally fermented and then fermented by the compound mycorrhizal fungi agent to prepare the biological organic fertilizer. The method has the advantages of improving the nutrient absorption capacity of the Chinese chestnut, adjusting the stable microbial community structure in the Chinese chestnut root system soil, comprehensively improving the yield and the quality of the Chinese chestnut, simultaneously contributing to improving the soil fertility, improving the soil function and realizing the circular sustainable development of the Chinese chestnut industry.
Description
Technical Field
The invention relates to the technical field of biological organic fertilizers, in particular to a biological organic fertilizer special for Chinese chestnut mycorrhiza and a preparation method thereof.
Background
Chinese chestnut (Castarea mollissima) is one of the main economic woods in China and is called woody grain. Chinese chestnut has wide distribution area, and is famous in Hebei Qianxi, Shaanxi Zhen' an, Hubei Huanggang and the like. Hubei Huanggang 'Lutian chestnut' is a national geographical sign protection variety. Chestnut tree is a chestnut plant of fagaceae, and the chestnut fruit is rich in nutrition, has higher vitamin content than tomato, and is more than ten times of apple. The minerals in chestnut contain nutrient elements such as potassium, zinc, iron, etc.
The chestnut is a mycorrhizal plant, and the root system of the chestnut can be symbiotic with ectomycorrhizal fungi to form typical ectomycorrhiza. After mycorrhiza is formed, epidermal cells of new roots of the Chinese chestnut are stimulated to be increased remarkably, the absorption area of the roots is increased, the absorption capacity is enhanced, the influence of ectomycorrhizal fungi on the growth of the Chinese chestnut is mainly realized by changing the soil environment, the ectomycorrhiza increases the contact area with the soil through special structures, and the epitaxial hypha and the mycorrhiza have certain influence on N, P circulation in the soil, so that the absorption and utilization of nutrient substances in the soil by plants are promoted. The secretion of the chestnut mycorrhiza can dissolve insoluble or insoluble minerals in soil, so that effective mineral nutrient elements are released for the root system of the chestnut to absorb. When the water content of the soil is reduced, the mycorrhiza can enhance the water absorption capacity, so that the drought resistance of the Chinese chestnut can be improved, and the exomycorrhizal fungi can improve the resistance of the Chinese chestnut to diseases and insect pests, heavy metals and other extreme environments, so that the survival capability of plants is improved. Promoting the formation of a large amount of Chinese chestnut mycorrhiza and playing an important role in improving the yield and the quality of Chinese chestnuts. How to strengthen the field planting and growth of the chestnut root system ectomycorrhizal fungi and how to adjust the stable microbial community structure in the chestnut root system soil has important significance for the formation and growth of a large number of chestnut root fungi.
Disclosure of Invention
The invention aims to provide the special biological organic fertilizer for the chestnut mycorrhiza, which strengthens the field planting and the growth of the chestnut root system ectomycorrhiza fungi, promotes the formation and the growth of a large amount of chestnut mycorrhiza, improves the nutrient absorption capacity of chestnuts, improves the stress resistance of chestnuts, regulates the stable microbial community structure in the chestnut root system soil, improves the yield and the quality of chestnuts all over, is beneficial to improving the soil fertility and the soil function, and realizes the circular sustainable development of the chestnut industry.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a special biological organic fertilizer for Chinese chestnut mycorrhiza, which is prepared from the following raw materials:
60-80 parts of chestnut mushroom dregs, 3-7 parts of bran, 1-5 parts of biochar, 8-12 parts of soybean meal and 0.1-1 part of composite mycorrhizal fungi agent;
the compound mycorrhizal fungi agent comprises cellulose degrading bacillus, phosphate solubilizing aspergillus niger, potassium bacteria bacillus licheniformis, mycorrhizal fusarium, bolete fusca and bolete edulis;
the raw materials are naturally fermented and then fermented by the compound mycorrhizal fungi agent to prepare the biological organic fertilizer.
Preferably, the cellulose degrading bacteria bacillus in the composite mycorrhizal agent: phosphate solubilizing bacteria aspergillus niger: potassium-decomposing bacterium bacillus licheniformis: fusarium mycorrhizae: lactobacillus fuscus: the volume ratio of the boletus edulis seed liquid is 0.5-1.5: 0.5-1.5: 0.5-1.5: 0.5-1.5: 1.5-4.5: 1.5-4.5.
Preferably, the cell concentrations of the single strain seed solutions of the phosphate solubilizing bacteria Aspergillus niger, the Lactobacillus brownii and the boletus edulis are respectively 0.4-0.8 g/mL.
Preferably, the cell concentrations in the single-strain seed solutions of the cellulose degrading bacteria bacillus, the potassium bacteria bacillus licheniformis and the mycorrhizal fusarium respectively are 6-8 multiplied by 10 9 one/mL.
Preferably, the preparation method of the compound mycorrhizal inoculant comprises the following steps:
and (2) mixing single strain seed solutions of cellulose degrading bacteria bacillus, phosphorus dissolving bacteria aspergillus niger, potassium bacteria bacillus licheniformis, mycorrhizal fusarium, boletus fuscogilus and boletus edulis according to a proportion, inoculating the mixture into a culture medium, culturing for 3-5 h, adding glycerol, and continuously culturing for 1-2 h to obtain the composite mycorrhizal agent.
Preferably, in the preparation method of the compound mycorrhizal inoculant, the inoculation amount of the mixed seed liquid accounts for 0.8-1.2% of the total volume.
Preferably, in the preparation method of the compound mycorrhizal fungi agent, the culture medium takes water as a solvent and comprises the following components: 180-220 g/L of potatoes, 15-25 g/L of glucose and 15-20 g/L of agar.
Preferably, in the preparation method of the compound mycorrhizal inoculant, the culture temperature is 26-30 ℃, and the culture rotation speed is 120-160 rpm.
The invention also provides a preparation method of the special biological organic fertilizer for the chestnut mycorrhiza, which comprises the following steps:
stacking chestnut mushroom dregs, bran, biochar and bean pulp, stacking and fermenting for 5-9 days, then spraying a compound mycorrhizal microbial inoculum on the surface, fermenting for 25-35 days at 22-28 ℃, and turning over once every 7 days.
Preferably, the stacking height is 0.8 to 1.5m and the width is 0.8 to 1.5 m.
The invention provides a special bio-organic fertilizer for Chinese chestnut mycorrhiza and a preparation method thereof, the bio-organic fertilizer adopts bolete fuscous, boletus edulis, fusarium mycorrhizae and phosphorus and potassium dissolving functional strains bacillus licheniformis and aspergillus niger to perform a specific fermentation process on fertilizer raw materials, and promotes the formation of Chinese chestnut mycorrhiza through the function of functional microorganisms, so that insoluble phosphorus and potassium elements in soil are converted into soluble phosphorus and potassium elements, nutrient elements are provided for the Chinese chestnut root system, and meanwhile, a stable soil biological environment for the Chinese chestnut root system in which Chinese chestnut microorganisms are in mutual beneficial symbiosis is provided for the Chinese chestnut root system. The special bio-organic fertilizer for promoting the growth of the chestnut mycorrhiza can obviously improve the growth nutrition condition of chestnuts, improve the resistance of the chestnuts, promote the structure and function of soil microorganisms and improve the yield and quality of the chestnuts by matching the compound microbial inoculum and fermenting raw materials.
Drawings
FIG. 1 is a view of the roots of chestnuts to which the bio-organic fertilizer described in example 1 is not applied;
FIG. 2 is a view of the roots of chestnuts to which the bio-organic fertilizer described in example 1 is applied;
FIG. 3 is a view of the roots of chestnut seedlings to which the bio-organic fertilizer described in example 1 is applied;
FIG. 4 shows the morphological characteristics of chestnut mycorrhiza of a Chinese chestnut to which the bio-organic fertilizer described in example 1 is applied under a stereomicroscope;
FIG. 5 is a paraffin section of mycorrhiza to which the bio-organic fertilizer described in example 1 is applied;
fig. 6 is a demonstration chart of the field application of the bio-organic fertilizer described in example 1.
Deposit description
Bacillus HM01, latin for Bacillus sp.hm01, deposited in the chinese collection of cultures at the address: the preservation date of Wuhan university in Wuhan, China is as follows: and 9 and 27 days 2017, wherein the preservation number is CCTCC NO: and M2017558.
Detailed Description
The invention provides a special biological organic fertilizer for Chinese chestnut mycorrhiza, which is prepared from the following raw materials:
60-80 parts of chestnut mushroom dregs, 3-7 parts of bran, 1-5 parts of biochar, 8-12 parts of soybean meal and 0.1-1 part of composite mycorrhizal fungi agent;
the compound mycorrhizal fungi agent comprises cellulose degrading bacillus, phosphate solubilizing aspergillus niger, potassium bacteria bacillus licheniformis, mycorrhizal fusarium, bolete fusca and bolete edulis;
the raw materials are naturally fermented and then fermented by the compound mycorrhizal fungi agent to prepare the biological organic fertilizer.
In the chestnut mushroom residue special biological organic fertilizer, the chestnut mushroom residue is 60-80 parts by weight, preferably 65-75 parts by weight, and preferably 68-72 parts by weight.
In the biological organic fertilizer special for the chestnut mycorrhiza, the weight parts of the bran are 3-7 parts, the further optimization is 4-6 parts, and the further optimization is 5 parts.
In the biological organic fertilizer special for the chestnut mycorrhiza, the weight part of the biochar is 1-5 parts, preferably 2-4 parts, and preferably 3 parts.
In the biological organic fertilizer special for the chestnut mycorrhiza, the weight part of the soybean meal is 8-12 parts, the preferable weight part is 9-11 parts, and the preferable weight part is 10 parts.
In the biological organic fertilizer special for Chinese chestnut mycorrhiza, the composite mycorrhiza microbial inoculum accounts for 0.1-1 part by weight, preferably 0.2-0.8 part by weight, and preferably 0.4-0.6 part by weight.
In the invention, the compound mycorrhizal inoculant preferably comprises cellulose degrading bacillus, phosphate solubilizing aspergillus niger, potassium bacteria bacillus licheniformis, mycorrhizal fusarium, boletus fuscogilus and boletus edulis; the cellulose degrading bacteria are bacillus: phosphate solubilizing bacteria aspergillus niger: potassium-decomposing bacterium bacillus licheniformis: fusarium mycorrhizae: lactobacillus fuscus: the volume ratio of the boletus edulis seed liquid is preferably 0.5-1.5: 0.5-1.5: 0.5-1.5: 0.5-1.5: 1.5-4.5: 1.5-4.5; more preferably 0.8 to 1.2: 0.8-1.2: 2.4-3.6; still more preferably 1:1:1:1:1.5: 1.5.
In the composite mycorrhizal inoculant, the cell concentrations of single strain seed solutions of the phosphorus-solubilizing bacteria Aspergillus niger, Lactobacillus fuscus and Boletus edulis are preferably 0.4-0.8 g/mL, more preferably 0.5-0.7 g/mL and even more preferably 0.6g/mL respectively; the cell concentrations in the single strain seed solutions of the cellulose degrading bacteria bacillus, the potassium decomposing bacteria bacillus licheniformis and the mycorrhizal fusarium are respectively preferably 6-8 multiplied by 10 9 One per mL, more preferably 6.5 to 7.5X 10 9 one/mL, still more preferably7×10 9 one/mL.
In the composite mycorrhizal inoculant provided by the invention, the preparation method of the seed liquid of cellulose degrading bacteria bacillus, potassium bacteria bacillus licheniformis and mycorrhizal fusarium preferably comprises the following steps:
respectively inoculating the single bacterial colonies of the strains in a culture medium, standing and culturing in the dark, centrifuging the cultured bacterial liquid, collecting precipitates, and then re-suspending the cells by using 0.9% of normal saline to obtain seed liquids, thereby respectively obtaining the seed liquids of cellulose degrading bacteria bacillus, potassium decomposing bacteria bacillus licheniformis and mycorrhizal fusarium.
In the preparation method of the seed liquid of the cellulose degrading bacteria bacillus, the potassium bacteria bacillus licheniformis and the mycorrhizal fungi fusarium, the culture medium takes water as a solvent and comprises the following components: 8-12 g/L of peptone, 10-20 g/L, NaCl 10-20 g/L of beef extract, further preferably 9-11 g/L of peptone, 12-18 g/L, NaCl 12-18 g/L of beef extract, further preferably 10g/L of peptone and 14-16 g/L, NaCl 14-16 g/L of beef extract; the pH of the culture medium is preferably 6.6 to 7.0, more preferably 6.7 to 6.9, and still more preferably 6.8.
In the preparation method of the seed liquid of the cellulose degrading bacteria bacillus, the potassium-decomposing bacteria bacillus licheniformis and the mycorrhizal fungi fusarium, the culture temperature is preferably 30-40 ℃, more preferably 34-36 ℃, and further preferably 35 ℃; the culture time is preferably 12 to 36 hours, more preferably 18 to 28 hours, and still more preferably 24 hours.
In the preparation method of the seed liquid of the cellulose degrading bacteria bacillus, the potassium bacteria bacillus licheniformis and the mycorrhizal fungi fusarium, the rotating speed of centrifugation after culture is preferably 2500-3500 rpm, more preferably 2800-3200 rpm, and still more preferably 3000 rpm; the centrifugation time is preferably 2-6 min, more preferably 3-5 min, and still more preferably 4 min.
In the composite mycorrhizal inoculant provided by the invention, the preparation method of the seed liquid of the phosphate solubilizing bacteria Aspergillus niger, Lactobacillus fuscogilus and boletus edulis preferably comprises the following steps:
respectively inoculating the single bacterial colonies of the strains in a culture medium, standing and culturing in the dark, centrifuging the cultured bacterial liquid, collecting precipitates, and then re-suspending the bacteria by using 0.9% of normal saline to obtain seed liquids, thereby respectively obtaining the seed liquids of phosphorus-solubilizing bacteria Aspergillus niger, boletus fuscogilus and boletus edulis.
In the preparation method of the seed liquid of the phosphate solubilizing bacteria Aspergillus niger, the Lactobacillus fuscus and the bolete, the culture medium takes water as a solvent, and comprises the following components of 180-220 g/L of potato and 10-30 g/L of glucose, more preferably 190-210 g/L of potato and 15-25 g/L of glucose, and further preferably 200g/L of potato and 20g/L of glucose; the pH of the culture medium is preferably 6.3-6.7, more preferably 6.4-6.6, and still more preferably 6.5.
In the method for preparing the seed liquid of the phosphate solubilizing bacteria Aspergillus niger, Lactobacillus fuscipus and boletus edulis, the culture temperature is preferably 25-35 ℃, more preferably 28-32 ℃, and still more preferably 30 ℃; the culture time is preferably 48 to 96 hours, more preferably 60 to 84 hours, and still more preferably 72 hours.
In the method for preparing the seed liquid of the phosphorus-solubilizing bacteria Aspergillus niger, Lactobacillus fusciparum and boletus edulis, the rotating speed of centrifugation after culture is preferably 2500-3500 rpm, more preferably 2800-3200 rpm, and even more preferably 3000 rpm; the centrifugation time is preferably 1 to 5min, more preferably 2 to 4min, and still more preferably 3 min.
In the present invention, the preparation method of the complex mycorrhizal fungi agent preferably comprises the following steps:
and (2) mixing single strain seed solutions of cellulose degrading bacteria bacillus, phosphorus dissolving bacteria aspergillus niger, potassium bacteria bacillus licheniformis, mycorrhizal fusarium, boletus fuscogilus and boletus edulis according to a proportion, inoculating the mixture into a culture medium, culturing for 3-5 h, adding glycerol, and continuously culturing for 1-2 h to obtain the composite mycorrhizal agent.
In the preparation method of the compound mycorrhizal fungi agent, the compound mycorrhizal fungi agent is inoculated in a culture medium for culture, and the inoculation amount of the mixed seed solution is preferably 0.8-1.2 percent of the total volume, more preferably 0.9-1.1 percent, and even more preferably 1 percent.
In the preparation method of the compound mycorrhizal fungi agent, a culture medium inoculated by the compound mycorrhizal fungi agent takes water as a solvent and comprises the following components: 180-220 g/L of potato, 15-25 g/L of glucose and 15-20 g/L of agar, more preferably 190-210 g/L of potato, 18-22 g/L of glucose and 16-19 g/L of agar, and still more preferably 200g/L of potato, 20g/L of glucose and 18g/L of agar.
In the preparation method of the compound mycorrhizal fungi agent, the compound mycorrhizal fungi agent is inoculated in a culture medium and cultured for a period of time, the culture time is preferably 3-5 h, more preferably 3.5-4.5 h, and even more preferably 4 h; and then adding glycerol, and continuing the culture, wherein the continuous culture time is preferably 1-2 h, more preferably 1.2-1.8 h, and still more preferably 1.4-1.6 h.
In the preparation method of the composite mycorrhizal inoculant, the glycerol and the culture medium are added according to the volume ratio of 1-1.4: 100, more preferably 1.1 to 1.3:100, and still more preferably 1.2: 100.
In the preparation method of the compound mycorrhizal inoculant, the compound mycorrhizal inoculant is inoculated in a culture medium for culture, the temperature is preferably 26-30 ℃, more preferably 27-29 ℃, and even more preferably 28 ℃; the rotation speed for culturing is preferably 120 to 160rpm, more preferably 130 to 150rpm, and still more preferably 140 rpm.
In the preparation method of the composite mycorrhizal inoculant, all culture media are disinfected and cooled for standby, and the preferred pressure of the disinfection treatment is 1.3-1.5 kg/cm 2 More preferably 1.4kg/cm 2 (ii) a The time for the disinfection treatment is preferably 2-2.5 h, and more preferably 2.25 h.
In the invention, the potassium bacteria, namely the bacillus licheniformis and the phosphorus solubilizing bacteria, namely the aspergillus niger strains are purchased from the China general microbiological culture collection center; the boletus fuscogilus, boletus edulis and fusarium mycorrhizae are purchased from the China agricultural microbial strain preservation management center; the cellulose degrading bacteria Bacillus is patent classical collection strain HM01, is named as Bacillus sp.HM01, is collected in China center for type culture collection in 2017, 9 and 27 months, and has a collection number of CCTCC NO: and M2017558.
The invention also provides a preparation method of the special biological organic fertilizer for the chestnut mycorrhiza, which comprises the following steps:
stacking chestnut mushroom dregs, bran, biochar and bean pulp, stacking and fermenting for 5-9 days, then spraying a compound mycorrhizal microbial inoculum on the surface, fermenting for 25-35 days at 22-28 ℃, and turning over once every 7 days.
According to the preparation method of the special bio-organic fertilizer for the chestnut mycorrhiza, chestnut mushroom dregs, bran, biochar and bean pulp are stacked, wherein the stacking height is preferably 0.8-1.5 m, more preferably 1-1.3 m, and further preferably 1.2 m; the stacking width is preferably 0.8 to 1.5m, more preferably 1 to 1.3m, and still more preferably 1.2 m.
According to the preparation method of the special bio-organic fertilizer for the chestnut mycorrhiza, the stack is stacked for fermentation, and the fermentation time is preferably 5-9 days, more preferably 6-8 days, and even more preferably 7 days; and then spraying the composite root bacteria agent on the surface, continuing fermentation, wherein the fermentation temperature is preferably 22-28 ℃, further preferably 24-26 ℃, further preferably 25 ℃, the fermentation time is preferably 25-35 days, further preferably 28-32 days, further preferably 30 days, and the pile is turned once every 7 days preferably in the fermentation process.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Respectively inoculating single colony (the formula of the culture medium is 10g of peptone, 15g of beef extract, 15 ml of NaCl, 15g of water and 1000ml of water, and the pH value is 6.8) to a culture medium, statically culturing at 35 ℃ in a dark place for 24 hours, centrifuging the cultured bacterium liquid at 3000rpm for 4 minutes, collecting precipitates, and then re-suspending the cells by using 0.9% physiological saline to respectively obtain the cellulose degrading bacteria bacillus, the potassium degrading bacteria bacillus licheniformis and the mycorrhizal fungi fusariumThe cell concentration of the seed liquid of the sword fungi is 0.6g/mL respectively; respectively inoculating single bacterial colonies (the formula of the culture medium is that potato 200g, glucose 20g and water 1000mL, and pH is 6.5) to a phosphorus-dissolving bacterium Aspergillus niger, Boletus luteus and Boletus edulis, standing and culturing at 30 ℃ in a dark place for 72h, centrifuging the cultured bacterial liquid at 3000rpm for 3min, collecting precipitates, and re-suspending the bacteria with 0.9% physiological saline to respectively obtain seed liquids of the phosphorus-dissolving bacterium Aspergillus niger, Boletus luteus and Boletus edulis, wherein the cell concentrations in the single bacterial seed liquids are respectively 7 × 10 9 one/mL.
The single strain seed liquid of cellulose degrading bacteria bacillus, phosphorus dissolving bacteria aspergillus niger, potassium bacteria bacillus licheniformis, mycorrhizal fusarium, lactobacillus brownii and bolete is mixed according to the volume ratio of 1:1:1:1:3:3, the inoculation amount of the mixed seed liquid is inoculated into a culture medium (the formula of the culture medium is that potato 200g, glucose 20g, agar 18g and water 1000mL) according to the inoculation ratio accounting for 1% of the total volume, the culture medium is cultured at 28 ℃ and 140rpm for 4 hours, then glycerol is added, the volume ratio of the glycerol to the culture medium is 1.2:100, and the culture is continued for 1.5 hours, so that the composite mycorrhizal microbial inoculum is obtained. All media were used at 1.4kg/cm 2 And (5) sterilizing for 2.25h under pressure.
Uniformly mixing 70 parts of chestnut mushroom dregs, 5 parts of bran, 3 parts of biochar and 10 parts of bean pulp, stacking for 7 days, wherein the stacking height is 1.2m and the width is 1.2m, spraying 0.5 part of composite mycorrhizal microbial inoculum on the surface, fermenting for 30 days at 25 ℃, and turning over once every 7 days.
Example 2
Respectively inoculating single bacterial colonies (the formula of the culture medium is 8g of peptone, 10g of beef extract, 10g of NaCl, 10g of water and 1000mL of water, and the pH value is 6.6) to cellulose-degrading bacteria bacillus, potassium-decomposing bacteria bacillus licheniformis and mycorrhizal fusarium in a culture medium, statically culturing for 12 hours at 30 ℃ in a dark place, centrifuging the cultured bacterial liquid for 2 minutes at 2500rpm, collecting precipitates, and then re-suspending the cells by using 0.9% physiological saline to respectively obtain seed liquids of the cellulose-degrading bacteria bacillus, the potassium-decomposing bacteria bacillus licheniformis and the mycorrhizal fusarium, wherein the cell concentrations in the single bacterial seed liquids are respectively 0.4 g/mL; respectively culturing Aspergillus niger, Lactobacillus brownii and Boletus edulisInoculating single colony (culture medium formula is potato 180g, glucose 10g, water 1000mL, pH6.3), standing at 25 deg.C in dark place for 48h, centrifuging the cultured bacterial solution at 2500rpm for 1min, collecting precipitate, re-suspending the bacteria with 0.9% normal saline to obtain seed solutions of phosphorus-solubilizing bacteria Aspergillus niger, Lactobacillus fuscogilus, and Boletus edulis, wherein the cell concentrations in the single-bacteria seed solutions are respectively 6 × 10 9 one/mL.
The single strain seed liquid of cellulose degrading bacteria bacillus, phosphorus dissolving bacteria aspergillus niger, potassium bacteria bacillus licheniformis, mycorrhizal fusarium, lactobacillus brownii and bolete is mixed according to the volume ratio of 0.5:0.5:0.5:0.5:1.5:1.5, the inoculation amount of the mixed seed liquid is inoculated into a culture medium (the formula of the culture medium is 180g of potato, 15g of glucose, 15g of agar and 1000mL of water) according to the inoculation ratio accounting for 0.8 percent of the total volume, the mixed seed liquid is cultured for 3 hours at 26 ℃ and 120rpm, then glycerol is added, the volume ratio of the glycerol to the culture medium is 1:100, and the composite mycorrhizal microbial inoculum is obtained after continuous culture for 1 hour. All media were used at 1.2kg/cm 2 And (5) sterilizing for 2 hours under pressure.
Uniformly mixing and stacking 60 parts of chestnut mushroom dregs, 3 parts of bran, 1 part of biochar and 8 parts of soybean meal, stacking for 5 days with the stacking height of 0.8m and the width of 0.8m, spraying 0.1 part of composite mycorrhizal inoculant on the surface, fermenting for 27 days at 22 ℃, and turning over once every 7 days.
Example 3
Respectively inoculating single bacterial colonies (the formula of the culture medium is that peptone 12g, beef extract 20g, NaCl20g, water 1000mL, and pH7.0) to cellulose-degrading bacteria bacillus, potassium-decomposing bacteria bacillus licheniformis and mycorrhizal fungi fusarium in a culture medium, statically culturing at 40 ℃ in a dark place for 28h, centrifuging the cultured bacterial liquid at 3500rpm for 6min, collecting precipitates, and then re-suspending the cells by using 0.9% physiological saline to respectively obtain seed liquids of the cellulose-degrading bacteria bacillus, the potassium-decomposing bacteria bacillus licheniformis and the mycorrhizal fungi fusarium, wherein the cell concentrations in the single bacterial seed liquids are respectively 0.8 g/mL; inoculating phosphate-solubilizing Aspergillus niger, Lactobacillus brownii and boletus edulis respectively to culture medium (the formula of the culture medium is 220g of potato, 30g of glucose and 1000mL of water, and the pH value is 6.7), standing at 35 deg.C in the dark for 96h, and culturingCentrifuging the bacterial solution at 3500rpm for 5min, collecting precipitate, re-suspending the bacteria with 0.9% physiological saline to obtain seed solutions of phosphorus-solubilizing bacteria Aspergillus niger, Lactobacillus fuscus and boletus edulis, wherein the cell concentrations in the seed solutions of single bacteria are respectively 8 × 10 9 one/mL.
The single strain seed liquid of cellulose degrading bacteria bacillus, phosphorus dissolving bacteria aspergillus niger, potassium bacteria bacillus licheniformis, mycorrhizal fusarium, lactobacillus brownii and bolete is mixed according to the volume ratio of 1:1:1:1:3:3, the inoculation amount of the mixed seed liquid is inoculated into a culture medium (the formula of the culture medium is 220g of potato, 25g of glucose, 20g of agar and 1000mL of water) according to the inoculation ratio accounting for 1.2 percent of the total volume, the culture medium is cultured for 5 hours at 30 ℃ and 160rpm, then glycerol is added, the volume ratio of the glycerol to the volume of the culture medium is 1.4:100, and the culture is continued for 2 hours to obtain the composite mycorrhizal microbial inoculum. All media were used at 1.2kg/cm 2 Sterilizing under pressure for 2.5 h.
Uniformly mixing 80 parts of chestnut mushroom dregs, 7 parts of bran, 5 parts of biochar and 12 parts of bean pulp, stacking for 9 days, wherein the stacking height is 1.5m and the width is 1.5m, spraying 1 part of compound mycorrhizal inoculant on the surface, fermenting for 35 days at 28 ℃, and turning over once every 7 days.
Examples of the experiments
In a Marble village Chinese chestnut garden base in a woodshop town of Macheng city, Hubei province, test lands with the width of 4 meters and the length of 30 meters are selected, 2-year-old Chinese chestnut seedlings are cultivated according to the interval of 1 meter, 10 Chinese chestnut seedlings in one row are applied with the special biological organic fertilizer as an experimental group, 10 Chinese chestnut seedlings in the other row are applied without fertilizer at the row spacing of 1 meter to serve as a blank group, and one row applied with a commercially available compound fertilizer is taken as a control group at the row spacing of 1 meter. After 1 year, the height, diameter and fresh weight of the overground part, dry weight of the overground part, fresh weight of the underground part and dry weight of the underground part of the Chinese chestnut are measured. The application method of the special bio-organic fertilizer in the embodiment 1 comprises the steps of digging a soil pit 50cm in length and 50cm in depth outside a position 50cm away from the root of the chestnut tree, applying the special bio-organic fertilizer for promoting the growth of the mycorrhiza of the chestnut according to the amount of 10-30 kg per pit, and covering the surface with 10cm of soil. The result shows that compared with the blank group, the effect of promoting the growth of the Chinese chestnut seedlings by applying the special organic fertilizer in the embodiment 1 is very obvious, wherein the plant height is increased by nearly 50%, the fresh weight of the underground part is increased by 64.7%, which shows that the growth of the Chinese chestnut root system is obviously promoted, as shown in table 1 (note: the numerical value is the average number +/-standard deviation, and the difference significance P is less than 0.05 for the lower case letters), in the aspect of promoting the growth of the Chinese chestnuts, the fertilizer efficiency of the special biological organic fertilizer for the Chinese chestnut mycorrhiza is similar to that of the compound fertilizer sold in the market, and the growth of the Chinese chestnuts can be obviously promoted.
TABLE 1 influence of applying special bio-organic fertilizer for Chinese chestnut mycorrhiza on growth characteristics of Chinese chestnut
Note: the abcd difference significance analysis shows that the differences are not significant if the same marked letters exist, and the differences are significant if the letters with different marks exist.
As can be seen from the figure 5, the Chinese chestnut seedlings using the special bio-organic fertilizer have significantly more mycorrhiza formation, and a typical mycorrhiza structure Harti's network can be seen through paraffin section, which indicates that the special bio-organic fertilizer can significantly promote the formation of mycorrhiza.
The above experiments were followed and the fruiting status of 10 chestnut trees in the experimental, blank and control groups was observed three years later, and the results are shown in table 2.
TABLE 2 Chinese chestnut tree after three years
The results in table 2 show that the application of the special bio-organic fertilizer for promoting the growth of the chestnut mycorrhiza has higher chestnut fruiting quantity and average single plant setting rate than the control, low empty rate and less plant diseases and insect pests, and the whole growth and setting conditions of chestnuts are stronger than those of the blank group and the control.
The special biological organic fertilizer for the chestnut mycorrhiza provided by the invention is named as 'Guanlv No. 5' and is tested in the chestnut base of Dongmu breed specialty Cooperation society in Ma City, and fig. 6 shows an application proof of test effect.
According to the embodiments, the biological organic fertilizer special for the chestnut mycorrhiza provided by the invention can strengthen the field planting and growth of chestnut root system ectomycorrhizal fungi, promote the formation and growth of a large number of chestnut mycorrhiza, improve the nutrient absorption capacity of chestnuts, improve the stress resistance of chestnuts, regulate the stable structure of microbial communities in the chestnut root system soil, comprehensively improve the yield and quality of chestnuts, and simultaneously contribute to improving the soil fertility, improving the soil function and realizing the circular sustainable development of the chestnut industry.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. The special bio-organic fertilizer for the chestnut mycorrhiza is characterized by being prepared from the following raw materials:
60-80 parts of chestnut mushroom dregs, 3-7 parts of bran, 1-5 parts of biochar, 8-12 parts of soybean meal and 0.1-1 part of composite mycorrhizal fungi agent;
the compound mycorrhizal fungi agent comprises cellulose degrading bacillus, phosphate solubilizing aspergillus niger, potassium bacteria bacillus licheniformis, mycorrhizal fusarium, bolete fusca and bolete edulis;
the raw materials are naturally fermented and then fermented by the compound mycorrhizal fungi agent to prepare the biological organic fertilizer.
2. The special bio-organic fertilizer for Chinese chestnut mycorrhiza of claim 1, wherein the cellulose degrading bacteria bacillus in the compound mycorrhiza microbial inoculum: phosphate solubilizing bacteria aspergillus niger: potassium-decomposing bacterium bacillus licheniformis: rhizobial fusarium: lactobacillus fuscus: the volume ratio of the boletus edulis seed liquid is 0.5-1.5: 0.5-1.5: 0.5-1.5: 0.5-1.5: 1.5-4.5: 1.5-4.5.
3. The special bio-organic fertilizer for chestnut mycorrhiza as claimed in claim 2, wherein the cell concentrations in the single strain seed liquid of the phosphate solubilizing bacteria aspergillus niger, lactobacillus fuscogilus and boletus edulis are respectively 0.4-0.8 g/mL.
4. The special bio-organic fertilizer for Chinese chestnut mycorrhiza as claimed in claim 2, wherein the cell concentration of the single strain seed liquid of the cellulose degrading bacteria bacillus, the potassium decomposing bacteria bacillus licheniformis and the mycorrhizal fusarium is 6-8 x 10 9 one/mL.
5. The special bio-organic fertilizer for Chinese chestnut mycorrhiza as claimed in claim 2, wherein the preparation method of the compound mycorrhiza microbial inoculum comprises the following steps:
the single strain seed solutions of cellulose degrading bacteria bacillus, phosphorus-solubilizing bacteria aspergillus niger, potassium-solubilizing bacteria bacillus licheniformis, mycorrhizal fusarium, boletus fuscogilus and boletus edulis are mixed in proportion, inoculated in a culture medium, cultured for 3-5 hours, added with glycerol and continuously cultured for 1-2 hours to obtain the composite mycorrhizal agent.
6. The special bio-organic fertilizer for Chinese chestnut mycorrhiza as claimed in claim 5, wherein the inoculation amount of the mixed seed solution in the preparation method of the compound mycorrhiza inoculant accounts for 0.8-1.2% of the total volume.
7. The biological organic fertilizer special for Chinese chestnut mycorrhiza as claimed in claim 6, wherein the culture medium in the preparation method of the compound mycorrhiza microbial inoculum takes water as a solvent, and comprises the following components: 180-220 g/L of potatoes, 15-25 g/L of glucose and 15-20 g/L of agar.
8. The biological organic fertilizer special for Chinese chestnut mycorrhiza as claimed in claim 7, wherein the culture temperature in the preparation method of the compound mycorrhiza microbial inoculum is 26-30 ℃, and the culture rotation speed is 120-160 rpm.
9. The preparation method of the special bio-organic fertilizer for chestnut mycorrhiza as claimed in any one of claims 1 to 8, is characterized by comprising the following steps:
stacking chestnut mushroom dregs, bran, biochar and soybean meal, stacking and fermenting for 5-9 days, then spraying a compound mycorrhizal microbial inoculum on the surface, fermenting for 25-35 days at 22-28 ℃, and turning over once every 7 days.
10. The preparation method of the special bio-organic fertilizer for chestnut mycorrhiza as claimed in claim 9, wherein the stacking height is 0.8-1.5 m, and the width is 0.8-1.5 m.
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| CN115537208A (en) * | 2022-09-30 | 2022-12-30 | 武汉工程大学 | Method for preparing soil conditioner by utilizing phosphorus potassium tailings and waste biomass |
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| CN101077078A (en) * | 2006-05-25 | 2007-11-28 | 秦岭 | Chinese chestnut wine and preparation method thereof Chinese chestnut fungus root preparations and preparation method thereof |
| CN106242713A (en) * | 2016-06-21 | 2016-12-21 | 章兴荣 | A kind of ecological organic composite solid is fertile and preparation method thereof |
| CN106748398A (en) * | 2016-11-18 | 2017-05-31 | 潘昱 | A kind of Chinese chestnut dedicated fertilizer and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101077078A (en) * | 2006-05-25 | 2007-11-28 | 秦岭 | Chinese chestnut wine and preparation method thereof Chinese chestnut fungus root preparations and preparation method thereof |
| CN106242713A (en) * | 2016-06-21 | 2016-12-21 | 章兴荣 | A kind of ecological organic composite solid is fertile and preparation method thereof |
| CN106748398A (en) * | 2016-11-18 | 2017-05-31 | 潘昱 | A kind of Chinese chestnut dedicated fertilizer and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115537208A (en) * | 2022-09-30 | 2022-12-30 | 武汉工程大学 | Method for preparing soil conditioner by utilizing phosphorus potassium tailings and waste biomass |
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