A kind of biological organic fertilizer special for chestnut mycorrhiza and preparation method thereof

Abstract

The invention provides a special bio-organic fertilizer for chestnut mycorrhiza and a preparation method thereof, which relates to the technical field of bio-organic fertilizer. The special bio-organic fertilizer is prepared from raw materials including the following components: 60-80 parts of chestnut mushroom residue, 3-7 parts of bran, 1-5 parts of biochar, 8-12 parts of soybean meal, and 0.1-1 part of composite mycorrhizal agent; the composite mycorrhizal agent includes cellulose-degrading bacteria Bacillus, phosphate-dissolving bacteria Aspergillus niger, potassium-dissolving bacteria Bacillus licheniformis, mycorrhizal fungi Fusarium, brown-ringed Suillus and Boletus edulis; the raw materials are naturally fermented and then fermented with the composite mycorrhizal agent to prepare bio-organic fertilizer. The ability of chestnut to absorb nutrients is improved, the stability of the microbial community structure in the soil of the chestnut root system is adjusted, the yield and quality of chestnut are comprehensively improved, and at the same time, it is helpful to improve soil fertility, improve soil function, and realize the circular and sustainable development of the chestnut industry.

Description

Technical Field

[0001] The invention relates to the technical field of bio-organic fertilizers, and in particular to a bio-organic fertilizer special for chestnut mycorrhiza and a preparation method thereof. Background Art

[0002] Chestnut (Castarcea mollissima) is one of the main economic trees in my country and is known as a woody food. The distribution area of ​​chestnuts in my country is vast, and Qianxi, Hebei, Zhen'an, Shaanxi, and Huanggang, Hubei are all famous chestnut producing areas. The "Luotian Chestnut" in Huanggang, Hubei is a national geographical indication protected variety. The chestnut tree is a plant of the genus Castanea in the Fagaceae family. Its fruit chestnuts are rich in nutrients, with a higher vitamin content than tomatoes and more than ten times that of apples. The minerals in chestnuts contain nutrients such as potassium, zinc, and iron.

[0003] Chestnut is a mycorrhizal plant, and its root system can coexist with ectomycorrhizal fungi to form a typical ectomycorrhiza. After the formation of mycorrhiza, the epidermal cells of the new chestnut roots are stimulated to increase significantly, the absorption area of ​​the roots increases, and the absorption capacity is enhanced. The effect of ectomycorrhizal fungi on the growth of chestnut is mainly through changing the soil environment. The ectomycorrhizae increase the contact area with the soil by virtue of their special structure, the extended hyphae and mycelium, and have a certain effect on the N and P cycles in the soil, thereby promoting the absorption and utilization of nutrients in the soil by plants. The secretions of chestnut mycorrhizae can dissolve the insoluble or insoluble minerals in the soil, so that they release effective mineral nutrients for absorption by chestnut roots. When the soil moisture content decreases, mycorrhizae can enhance the water absorption capacity, so it can improve the drought resistance of chestnuts. Ectomycorrhizal fungi can improve the resistance of chestnuts to extreme environments such as pests and diseases, heavy metals, etc., thereby improving the survival ability of plants. Promoting the formation of a large number of chestnut mycorrhizae plays a very important role in improving the yield and quality of chestnuts. How to enhance the colonization and growth of ectomycorrhizal fungi in chestnut root system and how to regulate the stability of microbial community structure in chestnut root soil are of great significance for the massive formation and growth of chestnut root fungi. Summary of the invention

[0004] The purpose of the present invention is to provide a biological organic fertilizer special for chestnut mycorrhiza, which can strengthen the colonization and growth of ectomycorrhizal fungi in the chestnut root system, promote the formation and growth of chestnut mycorrhiza in large quantities, improve the ability of chestnut to absorb nutrients, improve the stress resistance of chestnut, regulate the stability of the microbial community structure in the soil of the chestnut root system, comprehensively improve the yield and quality of chestnut, and at the same time help to improve soil fertility, improve soil function, and realize the circular and sustainable development of the chestnut industry.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides a special biological organic fertilizer for chestnut mycorrhiza, which is prepared from raw materials including the following components:

[0007] 60-80 parts of chestnut mushroom residue, 3-7 parts of bran, 1-5 parts of biochar, 8-12 parts of soybean meal, and 0.1-1 part of composite mycorrhizal agent;

[0008] The composite mycorrhizal agent includes cellulose degrading bacteria Bacillus, phosphate solubilizing bacteria Aspergillus niger, potassium solubilizing bacteria Bacillus licheniformis, mycorrhizal fungi Fusarium, Suillus edulis and Boletus edulis;

[0009] The raw materials are naturally fermented and then fermented with a composite mycorrhizal agent to prepare biological organic fertilizer.

[0010] Preferably, the seed liquid volume ratio of cellulose degrading bacteria Bacillus: phosphate solubilizing bacteria Aspergillus niger: potassium solubilizing bacteria Bacillus licheniformis: mycorrhizal fungus Fusarium: Suillus edulis: Boletus edulis in the composite mycorrhizal agent 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.

[0011] Preferably, the cell concentrations in the single-bacteria seed solutions of the phosphate-solubilizing bacteria Aspergillus niger, Suillus suillus, and Boletus edulis are 0.4-0.8 g / mL, respectively.

[0012] Preferably, the cell concentrations of the single bacterial seed solution of the cellulose degrading bacteria Bacillus, the potassium-dissolving bacteria Bacillus licheniformis, and the mycorrhizal fungus Fusarium are 6 to 8×10 9 Pieces / mL.

[0013] Preferably, the preparation method of the composite mycorrhizal agent comprises the following steps:

[0014] The single bacterial seed liquids of cellulose degrading bacteria Bacillus, phosphate-dissolving bacteria Aspergillus niger, potassium-dissolving bacteria Bacillus licheniformis, mycorrhizal fungi Fusarium, Suillus edulis and Boletus edulis are mixed in proportion, inoculated into a culture medium, cultured for 3 to 5 hours, and then glycerol is added and cultured for 1 to 2 hours to obtain a composite mycorrhizal agent.

[0015] Preferably, in the preparation method of the composite mycorrhizal agent, the inoculation amount of the mixed seed liquid accounts for 0.8% to 1.2% of the total volume.

[0016] Preferably, in the preparation method of the composite mycorrhizal agent, the culture medium uses water as a solvent and includes the following components: 180-220 g / L of potatoes, 15-25 g / L of glucose, and 15-20 g / L of agar.

[0017] Preferably, in the preparation method of the composite mycorrhizal agent, the culture temperature is 26-30° C. and the culture speed is 120-160 rpm.

[0018] The present invention also provides a method for preparing the special biological organic fertilizer for chestnut mycorrhiza, comprising the following steps:

[0019] Pile chestnut mushroom residue, bran, biochar and soybean meal, ferment for 5 to 9 days, then spray composite mycorrhizal agent on the surface, ferment at 22 to 28 ℃ for 25 to 35 days, and turn the pile over every 7 days.

[0020] Preferably, the stacking height is 0.8 to 1.5 m and the width is 0.8 to 1.5 m.

[0021] The present invention provides a special biological organic fertilizer for chestnut mycorrhiza and a preparation method thereof. The biological organic fertilizer of the present invention uses brown ring lactucas, boletus edulis, mycorrhizal fungi Fusarium and phosphorus-dissolving and potassium-dissolving functional strains Bacillus licheniformis and Aspergillus niger to carry out a specific fermentation process on the fertilizer raw materials, and promotes the formation of chestnut mycorrhiza through the action of functional microorganisms, so that insoluble phosphorus and potassium elements in the soil are converted into soluble phosphorus and potassium elements, providing nutrient elements for the chestnut root system, and at the same time providing the chestnut root system with a stable soil biological environment for mutual symbiosis of chestnut microorganisms. The special biological organic fertilizer for promoting chestnut mycorrhiza growth of the present invention can significantly improve the nutritional status of chestnut growth, improve chestnut resistance, promote the structure and function of soil microorganisms, and improve chestnut yield and quality through the combination of the composite bacterial agent and the fermentation of the raw materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a picture of the chestnut root without applying the biological organic fertilizer described in Example 1;

[0023] Figure 2 This is a picture of the chestnut root after applying the biological organic fertilizer described in Example 1;

[0024] Figure 3 This is a picture of the roots of chestnut seedlings after applying the biological organic fertilizer described in Example 1;

[0025] Figure 4 The chestnut mycorrhiza morphological characteristics of the biological organic fertilizer described in Example 1 were obtained under a stereo microscope;

[0026] Figure 5 Paraffin sections of mycorrhizae of the bio-organic fertilizer described in Example 1;

[0027] Figure 6 This is a diagram demonstrating the field application of the bio-organic fertilizer described in Example 1.

[0028] Collection Instructions

[0029] Bacillus sp. HM01 in Latin is deposited in China Center for Type Culture Collection, located at Wuhan University, Wuhan, China. The deposit date is September 27, 2017, and the deposit number is CCTCC NO: M2017558. DETAILED DESCRIPTION

[0030] The present invention provides a special biological organic fertilizer for chestnut mycorrhiza, which is prepared from raw materials including the following components:

[0031] 60-80 parts of chestnut mushroom residue, 3-7 parts of bran, 1-5 parts of biochar, 8-12 parts of soybean meal, and 0.1-1 part of composite mycorrhizal agent;

[0032] The composite mycorrhizal agent includes cellulose degrading bacteria Bacillus, phosphate solubilizing bacteria Aspergillus niger, potassium solubilizing bacteria Bacillus licheniformis, mycorrhizal fungi Fusarium, Suillus edulis and Boletus edulis;

[0033] The raw materials are naturally fermented and then fermented with a composite mycorrhizal agent to prepare biological organic fertilizer.

[0034] In the biological organic fertilizer dedicated to chestnut mycorrhiza of the present invention, the weight proportion of chestnut mushroom residue is 60 to 80 parts, more preferably 65 to 75 parts, and even more preferably 68 to 72 parts.

[0035] In the chestnut mycorrhiza-specific biological organic fertilizer of the present invention, the weight portion of bran is 3 to 7 parts, more preferably 4 to 6 parts, and even more preferably 5 parts.

[0036] In the chestnut mycorrhiza-specific biological organic fertilizer of the present invention, the weight portion of biochar is 1 to 5 parts, more preferably 2 to 4 parts, and even more preferably 3 parts.

[0037] In the biological organic fertilizer dedicated to chestnut mycorrhizae of the present invention, the weight portion of soybean meal is 8 to 12 parts, more preferably 9 to 11 parts, and even more preferably 10 parts.

[0038] In the chestnut mycorrhiza special biological organic fertilizer of the present invention, the weight portion of the composite mycorrhizal agent is 0.1 to 1 part, more preferably 0.2 to 0.8 part, and even more preferably 0.4 to 0.6 part.

[0039] In the present invention, the composite mycorrhizal agent preferably includes cellulose degrading bacteria Bacillus, phosphate-soluble bacteria Aspergillus niger, potassium-solubilizing bacteria Bacillus licheniformis, mycorrhizal fungi Fusarium, brown-ringed boletus and boletus edulis; the seed liquid volume ratio of the cellulose degrading bacteria Bacillus: phosphate-soluble bacteria Aspergillus niger: potassium-solubilizing bacteria Bacillus licheniformis: mycorrhizal fungi Fusarium: brown-ringed boletus: boletus edulis 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; further preferably 0.8-1.2: 0.8-1.2: 0.8-1.2: 0.8-1.2: 2.4-3.6: 2.4-3.6; and further preferably 1: 1: 1: 1: 1.5: 1.5.

[0040] In the composite mycorrhizal agent of the present invention, the cell concentrations of the single bacterial seed solution of the phosphate-solubilizing bacteria Aspergillus niger, Suillus fusae, and Boletus edulis are preferably 0.4-0.8 g / mL, more preferably 0.5-0.7 g / mL, and even more preferably 0.6 g / mL; the cell concentrations of the single bacterial seed solution of the cellulose-degrading bacteria Bacillus, the potassium-solubilizing bacteria Bacillus licheniformis, and the mycorrhizal fungus Fusarium are preferably 6-8×10 9 pcs / mL, more preferably 6.5 to 7.5×10 9 / mL, and more preferably 7×10 9 Pieces / mL.

[0041] In the composite mycorrhizal agent of the present invention, the method for preparing the seed solution of the cellulose degrading bacteria Bacillus, the potassium-solubilizing bacteria Bacillus licheniformis, and the mycorrhizal fungus Fusarium preferably comprises the following steps:

[0042] The culture medium was inoculated with single colonies of the strains, cultured statically in the dark, the cultured bacterial solution was centrifuged, the precipitate was collected, and the cells were resuspended in 0.9% saline to obtain seed solution, thereby obtaining seed solutions of cellulose-degrading bacteria Bacillus, potassium-solubilizing bacteria Bacillus licheniformis, and mycorrhizal fungus Fusarium.

[0043] In the method for preparing the seed solution of the cellulose degrading bacteria Bacillus, the potassium-solubilizing bacteria Bacillus licheniformis, and the mycorrhizal fungus Fusarium of the present invention, the culture medium uses water as a solvent and includes the following components: 8-12 g / L of peptone, 10-20 g / L of beef extract, and 10-20 g / L of NaCl, more preferably 9-11 g / L of peptone, 12-18 g / L of beef extract, and 12-18 g / L of NaCl, and further preferably 10 g / L of peptone, 14-16 g / L of beef extract, and 14-16 g / L of NaCl; the pH of the culture medium is preferably 6.6-7.0, more preferably 6.7-6.9, and further preferably 6.8.

[0044] In the method for preparing the seed solution of the cellulose degrading bacteria Bacillus, the potassium-solubilizing bacteria Bacillus licheniformis, and the mycorrhizal fungus Fusarium of the present invention, the culture temperature is preferably 30-40°C, more preferably 34-36°C, and further preferably 35°C; the culture time is preferably 12-36h, more preferably 18-28h, and further preferably 24h.

[0045] In the method for preparing the seed solution of the cellulose degrading bacteria Bacillus, the potassium-solubilizing bacteria Bacillus licheniformis, and the mycorrhizal fungus Fusarium of the present invention, the centrifugation speed after cultivation is preferably 2500-3500 rpm, more preferably 2800-3200 rpm, and further preferably 3000 rpm; the centrifugation time is preferably 2-6 min, more preferably 3-5 min, and further preferably 4 min.

[0046] In the composite mycorrhizal agent of the present invention, the method for preparing the seed solution of the phosphate-solubilizing bacteria Aspergillus niger, Suillus fusca, and Boletus edulis preferably comprises the following steps:

[0047] The single colonies of the above strains were inoculated into the culture medium respectively, and cultured statically in the dark. The cultured bacterial solution was centrifuged, the precipitate was collected, and the bacterial body was resuspended with 0.9% physiological saline to obtain seed solution, thereby obtaining seed solutions of phosphate-solubilizing bacteria Aspergillus niger, Suillus suillus and Boletus edulis respectively.

[0048] In the method for preparing the seed solution of the phosphate-solubilizing bacteria Aspergillus niger, Suillus fusca and Boletus edulis of the present invention, the culture medium uses water as a solvent and includes the following components: 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 even more preferably 200 g / L of potato and 20 g / L of glucose; the pH of the culture medium is preferably 6.3-6.7, more preferably 6.4-6.6, and even more preferably 6.5.

[0049] In the seed liquid preparation method of the phosphate-solubilizing bacteria Aspergillus niger, Suillus edulis and Boletus edulis described in the present invention, the culture temperature is preferably 25-35°C, more preferably 28-32°C, and further preferably 30°C; the culture time is preferably 48-96h, more preferably 60-84h, and further preferably 72h.

[0050] In the seed liquid preparation method of the phosphate-solubilizing bacteria Aspergillus niger, Suillus edulis, and Boletus edulis described in the present invention, the centrifugation speed after cultivation is preferably 2500-3500 rpm, more preferably 2800-3200 rpm, and further preferably 3000 rpm; the centrifugation time is preferably 1-5 min, more preferably 2-4 min, and further preferably 3 min.

[0051] In the present invention, the preparation method of the composite mycorrhizal agent preferably comprises the following steps:

[0052] The single bacterial seed liquids of cellulose degrading bacteria Bacillus, phosphate-dissolving bacteria Aspergillus niger, potassium-dissolving bacteria Bacillus licheniformis, mycorrhizal fungi Fusarium, Suillus edulis and Boletus edulis are mixed in proportion, inoculated into a culture medium, cultured for 3 to 5 hours, and then glycerol is added and cultured for 1 to 2 hours to obtain a composite mycorrhizal agent.

[0053] In the preparation method of the composite mycorrhizal agent of the present invention, the composite mycorrhizal agent is inoculated in a culture medium for culture, and the inoculation amount of the mixed seed liquid is preferably 0.8% to 1.2% of the total volume, more preferably 0.9 to 1.1%, and even more preferably 1%.

[0054] In the preparation method of the composite mycorrhizal agent of the present invention, the culture medium for inoculation of the composite mycorrhizal agent uses water as a solvent and includes the following components: 180-220 g / L of potatoes, 15-25 g / L of glucose, and 15-20 g / L of agar, more preferably 190-210 g / L of potatoes, 18-22 g / L of glucose, and 16-19 g / L of agar, and further preferably 200 g / L of potatoes, 20 g / L of glucose, and 18 g / L of agar.

[0055] In the preparation method of the composite mycorrhizal agent of the present invention, the composite mycorrhizal agent is inoculated in a culture medium and cultured for a period of time, preferably 3 to 5 hours, more preferably 3.5 to 4.5 hours, and further preferably 4 hours; then glycerol is added and culture is continued, and the continued culture time is preferably 1 to 2 hours, more preferably 1.2 to 1.8 hours, and further preferably 1.4 to 1.6 hours.

[0056] In the preparation method of the composite mycorrhizal agent of the present invention, the volume ratio of glycerol added to the culture medium is 1-1.4:100, more preferably 1.1-1.3:100, and even more preferably 1.2:100.

[0057] In the preparation method of the composite mycorrhizal agent of the present invention, the temperature at which the composite mycorrhizal agent is inoculated into the culture medium for cultivation is preferably 26-30°C, more preferably 27-29°C, and further preferably 28°C; the rotation speed for cultivation is preferably 120-160rpm, more preferably 130-150rpm, and further preferably 140rpm.

[0058] In the preparation method of the composite mycorrhizal agent of the present invention, the culture medium used is sterilized and cooled for standby use. The pressure of the sterilization treatment is preferably 1.3-1.5 kg / cm 2, more preferably 1.4 kg / cm 2 The disinfection treatment time is preferably 2 to 2.5 hours, more preferably 2.25 hours.

[0059] In the present invention, the potassium-dissolving bacteria Bacillus licheniformis and the phosphate-dissolving bacteria Aspergillus niger strains were purchased from the China General Microorganism Collection Center; the brown-ringed boletus, the delicious boletus, and the mycorrhizal fungus Fusarium were purchased from the China Agricultural Microorganism Culture Collection Center; the cellulose-degrading bacteria Bacillus is the patent collection strain HM01, named Bacillus sp. HM01, which was deposited in the China Center for Type Culture Collection on September 27, 2017, with a deposit number of CCTCC NO: M2017558.

[0060] The present invention also provides a method for preparing the special biological organic fertilizer for chestnut mycorrhiza, comprising the following steps:

[0061] Pile chestnut mushroom residue, bran, biochar and soybean meal, ferment for 5 to 9 days, then spray composite mycorrhizal agent on the surface, ferment at 22 to 28 ℃ for 25 to 35 days, and turn the pile over every 7 days.

[0062] In the preparation method of the chestnut mycorrhiza-specific biological organic fertilizer of the present invention, chestnut mushroom residue, bran, biochar and soybean meal are first stacked, and the stacking height is preferably 0.8-1.5m, more preferably 1-1.3m, and further preferably 1.2m; the stacking width is preferably 0.8-1.5m, more preferably 1-1.3m, and further preferably 1.2m.

[0063] In the preparation method of the special biological organic fertilizer for chestnut mycorrhiza of the present invention, the pile is piled for fermentation, and the fermentation time is preferably 5 to 9 days, more preferably 6 to 8 days, and further preferably 7 days; then the composite root fungus agent is sprayed on the surface, and the fermentation is continued, the fermentation temperature is preferably 22 to 28° C., more preferably 24 to 26° C., and further preferably 25° C., the fermentation time is preferably 25 to 35 days, more preferably 28 to 32 days, and further preferably 30 days, and the pile is preferably turned every 7 days during the fermentation process.

[0064] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0065] Example 1

[0066] The cellulose-degrading bacteria Bacillus, potassium-solubilizing bacteria Bacillus licheniformis, and mycorrhizal fungus Fusarium were inoculated with single colonies in a culture medium (the culture medium formula was: 10 g peptone, 15 g beef extract, 15 g NaCl, 1000 ml water, pH 6.8), and cultured statically at 35°C in the dark for 24 h. The cultured bacterial solution was centrifuged at 3000 rpm for 4 min, the precipitate was collected, and the cells were resuspended with 0.9% saline to obtain seed solutions of cellulose-degrading bacteria Bacillus, potassium-solubilizing bacteria Bacillus licheniformis, and mycorrhizal fungus Fusarium, and the cell concentrations in the single bacterial seed solutions were 0.6 g / mL, respectively; the phosphate-solubilizing bacteria Aspergillus niger, Suillus fusus, and Boletus edulis were inoculated with single colonies in a culture medium (the culture medium formula was: 200 g potato, 20 g glucose, 1000 ml water, The culture solution was centrifuged at 3000 rpm for 3 min, the precipitate was collected, and the bacteria were resuspended in 0.9% saline to obtain the seed solution of phosphate-soluble bacteria Aspergillus niger, Suillus chrysogenus, and Boletus edulis. The cell concentrations in the single bacterial seed solution were 7×10 9 Pieces / mL.

[0067] The single bacterial seed liquid of cellulose degrading bacteria Bacillus, phosphate-dissolving bacteria Aspergillus niger, potassium-dissolving bacteria Bacillus licheniformis, mycorrhizal fungi Fusarium, brown ring boletus, and delicious boletus were mixed in a volume ratio of 1:1:1:1:3:3, and the inoculation amount of the mixed seed liquid was inoculated into the culture medium (the culture medium formula was: 200g potato, 20g glucose, 18g agar, 1000mL water) at an inoculation ratio of 1% of the total volume, and cultured at 28°C, 140rpm for 4h, and then glycerol was added, and the volume ratio of glycerol to culture medium was 1.2:100, and the culture was continued for 1.5h to obtain a composite mycorrhizal agent. All culture media were heated at 1.4kg / cm before use. 2 Sterilize under pressure for 2.25 hours.

[0068] Mix 70 parts of chestnut mushroom residue, 5 parts of bran, 3 parts of biochar and 10 parts of soybean meal evenly and pile them up with a height of 1.2m and a width of 1.2m. Ferment them for 7 days, then spray 0.5 parts of composite mycorrhizal agent on the surface. Ferment them at 25℃ for 30 days and turn the pile over every 7 days.

[0069] Example 2

[0070] The cellulose-degrading bacteria Bacillus, the potassium-solubilizing bacteria Bacillus licheniformis, and the mycorrhizal fungus Fusarium were inoculated with single colonies in the culture medium (the culture medium formula was: 8g peptone, 10g beef extract, 10g NaCl, 1000ml water, pH6.6), and statically cultured at 30°C in the dark for 12h. The cultured bacterial solution was centrifuged at 2500rpm for 2min, the precipitate was collected, and the cells were resuspended with 0.9% physiological saline to obtain the seed liquid of the cellulose-degrading bacteria Bacillus, the potassium-solubilizing bacteria Bacillus licheniformis, and the mycorrhizal fungus Fusarium, and the cell concentration in the single bacterial seed liquid was 0.4g / mL; the phosphate-solubilizing bacteria Aspergillus niger, Suillus suillus, and Boletus edulis were inoculated with single colonies in the culture medium (the culture medium formula was: 180g potato, 10g glucose, 1000mL water, The culture solution was centrifuged at 2500 rpm for 1 min, the precipitate was collected, and the bacteria were resuspended in 0.9% saline to obtain the seed solution of phosphate-soluble bacteria Aspergillus niger, Suillus chrysogenus, and Boletus edulis. The cell concentrations in the single bacterial seed solution were 6×10 9 Pieces / mL.

[0071] The single bacterial seed liquid of cellulose degrading bacteria Bacillus, phosphate-dissolving bacteria Aspergillus niger, potassium-dissolving bacteria Bacillus licheniformis, mycorrhizal fungi Fusarium, brown-ringed boletus, and delicious boletus were mixed in a volume ratio of 0.5:0.5:0.5:0.5:1.5:1.5, and the inoculation amount of the mixed seed liquid was inoculated into the culture medium (the culture medium formula was: 180g potato, 15g glucose, 15g agar, 1000mL water) at an inoculation ratio of 0.8% of the total volume, and cultured at 26°C, 120rpm for 3h, and then glycerol was added, and the ratio of glycerol to culture medium volume was 1:100, and cultured for 1h to obtain a composite mycorrhizal agent. All culture media were heated at 1.2kg / cm before use. 2 Sterilize under pressure for 2 hours.

[0072] Mix 60 parts of chestnut mushroom residue, 3 parts of bran, 1 part of biochar and 8 parts of soybean meal evenly and pile them up with a height of 0.8m and a width of 0.8m. Ferment them for 5 days, then spray 0.1 parts of composite mycorrhizal agent on the surface and ferment them at 22℃ for 27 days, turning the pile every 7 days.

[0073] Example 3

[0074] The cellulose-degrading bacteria Bacillus, potassium-solubilizing bacteria Bacillus licheniformis, and mycorrhizal fungus Fusarium were inoculated with single colonies in a culture medium (the culture medium formula was: 12 g peptone, 20 g beef extract, 20 g NaCl, 1000 ml water, pH 7.0), and statically cultured at 40°C in the dark for 28 h. The cultured bacterial solution was centrifuged at 3500 rpm for 6 min, the precipitate was collected, and the cells were resuspended with 0.9% saline to obtain seed solutions of cellulose-degrading bacteria Bacillus, potassium-solubilizing bacteria Bacillus licheniformis, and mycorrhizal fungus Fusarium, and the cell concentrations in the single bacterial seed solutions were 0.8 g / mL, respectively; the phosphate-solubilizing bacteria Aspergillus niger, Suillus fusus, and Boletus edulis were inoculated with single colonies in a culture medium (the culture medium formula was: 220 g potato, 30 g glucose, 1000 ml water, The culture solution was centrifuged at 3500 rpm for 5 min, the precipitate was collected, and the bacteria were resuspended in 0.9% saline to obtain the seed solution of phosphate-soluble bacteria Aspergillus niger, Suillus chrysogenus, and Boletus edulis. The cell concentrations in the single bacterial seed solution were 8 × 10 9 Pieces / mL.

[0075] The single bacterial seed liquid of cellulose degrading bacteria Bacillus, phosphate-dissolving bacteria Aspergillus niger, potassium-dissolving bacteria Bacillus licheniformis, mycorrhizal fungi Fusarium, brown-ringed boletus, and delicious boletus were mixed in a volume ratio of 1:1:1:1:3:3, and the inoculation amount of the mixed seed liquid was inoculated into the culture medium (the culture medium formula was: 220g potato, 25g glucose, 20g agar, 1000mL water) at an inoculation ratio of 1.2% of the total volume, and cultured at 30°C, 160rpm for 5h, and then glycerol was added, and the volume ratio of glycerol to culture medium was 1.4:100, and the culture was continued for 2h to obtain a composite mycorrhizal agent. All culture media were heated at 1.2kg / cm before use. 2 Sterilize under pressure for 2.5 hours.

[0076] Mix 80 parts of chestnut mushroom residue, 7 parts of bran, 5 parts of biochar and 12 parts of soybean meal evenly and pile them up with a height of 1.5m and a width of 1.5m. Ferment them for 9 days, then spray 1 part of composite mycorrhizal agent on the surface. Ferment them at 28℃ for 35 days and turn the pile over every 7 days.

[0077] Experimental example

[0078] At the chestnut garden base in Shangmashi Village, Muzidian Town, Macheng City, Hubei Province, a test site with a width of 4 meters and a length of 30 meters was selected, and 2-year-old chestnut seedlings were cultivated at a spacing of 1 meter. A row of 10 chestnut seedlings was used to apply the special biological organic fertilizer described in Example 1 as the experimental group, another row of 10 chestnut seedlings with a row spacing of 1 meter was taken without fertilization as the blank group, and another row with a row spacing of 1 meter was taken to apply commercially available compound fertilizer as the control group. After one year, the height, diameter, fresh weight of the aboveground part, dry weight of the aboveground part, fresh weight of the underground part, and dry weight of the underground part of the chestnut were measured. The application method of the special biological organic fertilizer described in Example 1 is to dig a pit with a side length of 50 cm and a depth of 50 cm 50 cm away from the roots of the chestnut tree, and apply 10-30 kg of special biological organic fertilizer for promoting chestnut mycorrhiza growth per pit, and cover the surface with 10 cm of soil. The results showed that, compared with the blank group, the application of the special organic fertilizer described in Example 1 had an extremely obvious effect on promoting the growth of chestnut seedlings, among which the plant height increased by nearly 50%, and the fresh weight of the underground part increased by 64.7%, indicating that the growth of the chestnut root system was significantly promoted, as shown in Table 1 (Note: the values ​​are mean ± standard deviation, and lowercase letters represent significant differences P<0.05). In terms of promoting chestnut growth, the chestnut mycorrhiza special biological organic fertilizer has a fertilizer effect similar to that of commercially available compound fertilizers, and can significantly promote chestnut growth.

[0079] Table 1 Effects of applying chestnut mycorrhiza-specific bio-organic fertilizer on chestnut growth characteristics

[0080]

[0081] Note: In the abcd difference significance analysis, those with the same letter are not significantly different, and those with different letters are significantly different.

[0082] according to Figure 5 It can be seen that the chestnut seedlings treated with special biological organic fertilizer had significantly more mycorrhiza formation, and the typical mycorrhizal structure of Hatty's network could be seen through paraffin sections, indicating that special biological organic fertilizer can significantly promote the formation of mycorrhiza.

[0083] Following up on the above experiment, the fruit bearing of 10 chestnut trees in the experimental group, blank group and control group were observed three years later. The results are shown in Table 2.

[0084] Table 2 Fruiting situation of chestnut trees after three years

[0085]

[0086]

[0087] The results in Table 2 show that the application of chestnut mycorrhizal growth-promoting biological organic fertilizer can increase the number of chestnut fruits and the average fruit setting rate per plant compared with the control, and reduce the empty fruit rate, and reduce the number of diseases and insect pests. The overall growth and fruit setting of chestnut are better than those of the blank group and the control.

[0088] The chestnut mycorrhiza special biological organic fertilizer of the present invention is provided with the name of "Guanlu No. 5" chestnut chestnut mycorrhiza special biological organic fertilizer for promoting chestnut mycorrhiza growth, and is tested at the chestnut base of the Dongmu Seed Breeding Professional Cooperative in Macheng City. Figure 6 The figure shows the application proof of the test effect.

[0089] It can be seen from the above embodiments that the present invention provides a biological organic fertilizer specifically for chestnut mycorrhiza, which strengthens the colonization and growth of ectomycorrhizal fungi in the chestnut root system, promotes the formation and growth of a large number of chestnut mycorrhizae, improves the ability of chestnuts to absorb nutrients, improves the stress resistance of chestnuts, regulates the stability of the microbial community structure in the soil of the chestnut root system, comprehensively improves the yield and quality of chestnuts, and at the same time helps to improve soil fertility, improve soil function, and realize the circular and sustainable development of the chestnut industry.

[0090] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A biological organic fertilizer specially used for chestnut mycorrhiza, characterized in that: It is prepared from raw materials including the following components: 60-80 parts of chestnut mushroom residue, 3-7 parts of bran, 1-5 parts of biochar, 8-12 parts of soybean meal, and 0.1-1 part of composite mycorrhizal agent; The composite mycorrhizal agent comprises cellulose degrading bacteria, phosphate-dissolving bacteria Aspergillus niger, potassium-dissolving bacteria Bacillus licheniformis, mycorrhizal fungi, Suillus edulis and Boletus edulis, the cellulose-degrading bacteria is Bacillus HM01, and the mycorrhizal fungi is Fusarium; The raw materials are naturally fermented and then fermented with a composite mycorrhizal agent to prepare biological organic fertilizer; The volume ratio of the seed liquid of Bacillus HM01: phosphate-solubilizing bacteria Aspergillus niger: potassium-solubilizing bacteria Bacillus licheniformis: Fusarium: Suillus edulis: Boletus edulis in the composite mycorrhizal agent 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; The preparation method of the composite mycorrhizal agent comprises the following steps: The single bacterial seed liquids of Bacillus HM01, phosphate-dissolving bacteria Aspergillus niger, potassium-dissolving bacteria Bacillus licheniformis, Fusarium, Suillus edulis, and Boletus edulis are mixed in proportion, inoculated into a culture medium, cultured for 3 to 5 hours, and then glycerol is added, and cultured for 1 to 2 hours to obtain a composite mycorrhizal agent; In the preparation method of the composite mycorrhizal agent, the inoculation amount of the mixed seed liquid accounts for 0.8% to 1.2% of the total volume.

2. The special biological organic fertilizer for chestnut mycorrhiza as claimed in claim 1, characterized in that: The cell concentrations in the single-bacteria seed liquid of the phosphate-dissolving bacteria Aspergillus niger, Suillus fusca and Boletus edulis are 0.4-0.8 g / mL respectively.

3. The special biological organic fertilizer for chestnut mycorrhiza as claimed in claim 1, characterized in that: The cell concentrations in the single bacterial seed liquid of Bacillus HM01, Bacillus licheniformis, and Fusarium are 6 to 8×10 9 Pieces / mL.

4. The special biological organic fertilizer for chestnut mycorrhiza as claimed in claim 1, characterized in that: In the preparation method of the composite mycorrhizal agent, the culture medium uses 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.

5. The special biological organic fertilizer for chestnut mycorrhiza as claimed in claim 4, characterized in that: In the preparation method of the composite mycorrhizal agent, the culture temperature is 26-30° C. and the culture speed is 120-160 rpm.

6. The method for preparing the biological organic fertilizer for chestnut mycorrhizae according to any one of claims 1 to 5, characterized in that: The following steps are involved: Pile chestnut mushroom residue, bran, biochar and soybean meal, ferment for 5 to 9 days, then spray composite mycorrhizal agent on the surface, ferment at 22 to 28 ℃ for 25 to 35 days, and turn the pile over every 7 days.

7. The method for preparing the special biological organic fertilizer for chestnut mycorrhiza as claimed in claim 6, characterized in that: The stacking height is 0.8~1.5m and the width is 0.8~1.5m.

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