1. The formula of the cow and sheep manure biological organic fertilizer containing beneficial bacteria is characterized in that the organic fertilizer comprises: cow dung or sheep dung, bean pulp, wheat bran or straw, calcium superphosphate, calcium carbonate, beneficial bacteria, and loam or sandy loam or clay;

the beneficial bacteria comprise: any one or more than two of bacillus amyloliquefaciens, bacillus mucilaginosus, bacillus subtilis, 5406 actinomycetes, trichoderma harzianum and fusarium oxysporum wilt antagonistic bacteria LCQ 1801; the effective viable count of the bacillus amyloliquefaciens is 300-700 hundred million/g; the effective viable count of the bacillus mucilaginosus is 300-500 hundred million/g; the effective viable count of the bacillus subtilis is 100-600 hundred million/g; the effective viable count of the 5406 actinomycetes is 4-6 hundred million/g; the effective viable count of the trichoderma harzianum is 3-6 hundred million/gram; the number of effective viable bacteria of the fusarium oxysporum wilt antagonistic bacteria LCQ1801 is 300-600 hundred million/g;

the preservation number of the fusarium oxysporum wilt antagonistic bacteria LCQ1801 is as follows: GDMCC No.61431, with a preservation date of 14/1/2021, which belongs to Bacillus velezensis.

2. The formula of the cow and sheep manure biological organic fertilizer containing beneficial bacteria as claimed in claim 1, wherein the organic fertilizer comprises: 48-58 parts of cow dung or sheep dung, 10-15 parts of soybean meal, 10-20 parts of wheat bran or straw, 1 part of calcium superphosphate, 1 part of calcium carbonate, beneficial bacteria and 15-20 parts of loam or sandy loam or clay.

3. The formula of the biological organic fertilizer containing beneficial bacteria for cattle and sheep manure as claimed in claim 1 or 2, wherein animal manure in the organic fertilizer is cattle manure, the beneficial bacteria are Bacillus amyloliquefaciens, Bacillus mucilaginosus, Bacillus subtilis and Fusarium oxysporum wilt-antagonistic bacteria LCQ1801, the effective viable count of the Bacillus amyloliquefaciens is 500-700 hundred million/gram, the effective viable count of the Bacillus mucilaginosus is 350-500 hundred million/gram, the effective viable count of the Bacillus subtilis is 350-600 hundred million/gram, and the effective viable count of the Fusarium oxysporum wilt-antagonistic bacteria LCQ1801 is 400-550 hundred million/gram.

4. The formula of the biological organic fertilizer containing beneficial bacteria for cattle and sheep manure as claimed in claim 1 or 2, wherein animal manure in the organic fertilizer is cattle manure, the beneficial bacteria are 5406 actinomycetes, and the effective viable count of the 5406 actinomycetes is 4-6 hundred million/g.

5. The formula of the cow and sheep manure biological organic fertilizer containing beneficial bacteria as claimed in claim 1 or 2, wherein animal manure in the organic fertilizer is cow manure, the beneficial bacteria are trichoderma harzianum, and the number of effective viable bacteria of the trichoderma harzianum is 3-6 hundred million/g.

6. The formula of the biological organic fertilizer containing beneficial bacteria for cattle and sheep manure as claimed in claim 1 or 2, wherein animal manure in the organic fertilizer is sheep manure, the beneficial bacteria are Bacillus amyloliquefaciens, Bacillus mucilaginosus, Bacillus subtilis and Fusarium oxysporum wilt-antagonistic bacteria LCQ1801, the effective viable count of the Bacillus amyloliquefaciens is 300-400 hundred million/g, the effective viable count of the Bacillus mucilaginosus is 400-500 hundred million/g, the effective viable count of the Bacillus subtilis is 100-200 hundred million/g, and the effective viable count of the Fusarium oxysporum wilt-antagonistic bacteria LCQ1801 is 300-450 hundred million/g.

7. The formula of the biological organic fertilizer containing beneficial bacteria for cattle and sheep manure as claimed in claim 1 or 2, wherein animal manure in the organic fertilizer is sheep manure, the beneficial bacteria are 5406 actinomycetes, and the effective viable count of the 5406 actinomycetes is 4-6 hundred million/g.

8. The formula of the biological organic fertilizer containing beneficial bacteria for cattle and sheep manure as claimed in claim 1 or 2, wherein animal manure in the organic fertilizer is sheep manure, the beneficial bacteria are trichoderma harzianum, and the number of effective viable bacteria of the trichoderma harzianum is 4-5 hundred million/g.

9. The use of the bio-organic fertilizer formula of cow and sheep manure containing beneficial bacteria as defined in any one of claims 1-8 for planting, wherein the bio-organic fertilizer can inhibit blight.

10. The use according to claim 9, wherein the bio-organic fertilizer is capable of inhibiting banana wilt.

Background

The biological organic fertilizer is prepared by taking livestock and poultry manure, municipal domestic waste, crop straws, agricultural and sideline products and organic wastes generated by food processing as raw materials, matching with multifunctional fermentation strains to quickly deodorize, decompose and dehydrate the raw materials, and then adding functional microorganisms. Compared with chemical fertilizers, the biological organic fertilizer has more complete nutrient elements, can effectively improve soil, regulate and control the micro-ecological balance of soil and rhizosphere, improve the disease and insect resistance of crops and improve the product quality after being used for a long time. Compared with farmyard manure, the biological organic fertilizer has the following basic advantages: the functional bacteria in the biological organic fertilizer have specific effects on improving soil fertility and promoting crop growth, and the farmyard manure is generated by natural fermentation and has no special effect of dominant functional bacteria.

The bio-organic fertilizer has important significance for realizing resource-saving and environment-friendly society, and is a necessary choice for realizing sustainable development of agriculture. However, the existing biological organic fertilizer generally has the problems of insufficient fermentation, single effective strain, difficult colonization and the like.

Disclosure of Invention

The invention aims to provide a formula and application of a cattle and sheep manure biological organic fertilizer containing beneficial bacteria, which solves the problem of difficult colonization of the beneficial bacteria.

In order to achieve the aim, the invention provides a formula of a cow and sheep manure biological organic fertilizer containing beneficial bacteria, which comprises the following components: cow dung or sheep dung, bean pulp, wheat bran or straw, calcium superphosphate, calcium carbonate, beneficial bacteria, and loam or sandy loam or clay; the beneficial bacteria comprise: any one or more than two of bacillus amyloliquefaciens, bacillus mucilaginosus, bacillus subtilis, 5406 actinomycetes, trichoderma harzianum and fusarium oxysporum wilt antagonistic bacteria LCQ 1801; the effective viable count of the bacillus amyloliquefaciens is 300-700 hundred million/g; the effective viable count of the bacillus mucilaginosus is 300-500 hundred million/g; the effective viable count of the bacillus subtilis is 100-600 hundred million/g; the effective viable count of the 5406 actinomycetes is 4-6 hundred million/g; the effective viable count of the trichoderma harzianum is 3-6 hundred million/gram; the number of effective viable bacteria of the fusarium oxysporum wilt antagonistic bacteria LCQ1801 is 300-600 hundred million/g; the preservation number of the fusarium oxysporum wilt antagonistic bacteria LCQ1801 is as follows: GDMCC No.61431, with a preservation date of 14/1/2021, which belongs to Bacillus velezensis.

Preferably, the organic fertilizer comprises: 48-58 parts of cow dung or sheep dung, 10-15 parts of soybean meal, 10-20 parts of wheat bran or straw, 1 part of calcium superphosphate, 1 part of calcium carbonate, beneficial bacteria and 15-20 parts of loam or sandy loam or clay.

Preferably, animal manure in the organic fertilizer is cow manure, beneficial bacteria are Bacillus amyloliquefaciens, Bacillus mucilaginosus, Bacillus subtilis and Fusarium oxysporum wilt antagonistic bacteria LCQ1801, the number of effective live bacteria of the Bacillus amyloliquefaciens is 500-700 hundred million/gram, the number of effective live bacteria of the Bacillus mucilaginosus is 350-500 hundred million/gram, the number of effective live bacteria of the Bacillus subtilis is 350-600 hundred million/gram, and the number of effective live bacteria of the Fusarium oxysporum wilt antagonistic bacteria LCQ1801 is 400-550 hundred million/gram.

Preferably, animal manure in the organic fertilizer is cow manure, 5406 actinomycetes are selected as beneficial bacteria, and the effective viable count of the 5406 actinomycetes is 4-6 hundred million/gram.

Preferably, animal manure in the organic fertilizer is cow manure, beneficial bacteria are Trichoderma harzianum, and the effective viable count of the Trichoderma harzianum is 3-6 hundred million/g.

Preferably, animal manure in the organic fertilizer is sheep manure, beneficial bacteria are Bacillus amyloliquefaciens, Bacillus mucilaginosus, Bacillus subtilis and Fusarium oxysporum wilt antagonistic bacteria LCQ1801, the effective viable count of the Bacillus amyloliquefaciens is 300-400 hundred million/gram, the effective viable count of the Bacillus mucilaginosus is 400-500 hundred million/gram, the effective viable count of the Bacillus subtilis is 100-200 hundred million/gram, and the effective viable count of the Fusarium oxysporum wilt antagonistic bacteria LCQ1801 is 300-450 hundred million/gram.

Preferably, animal manure in the organic fertilizer is sheep manure, the beneficial bacteria are 5406 actinomycetes, and the effective viable count of the 5406 actinomycetes is 4-6 hundred million/gram.

Preferably, animal manure in the organic fertilizer is sheep manure, beneficial bacteria are trichoderma harzianum, and the effective viable count of the trichoderma harzianum is 4-5 hundred million/gram.

The invention also aims to provide application of the formula of the cattle and sheep manure biological organic fertilizer containing beneficial bacteria in planting, and the biological organic fertilizer can inhibit the blight.

Preferably, the bio-organic fertilizer can inhibit the blight of bananas.

The beneficial bacterium-containing cow and sheep manure biological organic fertilizer formula and the application thereof solve the problem of difficult colonization of beneficial bacteria, and have the following advantages:

the cattle and sheep manure biological organic fertilizer can well grow various functional bacteria and antagonistic bacteria, comprises bacillus amyloliquefaciens, bacillus mucilaginosus, bacillus subtilis, 5406 actinomycetes, trichoderma harzianum and fusarium oxysporum wilt antagonistic bacteria LCQ1801, is not easy to inactivate after being stored for a long time, and solves the problems of single effective bacteria and difficult colonization.

Compared with other cattle and sheep manure, the invention takes cheap cattle manure or sheep manure, soybean meal, wheat bran or straw and other agricultural product leftovers and fertile soil as main substrates, realizes the maximum utilization of resources, reduces the production cost and changes waste into valuable.

The cow and sheep manure biological organic fertilizer can improve soil fertility, inhibit growth and reproduction of harmful microorganism bacteria, inhibit blight, reduce the occurrence of the blight in banana planting, has no residue, reduces the use of chemical fertilizers and pesticides, and reduces and lowers environmental pollution.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

A cow dung biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: inoculating bacillus amyloliquefaciens, bacillus mucilaginosus, bacillus subtilis and fusarium oxysporum wilt antagonistic bacteria LCQ1801 on an LB solid culture medium in a streaking manner, and culturing for 1-2 d at 28-32 ℃;

(2) preparation of seed suspension: picking single colonies from the bacteria in the step (1), respectively inoculating the single colonies on an LB liquid culture medium, and performing shake culture at 28-32 ℃ and 200rpm for 1-2 days;

(3) preparing cattle manure: putting cow dung fertilizer weighed according to the proportion of 58 parts of cow dung, 15 parts of soybean meal, 10 parts of wheat bran or straw, 15 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, charging 1/4-1/3, adding about 30% (volume percentage) of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: and (3) when the temperature of the culture bottle is reduced to about 30 ℃ in the step (3), respectively adding the bacterial liquid with the volume percentage of about 30 percent in the step (2) under the aseptic condition, uniformly stirring, culturing for 3-4 days at 30-32 ℃, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly.

The colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on an LB solid medium plate, and cultured at 28 to 32 ℃ for 5 to 7 days and 2 months, respectively, and then the number of colonies was counted, and the results are shown in Table 1 below.

Example 2

A cow dung biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: streaking a 5406 actinomycete (streptomyces microflavus, also called Jingyang streptomyces) plate on a Gao's first solid culture medium, and culturing at 28-32 ℃ for 5-7 d;

(2) preparation of seed suspension: picking a single colony of the 5406 actinomycetes in the step (1), inoculating the single colony to a PBS liquid culture medium, and performing shake culture at 28-32 ℃ and 200rpm for 5-7 days;

(3) preparing cattle manure: putting cow dung fertilizer weighed according to the proportion of 58 parts of cow dung, 15 parts of soybean meal, 10 parts of wheat bran or straw, 15 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, charging 1/4-1/3, adding about 30% (volume percentage) of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: and (3) when the temperature of the culture bottle is reduced to about 30 ℃ in the step (3), respectively adding the bacterial liquid with the volume percentage of about 30% in the step (2) under an aseptic condition, uniformly stirring, culturing for 5-7 d in an incubator with the temperature of 28-32 ℃, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly.

The colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on a Gao's first solid medium, cultured at 28 to 32 ℃ for 5 to 7 days, and the number of colonies was counted after 2 months, and the results are shown in Table 1 below.

Example 3

A cow dung biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: inoculating a Trichoderma harzianum plate on a PDA culture medium (glucose potato agar culture medium) in a streak manner, and culturing at the temperature of 28-32 ℃ for 5-7 d;

(2) preparation of seed suspension: and (2) selecting a single colony of trichoderma harzianum in the step (1), inoculating the single colony on a PDB culture medium (glucose potato liquid culture medium), and performing shake culture at 28-32 ℃ and 200rpm for 5-7 days.

(3) Preparing cattle manure: putting cow dung fertilizer weighed according to the proportion of 58 parts of cow dung, 15 parts of soybean meal, 10 parts of wheat bran or straw, 15 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, charging 1/4-1/3, adding about 30% (volume percentage) of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: and (3) when the temperature of the culture bottle is reduced to about 30 ℃ in the step (3), respectively adding the bacterial liquid with the volume percentage of about 30% in the step (2) under an aseptic condition, uniformly stirring, culturing for 5-7 d in an incubator with the temperature of 28-32 ℃, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly.

The colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on a PDA solid medium and cultured at 28 to 32 ℃ for 5 to 7 days and 2 months, respectively, and then the colony count was calculated, and the results are shown in Table 1 below.

Table 1 shows the number of bacteria in the cow dung bioorganic fertilizer of examples 1-3 of the present invention

Note: the bacillus subtilis is bacillus subtilis; the jelly sample is bacillus mucilaginosus; the amylolytic bacteria are bacillus amyloliquefaciens.

Example 4

A sheep manure biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: inoculating bacillus amyloliquefaciens, bacillus mucilaginosus, bacillus subtilis and fusarium oxysporum wilt antagonistic bacteria LCQ1801 on an LB solid culture medium in a streaking manner, and culturing for 1-2 d at 28-32 ℃;

(2) preparation of seed suspension: picking single colonies from the bacteria in the step (1), respectively inoculating the single colonies on an LB liquid culture medium, and performing shake culture at 28-32 ℃ and 200rpm for 1-2 days;

(3) preparing sheep manure: loading sheep manure weighed according to the proportion of 58 parts of sheep manure, 15 parts of soybean meal, 10 parts of wheat bran or straw, 15 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, loading 1/4-1/3, adding about 30% (volume percentage) of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: and (3) when the temperature of the culture bottle is reduced to about 30 ℃ in the step (3), respectively adding the bacterial liquid with the volume percentage of about 30 percent in the step (2) under the aseptic condition, uniformly stirring, culturing for 3-4 days at 30-32 ℃, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly.

The colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on an LB solid medium plate, and the colony count was calculated after culturing at 28 ℃ to 32 ℃ for 5 to 7 days and 2 months, respectively, and the results are shown in Table 2 below.

Example 5

A sheep manure biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: streaking a 5406 actinomycete (streptomyces microflavus, also called Jingyang streptomyces) plate on a Gao's first solid culture medium, and culturing at 28-32 ℃ for 5-7 d;

(2) preparation of seed suspension: picking a single colony of the 5406 actinomycetes in the step (1), inoculating the single colony to a PBS liquid culture medium, and performing shake culture at 28-32 ℃ and 200rpm for 5-7 days;

(3) preparing sheep manure: loading sheep manure weighed according to the proportion of 58 parts of sheep manure, 15 parts of soybean meal, 10 parts of wheat bran or straw, 15 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, loading 1/4-1/3, adding about 30% (volume percentage) of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: when the temperature of the culture bottle in the step (3) is reduced to about 30 ℃, respectively adding about 30% (volume percentage) of the bacterial liquid in the step (2) under an aseptic condition, uniformly stirring, culturing for 5-7 d in an incubator at 28-32 ℃, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly;

the colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on a Gao's first solid medium, cultured at 28 to 32 ℃ for 5 to 7 days, and the number of colonies was counted after 2 months, and the results are shown in Table 2 below.

Example 6

A sheep manure biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: inoculating a Trichoderma harzianum plate on a PDA culture medium (glucose potato agar culture medium) in a streak manner, and culturing at the temperature of 28-32 ℃ for 5-7 d;

(2) preparation of seed suspension: picking a single colony of trichoderma harzianum in the step (1), inoculating the single colony on a PDB culture medium (glucose potato liquid culture medium), and performing shake culture at 28-32 ℃ and 200rpm for 5-7 days;

(3) preparing sheep manure: loading sheep manure weighed according to the proportion of 58 parts of sheep manure, 15 parts of soybean meal, 10 parts of wheat bran or straw, 15 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, loading 1/4-1/3, adding about 30% (volume percentage) of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: and (3) when the temperature of the culture bottle is reduced to about 30 ℃ in the step (3), respectively adding the bacterial liquid with the volume percentage of about 30% in the step (2) under an aseptic condition, uniformly stirring, culturing for 5-7 d in an incubator with the temperature of 28-32 ℃, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly.

The colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on a PDA solid medium and cultured at 28 to 32 ℃ for 5 to 7 days and 2 months, respectively, and then the colony count was calculated, and the results are shown in Table 2 below.

Table 2 shows the number of bacteria in the sheep manure bio-organic fertilizer of examples 4-6 of the present invention

Example 7

A cow dung biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: inoculating bacillus amyloliquefaciens, bacillus mucilaginosus, bacillus subtilis and fusarium oxysporum wilt antagonistic bacteria LCQ1801 on an LB solid culture medium in a streaking manner, and culturing for 1-2 d at 28-32 ℃;

(2) preparation of seed suspension: picking single colonies from the bacteria in the step (1), respectively inoculating the single colonies on an LB liquid culture medium, and performing shake culture at 28-32 ℃ and 200rpm for 1-2 days;

(3) preparing cattle manure: putting cow dung fertilizer weighed according to the proportion of 48 parts of cow dung, 10 parts of bean pulp, 20 parts of wheat bran or straw, 20 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, charging 1/4-1/3, adding about 30% (volume percentage) of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: and (3) when the temperature of the culture bottle is reduced to about 30 ℃ in the step (3), respectively adding the bacterial liquid with the volume percentage of about 30 percent in the step (2) under the aseptic condition, uniformly stirring, culturing for 3-4 days at 30-32 ℃, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly.

The colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on an LB solid medium plate, and cultured at 28 to 32 ℃ for 5 to 7 days and 2 months, respectively, and then the number of colonies was counted, and the results are shown in Table 3 below.

Example 8

A cow dung biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: streaking a 5406 actinomycete (streptomyces microflavus, also called Jingyang streptomyces) plate on a Gao's first solid culture medium, and culturing at 28-32 ℃ for 5-7 d;

(2) preparation of seed suspension: picking a single colony of the 5406 actinomycetes in the step (1), inoculating the single colony to a PBS liquid culture medium, and performing shake culture at 28-32 ℃ and 200rpm for 5-7 days;

(3) preparing cattle manure: putting cow dung fertilizer weighed according to the proportion of 48 parts of cow dung, 10 parts of bean pulp, 20 parts of wheat bran or straw, 20 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, charging 1/4-1/3, adding about 30% (volume percentage) of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: and (3) when the temperature of the culture bottle is reduced to about 30 ℃ in the step (3), respectively adding the bacterial liquid with the volume percentage of about 30% in the step (2) under an aseptic condition, uniformly stirring, culturing for 5-7 d in an incubator with the temperature of 28-32 ℃, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly.

The colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on a Gao's first solid medium, cultured at 28 to 32 ℃ for 5 to 7 days, and the number of colonies was counted after 2 months, and the results are shown in Table 3 below.

Example 9

A cow dung biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(2) plate seed culture: inoculating a Trichoderma harzianum plate on a PDA culture medium (glucose potato agar culture medium) in a streak manner, and culturing at the temperature of 28-32 ℃ for 5-7 d;

(2) preparation of seed suspension: and (2) selecting a single colony of trichoderma harzianum in the step (1), inoculating the single colony on a PDB culture medium (glucose potato liquid culture medium), and performing shake culture at 28-32 ℃ and 200rpm for 5-7 days.

(3) Preparing cattle manure: putting cow dung fertilizer weighed according to the proportion of 48 parts of cow dung, 10 parts of bean pulp, 20 parts of wheat bran or straw, 20 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, charging 1/4-1/3, adding about 30% (volume percentage) of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: and (3) when the temperature of the culture bottle is reduced to about 30 ℃ in the step (3), respectively adding the bacterial liquid with the volume percentage of about 30% in the step (2) under an aseptic condition, uniformly stirring, culturing for 5-7 d in an incubator with the temperature of 28-32 ℃, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly.

The colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on a PDA solid medium and cultured at 28 to 32 ℃ for 5 to 7 days and 2 months, respectively, and then the colony count was calculated, and the results are shown in Table 3 below.

Table 3 shows the number of bacteria in the cow dung bioorganic fertilizer of examples 7-9 of the present invention

Example 10

A sheep manure biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: inoculating bacillus amyloliquefaciens, bacillus mucilaginosus, bacillus subtilis and fusarium oxysporum wilt antagonistic bacteria LCQ1801 on an LB solid culture medium in a streak manner, and culturing for 1-2 d at 28-32 ℃;

(2) preparation of seed suspension: picking single colonies from the bacteria in the step (1), respectively inoculating the single colonies on an LB liquid culture medium, and performing shake culture at 28-32 ℃ and 200rpm for 1-2 days;

(3) preparing sheep manure: putting sheep manure weighed by 48 parts of sheep manure, 10 parts of soybean meal, 20 parts of wheat bran or straw, 20 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, charging 1/4-1/3, adding about 30% of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: and (3) when the temperature of the culture bottle is reduced to about 30 ℃ in the step (3), respectively adding about 30% of the bacterial liquid in the step (2) under an aseptic condition, uniformly stirring, culturing for 3-4 days at 30-32 ℃, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly.

The colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on an LB solid medium plate, and the colony count was calculated after culturing at 28 ℃ to 32 ℃ for 5 to 7 days and 2 months, respectively, and the results are shown in Table 4 below.

Example 11

A sheep manure biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: streaking a 5406 actinomycete (streptomyces microflavus, also called Jingyang streptomyces) plate on a Gao's first solid culture medium, and culturing at 28-32 ℃ for 5-7 d;

(2) preparation of seed suspension: picking a single colony of the 5406 actinomycetes in the step (1), inoculating the single colony to a PBS liquid culture medium, and performing shake culture at 28-32 ℃ and 200rpm for 5-7 days;

(3) preparing sheep manure: putting sheep manure weighed by 48 parts of sheep manure, 10 parts of soybean meal, 20 parts of wheat bran or straw, 20 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, charging 1/4-1/3, adding about 30% of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: when the temperature of the culture bottle in the step (3) is reduced to about 30 ℃, respectively adding about 30% of the bacterial liquid in the step (2) under an aseptic condition, uniformly stirring, culturing in an incubator at 28-32 ℃ for 5-7 d, and shaking the culture bottle 1-2 times every day during the culture period to enable bacterial colonies to grow uniformly;

the colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100 mul of the mixture is taken and coated on Gao's number oneThe colony counts were calculated after culturing on solid medium at 28-32 ℃ for 5-7 days and 2 months, respectively, and the results are shown in Table 4 below.

Example 12

A sheep manure biological organic fertilizer containing beneficial bacteria is prepared by the following steps:

(1) plate seed culture: inoculating a Trichoderma harzianum plate on a PDA culture medium (glucose potato agar culture medium) in a streak manner, and culturing at the temperature of 28-32 ℃ for 5-7 d;

(2) preparation of seed suspension: picking a single colony of trichoderma harzianum in the step (1), inoculating the single colony on a PDB culture medium (glucose potato liquid culture medium), and performing shake culture at 28-32 ℃ and 200rpm for 5-7 days;

(3) preparing sheep manure: putting sheep manure weighed by 48 parts of sheep manure, 10 parts of soybean meal, 20 parts of wheat bran or straw, 20 parts of fertile soil, 1 part of calcium superphosphate and 1 part of calcium carbonate into a culture bottle, charging 1/4-1/3, adding about 30% of distilled water, uniformly stirring, and sterilizing at 121 ℃ for 45 min;

(4) solid fermentation of the strain: and (3) when the temperature of the culture bottle is reduced to about 30 ℃ in the step (3), respectively adding about 30% of the bacterial liquid in the step (2) under an aseptic condition, uniformly stirring, culturing in an incubator at 28-32 ℃ for 5-7 d, and shaking the culture bottle 1-2 times every day during the culture period so as to enable bacterial colonies to grow uniformly.

The colony counting is carried out by adopting a dilution plate counting method, which comprises the following steps:

weighing 1g of fermentation sample, adding into a 15mL centrifuge tube, and diluting with sterile water to 10% by adopting a gradient dilution method^-7Then 100. mu.L of the suspension was spread on a PDA solid medium and cultured at 28 to 32 ℃ for 5 to 7 days and 2 months, respectively, and then the number of colonies was counted, and the results are shown in Table 4 below.

Table 4 shows the number of bacteria in the sheep manure bio-organic fertilizer of examples 10 to 12 of the present invention

Examples of the applications

The banana planting test is carried out on the cow and sheep manure bio-organic fertilizer. A field plot experiment is carried out, the test area is about 0.5 mu, a random block design is adopted, cow dung bio-organic fertilizer of the invention example 1 (the fertilizing amount of the bio-organic fertilizer of the invention is 400 kg/mu + the compound fertilizer is 100 kg/mu + the urea is 50 kg/mu + the potassium chloride is 40 kg/mu), sheep dung bio-organic fertilizer of the invention example 4 (the fertilizing amount of the bio-organic fertilizer of the invention is 400 kg/mu + the compound fertilizer is 100 kg/mu + the urea is 50 kg/mu + the potassium chloride is 40 kg/mu), and a fertilizer (farmer habit) control is respectively applied, the soil background is used as a blank, and 3 times are respectively carried out.

When the soil is harvested, various indexes of the soil are measured, and the measuring method is carried out according to related national standards or industry standards, and specifically comprises the following steps:

and (3) soil pH determination: potentiometric method (NY-T1377) -2007);

and (3) measuring the organic matters of the soil: potassium dichromate volumetric method (GB 9834-88);

and (3) soil alkaline hydrolysis nitrogen determination: alkaline diffusion method (LY-T1229-1999);

and (3) determining the soil available phosphorus: sodium bicarbonate leach-molybdenum antimony anti-spectrophotometry (HJ 704-;

soil quick-acting potassium: ammonium acetate leaching-flame photometry (NY/T8892004);

and (3) measuring total nitrogen of the soil: kjeldahl method (HJ 717 2014);

and (3) soil total phosphorus determination: alkali fusion or acid fusion (LY/T1232-2015);

and (3) soil total potassium determination: alkali fusion or acid fusion (LY/T-1234-2015);

number of pathogenic spores: by plate counting method, the incidence of blight is the number of diseased plants/total number of plants.

The results are shown in table 5, the fertilizer has high alkaline hydrolysis N, quick-acting phosphorus and quick-acting potassium, is excessive, and is easy to cause soil hardening and fertilizer pollution, the cow and sheep manure bio-organic fertilizer can improve soil fertility, reduce fertilizer application, prevent and control diseases (inhibit harmful microorganisms), effectively inhibit blight, and avoid soil acidification.

TABLE 5 determination of various indexes of banana planting test

While the present invention has been described in detail with reference to the preferred embodiments, it should be understood that the above description should not be taken as limiting the invention. Various modifications and alterations to this invention will become apparent to those skilled in the art upon reading the foregoing description. Accordingly, the scope of the invention should be determined from the following claims.