CN111849829A - Compound microbial agent and preparation method and application thereof - Google Patents
Compound microbial agent and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a compound microbial agent and a preparation method and application thereof, wherein the compound microbial agent is prepared by uniformly mixing a cellulose decomposition strain, a lignin decomposition strain, a protein degradation strain and a starch degradation strain; the cellulose decomposition strain is one or more of bacillus ginseng, bacillus megaterium and bacillus aryabhattai; the lignin decomposing strain is Klebsiella; the protein and starch degrading strain is one or more of bacillus thuringiensis, lysine bacillus borate, bacillus licheniformis and bacillus acidophilus. The invention selects the microbial inoculum to prevent and control the continuous cropping obstacles of the greenhouse vegetables, and regulates the nutrition status and the microbial structure of the continuous cropping soil by using related functional microorganisms, thereby improving the stress resistance of crops and achieving the purpose of eliminating the continuous cropping obstacles.
Description
Technical Field
The invention belongs to the technical field of microorganisms, relates to the field of compound microbial agents for eliminating continuous cropping obstacles of greenhouse vegetables, and particularly relates to a compound microbial agent as well as a preparation method and application thereof.
Background
In greenhouse vegetable planting, due to the pursuit of high yield, continuous cropping and continuous cropping for a long time and abuse of chemical fertilizers and pesticides cause the deterioration of soil quality, and the problem of continuous cropping obstacle is also derived. The continuous cropping obstacle refers to the condition that the abnormal growth and development of crops are caused by planting the same variety or crops with closer relativity on the same land. Continuous cropping obstacles often result in reduced crop yield and quality, low rate of emergence, and even no emergence or death. Continuous cropping obstacles have been the major factor that has been responsible for restricting the healthy development of the vegetable industry in facilities.
The continuous cropping obstacle is caused by the deterioration of soil physicochemical properties, aggravation of soil-borne diseases and accumulation of autotoxic substances in continuous cropping due to the long-term continuous cropping planting of the existing land and the excessive use of chemical fertilizers.
The method for preventing and treating continuous cropping obstacles mainly comprises the following steps:
(1) chemical pesticide control;
(2) cultivating crop varieties with disease and pest resistance;
(3) adopting a planting mode of crop rotation and stubble replacement;
(4) disinfecting the soil;
(5) microbial agents and related derivative products.
The existing traditional method can play a role in preventing continuous cropping obstacles, but has respective disadvantages. For example, chemical pesticides may cause additional pollution; the development cost of crops with disease and pest resistance is high; the rotation of crop and stubble replacement is complicated and time-consuming; the disinfection of the soil also destroys beneficial microbial communities, and destroys the ecological structural balance of the soil. Therefore, the technology of preventing and treating the microbial inoculum and related derivative products becomes a research hotspot and an advantageous industrial direction in the field of prevention and treatment of continuous cropping obstacles.
In summary, a new compound microbial agent and a preparation method thereof are urgently needed in combination with the reasons of continuous cropping obstacles and the advantages and disadvantages of various prevention and treatment methods.
Disclosure of Invention
The invention aims to provide a compound microbial agent, a preparation method and application thereof, so as to solve one or more technical problems. The invention selects the microbial inoculum to prevent and control the continuous cropping obstacles of the greenhouse vegetables, and regulates the nutrition status and the microbial structure of the continuous cropping soil by using related functional microorganisms, thereby improving the stress resistance of crops and achieving the purpose of eliminating the continuous cropping obstacles.
In order to achieve the purpose, the invention adopts the following technical scheme:
the compound microbial agent is prepared by uniformly mixing a cellulose decomposition strain, a lignin decomposition strain and a protein and starch degradation strain;
the cellulose decomposition strain is one or more of bacillus ginseng, bacillus megaterium and bacillus aryabhattai;
the lignin decomposing strain is Klebsiella;
the protein and starch degrading strain is one or more of bacillus thuringiensis, lysine bacillus borate, bacillus licheniformis and bacillus acidophilus.
The invention further improves the method and also comprises the following steps: attaching a carrier; the attaching carrier is zeolite or sawdust.
The invention is further improved in that the total number of viable bacteria contained in each gram of the attaching carrier is more than or equal to 5 multiplied by 109And (4) respectively.
The compound microbial agent is used for eliminating continuous cropping obstacles of greenhouse vegetables.
The application of the compound microbial agent comprises the following steps: and when the compound microbial agent is used for plowing, the compound microbial agent is uniformly spread according to the dosage of 20-25kg per mu.
The preparation method of the compound microbial agent comprises the following steps:
step 1, respectively activating and culturing cellulose decomposing strains, lignin decomposing strains and protein and starch degrading strains to ensure that the bacteria content of each strain is more than or equal to 4 multiplied by 109CFU/mL, respectively obtaining bacterial liquids of cellulose decomposition strains, lignin decomposition strains and protein and starch degradation strains;
and 2, preparing a bacterial solution of the cellulose decomposition strain, the lignin decomposition strain and the protein and starch degradation strain obtained in the step 1 according to a volume ratio of (1-3): (2-6): and (1) mixing the components in the ratio of (1) to (3) to obtain the liquid compound microbial agent.
The invention further improves the method and also comprises the following steps: and 3, adding an attachment carrier into the liquid compound microbial agent obtained in the step 2, uniformly mixing, and airing to obtain the solid compound microbial agent.
In a further development of the invention, the attachment support is a zeolite; wherein, 0.5 kg zeolite is added into each liter of liquid compound microorganism bacterium agent.
The preparation method of the compound microbial agent comprises the following steps:
respectively activating 8 strains of bacillus ginseng, bacillus megaterium, bacillus aryabhattai, klebsiella, bacillus thuringiensis, lysine bacillus borate, bacillus licheniformis and bacillus acidophilus in an NA solid slant culture medium; then respectively inoculating into LBCulturing in a liquid culture medium at 30-37 deg.C for 20-24 hr to make the bacteria content of each strain greater than or equal to 4 × 109CFU/mL; then the 8 bacterial liquids are combined according to the volume ratio of 1: 3: 1: 3: 1 to obtain the liquid compound microbial inoculum.
The invention further improves the method and also comprises the following steps: and adding an attachment carrier into the obtained liquid compound microbial agent, uniformly mixing, and airing to obtain the solid compound microbial agent.
Compared with the prior art, the invention has the following beneficial effects:
the invention selects the microbial inoculum to prevent and control the continuous cropping obstacles of the greenhouse vegetables, and regulates the nutrition status and the microbial structure of the continuous cropping soil by using related functional microorganisms, thereby improving the stress resistance of crops and achieving the purpose of eliminating the continuous cropping obstacles. According to the invention, the cellulose and lignin degrading strains can rapidly decompose plant residues in the continuous cropping soil, generate nutrient substances which can be absorbed and utilized by crops, and avoid the damage to crop roots after the plant residues are naturally rotten; the starch and protein degrading strain can kill pests, worm eggs and the like hidden in the continuous cropping soil, and effectively reduce the occurrence of related crop soil-borne diseases. The multifunctional microorganism composition can effectively antagonize germs, improve the microbial community structure of soil and reduce the damage of bacteria and fungal diseases to the roots of plants; can enhance the capability of resisting diseases and insect pests of crops, improve the yield of crops and eliminate continuous cropping obstacles.
Detailed Description
The invention provides a compound microbial agent for eliminating continuous cropping obstacles of greenhouse vegetables, which comprises the following components in part by weight: microbial agents and attachment carriers; the microbial agent is prepared by uniformly mixing cellulose, a lignin decomposition strain, a protein and a starch degradation strain; the cellulose and lignin decomposing strains comprise: bacillus ginseng ginsengisoli, Bacillus megaterium, Bacillus aryabhattai, Klebsiella; the protein and starch degrading strain comprises: bacillus thuringiensis, lysine Bacillus boronate, Bacillus licheniformis and Bacillus acidophilus acidocola strain, wherein the strains can be purchased from Shanxi Evernicity soil environment technology, but the limited liability company; the attachment support is zeolite.
In the embodiment of the invention, the ratio of the microbial agent to the attachment carrier is 1L corresponding to 500 g.
The method for preparing the composite microbial agent for eliminating the continuous cropping obstacles of the greenhouse vegetables, disclosed by the embodiment of the invention, can be realized by the following steps:
(1) the strains used were 8 species, respectively: bacillus ginseng ginsengisoli, Bacillus megaterium, Bacillus aryabhattai, Klebsiella Klebsiella, Bacillus thuringiensis, lysine Bacillus boronicum, Bacillus licheniformis, Bacillus acidophilus strain;
(2) the mixing volume ratio of the microorganisms is as follows: the concentration of the bacterial liquid is more than or equal to 4 multiplied by 109CFU/mL；
(3) Culture medium
1) The NA solid culture medium is used for strain activation and has the following formula:
10g of tryptone, 5g of yeast extract powder, 10g of sodium chloride, 1000mL of deionized water, 15-20g of agar and pH 7.0.
2) LB liquid culture medium, used for strain fermentation, the formulation is:
10g of tryptone, 5g of yeast extract powder, 10g of sodium chloride, 1000mL of deionized water and pH 7.0.
The attachment carrier used by the microbial agent is sterilized zeolite, and the ratio of the microbial agent to the attachment carrier is 1L corresponding to 500 g.
(4) Preparation method of microbial inoculum
1) Respectively activating 8 strains in a NA solid slant culture medium,
2) the activated strains are respectively inoculated into sterilized LB liquid culture medium and cultured in a constant temperature incubator at 37 ℃ for 24 hours.
3) Adjusting the bacterial content of 8 strains of bacteria to be 4 × 10 or more9CFU/mL, and then mixing 8 bacterial liquids according to the volume ratio (1: 3: 1: 3: 1) to obtain the liquid compound microbial agent.
4) The sterilized and dried zeolite is quantified and used as an attachment carrier for standby.
5) Adding 0.5 kg of zeolite into each liter of mixed bacteria liquid and mixing with the zeolite uniformly.
6) And (3) airing the mixture in a dark and ventilated place to finish the preparation of the compound microbial agent.
The application of the compound microbial agent provided by the embodiment of the invention comprises the following steps: when in cultivation, the microbial inoculum is uniformly spread according to the dosage of 20-25kg per mu; or applied to soil together with farmyard manure or biological organic fertilizer.
In conclusion, the invention belongs to the technical field of microorganisms, and particularly relates to the technical field of elimination of continuous cropping obstacles of greenhouse vegetables. The compound microbial agent is prepared by uniformly mixing cellulose, lignin decomposing strains, protein and starch degrading strains; the cellulose and lignin decomposing strains comprise: bacillus ginseng ginsengisoli, Bacillus megaterium, Bacillus aryabhattai, Klebsiella pneumoniae; the protein and starch degrading strain comprises: bacillus thuringiensis, Lysinibacillus boronicus, Bacillus licheniformis, Bacillus acidophilus acidocola strain; the invention also discloses a preparation method of the compound microbial agent and application of the compound microbial agent; in addition, the invention adopts the composition of various strains, so that the functions can be mutually complemented, and the microbial inoculum has strong stability; the fertilizer can be applied to soil to antagonize germs, improve the microbial community structure of the soil, reduce the damage of bacteria and fungal diseases to the roots of plants, decompose plant residues in continuous cropping soil, kill pests and ova in the soil and reduce the occurrence of soil-borne diseases; enhance the capability of resisting diseases and insect pests of crops and improve the yield of the crops, and play a role in eliminating continuous cropping obstacles.
Example 1
The preparation method of the compound microbial agent comprises the following steps
(1) Activating 8 strains in a sterilized NA solid culture medium, wherein the formulation of the NA solid culture medium comprises 10g of tryptone, 5g of yeast extract powder, 10g of sodium chloride, 1000mL of deionized water, 15-20g of agar and pH 7.0.
(2) The activated 8 strains were inoculated into LB liquid medium, respectively, and cultured in a 37 ℃ incubator for 24 hours. The LB liquid medium formula is tryptone 10g, yeast extract 5g, sodium chloride 10g, deionized water 1000mL, pH7.0.
(3) Adjusting the cultured bacterial liquid to make the bacterial content of each strain be not less than 4 × 109CFU/mL, and then mixing 8 bacterial liquids according to the volume ratio (1: 3: 1: 3: 1) to obtain the liquid compound microbial agent.
(4) Adding 0.5 kg of zeolite into each liter of mixed bacteria liquid and mixing with the zeolite uniformly. And (3) airing the mixture in a dark and ventilated place to finish the preparation of the compound microbial agent.
Example 2
The application of the compound microbial agent provided by the embodiment of the invention comprises the following steps:
the experimental site is located in a greenhouse (continuous cropping of tomatoes for more than 4 years) of a Shenmuwusu desertification control forestation base, and four treatments are set: control group: original continuous cropping soil is not added with other substances; treatment group 1: the microbial inoculum is evenly mixed with continuous cropping soil according to 20kg per mu; treatment group 2: mixing a biological organic fertilizer and zeolite serving as base fertilizers and applying the mixture to continuous cropping soil, wherein the dosage of the zeolite is 20kg per mu; treatment group 3: the biological organic fertilizer and the composite microbial inoculum are used as base fertilizers to be mixed and applied to the continuous cropping soil, and the dosage of the microbial inoculum is 20kg per mu. In the four tests, the tomato plant spacing is 40cm, the tomato row spacing is 50cm, various management measures are consistent, the number of diseased and withered plants of the plants and the number of soil microorganisms in the mature period are recorded, and finally the total yield of the tomatoes is counted.
TABLE 1 Total tomato yield and disease statistics
As can be seen from table 1, compared with the control group 1, the tomato prevalence rate decreased by 24% and the yield increased by 19.37% in the treatment group 1, i.e. the group to which only the fungicide was applied, and the fungicide could be seen to have an effect of alleviating continuous cropping disorders; compared with the control group, the tomato prevalence rate is reduced by 30% and the yield is increased by 24.22% in the treatment group 2, so that the application of the bio-organic fertilizer has a quite good effect on continuous cropping obstacles; compared with a control group, the morbidity of the tomatoes in the treatment group 3 is reduced by 40%, the yield is increased by 29.33%, and the effect of the biological organic fertilizer used in combination with the microbial inoculum is far greater than the effect of the microbial inoculum used alone or the effect of the organic fertilizer used alone.
TABLE 2 soil microbial Condition
As can be seen from the number of microorganisms in each group shown in Table 2, the microbial community structure of soil can be improved by applying a microbial inoculum, applying a bio-organic fertilizer and applying the microbial inoculum in combination with the bio-organic fertilizer, wherein the microbial inoculum in combination with the bio-organic fertilizer can significantly improve the number of microorganisms in the soil, enhance the microbial activity and the related enzyme activity of the soil, and improve the conversion process of continuous cropping soil substances, thereby being beneficial to the growth and development of crops.
Example 3
The compound microbial agent is prepared by uniformly mixing a cellulose decomposition strain, a lignin decomposition strain and a protein and starch degradation strain;
the cellulose decomposition strain is bacillus ginseng;
the lignin decomposing strain is Klebsiella;
the protein and starch degrading strain is bacillus thuringiensis.
Example 4
The compound microbial agent is prepared by uniformly mixing a cellulose decomposition strain, a lignin decomposition strain and a protein and starch degradation strain; the cellulose decomposition bacterial strain is bacillus ginseng and bacillus megaterium; the lignin decomposing strain is Klebsiella; the protein and starch degrading strain is bacillus thuringiensis and lysine borate bacillus.
Example 5
The compound microbial agent is prepared by uniformly mixing a cellulose decomposition strain, a lignin decomposition strain and a protein and starch degradation strain;
the cellulose decomposition bacterial strain is bacillus ginseng, bacillus megaterium and bacillus aryabhattai; the lignin decomposing strain is Klebsiella; the protein and starch degrading strains are bacillus thuringiensis, lysine bacillus borate, bacillus licheniformis and bacillus acidophilus.
Further comprising: attaching a carrier; the attachment support is zeolite. The total viable count of each gram of the attached carrier is more than or equal to 5 multiplied by 109And (4) respectively.
Example 6
The compound microbial agent provided by the embodiment of the invention is used for eliminating continuous cropping obstacles of greenhouse vegetables; the method specifically comprises the following steps: and when the compound microbial agent is used for plowing, the compound microbial agent is uniformly spread according to the dosage of 20-25kg per mu.
Example 7
The preparation method of the compound microbial agent provided by the embodiment of the invention comprises the following steps:
step 1, respectively activating and culturing cellulose decomposing strains, lignin decomposing strains and protein and starch degrading strains to ensure that the bacteria content of each strain is more than or equal to 4 multiplied by 109CFU/mL, respectively obtaining bacterial liquids of cellulose decomposition strains, lignin decomposition strains and protein and starch degradation strains;
step 2, the bacterial liquid of the cellulose decomposition strain, the lignin decomposition strain and the protein and starch degradation strain obtained in the step 1 is mixed according to the volume ratio of 1: 2: 1 to obtain the liquid compound microbial agent.
Example 8
The preparation method of the compound microbial agent provided by the embodiment of the invention comprises the following steps:
step 1, respectively activating and culturing cellulose decomposing strains, lignin decomposing strains and protein and starch degrading strains to ensure that the bacteria content of each strain is more than or equal to 4 multiplied by 109CFU/mL, respectively obtaining bacterial liquids of cellulose decomposition strains, lignin decomposition strains and protein and starch degradation strains;
step 2, the bacterial liquid of the cellulose decomposition strain, the lignin decomposition strain and the protein and starch degradation strain obtained in the step 1 is mixed according to the volume ratio of 3: 5: 3 to obtain the liquid compound microbial agent.
Example 9
The preparation method of the compound microbial agent provided by the embodiment of the invention comprises the following steps:
step 1, respectively activating and culturing cellulose decomposing strains, lignin decomposing strains and protein and starch degrading strains to ensure that the bacteria content of each strain is more than or equal to 4 multiplied by 109CFU/mL, respectively obtaining bacterial liquids of cellulose decomposition strains, lignin decomposition strains and protein and starch degradation strains;
step 2, the bacterial liquid of the cellulose decomposition strain, the lignin decomposition strain and the protein and starch degradation strain obtained in the step 1 is mixed according to the volume ratio of 2: 6: 1 to obtain the liquid compound microbial agent.
Example 10
The preparation method of the compound microbial agent provided by the embodiment of the invention comprises the following steps:
step 1, respectively activating and culturing cellulose decomposing strains, lignin decomposing strains and protein and starch degrading strains to ensure that the bacteria content of each strain is more than or equal to 4 multiplied by 109CFU/mL, respectively obtaining bacterial liquids of cellulose decomposition strains, lignin decomposition strains and protein and starch degradation strains;
step 2, the bacterial liquid of the cellulose decomposition strain, the lignin decomposition strain and the protein and starch degradation strain obtained in the step 1 is mixed according to the volume ratio of 2: 3: 2 to obtain the liquid compound microbial agent.
Example 11
The preparation method of the compound microbial agent provided by the embodiment of the invention is the same as that of the embodiment 10;
further comprising: and 3, adding an attachment carrier into the liquid compound microbial agent obtained in the step 2, uniformly mixing, and airing to obtain the solid compound microbial agent.
The attachment carrier is zeolite; wherein, 0.5 kg zeolite is added into each liter of liquid compound microorganism bacterium agent.
Example 12
The preparation method of the compound microbial agent comprises the following steps:
respectively activating 8 strains of bacillus ginseng, bacillus megaterium, bacillus aryabhattai, klebsiella, bacillus thuringiensis, lysine bacillus borate, bacillus licheniformis and bacillus acidophilus in an NA solid slant culture medium; then respectively inoculating the strains into LB liquid culture medium, and then culturing for 20-24 hours in a constant temperature incubator at 30-37 ℃ so that the bacteria content of each strain is more than or equal to 4 multiplied by 109CFU/mL; then the 8 bacterial liquids are combined according to the volume ratio of 1: 3: 1: 3: 1 to obtain the liquid compound microbial inoculum.
Example 13
The preparation method of the compound microbial agent provided by the embodiment of the invention is the same as that in the embodiment 12;
further comprising: and adding an attachment carrier into the obtained liquid compound microbial agent, uniformly mixing, and airing to obtain the solid compound microbial agent.
Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art can make modifications and equivalents to the embodiments of the present invention without departing from the spirit and scope of the present invention, which is set forth in the claims of the present application.
Claims (10)
1. The compound microbial agent is characterized by being prepared by uniformly mixing a cellulose decomposition strain, a lignin decomposition strain and a protein and starch degradation strain;
the cellulose decomposition strain is one or more of bacillus ginseng, bacillus megaterium and bacillus aryabhattai;
the lignin decomposing strain is Klebsiella;
the protein and starch degrading strain is one or more of bacillus thuringiensis, lysine bacillus borate, bacillus licheniformis and bacillus acidophilus.
2. The complex microbial inoculant of claim 1, further comprising: attaching a carrier;
the attaching carrier is zeolite or sawdust.
3. The complex microbial inoculant according to claim 2, wherein the total viable count per gram of the carrier is 5 x 10 or more9And (4) respectively.
4. The compound microbial inoculant as claimed in claim 1 is used for eliminating continuous cropping obstacles of greenhouse vegetables.
5. The use of the complex microbial inoculant of claim 1, comprising: and when the compound microbial agent is used for plowing, the compound microbial agent is uniformly spread according to the dosage of 20-25kg per mu.
6. A method for preparing the compound microbial agent of claim 1, which comprises the following steps:
step 1, respectively activating and culturing cellulose decomposing strains, lignin decomposing strains and protein and starch degrading strains to ensure thatThe bacteria content of each strain is 4 × 10 or more9CFU/mL, respectively obtaining bacterial liquids of cellulose decomposition strains, lignin decomposition strains and protein and starch degradation strains;
and 2, preparing a bacterial solution of the cellulose decomposition strain, the lignin decomposition strain and the protein and starch degradation strain obtained in the step 1 according to a volume ratio of (1-3): (2-6): and (1) mixing the components in the ratio of (1) to (3) to obtain the liquid compound microbial agent.
7. The method of claim 6, further comprising:
and 3, adding an attachment carrier into the liquid compound microbial agent obtained in the step 2, uniformly mixing, and airing to obtain the solid compound microbial agent.
8. The method according to claim 7, wherein the attachment support is zeolite; wherein, 0.5 kg zeolite is added into each liter of liquid compound microorganism bacterium agent.
9. The preparation method of the compound microbial agent is characterized by comprising the following steps of:
respectively activating 8 strains of bacillus ginseng, bacillus megaterium, bacillus aryabhattai, klebsiella, bacillus thuringiensis, lysine bacillus borate, bacillus licheniformis and bacillus acidophilus in an NA solid slant culture medium; then respectively inoculating the strains into LB liquid culture medium, and then culturing for 20-24 hours in a constant temperature incubator at 30-37 ℃ so that the bacteria content of each strain is more than or equal to 4 multiplied by 109CFU/mL; then the 8 bacterial liquids are combined according to the volume ratio of 1: 3: 1: 3: 1 to obtain the liquid compound microbial inoculum.
10. The method of manufacturing according to claim 9, further comprising:
and adding an attachment carrier into the obtained liquid compound microbial agent, uniformly mixing, and airing to obtain the solid compound microbial agent.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110643545A (en) * | 2018-11-05 | 2020-01-03 | 山西农业大学 | Bacillus aryabhattai and application thereof in hay blending |
| CN112501084A (en) * | 2020-12-22 | 2021-03-16 | 山东农业大学 | Rhizosphere probiotic Klebsiella ZH07 and application thereof |
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| CN112501084A (en) * | 2020-12-22 | 2021-03-16 | 山东农业大学 | Rhizosphere probiotic Klebsiella ZH07 and application thereof |
| CN112501084B (en) * | 2020-12-22 | 2022-04-08 | 山东农业大学 | Rhizosphere probiotic Klebsiella ZH07 and application thereof |
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