CN111378590A - Compound microbial agent and method for soil fertility improvement and ecological improvement by using same - Google Patents
Compound microbial agent and method for soil fertility improvement and ecological improvement by using same Download PDFInfo
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12R2001/38—Pseudomonas
Abstract
The invention provides a compound microbial agent, which is prepared by mixing the following microorganisms in equal proportion: bacillus megaterium GRINML5, Azotobacter salinistris strain GRINML6, Pseudomonas sp GRINML7, Bacillus licheniformis GRINML8, Stenotrophoromonas sp GRINML9 and Brevibacterium sp GRINM L2. The invention also provides a method for soil fertility improvement and ecological improvement by using the compound microbial agent. The invention separates a plurality of strains of microorganisms which are beneficial to plant growth from the environment, and the microorganisms are artificially compounded to form the microbial fertilizer. The plant growth is promoted by artificially supplementing microbial fertilizers to soil and building a microbial ecological system for promoting the solidification, release and transformation of vegetation nutrients, so that the virtuous cycle of promoting the plant growth by the microbes, promoting the microbial growth by the plants and mutually promoting the plants and the microbes is gradually formed, and the soil ecology is fundamentally improved.
Description
Technical Field
The invention belongs to the field of soil fertility enhancement and ecological environment restoration, and relates to a compound microbial agent and a method for soil fertility enhancement and ecological improvement by using the same.
Background
Soil impoverishment is also called soil degradation and is a comprehensive characterization of soil environment and soil physical, chemical and biological characteristic deterioration, such as the reduction of organic matter content, the deficiency of nutrient elements, the destruction of soil structure, the erosion of soil, the thinning of soil layer, the hardening of soil, the acidification, alkalization and desertification of soil, and the like. In these characterizations, the decrease in organic matter content can be an important indicator of soil degradation, which is associated with many attributes of the soil. For example, the content of organic matters in black soil reclaimed primarily in northeast China is 7-10%, but the content of organic matters in the soil is reduced to 3-4% and even about 2% in some cases, which is less than one hundred years of reclamation. In the farmland of Huang-Huai-Hai area, large areas of drought and waterlogging saline-alkali soil are not improved, irrigation is carried out in the past to improve the yield without considering drainage conditions, and serious secondary salinization is caused as a result. After the forests in southern areas are cut down, the content of organic matters in the soil is reduced from 5-8% to 1-2%, and the soil fertility is obviously reduced. Therefore, how to recover the soil fertility and improve the ecological environment is one of the major problems facing the environmental protection work.
Disclosure of Invention
In order to solve the problems, the invention aims to provide a compound microbial agent which is an organic combination of various microbial strains such as azotobacter, phosphate solubilizing bacteria, carbon fixing bacteria, potassium fixing and potassium promoting bacteria, humus degrading bacteria and the like and can build primary environmental microbial ecology for soil. The established microbial ecological environment creates conditions for the recovery and growth of vegetation and promotes the recovery of the vegetation.
Another object of the present invention is to provide a method for soil fertility and ecological improvement.
In order to achieve the purpose, the invention provides a compound microbial agent which is prepared by mixing the following microorganisms in equal proportion:
bacillus megaterium GRINML5, Azotobacter salinistris strain GRINML6, Pseudomonas sp GRINML7, Bacillus licheniformis GRINML8, Stenotrophoromon p.GRINML9 and Brevibacterium sp GRINM L2;
wherein, the Bacillus megaterium GRINML5 is named as follows: bacillus megaterium (Bacillus megaterium) GRINML5, the collection unit is: china center for type culture Collection, Address: the preservation date of Wuhan university in Wuhan, China is as follows: in 2018, 10 and 15 days, the preservation number is: CCTCC NO: m2018672;
the Azotobacter salinestris strain GRINML6 classification name is: azotobacteroidelinestis strain GRINML6, with the collection unit of China center for type culture Collection, address: wuhan university in Wuhan, China, preservation date: year 2018, 10 month 15, deposit accession number: CCTCC NO: m2018673;
pseudomonas sp GRINML7 is named after classification: pseudomonas sp GRINML7, with the collection unit: china center for type culture Collection, Address: the preservation date of Wuhan university in Wuhan, China is as follows: in 2018, 10 and 15 days, the preservation number is: CCTCC NO: m2018674;
the Bacillus licheniformis GRINML8 is named after classification: bacillus licheniformis (Bacillus licheniformis) GRINML8, the preservation unit is: china center for type culture Collection, Address: wuhan university in Wuhan, China, preservation date: year 2018, 10 month 15, deposit accession number: CCTCC NO: m2018675;
grinml9 classification was named as: grinml9, deposited in units of: china center for type culture Collection, addresses: the preservation date of Wuhan university in Wuhan, China is as follows: in 2018, 10 and 15 days, the preservation number is: CCTCC NO: m2018676;
brevibacterium sp GRINM L2 is classified and named as: brevibacterium sp GRINML2 with the collection unit: china general microbiological culture Collection center, addresses are: west road No.1, north west of the morning area, beijing, institute of microbiology, china academy of sciences, date of deposit: 2016, 9, 29 days, accession number: CGMCC No. 13064.
The invention also provides a method for increasing the fertilizer and improving the ecology of the soil, which comprises the following steps:
1) mixing Bacillus megaterium GRINML5, Azotobacter salinistris strain GRINML6, Pseudomonas sp GRINML7, Bacillus licheniformis GRINML8, Stenotrophoromonas sp GRINML9 and Brevibacterium sp GRINM L2 in equal proportion;
2) preparation of culture Medium
The formula of the culture medium is as follows: glucose 0-3g/L, K2HPO40-3g/L，KH2PO40-3g/L, peptone 0-5g/L, (NH)4)2CO30-3g/L，CaCl20-0.5g/L，MgSO40-1g/L of yeast extract, 0-5g/L of sodium lactate and 0-5g/L of sodium lactate, adding the above components into 1L of distilled water, sterilizing at 121 deg.C for 20min, and cooling to room temperature, wherein the components are not zero at the same time;
3) adding the bacterial liquid compounded in the step 1) into the culture medium in the step 2), adding the bacterial liquid with the volume being 1-10% of the volume of the culture medium, and standing and culturing at the temperature of 20-38 ℃ for 1-15 days to logarithmic phase to prepare a microbial liquid;
4a) crushing crop straws or fallen leaves to 1-20mm, spraying the microbial inoculum prepared in the step 3) into the crushed crop straws or fallen leaves according to the solid-liquid volume ratio of 3:1, standing and culturing at 20-38 ℃ for 5-30 days to prepare a solid culture, and then culturing the solid culture according to the solid-liquid volume ratio of 2-50kg/m2The amount of (a) is added to the soil;
or 4b) directly adding the microbial liquid prepared in the step 3) to the solution according to the total amount of 5-100L/m2The proportion of the components is sprayed into the soil in 1-20 batches.
Because the soil to be fertilized is in different geographical environments, has different conditions such as planes or slopes, and the water and soil retention capacity of different soils is different, solid cultures or liquid microbial inoculum added at one time or lost under some conditions (such as slope soil) causes waste, so that the solid cultures or the liquid microbial inoculum is averagely added into target soil in batches under the condition of calculating the total dosage, and the solid cultures or the liquid microbial inoculum can also be added at one time if the soil condition is good.
Preferably, the culture temperature in step 3) is 25-38 ℃.
The invention firstly builds primary environmental microbial ecology on the site by artificially supplementing environmental beneficial microbes (organic combination of various microbial strains such as azotobacter, phosphate solubilizing bacteria, humus degrading bacteria, carbon fixing bacteria and the like) and nutrient sources and by artificial intervention for a period of time.
The Bacillus megaterium GRINML5 has the functions of dissolving phosphorus, fixing potassium and promoting potassium, the Azotobacter salinistis strain GRINML6 has the function of fixing nitrogen, the Pseudomonas sp GRINML7 has the strong capability of decomposing organic matters and is beneficial to the decomposition of soil organic matters, the Bacillus licheniformis GRINML8 has the functions of promoting the decomposition of organic matters and fixing carbon, can inhibit the propagation of pathogenic bacteria in soil and the attack on the roots of plants, reduce soil-borne diseases of plants, promote the growth of crops and improve the germination rate and seedling preservation rate of seeds, the Stenotrophoromonas sp GRINML9 has the functions of dissolving phosphorus and fixing nitrogen, and the Brevibacterium sp GRINM L2 has the functions of degrading humus and antagonizing plant diseases and insect pests.
The established microbial ecological environment then creates conditions for the recovery and growth of vegetation, promotes the recovery of vegetation, for example, nitrogen in the air is solidified by nitrogen fixing microorganisms to become a nitrogen source which can be used by plants, phosphorus is provided for plants by phosphorus dissolving microorganisms in soil, potassium is provided for plants by potassium dissolving microorganisms, and the existence of the microorganisms is beneficial to the growth of plants. The microbial ecosystem constructed by the invention creates conditions for the growth of plants, and the indigenous plants can efficiently and rapidly propagate on bare rock cracks with less soil and the surface of sandy soil, thereby promoting the natural greening of the bare rock and the sandy soil. After the vegetation is recovered, fallen leaves and the like of the plants can be decomposed into nutrients required for the growth of the plants and the microorganisms by the humus degradation microorganisms, so that the growth of the plants and the microorganisms is promoted. Meanwhile, the mountain vegetation can be recovered to shield the rocks from sunlight, so that evaporation of rock moisture is reduced, and growth of microorganisms and plants is facilitated.
The invention separates a plurality of strains of microorganisms which are beneficial to plant growth from the environment, and the microorganisms are artificially compounded to form the microbial fertilizer. A microbial ecological system for promoting vegetation nutrient solidification, release and conversion is built by artificially supplementing microbial fertilizers to soil, plant growth is promoted, virtuous circle that the microorganisms promote plant growth, the plants promote microbial growth and the plants and the microorganisms mutually promote is gradually formed, and soil ecology is fundamentally improved.
The invention has the beneficial effects that:
1. the microorganism is a compound preparation prepared by natural microorganisms separated from the nature through compounding and specific conditions, and has no ecological risk.
2. The microbial soil fertility increasing technology is characterized in that the soil structure and the ecology are fundamentally improved through the combination of various microbes, the enrichment and the transformation of nutrients are continuously realized through the action of the microbes, the soil fertility is continuously improved, the microbial soil fertility increasing technology is effective for a long time after being used once, and the microbial soil fertility increasing technology is different from chemical fertilizers and the like and has short effect taking time.
3. The microorganisms and the plants have the mutual promotion effect, and the restoration of the microbial ecosystem can promote the growth of the plants.
Drawings
FIG. 1A is a photograph showing the vegetation growth before 5 months of maintenance in the mountain plant maintenance area in example 1 of the present invention.
FIG. 1B is a photograph showing the vegetation growth after 5 months of maintenance in the mountain plant maintenance area in example 1 of the present invention.
FIG. 2 is a photograph comparing the vegetation growth in the area of the mountain body plants and the area of the control area in example 1 of the present invention.
FIG. 3 is a photograph comparing the growth of the maintained area and the control area after 2 months of sand soil maintenance in the mountain in example 1 of the present invention.
FIG. 4 is a photograph comparing the growth of the maintained area and the control area after 5 months of mountain sandy soil maintenance in example 1 of the present invention.
FIG. 5A is a photograph showing the vegetation growth in the area before 5 months of sand curing in the mountain in example 1 of the present invention.
FIG. 5B is a photograph showing the vegetation growth in the area after 5 months of sand curing in the mountain in example 1 of the present invention.
FIG. 6A is a photograph showing the vegetation growth in the area under maintenance 6 months before the area under maintenance of the greensward in the small area in example 2 of the present invention.
FIG. 6B is a photograph showing the vegetation growth in the area under maintenance 6 months after the greensward is maintained in the area under embodiment 2.
FIG. 7A is a photograph showing the vegetation growth in the area to be maintained before 6 months for maintaining another area of greenery turf in a small area according to example 2 of the present invention.
FIG. 7B is a photograph showing the vegetation growth in the area under maintenance 6 months after the greensward is maintained in another area in the small area in example 2 of the present invention.
Detailed Description
The compound microbial agent used by the invention is prepared by mixing the following microorganisms in equal proportion:
bacillus megaterium GRINML5, Azotobacter salinistris strain GRINML6, Pseudomonas sp GRINML7, Bacillus licheniformis GRINML8, Stenotrophoromon p.GRINML9 and Brevibacterium sp GRINM L2;
wherein, the Bacillus megaterium GRINML5 is named as follows: bacillus megaterium (Bacillus megaterium) GRINML5, the collection unit is: china center for type culture Collection, Address: the preservation date of Wuhan university in Wuhan, China is as follows: in 2018, 10 and 15 days, the preservation number is: CCTCC NO: m2018672;
the Azotobacter salinestris strain GRINML6 classification name is: azotobacteroidelinestis strain GRINML6, with the collection unit of China center for type culture Collection, address: wuhan university in Wuhan, China, preservation date: year 2018, 10 month 15, deposit accession number: CCTCC NO: m2018673;
pseudomonas sp GRINML7 is named after classification: pseudomonas sp GRINML7, with the collection unit: china center for type culture Collection, Address: the preservation date of Wuhan university in Wuhan, China is as follows: in 2018, 10 and 15 days, the preservation number is: CCTCC NO: m2018674;
the Bacillus licheniformis GRINML8 is named after classification: bacillus licheniformis (Bacillus licheniformis) GRINML8, the preservation unit is: china center for type culture Collection, Address: wuhan university in Wuhan, China, preservation date: year 2018, 10 month 15, deposit accession number: CCTCC NO: m2018675;
grinml9 classification was named as: grinml9, deposited in units of: china center for type culture Collection, addresses: the preservation date of Wuhan university in Wuhan, China is as follows: in 2018, 10 and 15 days, the preservation number is: CCTCC NO: m2018676;
brevibacterium sp GRINM L2 is classified and named as: brevibacterium (Brevibacterium sp.) GRINML2, depository: china general microbiological culture Collection center, addresses are: west road No.1, north west of the morning area, beijing, institute of microbiology, china academy of sciences, date of deposit: 2016, 9, 29 days, accession number: CGMCC No. 13064.
Example 1 demonstration of microbial conservation technology in mountain III of Sansui Baoqinling Ridge and its bottom
The mountain III of the third Bao slope ridge of the third City is a bare stone mountain, V-shaped groove construction and vegetation cultivation are already finished, the soil in the V-shaped groove of the mountain is more sand, less clay and nutrient-poor, and plants are difficult to grow. Most of the bottom of the mountain is building garbage and sand, the nutrition is insufficient, and the plant growth is difficult.
The method for increasing the fertilizer and improving the ecology of the concrete soil in the area comprises the following steps:
1. the microorganisms are compounded according to equal proportion to form the compound microbial fertilizer.
2. Preparing a culture medium: the formula of the culture medium is as follows: glucose 1.2g/L, K2HPO40.6g/L, peptone 0.5g/L, (NH)4)2CO30.5g/L，CaCl20.1g/L，MgSO40.1g/L yeast extract 0.25g/L sodium lactate 0.5g/L, adding into 1L distilled water, sterilizing at 121 deg.C for 20min, and cooling to room temperature;
3. adding the compound microbial inoculum into a culture medium, wherein the addition amount of the compound microbial inoculum accounts for 10% of the volume of the culture medium, and standing and culturing at 25 ℃ for 15 days to logarithmic phase to prepare a microbial inoculum;
4. directly adding the microbial liquid according to the total amount of 80L/m2The dosage of the fertilizer is sprayed into soil in 10 batches.
Because the mountain body has a slope, water and soil are not easy to keep, and a small amount of times of modes are adopted.
Effects of the implementation
1.3000 demonstration of a maintenance technique of microorganisms on the green-recovered side wall of a square rice
3000 square meters are selected as a maintenance area, the same area is used as a comparison area, as shown in a picture 1A and a picture 1B, a comparison photo is taken before and after 5 months of mountain plant maintenance area maintenance, and as can be seen from the comparison between the picture 1A (before maintenance) and the picture 1B (after maintenance), the vegetation growth is vigorous in the area maintained by the soil fertility increasing and ecological improving method provided by the invention. As shown in figure 2, the photo of the area is compared with the control area, the control area is not maintained, after 5 months, the vegetation in the control area is rare, the growth of plants is difficult, and the vegetation in the area is flourishing.
2.4500 demonstration of quick greening technique for flat rice sandy soil microorganisms
4500 square meters of sandy soil are selected as a maintenance area, and the same area is selected as a control area. As shown in FIG. 3, which is a graph comparing the curing area and the control area after 2 months of sandy soil curing, it can be seen from FIG. 3 that the change of the curing area and the control area is already obvious after 2 months of sandy soil curing. Fig. 4 is a comparison graph of the maintenance area and the control area after 5 months of maintenance, from which it can be seen that the vegetation in the maintenance area is flourishing, while the vegetation in the control area is still sparse, and the vegetation growth difference between the maintenance area and the control area is significant.
Fig. 5A and 5B are a comparison of a small area of the maintenance area before and after 5 months, and it can be seen that the vegetation grows vigorously after maintenance, and the bare soil is covered completely.
Example 2 microbial conservation demonstration of turf greenery areas
The area is a newly-built residential green turf, and due to the fact that much garbage and sand are built in the field, vegetation grows unevenly, and partial areas are dry and withered.
The method for increasing the fertilizer and improving the ecology of the concrete soil in the area comprises the following steps:
1. the microorganisms are compounded according to equal proportion to form the compound microbial fertilizer.
2. Preparing a culture medium: the formula of the culture medium is as follows: glucose 0.8g/L, K2HPO40.5g/L, peptone 0.5g/L, (NH)4)2CO30.5g/L，CaCl20.1g/L sodium lactate 0.2g/L, adding into 1L distilled water, sterilizing at 121 deg.C for 20min, and cooling to room temperature;
3. adding the compound microbial inoculum into a culture medium, wherein the addition amount of the compound microbial inoculum accounts for 5% of the volume of the culture medium, and culturing for 5 days at 30 ℃ and 200rpm of a shaking table to logarithmic phase to prepare a microbial inoculum;
4. crushing fallen leaves to 10mm, spraying the microbial inoculum into the crushed crop straws or fallen leaves according to the solid-liquid volume ratio of 3:1, standing and culturing at 30 ℃ for 10 days to prepare a solid culture, and then culturing the solid culture according to the weight of 15kg/m2The amount of (2) is added to the soil, and the addition is carried out 3 times.
Effects of the implementation
As shown in fig. 6A and 6B, which are comparative graphs of the effect of the greened grass in one area of the residential area before and after 6 months of maintenance, and as shown in fig. 7A and 7B, which are comparative graphs of the effect of the greened grass in another area of the residential area before and after 6 months of maintenance, it can be seen from fig. 6A and 7A that the greened grass in the residential area before maintenance is mottled and the greened grass in some areas is nearly dry, and after 6 months of maintenance, the bare portion is covered with vegetation as shown in fig. 6B and 7B.
From the above examples, it can be seen that after a period of artificial intervention, a virtuous cycle of microbial growth promotion by the plant, and mutual promotion by the plant and the microorganism develops. With the virtuous cycle, more and more humus is formed by the plants, and the soil fertility is greatly increased. Meanwhile, due to the existence of root systems and humus of the vegetation, the water storage capacity of the environment is gradually enhanced, and an ecological environment capable of self-maintaining and self-balancing under the conditions of illumination, rainfall and the like is gradually formed.
The invention provides a compound microbial agent and a method for soil fertility improvement and ecological improvement by using the same, which are mainly oriented to nutrient barren soil, soil destruction and ecological restoration in engineering construction sites, and build a micro-ecological environment for plant growth by using a combination of various microorganisms, so that the soil fertility is improved, the soil quality is fundamentally improved, and ecological vegetation and microbial ecological restoration which are destroyed are promoted.
The microorganism used in the microbial ecological restoration technology is an environment-friendly microorganism, so the invention is an environment-friendly restoration technology. The invention carries out restoration from the restoration of the ecology of microorganisms and plants in the field, and can fundamentally improve the ecology of the field environment.
Claims (3)
1. The compound microbial agent is characterized by being prepared by mixing the following microorganisms in equal proportion:
bacillus megaterium GRINML5, Azotobacter salinistris strain GRINML6, Pseudomonas sp GRINML7, Bacillus licheniformis GRINML8, Stenotrophoromon p.GRINML9 and Brevibacterium sp GRINM L2;
wherein, the Bacillus megaterium GRINML5 is named as follows: bacillus megaterium (Bacillus megaterium) GRINML5, the collection unit is: china center for type culture Collection, Address: the preservation date of Wuhan university in Wuhan, China is as follows: in 2018, 10 and 15 days, the preservation number is: CCTCC NO: m2018672;
the Azotobacter salinestris strain GRINML6 classification name is: azotobacter salinestristrain GRINML6, with a collection unit: china center for type culture Collection, Address: wuhan university in Wuhan, China, preservation date: year 2018, 10 month 15, deposit accession number: CCTCC NO: m2018673;
pseudomonas sp GRINML7 is named after classification: pseudomonas sp GRINML7, with the collection unit: china center for type culture Collection, Address: the preservation date of Wuhan university in Wuhan, China is as follows: in 2018, 10 and 15 days, the preservation number is: CCTCC NO: m2018674;
the Bacillus licheniformis GRINML8 is named after classification: the classification is named as: bacillus licheniformis (Bacillus licheniformis) GRINML8, the preservation unit is: china center for type culture Collection, Address: wuhan university in Wuhan, China, preservation date: year 2018, 10 month 15, deposit accession number: CCTCC NO: m2018675;
grinml9 classification was named as: grinml9, deposited in units of: china center for type culture Collection, addresses: the preservation date of Wuhan university in Wuhan, China is as follows: in 2018, 10 and 15 days, the preservation number is: CCTCC NO: m2018676;
brevibacterium sp GRINM L2 is classified and named as: brevibacterium sp GRINM L2 with the collection unit: china general microbiological culture Collection center, addresses are: west road No.1, north west of the morning area, beijing, institute of microbiology, china academy of sciences, date of deposit: 2016, 9, 29 days, accession number: CGMCC No. 13064.
2. A method for soil fertility improvement and ecological improvement is characterized by comprising the following steps:
1) the method of claim 1, wherein: bacillus megaterium GRINML5, Azotobacter salinistis strain GRINML6, Pseudomonas sp GRINML7, Bacillus licheniformis GRINML8, Stenotrophoromonas sp GRINML9 and Brevibacterium sp GRINM L2 are mixed in equal proportions;
2) preparing a culture medium: the formula of the culture medium is as follows: glucose 0-3g/L, K2HPO40-3g/L，KH2PO40-3g/L, peptone 0-5g/L, (NH)4)2CO30-3g/L，CaCl20-0.5g/L，MgSO40-1g/L of yeast extract, 0-5g/L of sodium lactate and 0-5g/L of sodium lactate, adding the above components into 1L of distilled water, sterilizing at 121 deg.C for 20min, and cooling to room temperature, wherein the components are not zero at the same time;
3) adding the bacterial liquid compounded in the step 1) into the culture medium in the step 2), adding the bacterial liquid with the volume being 1-10% of the volume of the culture medium, and standing and culturing at the temperature of 20-38 ℃ for 1-15 days to logarithmic phase to prepare a microbial liquid;
4a) crushing crop straws or fallen leaves to 1-20mm, spraying the microbial inoculum prepared in the step 3) into the crushed crop straws or fallen leaves according to the solid-liquid volume ratio of 3:1, standing and culturing at 20-38 ℃ for 5-30 days to prepare a solid culture, and then culturing the solid culture according to the weight ratio of 2-50kg/m2The amount of (a) is added to the soil;
or 4b) directly adding the microbial liquid prepared in the step 3) to the solution according to the total amount of 5-100L/m2The proportion of the components is sprayed into the soil in 1-20 batches.
3. The method for soil fertility and ecological improvement according to claim 2, wherein the cultivation temperature in the step 3) is 25 to 38 ℃.
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| CN114616942A (en) * | 2022-04-09 | 2022-06-14 | 中南林业科技大学 | Carbon sink system plants and uses soil treatment device |
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