Emission reduction and efficiency improvement application method of leguminous green manure matched with growth-promoting bacteria

Abstract

The invention discloses an application method of leguminous green manure in cooperation with growth-promoting bacteria for emission reduction and efficiency improvement, and relates to the technical field of crop planting. The method comprises the following steps that rye bean seeds are sown in soil inoculated with bacillus velezensis, rye bean plants are cultured to the full-bloom stage, and then the rye bean plants serve as green manure to be subjected to smashing and turning and pressing treatment. A special bacillus velezensis strain is obtained through separation, has the effects of inhibiting bacteria, fixing nitrogen, dissolving phosphorus, dissolving potassium, forming a biological membrane and generating auxin, has excellent biological characteristics of stress resistance, rapid growth, easiness in separation and the like, further has the advantages of easiness in culture, safety, harmlessness, no pollution and the like, can be fermented to generate rich metabolites, and has a good application prospect. The antibacterial activity and the anti-stress capability are good. The bacillus velezensis is particularly suitable for being combined with rye beans for green manure cultivation, and the effects of fertilizing green manure soil and reducing gas emission can be maximized.

Description

Technical Field

[0001] The present invention relates to the technical field of crop planting, and in particular to an application method of leguminous green manure combined with growth-promoting bacteria to reduce emissions and increase efficiency. Background Art

[0002] In recent years, my country has proposed a series of policies to accelerate the comprehensive green transformation of agriculture, protect and improve the quality of cultivated land, and so on. People's requirements for the safety and quality of agricultural products have also been continuously improved. In addition to increasing the output, agricultural production should also focus on the transformation to high quality, diversification and safety. Long-term biased application of chemical fertilizers and excessive reliance on pesticides will lead to the degradation of cultivated land quality and affect the quality and safety of agricultural products. Green manure, the essence of traditional Chinese agriculture, is a clean organic fertilizer source that can not only improve soil fertility, improve soil environmental quality, prevent soil erosion, and improve the ecological environment, but also increase crop yield and quality. It is an important measure to reduce the use of chemical fertilizers, green prevention and control of pests and diseases, and strengthen the quality of cultivated land. However, in the current application mode of green manure in dryland my country, there are still practical production problems such as the lack of obvious comprehensive effect of green manure and unstable benefits, which restrict the sustainable development of the green manure industry.

[0003] In order to better give full play to the ecological effects of green manure, it must be planted reasonably. The method of combining green manure with synergistic products can solve the problems of the current immature cultivation and utilization model of green manure in summer in my country and the unclear comprehensive benefits. It can provide a scientific basis and theoretical reference for improving the contribution of green manure crops to the development of dryland agriculture in my country, rationally utilizing green manure resources, and promoting the development of the green manure industry system. However, the research on the screening of different green manure + synergistic product combinations is still very imperfect. The present invention intends to develop a green manure cultivation and utilization model of ryegrass green manure + Bacillus Velez, in order to maximize the effects of green manure soil fertilization and reduction of gas emissions. Summary of the invention

[0004] The purpose of the present invention is to provide a method for reducing emissions and increasing efficiency by combining leguminous green manure with growth-promoting bacteria to solve the problems of the above-mentioned prior art. The method can maximize the effect of green manure soil fertilization and gas emission reduction by combining Bacillus Velezii with rye beans for green manure cultivation.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides a Bacillus velezensis for promoting plant growth. The Bacillus velezensis has been deposited in the China Center for Type Culture Collection on June 6, 2024, with the deposit address being Wuhan University, Wuhan, China, and the deposit number being CCTCC NO:M 20241160.

[0007] The present invention also provides the use of the above-mentioned Bacillus Velezii in preparing a microbial agent for promoting plant growth.

[0008] The present invention also provides a microbial agent for promoting plant growth, wherein the active ingredient comprises the above-mentioned Bacillus Velezii.

[0009] The present invention also provides a method for preparing the above-mentioned microbial agent, comprising the following steps: inoculating the above-mentioned Bacillus Velezii into a fermentation medium for fermentation treatment to obtain a fermentation liquid, then separating the bacterial cells, and then freeze-drying the cells to obtain the microbial agent.

[0010] Furthermore, the fermentation medium is beef extract peptone medium.

[0011] Furthermore, the fermentation treatment is carried out at a temperature of 27° C. and for a period of 2-3 days.

[0012] The present invention also provides an application method of leguminous green manure in combination with growth-promoting bacteria for emission reduction and efficiency enhancement, comprising the steps of sowing rye bean seeds in soil inoculated with the above-mentioned Bacillus Velez, culturing the rye bean plants to the full flowering stage, and then using the rye bean plants as green manure for crushing and turning over treatments.

[0013] Furthermore, the inoculation of the Bacillus Velez subtilis is achieved by seeding the above-mentioned microbial agent into the soil.

[0014] Furthermore, the inoculation amount of the microbial agent is 2.5% of the sowing amount of the rye bean seeds.

[0015] Furthermore, the pressing depth of the pressing treatment is 10-15 cm.

[0016] The present invention discloses the following technical effects:

[0017] The present invention separates and obtains a special Velez subspecies bacillus, which has the functions of antibacterial, nitrogen fixation, phosphorus solubilization, potassium solubilization, biofilm formation and auxin production, and has excellent biological characteristics such as stress resistance, rapid growth and easy separation, and also has the advantages of easy cultivation, safety, harmlessness and pollution-free, and can ferment to produce rich metabolites, and has good antibacterial activity and stress resistance.

[0018] Rye beans have the advantages of strong adaptability and stress resistance, short growth cycle and strong nitrogen fixation ability. The Velez bacillus isolated by the present invention is particularly suitable for green manure cultivation in combination with rye beans, which can maximize the effect of green manure soil fertilization and gas emission reduction. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0020] Figure 1 is a single colony image of strain ANB1;

[0021] Figure 2 This is the phylogenetic tree of strain ANB1. DETAILED DESCRIPTION

[0022] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0023] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0024] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0025] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0026] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0027] Example 1 Isolation, Identification and Preservation of Bacillus Velez

[0028] 1. Separation

[0029] A strain was isolated from the soil of Inner Mongolia and named ANB1.

[0030] 2. Identification

[0031] The strain ANB1 was identified as Bacillus velezensis. The 16SrDNA sequence of strain ANB1 is shown in SEQ ID NO.1, and the single colony map is shown in Figure 1 , phylogenetic tree see Figure 2 .

[0032] SEQ ID NO.1:

[0033] GTGGCGTCTGCTATAATGCAGTCGAGCGGACAGATGGGAGCTTGCTCCCTGATGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAAACCGGGGCTAATACCGGATGGTTGTTTGAACCGCATGGTTCAGACATAAAAGGTGGCTTCGGCTACCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTTGTTAGGGAAGAACAAGTGCCGTTCAAATAGGGCGGCACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCTCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACAATCCTAGAGATAGGACGTCCCTTCGGGGGCAAAGGGCAGGGTGGGGATGGTTGCCCCAACTCCGGCCGGAAATT.

[0034] 3..Strain preservation

[0035] Bacillus Velez ANB1 was deposited in the China Center for Type Culture Collection on June 6, 2024, with the deposit address at Wuhan University, Wuhan, China, and the deposit number is CCTCC NO:M 20241160.

[0036] Example 2

[0037] Preparation of Bacillus Velez inoculum:

[0038] The Bacillus Velez subtilis ANB1 isolated in Example 1 was inoculated into a fermentation medium and cultured at 27° C. for 2 days (up to 3 days) to obtain a fermentation broth; the fermentation broth was centrifuged to obtain a precipitate, which was then freeze-dried to obtain a Bacillus Velez subtilis agent.

[0039] The fermentation medium used was beef extract peptone medium.

[0040] Example 3 Potted Plant Test

[0041] The pot experiment was divided into two groups, namely the control group and the black bean + Bacillus Velezii group, and the planting was carried out according to the method shown in Table 1.

[0042] Table 1 Grouping and treatment methods of potted plant experiments

[0043]

[0044] After 45 days of rye bean planting, the biomass dry weight, plant total N, plant total P, plant total K and soil pH of the rye bean plants in the control group and the rye bean + Bacillus Velez group were tested, and the results are shown in Table 2. The results showed that compared with the single rye bean treatment, the biomass of the rye bean + Bacillus Velez treatment increased significantly by 44%.

[0045] Table 2 Effects of potted inoculation with Bacillus Velez on rye beans

[0046] index Control group Rye bean + Bacillus velez Biomass dry weight (g / pot) 1.84 3.69 pH 8.26 8.24

[0047] Example 4 Field Test

[0048] Field trials were conducted in Yongshou and Yangling respectively, and the methods are as follows:

[0049] Sowing: In early July 2023, the Bacillus Velez microbial agent prepared in Example 2 was inoculated into the planting position of rye beans by strip sowing, and then the rye beans were sown, wherein the inoculation amount of the Bacillus Velez microbial agent was 2.5% of the mass of the rye bean seeds, and the sowing amount of the rye beans was 8 kg / mu. The group of single rye beans (i.e., not inoculated with the Bacillus Velez microbial agent) was used as the control.

[0050] Green manure turning: When the black bean plants grow to the peak flowering stage (about 60 days of growth), they are directly turned over and used as green manure using a local crushing and returning machine. The turning depth of the green manure crop is 10-15cm, and the turning time is 30-40 days before the sowing of the next winter wheat crop.

[0051] Each group was tested for indicators including plant biomass, nitrogen return to the field after green manure turning, soil pH, soil available phosphorus, N 2 O accumulation, CH 4 Cumulative amount, CO 2 Cumulative amount, global warming potential GWP and greenhouse gas emission intensity, the results are shown in Table 3. The results showed that in the Yongshou experiment, the biomass of rye beans + Bacillus Velez significantly increased by 11.5% and the amount of nitrogen returned to the field increased by 13% compared with the control group of single rye beans; in the Yangling field experiment, the biomass of rye beans + Bacillus Velez increased by 11% and the amount of nitrogen returned to the field increased by 30% compared with the control group of single rye beans. The technology of inoculating Bacillus Velez bacteria can significantly increase the biomass of rye beans. Compared with single rye beans, inoculation of bacteria can enhance the ability to reduce soil pH; after turning green manure in Yongshou, the soil available phosphorus of rye beans + Bacillus Velez significantly increased by 67.4% compared with single rye beans; after turning green manure in Yangling, the soil available phosphorus of rye beans + Bacillus Velez increased by 8.4% compared with single rye beans.

[0052] Table 3 Effects of field inoculation with Bacillus Velez on rye beans

[0053]

[0054] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A plant growth promoting Bacillus velezensis, characterized in that: The Bacillus Velezii was deposited in the China Center for Type Culture Collection on June 6, 2024, with the deposit address being Wuhan University, Wuhan, China, and the deposit number being CCTCC NO:M 20241160.

2. Use of the Bacillus Velezii strain as claimed in claim 1 in the preparation of a microbial agent for promoting plant growth.

3. A microbial agent for promoting plant growth, characterized in that: The active ingredient comprises the Bacillus Velezii described in claim 1.

4. A method for preparing the microbial agent according to claim 3, characterized in that: The method comprises the following steps: inoculating the Bacillus Velez subtilis described in claim 1 into a fermentation medium for fermentation treatment to obtain a fermentation liquid, then separating the bacterial bodies, and then freeze-drying the bacterial bodies to obtain the microbial agent.

5. The preparation method according to claim 4, characterized in that: The fermentation medium is beef extract peptone medium.

6. The preparation method according to claim 4, characterized in that: The temperature of the fermentation treatment is 27° C. and the time is 2-3 days.

7. A method for using leguminous green manure in combination with growth-promoting bacteria to reduce emissions and increase efficiency, characterized in that: The method comprises the steps of sowing rye bean seeds in the soil inoculated with the Bacillus Velez subtilis according to claim 1, cultivating the rye bean plants to the blooming stage, and then using the rye bean plants as green manure for crushing and turning over treatment.

8. The application method according to claim 7, characterized in that: The inoculation of the Bacillus Velez is achieved by broadcasting the microbial agent according to claim 3 into the soil.

9. The application method according to claim 8, characterized in that: The inoculation amount of the microbial agent is 2.5% of the sowing amount of the rye bean seeds.

10. The application method according to claim 7, characterized in that: The pressing depth of the pressing treatment is 10-15 cm.

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