CN117778252A - Soil remediation microbial inoculum for preventing and treating bighead atractylodes rhizome continuous cropping obstacle as well as preparation method and application thereof - Google Patents
Soil remediation microbial inoculum for preventing and treating bighead atractylodes rhizome continuous cropping obstacle as well as preparation method and application thereof Download PDFInfo
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Abstract
The application relates to the technical field of plant planting, and particularly discloses a soil restoration microbial inoculum for preventing and treating bighead atractylodes rhizome continuous cropping obstacle, and a preparation method and application thereof. The soil restoration microbial agent for preventing and treating bighead atractylodes rhizome continuous cropping obstacle provided by the application comprises bacillus bailii, bacillus amyloliquefaciens, pseudomonas, bacillus atrophaeus and zoogloea; further, the soil remediation microbial inoculum comprises the following components in parts by weight: 1.5 to 2.3 parts of bacillus belicus, 2.7 to 3.1 parts of bacillus amyloliquefaciens, 1.0 to 2.3 parts of pseudomonas, 0.4 to 1.0 part of bacillus atrophaeus and 0.8 to 1.2 parts of zoogloea; the application also provides a preparation method and application of the soil remediation microbial inoculum. On one hand, the soil remediation microbial inoculum can antagonize pathogenic bacteria and inhibit the propagation and growth of pathogenic bacteria in soil; on the other hand, the autotoxic phenomenon of the bighead atractylodes rhizome can be prevented, and the effective prevention and treatment of bighead atractylodes rhizome continuous cropping obstacle can be realized.
Description
Technical Field
The application relates to the technical field of plant planting, in particular to a soil restoration microbial inoculum for preventing and treating bighead atractylodes rhizome continuous cropping obstacle, and a preparation method and application thereof.
Background
The bighead atractylodes rhizome is a perennial herb of the Compositae, is a famous traditional common medicinal material in China, and the rhizome of the bighead atractylodes rhizome contains a plurality of effective medicinal components such as atractylenolide substances, atractylenolide polysaccharide, atractylone volatile oil and the like, so the bighead atractylodes rhizome has a large share in the traditional Chinese medicine market. In recent years, along with the continuous expansion of the application field of the bighead atractylodes rhizome, the market demand for the bighead atractylodes rhizome is also increasing. However, bighead atractylodes rhizome has serious continuous cropping obstacle problem and cannot be planted continuously on the same land; even under the normal management condition, the continuous cropping bighead atractylodes rhizome can also have the phenomena of growth retardation, serious diseases and insect pests, reduced yield, poor quality and the like.
At present, aiming at the problem of continuous cropping obstacle of bighead atractylodes rhizome, the main treatment measures are as follows; firstly, screening and improving varieties of the white atractylodes rhizome, and breeding high-resistance varieties or detoxified plants; secondly, performing damp-heat sterilization on the white atractylodes rhizome planting soil, and reducing the content of soil-borne bacteria in the white atractylodes rhizome planting soil; thirdly, applying biological bacterial fertilizer to balance soil microbial flora; fourthly, various continuous cropping fertilizers are applied to overcome continuous cropping obstacles of bighead atractylodes rhizome planting. However, the method has one or more problems of high operation difficulty, unobtrusive effect, slow effect and the like in practical application. Therefore, farmers still adopt a mode of 'alternate cropping' to solve the problem of continuous cropping obstacle of bighead atractylodes rhizome at present, which causes great waste of land resources and seriously affects sustainable development of bighead atractylodes rhizome production.
Disclosure of Invention
In order to realize effective prevention and treatment of continuous cropping obstacle of bighead atractylodes rhizome and reduce incidence of continuous cropping bighead atractylodes rhizome, the application provides a soil restoration microbial inoculum for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome, and a preparation method and application thereof.
In a first aspect, the application provides a soil remediation microbial inoculum for preventing and treating bighead atractylodes rhizome continuous cropping obstacle, which adopts the following technical scheme:
a soil restoration microbial agent for preventing and treating continuous cropping obstacle of Atractylodis rhizoma comprises Bacillus bailii, bacillus amyloliquefaciens, pseudomonas, bacillus atrophaeus and Acinetobacter.
The application prepares the soil restoration microbial inoculum for preventing and treating the bighead atractylodes rhizome continuous cropping obstacle by utilizing bacillus bailii, bacillus amyloliquefaciens, pseudomonas, bacillus atrophaeus and zoogloea, and the soil restoration microbial inoculum can antagonize pathogenic bacteria in soil and prevent the growth and reproduction of the pathogenic bacteria on one hand; the production of terpenoid substances such as beta-eucalyptol in the rhizome of the bighead atractylodes rhizome can be inhibited, and the autotoxicity of the bighead atractylodes rhizome is reduced. Therefore, the soil remediation microbial inoculum provided by the application can improve the flora environment of the white atractylodes rhizome planting soil, promote the absorption and utilization of the bighead atractylodes rhizome root system to mineral nutrient elements in the soil, and further realize the effective prevention and treatment of bighead atractylodes rhizome continuous cropping obstacles.
In this application, bighead atractylodes rhizome is a major cause of continuous cropping obstacle: firstly, the large quantity of pathogenic bacteria in the bighead atractylodes rhizome continuous cropping soil, such as rhizoctonia solani, fusarium, yellow mold and the like, and the pathogenic bacteria and parasitic pests with strong reproductive capacity are in dominant positions in bighead atractylodes rhizome root systems, so that bighead atractylodes rhizome is easy to generate soil-borne diseases and insect pests. Secondly, the bighead atractylodes rhizome has an autotoxicity phenomenon, namely, the rhizome of bighead atractylodes rhizome contains a large amount of terpenoid substances such as beta-eucalyptol, and the terpenoid substances such as beta-eucalyptol with high concentration weaken the activity of phytohormone on one hand and restrict the growth of plants; on the other hand, the absorption of nutrients by plant roots is also hindered. In the soil restoration microbial agent, bacillus belicus, bacillus amyloliquefaciens and bacillus atrophaeus can generate phenolphthalein antibiotics which can antagonize pathogenic bacteria and mould which can regulate plant growth and development and defend diseases and insect pests, such as chitinase, cellulase and the like, the enzymes can dissolve cell walls of pathogenic fungus cells and damage cell structures of pathogenic microorganisms, so that the growth and reproduction of the pathogenic bacteria are inhibited and the plant growth is promoted; pseudomonas can secrete one or more extracellular hydrolases, and the extracellular hydrolases have the effects of inhibiting the germination of pathogenic bacteria spores and the growth of germ tubes, and finally inhibiting the growth and reproduction of pathogenic bacteria. The zoogloea has good degradation effect on organic toxic substances in white soil, and extracellular enzymes secreted by the zoogloea can damage the cell structure of pathogenic microorganisms, so that diseases and insect pests are prevented.
Optionally, the soil restoration microbial inoculum for preventing and treating bighead atractylodes rhizome continuous cropping obstacle comprises the following components in parts by weight: 1.5 to 2.3 parts of bacillus belicus, 2.7 to 3.1 parts of bacillus amyloliquefaciens, 1.0 to 2.3 parts of pseudomonas, 0.4 to 1.0 part of bacillus atrophaeus and 0.8 to 1.2 parts of zoogloea.
In the application, the addition of each component in the soil restoration microbial agent for preventing and treating bighead atractylodes rhizome continuous cropping obstacle can influence the antibacterial effect of the soil restoration microbial agent. The inventor of the application discovers through experiments that the addition amount of each component in the soil remediation microbial inoculum is controlled within the range, and the obtained soil remediation microbial inoculum has good inhibition effect on three fungi of rhizoctonia solani, fusarium and flavum in soil, and can reduce the content of rhizoctonia solani from 9.83 to 3.24-4.32 ten thousand cfu/g, the content of fusarium from 9.76 to 1.62-3.18 ten thousand cfu/g and the content of flavum from 10.12 ten thousand cfu/g to 2.34-4.20 ten thousand cfu/g.
In some embodiments, the bacillus belicus may be 1.5 to 1.8 parts or 1.8 to 2.3 parts by weight.
In a specific embodiment, the bacillus belicus may also be 1.5 parts, 1.8 parts or 2.3 parts by weight.
In some embodiments, the bacillus amyloliquefaciens may be 2.7-2.9 parts or 2.9-3.1 parts by weight.
In a specific embodiment, the bacillus amyloliquefaciens may also be 2.7 parts, 2.9 parts, or 3.1 parts by weight.
In some embodiments, the pseudomonas may be 1.5 to 1.8 parts, 1.5 to 2.0 parts, 1.8 to 2.3 parts, or 2.0 to 2.3 parts by weight.
In a specific embodiment, the parts by weight of the pseudomonas may also be 1.5 parts, 1.8 parts, 2.0 parts, or 2.3 parts.
In some embodiments, the bacillus atrophaeus may be 0.4-0.6 parts or 0.6-1.0 parts by weight.
In a specific embodiment, the bacillus atrophaeus may also be 0.4 parts, 0.6 parts or 1.0 parts by weight.
In some embodiments, the zoogloea may be 0.8 to 1.0 parts by weight or 1.0 to 1.2 parts by weight.
In a specific embodiment, the weight part of the zoogloea can be 0.8 part, 1.0 part or 1.2 parts.
Optionally, 1.5 to 2.3 parts of bacillus belicus, 2.7 to 3.1 parts of bacillus amyloliquefaciens, 1.5 to 2.0 parts of pseudomonas, 0.4 to 1.0 part of bacillus atrophaeus and 0.8 to 1.2 parts of zoogloea.
Alternatively, the pseudomonas is selected from the group consisting of pseudomonas aeruginosa, pseudomonas fluorescens, pseudomonas syringae, and pseudomonas rhodochrous.
In a specific embodiment, the pseudomonas is pseudomonas aeruginosa, pseudomonas fluorescens or pseudomonas syringae.
In a specific embodiment, the pseudomonas is a mixture of 1 by weight: 0.6 of Pseudomonas fluorescens and Pseudomonas syringae in a weight ratio of 1:0.6 of Pseudomonas syringae and Pseudomonas aeruginosa in a weight ratio of 1:0.6 of Pseudomonas fluorescens and Pseudomonas rhodochrous.
In some embodiments, the pseudomonas is a mixture of pseudomonas fluorescens and pseudomonas aeruginosa.
Optionally, the pseudomonas is a mixture with a weight ratio of 1: (0.2-1) Pseudomonas fluorescens and Pseudomonas aeruginosa.
In some embodiments, the weight ratio of pseudomonas fluorescens to pseudomonas aeruginosa can be 1: (0.2-0.4), 1: (0.2-0.6), 1: (0.2-0.8), 1: (0.2-1), 1: (0.4-0.6), 1: (0.4-0.8), 1:
(0.4-1), 1: (0.6-0.8), 1: (0.6-1) or 1: (0.8-1).
In a specific embodiment, the weight ratio of Pseudomonas fluorescens to Pseudomonas aeruginosa may also be 1:0.2, 1:0.4, 1:0.6, 1:0.8 or 1:1.
in the application, experiments find that the two kinds of pseudomonas which are fluorescent pseudomonas and pseudomonas aeruginosa are further adopted for compounding and use, the obtained soil remediation microbial inoculum has better inhibition effect on pathogenic bacteria in soil, and the soil remediation microbial inoculum is used in bighead atractylodes rhizome planting soil, so that the prevention and treatment effect of bighead atractylodes rhizome planting soil continuous cropping obstacle is more remarkable.
According to the application, the weight ratio of the fluorescent pseudomonas to the pseudomonas aeruginosa is controlled within the range, the obtained soil remediation microbial inoculum can enable the content of rhizoctonia solani in soil to be reduced from 9.83 ten thousand cfu/g to less than 4.0 ten thousand cfu/g, the content of fusarium is reduced from 9.76 ten thousand cfu/g to less than 2.0 ten thousand cfu/g, and the content of yellow mould is reduced from 10.12 ten thousand cfu/g to less than 3.0 ten thousand cfu/g.
Optionally, the effective viable count of bacillus bailii is more than or equal to 20 hundred million cfu/g, the effective viable count of bacillus amyloliquefaciens is more than or equal to 20 hundred million cfu/g, the effective viable count of pseudomonas is more than or equal to 20 hundred million cfu/g, the effective viable count of bacillus atrophaeus is more than or equal to 20 hundred million cfu/g, and the effective viable count of zoogloea is more than or equal to 20 hundred million cfu/g.
In a second aspect, the application provides a preparation method of a soil remediation microbial inoculum for preventing and treating bighead atractylodes rhizome continuous cropping obstacle.
A preparation method of a soil remediation microbial inoculum for preventing and treating bighead atractylodes rhizome continuous cropping obstacle comprises the following steps: and uniformly mixing all the strains to obtain the soil restoration microbial inoculum for preventing and treating continuous cropping obstacle of the bighead atractylodes rhizome.
In a second aspect, the present application provides a method for preventing and treating bighead atractylodes rhizome continuous cropping obstacle.
A method for preventing and treating bighead atractylodes rhizome continuous cropping obstacle, which comprises the following steps: the soil restoration microbial inoculum for preventing and treating the bighead atractylodes rhizome continuous cropping obstacle is mixed with water and then applied to bighead atractylodes rhizome planting soil.
Optionally, the dosage of the soil restoration microbial inoculum is 2-4 kg/mu.
In some embodiments, the soil remediation agent may be used in an amount of 2-3 kg/mu or 3-4 kg/mu.
In a specific embodiment, the soil remediation agent may be used in an amount of 2 kg/mu, 3 kg/mu or 4 kg/mu.
In summary, the present application has the following beneficial effects:
1. the application provides a soil restoration microbial inoculum prepared by mixing bacillus bailii, bacillus amyloliquefaciens, pseudomonas, bacillus atrophaeus and zoogloea, which can antagonize pathogenic bacteria such as rhizoctonia solani, fusarium, yellow mold and the like in soil, inhibit growth and propagation of the pathogenic bacteria, realize improvement of soil flora environment, reduce occurrence and propagation of soil diseases and insect pests and ensure healthy growth of bighead atractylodes rhizome plants.
2. The soil remediation microbial inoculum provided by the application can inhibit the generation of terpenoid substances such as beta-eucalyptol in rhizome of bighead atractylodes rhizome, reduce the autotoxic phenomenon of bighead atractylodes rhizome, ensure the absorption of bighead atractylodes rhizome root system to nutrients, and realize the effective prevention and treatment of bighead atractylodes rhizome continuous cropping obstacle.
3. The application controls the addition amount of each component in the soil remediation microbial inoculum within the following range: 1.5 to 2.3 parts of bacillus belius, 2.7 to 3.1 parts of bacillus amyloliquefaciens, 1.0 to 2.3 parts of pseudomonas, 0.4 to 1.0 part of bacillus atrophaeus and 0.8 to 1.2 parts of zoogloea, the content of rhizoctonia solani in soil can be reduced from 9.83 to 3.24 to 4.32 ten thousand cfu/g, the content of fusarium is reduced from 9.76 to 1.62 to 3.18 ten thousand cfu/g, and the content of yellow mold is reduced from 10.12 to 2.34 to 4.20 ten thousand cfu/g.
4. The application further adopts the weight ratio of 1: the Pseudomonas fluorescens and the Pseudomonas aeruginosa (0.2-1) are compounded, the prepared soil remediation microbial inoculum has better effect of inhibiting pathogenic bacteria in soil, the content of rhizoctonia solani in the soil can be reduced from 9.83 to below 4.0 ten thousand cfu/g, the content of fusarium is reduced from 9.76 to below 2.0 ten thousand cfu/g, and the content of flavomycetes is reduced from 10.12 to below 3.0 ten thousand cfu/g.
Detailed Description
The application provides a soil restoration microbial agent for preventing and treating bighead atractylodes rhizome continuous cropping obstacle, which comprises bacillus bailii, bacillus amyloliquefaciens, pseudomonas, bacillus atrophaeus and zoogloea. Further, the soil remediation microbial inoculum comprises the following components in parts by weight: 1.5 to 2.3 parts of bacillus belicus, 2.7 to 3.1 parts of bacillus amyloliquefaciens, 1.0 to 2.3 parts of pseudomonas, 0.4 to 1.0 part of bacillus atrophaeus and 0.8 to 1.2 parts of zoogloea. Still further, the soil remediation microbial inoculum comprises the following components in parts by weight: 1.5 to 2.3 parts of bacillus bailii, 2.7 to 3.1 parts of bacillus amyloliquefaciens, 1.5 to 2.0 parts of pseudomonas, 0.4 to 1.0 part of bacillus atrophaeus and 0.8 to 1.2 parts of zoogloea.
In this application, pseudomonas is selected from Pseudomonas aeruginosa, pseudomonas fluorescens, pseudomonas syringae, and Pseudomonas rhodopseudomonas. Further, the pseudomonas is a mixture of pseudomonas fluorescens and pseudomonas aeruginosa; still further, the pseudomonas is 1 by weight: (0.2-1) Pseudomonas fluorescens and Pseudomonas aeruginosa.
In the application, each strain is bacterial powder, the effective viable count of bacillus bailii is more than or equal to 20 hundred million cfu/g, the effective viable count of bacillus amyloliquefaciens is more than or equal to 20 hundred million cfu/g, the effective viable count of pseudomonas is more than or equal to 20 hundred million cfu/g, the effective viable count of bacillus atrophaeus is more than or equal to 20 hundred million cfu/g, and the effective viable count of zoogloea is more than or equal to 20 hundred million cfu/g.
In the specific embodiment of the application, the bacillus beljalis is bacillus beljalis BGB-89R, and the preservation number is CGMCC No 28824; the bacillus amyloliquefaciens is bacillus amyloliquefaciens BGB-95R, and the preservation number is CGMCC No 24545; pseudomonas aeruginosa is Pseudomonas aeruginosa SU8, and the preservation number is CCTCC NO: m2013178; pseudomonas fluorescens is Pseudomonas fluorescens G7, and the preservation number is CCTCC NO: m2018359; pseudomonas syringae is Pseudomonas syringae B-1, and the preservation number is CCTCC NO: M2015813; the rhodopseudomonas is rhodopseudomonas DC-2, and the preservation number is CCTCC NO: m2016677; the bacillus atrophaeus is bacillus atrophaeus BGB-98R with the preservation number of CGMCC No 27752; the zoogloea is zoogloea-like Shen fungus B230 with a preservation number of CGMCC No 9280; the strains, solvents, etc. in this application are all commercially available.
The present application is described in further detail below in connection with preparation examples, comparative examples and detection tests.
Preparation examples 1 to 13
Preparation examples 1-13 respectively provide a soil remediation microbial inoculum.
The above preparation example is different in that: the addition amounts of the components in the soil remediation microbial inoculum are shown in the following table 1.
The preparation method of the soil remediation microbial inoculum of preparation examples 1-13 comprises the following steps:
(1) Activating and culturing strains: test strains stored at-80 ℃): bacillus subtilis, bacillus amyloliquefaciens, streptomyces microflavus, fiber micro-bacteria, stenotrophomonas maltophilia and fast-growing bacillus indicus are respectively subjected to 3 times of activation and then are transferred to a nutrient liquid culture medium for culture until the logarithmic phase, and the culture is performed by using a dilution coating plate for counting;
(2) Preparing each bacterial powder: centrifuging the bacterial solutions by using a centrifugal machine, adding a protective agent into each bacterial solution, preparing bacterial powder by using a spray dryer, and adjusting the bacterial content of the bacterial powder to 20 hundred million cfu/g for later use; wherein the protective agent is prepared from the following components in percentage by mass: 4.5:1, dextrin, anhydrous sodium sulfate and sodium metabisulfite, wherein the addition amount of a protective agent is 20% of the mass of the bacterial liquid; obtaining bacillus behenii powder, bacillus amyloliquefaciens powder, pseudomonas fluorescens powder, bacillus atrophaeus powder and zoogloea powder.
(3) Preparing a soil remediation microbial inoculum: the bacillus behenii powder, bacillus amyloliquefaciens powder, pseudomonas fluorescens powder, bacillus atrophaeus powder and zoogloea powder are respectively weighed according to the addition amount of the table 1, and then uniformly mixed to obtain a soil restoration microbial inoculum, and the soil restoration microbial inoculum is stored at 4 ℃ for standby.
TABLE 1 addition amount of each component in soil remediation microbial inoculum provided in preparation examples 1-13
Preparation examples 14 to 23
Preparation examples 14-23 respectively provide a soil remediation microbial inoculum.
The above preparation example differs from preparation example 2 in that: the type and the proportion of the pseudomonas in the soil remediation microbial inoculum are shown in the following table 2.
TABLE 2 types and proportions of Pseudomonas in soil remediation microbial agents provided in PREPARATION EXAMPLE 2 and PREPARATION EXAMPLES 14-23
| Preparation example | Type and proportion of pseudomonas |
| 2 | Pseudomonas fluorescens |
| 14 | Pseudomonas aeruginosa |
| 15 | Pseudomonas syringae |
| 16 | The weight ratio is 1:0.2 mixture of Pseudomonas fluorescens and Pseudomonas aeruginosa |
| 17 | The weight ratio is 1:0.4 mixture of Pseudomonas fluorescens and Pseudomonas aeruginosa |
| 18 | The weight ratio is 1:0.6 mixture of Pseudomonas fluorescens and Pseudomonas aeruginosa |
| 19 | The weight ratio is 1:08 mixture of Pseudomonas fluorescens and Pseudomonas aeruginosa |
| 20 | The weight ratio is 1:1 and Pseudomonas aeruginosa |
| 21 | The weight ratio is 1:0.6 mixture of Pseudomonas fluorescens and Pseudomonas syringae |
| 22 | The weight ratio is 1:0.6 mixture of Pseudomonas syringae and Pseudomonas aeruginosa |
| 23 | The weight ratio is 1:0.6 mixture of Pseudomonas fluorescens and Pseudomonas rhodochrous |
Comparative preparation examples 1 to 5
Comparative preparation examples 1 to 5 each provide a soil restoration microbial agent.
The above comparative preparation example differs from preparation example 2 in that: the amounts of each strain added to the soil remediation microbial inoculum are shown in Table 3.
TABLE 3 addition amount of each strain in soil restoration microbial agents provided in PREPARATIVE EXAMPLE 2 and COMPARATIVE PREPARATIVE EXAMPLES 1-5
Comparative preparation example 6
Comparative preparation 6 provides a soil remediation microbial agent.
The above comparative preparation example differs from preparation example 2 in that: the components and the addition amount of the soil remediation microbial inoculum.
The soil remediation microbial inoculum provided in comparative preparation example 6 comprises the following components in addition: bacillus bailii 1.8g, bacillus amyloliquefaciens 3.0g, bacillus licheniformis 1.8g, bacillus atrophaeus 0.6g and zoogloea 1.0g.
The bacillus licheniformis is bacillus licheniformis BGB-83R, and the preservation number is CGMCC No 24182.
Comparative preparation example 7
Comparative preparation 7 provides a soil remediation microbial agent.
The above comparative preparation example differs from preparation example 2 in that: the components and the addition amount of the soil remediation microbial inoculum.
The soil remediation microbial inoculum provided in comparative preparation example 7 comprises the following components in addition: bacillus bailii 1.8g, bacillus amyloliquefaciens 3.0g, pseudomonas 1.8g, bacillus atrophaeus 0.6g and Bacillus licheniformis 1.0g.
The bacillus licheniformis is bacillus licheniformis BGB-83R, and the preservation number is CGMCC No 24182.
Antibacterial effect detection
The antibacterial effect of the soil restoration microbial agents provided in preparation examples 1 to 23 and comparative preparation examples 1 to 7 was tested. The test procedure was as follows:
(1) Fungus culture: firstly, respectively activating rhizoctonia solani, fusarium and flavum strains stored at the temperature of minus 80 ℃ by using a PDA culture medium, and carrying out plate culture for 5-7d at the temperature of 28 ℃; transferring the activated strain to PD liquid culture medium for shaking culture under the following conditions: 28 ℃,180r/min and 36h; observing that the mycelium of the culture medium is more and after the mycelium grows into spores, finishing the culture, and detecting the viable bacteria by using a dilution coating plate method; finally, the live bacteria amount of the liquid culture is diluted into bacterial suspension of 100 ten thousand cfu/ml by sterile water, so as to obtain bacterial suspension of 3 potential fungi.
(2) Preparing simulated soil: adding rhizoctonia solani suspension into soil, adding sterile water, and adjusting the bacterial concentration in the soil to 10 ten thousand cfu/g to obtain the simulated soil containing rhizoctonia solani. The simulated soil containing fusarium and the simulated soil containing flavomycetes are prepared in the same method. Detecting the bacterial content of the simulated soil by a dilution coating flat plate method, wherein the bacterial content of the simulated soil containing rhizoctonia solani is 9.83 ten thousand cfu/g; the bacteria content of the simulated soil containing fusarium is 9.76 ten thousand cfu/g; the fungus content of the simulated soil containing the flavomycetes is 10.12 ten thousand cfu/g.
(3) Fungal inhibition assay:
test group: taking 10g of the simulated soil containing rhizoctonia solani, adding 20mg of the soil restoration microbial inoculum of the example or the comparative example, uniformly mixing, placing in a constant temperature incubator at 30 ℃ for culturing for 48 hours, and setting 3 repetitions; fusarium simulated soil and yellow mold simulated soil were treated in the same manner as described above.
Blank control group: soil without soil restoration bacteria was used as a blank control group.
(4) After the culture of the experimental group and the blank control group was completed, the number of viable bacteria of 3 fungi in the soil was detected by a dilution-coating plate method, and the detection results are shown in table 4.
TABLE 4 antibacterial effect test results of soil restoration microbial agents of preparation examples 1 to 23 and comparative preparation examples 1 to 7
According to the detection results of Table 4, it is known that the soil restoration microbial inoculum prepared by using Bacillus bailii, bacillus amyloliquefaciens, pseudomonas fluorescens, bacillus atrophicus and Acinetobacter in preparation examples 1-23 of the application can reduce the content of rhizoctonia solani in simulated soil from 9.83 to 3.24 to 4.32 (less than 4.5) ten thousand cfu/g, the content of fusarium from 9.76 to 1.62 to 3.18 (less than 3.5) ten thousand cfu/g and the content of flavomycetes from 10.12 to 2.34 to 4.20 ten thousand cfu/g (less than 4.5) ten thousand cfu/g).
The soil restoration microbial inoculum prepared by adopting any four bacteria of bacillus belicus, bacillus amyloliquefaciens, pseudomonas fluorescens, bacillus atrophaeus and zoogloea in comparative preparation examples 1-5 can reduce the content of rhizoctonia solani in simulated soil from 9.83 to 5.88-7.30 ten thousand cfu/g, reduce the content of fusarium from 9.76 to 3.15-6.20 ten thousand cfu/g and reduce the content of flavomycetes from 10.12 to 3.96-5.55 ten thousand cfu/g.
Comparative preparation 6 soil remediation microbial inoculum prepared from bacillus belicus, bacillus amyloliquefaciens, bacillus licheniformis, bacillus atrophaeus and zoogloea can reduce the content of rhizoctonia solani in simulated soil from 9.83 to 6.17 ten thousand cfu/g, and the content of fusarium from 9.76 to 4.66 ten thousand cfu/g.
Comparative preparation example 7 soil remediation microbial inoculum prepared from bacillus belicus, bacillus amyloliquefaciens, pseudomonas fluorescens, bacillus atrophaeus and bacillus licheniformis can reduce the content of rhizoctonia solani in simulated soil from 9.83 to 5.29 ten thousand cfu/g, and the content of fusarium from 9.76 to 5.18 ten thousand cfu/g.
The blank control group does not have any intervention, the rhizoctonia solani content in the simulated soil is increased from 9.83 to 1834 ten thousand cfu/g, the fusarium content is increased from 9.76 to 2549 ten thousand cfu/g, and the yellow mould content is increased from 10.12 to 4587 ten thousand cfu/g.
In summary, 5 bacteria of bacillus bailii, bacillus amyloliquefaciens, pseudomonas fluorescens, bacillus atrophaeus and sericin are adopted for preparation examples 1-23, and the addition amount of each component is controlled within the following range: 1.5 to 2.3 parts of bacillus belicus, 2.7 to 3.1 parts of bacillus amyloliquefaciens, 1.0 to 2.3 parts of pseudomonas, 0.4 to 1.0 parts of bacillus atrophaeus and 0.8 to 1.2 parts of zoogloea, and the prepared soil restoration microbial inoculum has good inhibition effect on three fungi of rhizoctonia solani, fusarium and yellow mould.
As can be seen from the detection results of preparation example 2 and preparation examples 14-23, the soil remediation agent prepared by adopting single pseudomonas for preparation example 2, preparation examples 14-15 and compounding pseudomonas fluorescens and pseudomonas syringae for preparation example 21, pseudomonas syringae and pseudomonas green for preparation example 22 and compounding pseudomonas fluorescens and pseudomonas rhodochrous for preparation example 23 can reduce the content of rhizoctonia solani in simulated soil from 9.83 to 3.95-4.38 ten thousand cfu/g, the content of fusarium from 9.76 to 1.73-2.89 ten thousand cfu/g and the content of flavomycetes from 10.12 to 2.65-3.76 ten thousand cfu/g; the soil remediation microbial inoculum prepared by compounding pseudomonas fluorescens and pseudomonas aeruginosa in preparation examples 16-20 can reduce the content of rhizoctonia solani in simulated soil from 9.83 to 3.24-3.64 (less than 4.0) kilocfu/g, reduce the content of fusarium from 9.76 to 1.62-1.80 (less than 2.0) kilocfu/g and reduce the content of yellow mould from 10.12 to 2.34-2.77 (less than 3.0) kilocfu/g. Therefore, the application shows that the soil remediation microbial inoculum prepared by compounding the pseudomonas fluorescens and the pseudomonas aeruginosa has better comprehensive inhibition effect on three fungi of rhizoctonia solani, fusarium and yellow mould in soil.
Bighead atractylodes rhizome planting test
1. The test method comprises the following steps: in 2023, 3 and 25 days, selecting a test field with 2 years of planting years of bighead atractylodes rhizome in New wortmann county in Zhejiang pan, setting 9 test areas (test areas 1-9, 4m×4m each) in the test field, making labels in each test area, and applying base fertilizer (0.25 kg/m) to the test field 2 Urea + organic fertilizer 1kg/m 2 +calcium magnesium phosphate fertilizer 0.5kg/m 2 +borax 0.25kg/m 2 ). And then transplanting the presoaked bighead atractylodes rhizome seed balls into an experimental area according to row spacing of 20cm multiplied by plant spacing of 20cm multiplied by depth of 3cm, and irrigating roots of bighead atractylodes rhizome seedlings in each experimental area on 25 days of 2023, 5 days of 2023 and 5 days of 2023, and 15 days of 2023 (the treatment mode is as follows), and performing conventional agronomic management in other times without using pesticides during the experiment.
The treatment modes of the 9 test areas are respectively as follows:
test area 1: adding water to dilute 72g of the soil remediation microbial inoculum provided in preparation example 1 according to the proportion of 1:800, and irrigating to a test area 1; namely, the dosage of the soil restoration microbial inoculum is 3 kg/mu.
Test area 2: adding water to dilute 72g of the soil remediation microbial inoculum provided in preparation example 2 according to the proportion of 1:800, and irrigating to a test area 2; namely, the dosage of the soil restoration microbial inoculum is 3 kg/mu.
Test area 3: adding water to dilute 72g of the soil remediation microbial inoculum provided in preparation example 18 according to the proportion of 1:800, and irrigating to a test area 3; namely, the dosage of the soil restoration microbial inoculum is 3 kg/mu.
Test area 4: 48g of the soil remediation microbial inoculum provided in preparation example 18 is diluted by adding water according to the proportion of 1:800, and irrigated to a test area 4; namely, the dosage of the soil restoration microbial inoculum is 2 kg/mu.
Test area 5: 96g of the soil remediation microbial inoculum provided in preparation example 18 is diluted by adding water according to the proportion of 1:800, and irrigated to a test area 5; namely, the dosage of the soil restoration microbial inoculum is 4 kg/mu.
Test area 6: 120g of the soil remediation microbial inoculum provided in preparation example 18 is diluted by adding water according to the proportion of 1:800, and irrigated to a test area 6; namely, the dosage of the soil restoration microbial inoculum is 5 kg/mu.
Test area 7: adding water to dilute 72g of the soil remediation microbial inoculum provided in comparative preparation example 6 according to the proportion of 1:800, and pouring the soil remediation microbial inoculum to a test area 7; namely, the dosage of the soil restoration microbial inoculum is 3 kg/mu.
Test area 8: adding water to dilute 72g of the soil remediation microbial inoculum provided in comparative preparation example 7 according to the proportion of 1:800, and pouring the soil remediation microbial inoculum to a test area 8; namely, the dosage of the soil restoration microbial inoculum is 3 kg/mu.
Test area 9: the test area 9 is irrigated with clean water.
2. The detection method comprises the following steps: counting the emergence rate of the seedlings on the 4 th month and 15 th year of 2023, randomly selecting 10 plants in each test area on the 5 th month and 15 th year of 2023 and the 7 th month and 10 th year of 2023, and measuring the plant height, the stem thickness and the fresh weight; and (3) counting the dead seedling rate of the bighead atractylodes rhizome on 7 months and 10 days and 7 months and 20 days of 2023. The results are shown in Table 5 below.
Measuring the plant height of the bighead atractylodes rhizome seedlings by using a tape measure; and measuring the stem thickness of the bighead atractylodes rhizome seedlings by using a vernier caliper, wherein the measurement position of the stem thickness is 1cm away from the stem base. The whole bighead atractylodes rhizome seedlings are pulled out and cleaned, and the fresh weight is weighed. The calculation method of the death rate is as follows:
death rate (%) = (number of seedlings emergence on 15 days of 4 months-number of survival)/number of seedlings emergence on 15 days of 4 months
Table 5 results of detection of various indexes of Atractylodis rhizoma in Atractylodis rhizoma planting test areas 1 to 9
As can be seen from the detection results of Table 5, the emergence rate, plant height, stem thickness, fresh weight and death rate of the bighead atractylodes rhizome obtained in test areas 1 to 8 are superior to those of test area 9. The soil remediation microbial inoculum is used for irrigating white atractylodes plants, so that the flora environment of the planting soil of the white atractylodes rhizome can be improved, the autotoxicity of the white atractylodes rhizome is reduced, and the Crohn promotes the normal growth of the white atractylodes rhizome plants.
As can be seen from the detection results of the test areas 1-8, the indexes of the bighead atractylodes rhizome obtained in the test areas 1-6 are better than those of the bighead atractylodes rhizome obtained in the test areas 7-8, and the soil restoration microbial inoculum provided in the embodiments 1-2 and 18 of the application has better restoration effect on the white atractylodes planting soil. Further comparing the detection results of the test areas 1-6, the obtained bighead atractylodes rhizome in the test area 3-5 has a plant height of 48.1-49.2cm in 10 days of 7 months, a stem thickness of 7.01-7.09mm, a fresh weight of 25.13-26.26g and a death rate of 6.25-9.27% (< 10%) in 20 days of 7 months. Therefore, the application shows that the soil remediation microbial inoculum provided by the embodiment 18 is used for irrigation, the dosage of the soil remediation microbial inoculum is controlled within the range of 2-3 kg/mu, the dosage of the soil remediation microbial inoculum can be saved, meanwhile, the content of pathogenic bacteria in the soil is reduced, and the improvement effect of the bighead atractylodes rhizome planting soil is better and more remarkable.
In conclusion, the soil remediation microbial inoculum provided by the application can antagonize pathogenic bacteria in the bighead atractylodes rhizome planting soil on one hand and prevent the growth and propagation of the pathogenic bacteria, so that the flora environment of the bighead atractylodes rhizome planting soil is improved; on the other hand, the preparation method can inhibit the generation of terpenoid substances such as beta-eucalyptol in the rhizome of the bighead atractylodes rhizome, reduce the autotoxic phenomenon of the bighead atractylodes rhizome, reduce the death rate of the bighead atractylodes rhizome and finally realize the effective prevention and treatment of bighead atractylodes rhizome continuous cropping obstacle.
The foregoing examples illustrate the invention in detail, but are merely preferred embodiments of the invention and are not to be construed as limiting the scope of the invention. All equivalent changes and modifications within the scope of the present invention are intended to be covered by the present invention.
Claims (10)
1. The soil restoration microbial agent for preventing and treating the continuous cropping obstacle of the bighead atractylodes rhizome is characterized by comprising bacillus bailii, bacillus amyloliquefaciens, pseudomonas, bacillus atrophaeus and zoogloea.
2. The soil restoration microbial agent for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome according to claim 1, wherein the soil restoration microbial agent for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome comprises the following components in parts by weight: 1.5 to 2.3 parts of bacillus belicus, 2.7 to 3.1 parts of bacillus amyloliquefaciens, 1.0 to 2.3 parts of pseudomonas, 0.4 to 1.0 part of bacillus atrophaeus and 0.8 to 1.2 parts of zoogloea.
3. The soil restoration microbial agent for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome according to claim 1, wherein the soil restoration microbial agent for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome comprises the following components in parts by weight: 1.5 to 2.3 parts of bacillus bailii, 2.7 to 3.1 parts of bacillus amyloliquefaciens, 1.5 to 2.0 parts of pseudomonas, 0.4 to 1.0 part of bacillus atrophaeus and 0.8 to 1.2 parts of zoogloea.
4. The soil restoration microbial agent for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome according to claim 1, wherein the soil restoration microbial agent for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome comprises the following components in parts by weight: 1.8 parts of bacillus belicus, 3.0 parts of bacillus amyloliquefaciens, 1.8 parts of pseudomonas, 0.6 part of bacillus atrophaeus and 1.0 part of zoogloea.
5. The soil restoration microbial agent for preventing and treating continuous cropping obstacle of white atractylodes according to any one of claims 1 to 4, wherein the pseudomonas is selected from pseudomonas aeruginosa, pseudomonas fluorescens, pseudomonas syringae and pseudomonas rhodochrous.
6. The soil restoration microbial agent for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome according to claim 5, wherein the pseudomonas is characterized by having a weight ratio of 1: (0.2-1) Pseudomonas fluorescens and Pseudomonas aeruginosa.
7. The soil restoration microbial agent for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome according to claim 1, wherein the effective viable count of bacillus belicus is more than or equal to 20 hundred million cfu/g, the effective viable count of bacillus amyloliquefaciens is more than or equal to 20 hundred million cfu/g, the effective viable count of pseudomonas is more than or equal to 20 hundred million cfu/g, the effective viable count of bacillus atrophaeus is more than or equal to 20 hundred million cfu/g, and the effective viable count of zoogloea is more than or equal to 20 hundred million cfu/g.
8. The method for preparing a soil restoration microbial agent for preventing and treating continuous cropping obstacle of white atractylodes according to any one of claims 1 to 7, comprising the following steps: and uniformly mixing all the strains to obtain the soil restoration microbial inoculum for preventing and treating continuous cropping obstacle of the bighead atractylodes rhizome.
9. A method for preventing and treating bighead atractylodes rhizome continuous cropping obstacle, which is characterized by comprising the following steps: the soil restoration microbial agent for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome according to any one of claims 1 to 7 is mixed with water and then applied to bighead atractylodes rhizome planting soil.
10. The method for preventing and treating continuous cropping obstacle of bighead atractylodes rhizome according to claim 9, wherein the dosage of the soil restoration microbial inoculum is 2-4 kg/mu.
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