CN116200311A - Basil azospirillum, composite microbial inoculum, and preparation method and application thereof - Google Patents
Basil azospirillum, composite microbial inoculum, and preparation method and application thereof Download PDFInfo
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- CN116200311A CN116200311A CN202310278862.XA CN202310278862A CN116200311A CN 116200311 A CN116200311 A CN 116200311A CN 202310278862 A CN202310278862 A CN 202310278862A CN 116200311 A CN116200311 A CN 116200311A
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- agent
- composite microbial
- powder
- microbial inoculum
- azospirillum brasilense
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/05—Fruit crops, e.g. strawberries, tomatoes or cucumbers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The invention provides azospirillum brasilense (Azospirillum brasilense), a composite microbial inoculum, a preparation method and application thereof. The strain preservation number of the azospirillum brasilense is CGMCC 26716. The composite microbial inoculum comprises an agent A, an agent B and an agent C; the agent A comprises one or more than two of phosphorite powder, potassium mineral powder, iron silicate mineral powder and straw powder; the agent B comprises azospirillum brasilense; the agent C comprises algal polysaccharide and molasses. The azoospiram barcetii disclosed by the invention can promote the growth of tomato seedlings; the prepared composite microbial inoculum is inoculated in a tomato seedling raising stage, can obviously promote the growth and establishment of a tomato seedling root system, make the root system more developed, thicken stems, enlarge leaves and increase chlorophyll, enhance the resistance to soil-borne diseases, enhance the resistance to secondary salinization, and obviously improve the yield and quality of tomato fruits.
Description
Technical Field
The invention belongs to the technical fields of microbial resource utilization and ecological agriculture, and relates to azospirillum brasilense (Azospirillum brasilense), a composite microbial agent, a preparation method and application thereof; in particular to azospirillum brasilense and compound bacteria for promoting the stress-resistant growth of plants in a tomato seedling stage, and a preparation method and application thereof, and is suitable for the tomato seedling stage.
Background
With the increase of consumption, the planting area of tomatoes in the world is gradually increased, and China is the country with the widest planting area of tomatoes in the world, and particularly the planting area and scale of tomatoes in a greenhouse are increasingly enlarged, so that the tomatoes in the greenhouse become one of important agricultural products in many areas. However, long-term continuous cropping causes serious continuous cropping obstacles, which results in accumulation of soil-borne disease microorganisms and soil salinization. These can result in low tomato yields and limit the associated production. In recent years, as the planting scale of tomatoes in a greenhouse is enlarged, a large amount of pesticides have to be used in the planting process to prevent frequent occurrence of diseases and insect pests, so that the quality of the tomatoes is seriously influenced, and the tomatoes have great pressure on the ecological environment and do not accord with the sustainable agriculture development trend.
Improving plant nutrition supply, especially increasing potassium and silicate level, and contributing to drought resistance and salt stress resistance of plants. And microorganisms can promote the growth, yield and quality improvement of plants under stress through a series of functions and actions: various plant hormones such as auxin are produced to promote the development and establishment of plant root systems and increase the nutrition and moisture absorption; can also stimulate plants to synthesize soluble sugar and proline, and improve activity of antioxidant enzyme such as SOD to reduce adverse stress such as drought; improving photosynthetic performance and enhancing plant productivity; ACC deaminase production to inhibit premature senility or premature apoptosis of plants; inhibiting occurrence and development of diseases; etc. Nitrogen, phosphorus and potassium are macronutrients required for plant growth and microorganisms can increase supply by phosphate and potassium dissolving and nitrogen fixation functions and actions, especially for the tomato seedling stage. Stress such as drought has the greatest influence on plant emergence and seedling growth, and seeding at a seedling stage is an important technology for promoting plant seed germination and seedling growth.
At present, most of domestic related patent applications related to microbial agents relate to microbial agents of chemical substances and microorganisms which are applied at the later stage for growth promotion and disease prevention, and a small number of related patent applications relate to the intervention of the characteristics of the microorganisms in the seedling stage. For example, a patent application entitled "a microbial compound fertilizer for controlling tomato blight and a preparation method thereof" (application number "201810318677.8") describes mixing chemical fertilizers such as urea with microorganisms and trace elements to control tomato blight; the patent application entitled "a tomato seedling substrate and method of use thereof" (application number "201910798120.3") describes mixing distillers' grains and rotten chicken manure in a certain ratio as a seedling substrate; the patent application entitled "preparation method of excellent tomato seedling substrate" (application number is "2018101239761. X") describes that milk cow dung, mushroom dregs, decomposed microbial agents and wormcast are mixed in different proportions, so that the excellent tomato seedling substrate with strong fertilizer and water retention capacity and reduced plant diseases and insect pests can be obtained; the patent application named as ' a pseudomonas fluorescens strain, a microbial inoculum and application of the pseudomonas fluorescens strain and the microbial inoculum as a seedling raising matrix for preventing and treating tomato bacterial wilt ' (application number is 201010109162.0 ') shows that the occurrence of the tomato bacterial wilt can be effectively solved by directly combining mushroom residues after the cultivation of decomposed edible fungi with the pseudomonas fluorescens microbial inoculum; the patent application entitled "a seedling substrate and preparation method and application thereof" (application number "20110420456. X") describes a seedling substrate obtained by mixing cotton stalk and cow dung after respective fermentation with weathered coal and vermiculite, and the cultivated tomato seedling is superior to the conventional seedling substrate in the aspects of strong seedling index, root system activity, chlorophyll content of tomato leaves, yield after field planting and disease resistance; etc.
However, no effective single strain has been reported in promoting stress-tolerant growth of plants at the tomato seedling stage.
Disclosure of Invention
Based on the drawbacks of the prior art, an object of the present invention is to provide a azospirillum brasilense (Azospirillum brasilense); the second aim of the invention is to provide the application of the azospirillum brasilense in promoting the stress-resistant growth of plants in the tomato seedling stage; the third purpose of the invention is to provide a composite microbial inoculant containing the azoospiram brasiliensis, which can promote the stress-resistant growth of tomato seedlings after being directed against plants in the tomato seedling raising stage; the fourth object of the invention is to provide a preparation method of the composite microbial inoculum; the fifth aim of the invention is to provide the application of the composite microbial inoculum in promoting the stress-resistant growth of plants in the tomato seedling stage; the sixth object of the present invention is to provide a method for promoting seedling growth in tomato seedling raising period by using the composite microbial inoculum.
The aim of the invention is achieved by the following technical scheme:
in one aspect, the invention provides a azospirillum bassinense (Azospirillum brasilense), namely: the azoospiram bassinense JN25 (Azospirillum brasilense JN), and the bacterial preservation number of the azoospiram bassinense is CGMCC 26716; the preservation date is: 2023.2.28; the preservation units are as follows: china general microbiological culture Collection center (CGMCC); the preservation unit addresses are: post code of the institute of microbiology of the national academy of sciences of Beijing, china: 100101; taxonomies were named: azospirillum brasilense (Azospirillum brasilense).
The inventor collects the roots of field tomato seedlings and uses a coating and scribing technology to separate the functions of plant-promoting strains such as nitrogen fixation, phosphorus and potassium dissolution of rhizosphere, plant hormone production, siderophore production, ACC deaminase production and the like; the strain capable of promoting the cut-block stress-resistant growth of potato tuber planting is obtained through separation and purification, and is identified as azospirillum brasilense (Azospirillum brasilense) through species identification, and the strain is preserved.
The Bacillusazaspira JN25 (Azospirillum brasilense JN) has the functions of producing plant hormone auxin and gibberellin, decomposing calcium phosphate-phosphorus (phosphate rock powder-phosphorus), decomposing potassium (potassium mineral powder-potassium), autogenously fixing nitrogen, producing ACC (p-2, 4-diacetyl gamboge phenol) deaminase, producing siderophores, producing HCN (hydrocyanic acid) or producing DAPG (p-2, 4-diacetyl gamboge phenol) and the like; can obviously promote the growth and establishment of the root system of the tomato seedling, the root system is more developed, the stems are thickened, the leaves are enlarged, the chlorophyll is increased, the resistance to soil-borne diseases is enhanced, the resistance to drought, plant diseases and insect pests and salt stress is enhanced, the growth is promoted, and the yield and quality of the tomato are obviously improved.
On the other hand, the invention also provides application of the azospirillum brasilense in promoting the stress-resistant growth of plants in the tomato seedling stage.
In still another aspect, the present invention further provides a composite microbial agent, wherein the composite microbial agent includes an agent a, an agent B, and an agent C;
the agent A comprises one or more than two of phosphorite powder, potassium mineral powder, iron silicate mineral powder and straw powder;
the agent B comprises the azospirillum brasilense;
the agent C comprises algal polysaccharide and molasses.
The inventor researches find that the composite microbial inoculum can obviously promote the establishment of a tomato seedling root system, enhance the resistance to drought and diseases, promote growth and improve the yield and quality of tomatoes after being used for inoculating plants in a tomato seedling stage.
In the composite microbial agent, the weight ratio of the agent A to the agent B to the agent C is preferably (100-200 g): (5-25) g: (100-300) g; in the agent B, the effective viable count concentration of the azospirillum brasilense is 10 9 ～10 11 CFU/g or 10 9 ～10 11 CFU/ml。
In the composite microbial inoculum, preferably, the agent A consists of ground phosphate rock, potassium mineral powder, iron silicate mineral powder and straw powder;
wherein the weight ratio of the phosphate rock powder to the potassium mineral powder to the iron silicate mineral powder to the straw powder is (40-60): (40-50): (1-5): (0.01-0.05).
In the composite microbial inoculum, preferably, the weight ratio of the potassium mineral powder to the iron silicate mineral powder to the straw powder is 50:40:1:0.03.
In the composite microbial inoculum A, the phosphate rock powder, the potassium mineral powder, the iron silicate mineral powder and the like are all obtained in the market and are mixed according to a specific proportion, so that nutrition can be increased, and nutrition supply in the germination period can be improved; the straw powder is crushed from waste plants of crops such as corn and the like, has the granularity of 0.05-0.2 mm, can form a film with good ventilation together with polysaccharide and other materials produced by azospirillum brasilense, and is subjected to sterilization treatment in modes such as high-temperature steaming or ammonia fumigation before compounding.
In the above composite microbial agent, preferably, the agent B includes a liquid microbial agent or a solid powdery microbial agent.
In the above-mentioned composite microbial inoculum, preferably, the liquid microbial inoculum includes a protective agent and azospirillum brasilense; the effective viable count concentration of the azoospira brasiliensis in the liquid microbial inoculum is 10 9 ～10 11 CFU/mL。
In the composite microbial agent, the protective agent preferably comprises glycerol and/or K with pH of 7.2 2 HPO 4 -KH 2 PO 4 And (3) a buffer solution.
In the above composite microbial inoculum, preferably, the solid powdery microbial inoculum comprises azoospermia bassinensis and an additive for maintaining bacterial survival (which is a conventional additive in the field), and the effective viable count concentration of the azoospermia bassinensis in the solid powdery microbial inoculum is 10 9 ～10 11 CFU/g。
In the above-mentioned composite microbial agent, preferably, in the agent C, the algal polysaccharide is mixed with the molasses in equal mass.
In the composite microbial inoculant C, algal polysaccharide and molasses can be used as adhesives, and meanwhile, the molasses contains a large amount of carbon sources, vitamins and other large nutrients.
In still another aspect, the present invention also provides a method for preparing a composite microbial inoculant, which includes:
performing strain activation, shake flask culture and fermentation tank amplification culture on the azoospiram brazilian by using a modified R2A culture medium, and centrifugally collecting cells; freeze drying and adding additive to obtain solid powdery microbial agent, or directly adding liquid protective agent (such as glycerol, buffer solution, etc.) to obtain liquid microbial agent, to obtain the above agent B;
wherein the modified R2A medium has the formula: glucose 1.3g, bacteriological peptone 1.3g, acid casein 1.3g, yeast extract powder 1.3g, K 2 HPO 3 ·3H 2 O0.8 g, sodium pyruvate 0.8g, mgSO 4 ·7H 2 O0.15g, distilled water 1000mL; agar solid medium is additionally added with 15g of agar;
the additive is an additive for maintaining the survival of strains;
preparing the agent A and the agent C respectively according to the formulas of the agent A and the agent C; optionally, the composition may be used in combination with,
and packaging the agent A, the agent B and the agent C independently.
In the composite microbial agent, the agent A, the agent B and the agent C can be respectively and independently packaged, and when in use, water is added according to a proportion to mix and prepare, and the independent packaging is more beneficial to ensuring the survival of microorganisms, prolonging the preservation time and ensuring the coating effect. In addition, the material components and the proportion of the composite microbial inoculum have outstanding effects particularly for emergence of seedlings in tomato seedling stage, but can also be expanded to application for other crops.
In the preparation method, in the process of preparing the agent A, high-temperature steaming or ammonia fumigation sterilization is required for straw powder, and then the straw powder is mixed with other mineral powder according to a proportion.
In the above preparation method, preferably, the composite microbial inoculum further includes the following preparation steps:
adding the agent B into the agent C, adding sterile water (sterile water subjected to filtration sterilization or high-temperature sterilization), and uniformly stirring to obtain a first mixed solution;
and then adding the agent A into the first mixed solution, and uniformly mixing to obtain the liquid composite microbial inoculum.
In still another aspect, the invention also provides application of the composite microbial inoculum in promoting stress-resistant growth of plants in a tomato seedling stage.
In still another aspect, the present invention also provides a method for inoculating and planting a plant microbial inoculum at a tomato seedling stage, comprising:
uniformly stirring the agent B and the agent C by using sterile water to obtain a first mixed solution; then adding the agent A into the first mixed solution to obtain a liquid solution of the composite microbial inoculum to be used;
wherein, in the liquid solution of the composite microbial inoculum, the dosage ratio of the agent A to the sterile water is (200-400) g: (4-6) L; the effective viable count concentration of the azoospiram brasiliensis in the composite microbial inoculant liquid solution is 10 6 ～10 8 CFU/mL；
Uniformly spraying the liquid solution of the composite microbial inoculum into a tomato seedling pot, and inoculating once every 10 days;
optionally, an equal volume of clear water is used for sprinkling irrigation every morning;
wherein, the dosage ratio of the tomato seedling stage plants to the liquid solution of the composite microbial inoculum is (50-100) g: (1-5) L.
In the above planting method, preferably, the dosage ratio of the agent a to the sterile water is 200g:5L.
In the above planting method, preferably, the ratio of the plant seeds at the tomato seedling stage to the liquid solution of the composite microbial inoculum is 80g:4L.
In the planting method, preferably, 8L of the composite microbial inoculum is inoculated each time, and the bacterial load is ensured to reach the standard.
The invention has the beneficial effects that:
the azoospiram barcetii disclosed by the invention can promote the growth of seedling plants at the seedling raising stage of tomatoes; the prepared composite microbial inoculum is inoculated to the root system of a seedling plant in a tomato seedling raising stage, can obviously promote the growth and establishment of the root system of the tomato seedling, has more developed root system, thickens stems, enlarges leaves and increases chlorophyll, enhances the resistance to soil-borne diseases, enhances the resistance to drought and salt stress, and obviously improves the yield and quality of tomatoes.
Drawings
FIG. 1 shows tomato seedlings (seedlings inoculated with agent A, agent A+B and the complex microbial agent for 20 days in the order from left to right in FIG. 1A; seedlings inoculated with the complex microbial agent and agent A+B for 35 days and root systems thereof in the order from left to right in FIGS. 1B and 1C) after different treatments according to the embodiment 5 of the present invention.
FIG. 2 shows a composite microbial inoculum (A in FIG. 2 is a dry powder mixture of A and B in FIG. 2 is a liquid B agent containing azospirillum brasilense of the invention, and C in FIG. 2 is a liquid mixture of C agents).
Culture preservation for patent procedures:
the azospirillum bazedox JN25 (Azospirillum brasilense JN 25) of the invention:
preservation date: 2023.2.28
Preservation unit: china general microbiological culture Collection center (CGMCC);
deposit unit address: post code of the institute of microbiology of the national academy of sciences of Beijing, china: 100101;
preservation number: CGMCC 26716
Classification naming: azospirillum brasilense (Azospirillum brasilense).
Detailed Description
The technical solution of the present invention will be described in detail below for a clearer understanding of technical features, objects and advantageous effects of the present invention, but should not be construed as limiting the scope of the present invention.
The formulation in the following examples of the invention was:
the medium formulations used in the examples below were:
1. modified R2A medium: 1.3 to 1.5g of glucose, 1.3 to 1.5g of bacteriological peptone, 1.3 to 1.5g of acid casein, 1.3 to 1.5g of yeast extract powder and K 2 HPO 3 ·3H 2 0.8 to 1.0g of O and 0.8 to 1g of sodium pyruvate.0g，MgSO 4 ·7H 2 0.15-0.20 g of O and 1000-1500 mL of distilled water; adding 15-20 g of agar into the agar solid culture medium; .
2. Nitrogen fixation medium: KH (KH) 2 PO 4 0.2g，CaCO 3 5g，MgSO 4 ·7H 2 O0.2 g, glucose 10g, naCl1.2g, caSO 4 ·2H 2 O0.1 g, agar 20g, distilled water 1000mL, and the pH is natural.
3. Inorganic phosphorus medium: glucose 10g, (NH) 4 ) 2 SO 4 0.5g，NaCl 0.3g，KCl 0.3g，MgSO 4 ·7H 2 O0.3g，Ca 3 (PO 4 ) 2 25g，FeSO 4 ·7H 2 O 0.03g，MnSO 4 ·4H 2 O0.03 g, agar 20g, distilled water 1000mL, and the pH is natural.
4. Organophosphorus medium: glucose 10g, (NH) 4 ) 2 SO 4 0.5g，NaCl 0.3g，KCl 0.3g，MgSO 4 ·7H 2 0.3g of O, 25g of calcium phytate and FeSO 4 ·7H 2 O 0.03g，MnSO 4 ·4H 2 O0.03 g, agar 20g, distilled water 1000mL, and the pH is natural.
5. Potassium-dissolving culture medium: 5.0g of sucroses, NH 4 NO 3 1.7g，Na 2 HPO 4 2.0g，CaCO 3 0.1g，MgSO 4 0.5g，FeCl 3 0.005g of potassium feldspar 1.0g; distilled water 1000mL.
6. LB medium: 10g of tryptone, 5g of yeast extract, 10g of NaCl, 15g of agar, 1000mL of deionized water and pH 7.4.
7. DF medium: KH (KH) 2 PO 4 4g，Na 2 HPO 4 6g，MgSO 4 ·7H 2 O0.2 g, glucose 2g, gluconic acid 1.62mL, citric acid 2g, (NH) 4 ) 2 SO 4 2g, 1000mL of deionized water, pH 7.2.
8. ADF medium: (NH) in DF Medium 4 ) 2 SO 4 The ADF medium is obtained after removal.
Example 1: screening, separating and identifying strain
The root systems of field tomato seedlings are collected, the functions of the strains for fixing nitrogen, dissolving phosphorus, dissolving potassium, producing plant hormone, producing siderophore, producing ACC deaminase and other plant growth promoting characters of rhizosphere are separated by a coating and scribing technology, qualitative or quantitative determination of plant growth promoting capacity is carried out by using culture mediums for fixing nitrogen, dissolving phosphorus, dissolving potassium and the like, and 5 bacteria shown in table 1 are obtained through screening.
Table 1:
according to the experimental results of Table 1, the strain JN25 has various growth promoting abilities and is suitable for climates or geological conditions (large temperature difference, secondary salinization and the like) in northern areas, so that the strain is taken as an excellent strain obtained by screening.
The genomic DNA of the strain obtained by separation and purification was used as a template, the 16S rRNA gene of the strain was PCR amplified using 27F and 1492R as primers, and the amplified product was sequenced, and the obtained sequence was classified on line by NCBI (preliminary determination that the sequence similarity of the 16S rRNA gene was greater than 97% was the same species), and the results are shown in Table 2.
Table 2:
| strain numbering | The nearest species | Similarity (%) |
| JN25 | Azospirillum brasilense | 100 |
The results in Table 2 show that the similarity between the 16S rRNA gene sequence of the excellent strain JN25 obtained by screening and the model strain Azospirillum brasilense is 100%.
Example 2: bacteria activation, culture and collection
100 mu L of glycerol-retaining liquid (Bacillusazaspira JN25 of example 1) is sucked and coated on a modified R2A culture medium, and then the culture is carried out at 25 ℃, and a single colony with typical colony characteristics and rapid growth is picked and placed on the modified R2A corresponding liquid culture medium and then is carried out at 25 ℃; centrifuging at 8000g centrifugal force during late logarithmic phase, and culturing at K 2 HPO 4 -KH 2 PO 4 After repeated washing 3 times with buffer (pH 7.2), the cells were resuspended to a cell concentration of 10 with the same buffer 9 ～10 11 CFU/mL (as in FIG. 2B), agent B was obtained.
Example 3: optimization of abiotic components
Preparing a composite microbial inoculum liquid solution:
step one, adding 5g of the agent B in the embodiment 2 into the agent C (the agent C consists of algal polysaccharide, molasses and the like, and the mass concentration of the agent C in the liquid solution of the composite microbial agent is shown in the following table 3), and then adding 5L of sterile water to uniformly stir to obtain a first mixed solution; then adding 200g of the agent A into the first mixed solution to obtain a liquid solution of the composite microbial inoculum to be used;
wherein the agent A consists of phosphorite powder, potassium mineral powder, iron silicate mineral powder and straw powder; the effective viable count concentration of the azoospira brasiliensis in the composite microbial inoculant liquid solution is 10 6 ～10 8 CFU/mL。
Uniformly spraying the prepared composite microbial inoculum liquid solution into a tomato seedling pot, and inoculating once every 10 days; in addition, an equal volume of clear water was used for sprinkling every morning;
wherein the dosage ratio of the tomato seeds to the liquid solution of the composite microbial inoculum is 80g:4L.
Tomato seedlings under secondary salinization stress are simulated by pH=7.0, and on the basis of single factor study, the non-biological coating component proportion is comprehensively selected and optimized according to the component content of the agent A (shown in the following table 3), the concentration of the agent C, the emergence rate of tomato seedlings and the seedling state at 35 days, and the result is shown in the table 3.
Table 3:
note that: the ratio of non-biological components:
the mass ratio of the raw materials of the agent A is expressed as follows: phosphate rock powder: potassium mineral powder: iron silicate mineral powder: straw powder.
The mass concentration of the C agent is as follows: for example, 0.40% (about 20 g) represents a mass concentration of both seaweeds polysaccharide and molasses of 0.20% in agent C.
As can be seen from table 3: the combination 1 is excellent in coating effect and overall effect in promoting growth.
Example 4: effective bacterial count proportion optimization determination
Tomato seedlings were simulated at ph=7.0 under secondary salinization stress, fixing dose a: the weight ratio of the phosphorite powder to the potassium mineral powder to the iron silicate powder to the straw powder is 50:40:1:0.03, and the effective bacterial count proportion is comprehensively selected and optimized by the total viable count of the azoospermi brasiliensis JN25, the tomato seedling emergence rate and the strong seedling index at 30 days, and the result is shown in Table 4.
Table 4:
note that:
agent A: the weight ratio of the phosphorite powder to the potassium mineral powder to the iron silicate powder to the straw powder is 50:40:1:0.03;
and (2) a B agent: the effective viable count concentration of the azospirillum brasilense in the agent B is 10 9 ～10 11 CFU/mL；
And (C) agent: the mass concentration of the seaweed polysaccharide and the molasses in the liquid solution of the composite microbial inoculant is as follows: 0.4%;
effective viable count concentration range (CFU/mL) in the composite microbial inoculant liquid solution: the amount of bacteria when adding the agent B is maintained at 10 9 ～10 11 The effective viable count concentration in the liquid solution of the composite microbial agent prepared by adding the agent B in the CFU/mL range is shown in Table 4.
As can be seen from table 4: the combination 1 is excellent in coating effect and overall effect in promoting growth.
Example 5: comparative experiment of microbial agent on tomato seedling stage under secondary salinization stress
Preparing a composite microbial inoculum liquid solution:
10g of the agent B in the example 2 is added into 100g of the agent C (the agent C consists of algal polysaccharide, molasses and the like), and 5L of sterile water is added and stirred uniformly to obtain a first mixed solution; then adding 200g of the agent A into the first mixed solution to obtain a liquid solution of the composite microbial inoculum to be used;
wherein the agent A consists of phosphorite powder, potassium mineral powder, iron silicate mineral powder and straw powder; the effective viable count concentration of the azoospira brasiliensis in the composite microbial inoculant liquid solution is 10 6 ～10 8 CFU/mL。
Inoculating and raising seedlings: uniformly spraying the prepared composite microbial inoculum liquid solution into a tomato seedling pot, and inoculating once every 10 days; in addition, an equal volume of clear water was used for sprinkling every morning;
wherein the dosage ratio of the tomato seeds to the liquid solution of the composite microbial inoculum is 80g:4L.
Tomato seedlings were simulated under secondary salinization stress at ph=7.0 and the tomato seedling growth index was determined and the results are shown in table 5 and fig. 1.
Table 5:
note that:
JN25: the composite microbial inoculum liquid solution of the embodiment;
c: only strain JN25 was replaced with Azospirillum brasilense (ATCC 49958) strain, other procedures remained consistent with JN25 group;
CK: control group without microbial agent treatment.
From the experimental data in table 5, it can be seen that: the composite microbial inoculum prepared from the excellent strain Bacillusazotobacter JN25 preserved by the invention has better capability of promoting the growth and development of potatoes than the Bacillusazotobacter preserved by the same collection center.
As can be seen from fig. 1: compared with the tomato seedlings which are not sprayed, the tomato seedlings sprayed with the composite microbial inoculum of the embodiment have good growth vigor, thick and strong stems and obviously larger root system volume.
Claims (10)
1. A azoospiram bassinense (Azospirillum brasilense) has a strain preservation number of CGMCC 26716.
2. A composite microbial agent, wherein the composite microbial agent comprises an agent A, an agent B and an agent C;
the agent A comprises one or more than two of phosphorite powder, potassium mineral powder, iron silicate mineral powder and straw powder;
the agent B comprises azospirillum brasilense of claim 1;
the agent C comprises algal polysaccharide and molasses.
3. The composite microbial inoculant according to claim 2, wherein the weight ratio of the agent A to the agent B to the agent C is (100-200 g): (5-25) g: (100-300) g; in the agent B, the effective viable count concentration of the azospirillum brasilense is 10 9 ～10 11 CFU/g or 10 9 ～10 11 CFU/mL。
4. A composite microbial agent according to claim 2 or 3, wherein the agent a consists of ground phosphate rock, potassium ore powder, iron silicate ore powder and straw powder;
wherein the weight ratio of the phosphate rock powder to the potassium mineral powder to the iron silicate mineral powder to the straw powder is (40-60): (40-50): (1-5): (0.01-0.05);
preferably, the weight ratio of the phosphate rock powder to the potassium mineral powder to the iron silicate mineral powder to the straw powder is 50:40:1:0.03.
5. a composite microbial agent according to claim 2 or 3, wherein the agent B comprises a liquid microbial agent or a solid powdered microbial agent.
6. The composite microbial inoculant of claim 5, wherein the liquid microbial inoculant comprises a protectant and azospirillum brasilense; the effective viable count concentration of the azoospira brasiliensis in the liquid microbial inoculum is 10 9 ～10 11 CFU/mL；
Preferably, the protective agent comprises glycerol and/or K at pH7.2 2 HPO 4 -KH 2 PO 4 A buffer;
wherein the solid powdery microbial agent comprises azospirillum brasilense and an additive for maintaining the survival of strains, and the effective viable count concentration of the azospirillum brasilense in the solid powdery microbial agent is 10 9 ～10 11 CFU/g。
7. A composite microbial agent according to claim 2 or 3, wherein in the agent C, the algal polysaccharide is mixed with the molasses in equal mass.
8. A method for preparing a composite microbial inoculant, which comprises the following steps:
performing strain activation, shake flask culture and fermentation tank amplification culture on the azoospiram barceti according to claim 1 by using a modified R2A culture medium, and centrifugally collecting cells; freeze drying and adding additives to prepare solid powdery microbial agent, or directly adding liquid protective agent to prepare liquid microbial agent, namely preparing the agent B in any one of claims 2-7;
wherein the modified R2A medium has the formula: 1.3 to 1.5g of glucose, 1.3 to 1.5g of bacteriological peptone, 1.3 to 1.5g of acid casein, 1.3 to 1.5g of yeast extract powder and K 2 HPO 3 ·3H 2 0.8 to 1.0g of O, 0.8 to 1.0g of sodium pyruvate and MgSO 4 ·7H 2 O 0.15～0.20g, distilled water 1000-1500 mL; adding 15-20 g of agar into the agar solid culture medium;
the additive is an additive for maintaining the survival of strains;
the preparation of agent A and agent C according to the formulation of agent A and agent C in any one of claims 2-7, respectively.
9. Use of the azoospiram brasiliensis of claim 1 or the composite microbial inoculant of any one of claims 2-7 to promote stress-tolerant growth of plants in a tomato seedling stage.
10. A method for cultivating plants in a tomato seedling stage, comprising:
uniformly stirring the agent B and the agent C in any one of claims 2-7 with sterile water to obtain a first mixed solution; then adding the agent A into the first mixed solution to obtain a liquid solution of the composite microbial inoculum to be used;
wherein, in the liquid solution of the composite microbial inoculum, the dosage ratio of the agent A to the sterile water is (200-400) g: (4-6) L; the effective viable count concentration of the azoospiram brasiliensis in the composite microbial inoculant liquid solution is 10 6 ～10 8 CFU/mL；
Uniformly spraying the liquid solution of the composite microbial inoculum into a tomato seedling pot, and inoculating once every 10 days;
optionally, sprinkling water daily in the morning;
wherein, the dosage ratio of the plant seeds at the tomato seedling stage to the liquid solution of the composite microbial inoculum is (50-100) g: (1-5) L.
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| CN117736944A (en) * | 2024-02-20 | 2024-03-22 | 云南省农业科学院农业环境资源研究所 | Streptomyces griseus as well as microbial inoculum and application thereof |
| CN117736944B (en) * | 2024-02-20 | 2024-04-26 | 云南省农业科学院农业环境资源研究所 | Streptomyces griseus as well as microbial inoculum and application thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2941588A1 (en) * | 2009-02-03 | 2010-08-06 | Pierre Philippe Claude | AZOTOBACTERIAL BIOFERTILIZATION COMPRISING MOLYBDENUM AND TRYPTOPHAN |
| WO2011154963A1 (en) * | 2010-06-09 | 2011-12-15 | Patel, Babubhai C. | Advance material and method of preparation of bacterial formulation using phosphorus solubilizing bacteria that makes phosphorous available to plant which are unavailable due to higher soil ph |
| US20170020138A1 (en) * | 2013-06-26 | 2017-01-26 | Symbiota, Inc. | Agricultural endophyte-plant compositions, and methods of use |
| ES2789973A1 (en) * | 2019-04-26 | 2020-10-26 | Probelte S A U | LIQUID BIOFERTILIZER THAT INCLUDES AZOSPIRILLUM BRASILENSE STRAINS AND DISPERSE PANTOEA AND A METHOD OF OBTAINING THE SAME (Machine-translation by Google Translate, not legally binding) |
| CN112175840A (en) * | 2020-05-26 | 2021-01-05 | 兰州大学 | Ustilago esculenta and application thereof in promoting plant growth in normal and stress environments |
| CN116716225A (en) * | 2023-06-29 | 2023-09-08 | 内蒙古农业大学 | Radiation-resistant streptomycete and application thereof |
| US20240049727A1 (en) * | 2014-08-25 | 2024-02-15 | Christelle Martinez-Barbreau | Biostimulant and method for stimulating plant growth |
-
2023
- 2023-03-21 CN CN202310278862.XA patent/CN116200311A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2941588A1 (en) * | 2009-02-03 | 2010-08-06 | Pierre Philippe Claude | AZOTOBACTERIAL BIOFERTILIZATION COMPRISING MOLYBDENUM AND TRYPTOPHAN |
| WO2011154963A1 (en) * | 2010-06-09 | 2011-12-15 | Patel, Babubhai C. | Advance material and method of preparation of bacterial formulation using phosphorus solubilizing bacteria that makes phosphorous available to plant which are unavailable due to higher soil ph |
| US20170020138A1 (en) * | 2013-06-26 | 2017-01-26 | Symbiota, Inc. | Agricultural endophyte-plant compositions, and methods of use |
| US20240049727A1 (en) * | 2014-08-25 | 2024-02-15 | Christelle Martinez-Barbreau | Biostimulant and method for stimulating plant growth |
| ES2789973A1 (en) * | 2019-04-26 | 2020-10-26 | Probelte S A U | LIQUID BIOFERTILIZER THAT INCLUDES AZOSPIRILLUM BRASILENSE STRAINS AND DISPERSE PANTOEA AND A METHOD OF OBTAINING THE SAME (Machine-translation by Google Translate, not legally binding) |
| CN112175840A (en) * | 2020-05-26 | 2021-01-05 | 兰州大学 | Ustilago esculenta and application thereof in promoting plant growth in normal and stress environments |
| CN116716225A (en) * | 2023-06-29 | 2023-09-08 | 内蒙古农业大学 | Radiation-resistant streptomycete and application thereof |
Non-Patent Citations (7)
| Title |
|---|
| J. M. ELIAS等: "Role of ethylene and related gene expression in the interaction between strawberry plants and the plant growth-promoting bacterium Azospirillum brasilense", 《PLANT BIOLOGY》, 31 January 2018 (2018-01-31) * |
| LORENZO VETTORI等: "Improving micropropagation: effect of Azospirillum brasilense Sp245 on acclimatization of rootstocks of fruit tree", 《JOURNAL OF PLANT INTERACTIONS》, 15 September 2010 (2010-09-15) * |
| 屈璐璐;王俊杰;: "白三叶抗逆性研究进展", 中国草地学报, no. 02, 25 March 2020 (2020-03-25) * |
| 张晓军;赵宇龙;金一荻;冯福应;: "乌梁素海小口湖区沉积物细菌多样性及其系统发育分析", 内蒙古农业大学学报(自然科学版), no. 04, 15 October 2011 (2011-10-15) * |
| 檀国印;杨志玲;袁志林;杨旭;: "植物根际促生菌及其在克服连作障碍中的潜力", 热带作物学报, no. 01, 25 January 2013 (2013-01-25) * |
| 胡春锦;史国英;曾泉;岑贞陆;宋娟;杨丽涛;李杨瑞;: "两种不同土壤条件下可培养甘蔗内生固氮菌的多样性", 中国土壤与肥料, no. 02, 10 April 2017 (2017-04-10) * |
| 郑红丽;李少红;樊明寿;乌兰巴特尔;王曙光;: "四种牧草根际固氮细菌的分离及固氮活性的研究", 干旱区资源与环境, no. 04, 30 July 2007 (2007-07-30) * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117736944A (en) * | 2024-02-20 | 2024-03-22 | 云南省农业科学院农业环境资源研究所 | Streptomyces griseus as well as microbial inoculum and application thereof |
| CN117736944B (en) * | 2024-02-20 | 2024-04-26 | 云南省农业科学院农业环境资源研究所 | Streptomyces griseus as well as microbial inoculum and application thereof |
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