CN113046260B - Microbial mixed microbial agent for promoting soybean growth and application thereof - Google Patents
Microbial mixed microbial agent for promoting soybean growth and application thereof Download PDFInfo
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 57
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- 241000589938 Azospirillum brasilense Species 0.000 claims abstract description 37
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- 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
- A01G22/40—Fabaceae, e.g. beans or peas
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- 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
- A01G7/00—Botany in general
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Abstract
The invention relates to the technical field of agricultural microorganisms, in particular to a microorganism mixture for promoting soybean growthMicrobial inoculum and application thereof. The microbial mixed microbial inoculum comprises fermentation liquor of azospirillum brasilense and hydrogenophila praecox, and the concentration of azospirillum brasilense is 1 multiplied by 10 6 cfu/ml～1×10 10 cfu/ml; the concentration of the Paraffin hydrogen bacteria is 1 multiplied by 10 6 cfu/ml～1×10 10 cfu/ml. The microbial mixed bacterial agent can stimulate plant cells to secrete various growth hormones, strongly promote root growth, rooting, root strengthening and capillary root growth, has the length or width increased, enhances the water and fertilizer absorbing capacity of crops, has the effects of growing and thickening stems, resisting lodging and the like, further increases the yield of crops, and solves the problem of abusing or using chemical fertilizers for a long time by farmers.
Description
Technical Field
The invention relates to the technical field of agricultural microorganisms, in particular to a microbial mixed microbial agent for promoting soybean growth and application thereof.
Background
Soybean is one of the most important grains in China, the soybean yield in China is in the fourth world, and the soybean planting area in 2017/18 is increased. From regional subareas, the soybean production and planting regions in China mainly comprise four large areas, namely, a Heilongjiang planting area, an Anhui planting area, an inner Mongolian planting area and a Jilin province planting area, and the areas respectively account for 36%, 11%, 8% and 4% of the national planting area. The average unit yield of soybeans in China is 120 kg/mu, which is far lower than the international level of 200 kg/mu, and the unit yield of domestic soybeans is to be improved.
At present, the cultivation management of soybeans mainly comprises main production links such as soybean variety selection, seed treatment, soil preparation, sowing, field management and the like. The current popular technique is mainly chemical fertilizer farming technique, and five kinds of chemical fertilizers are mainly added, namely potassium sulfate (K 2 SO 4 ) Potassium chloride (KCl), ammonium Nitrate (NH) 4 NO 3 ) Ammonium chloride (NH) 4 Cl) and ammonium bicarbonate (NH) 4 HCO 3 ). The fertilizer is mainly used for achieving the effect of increasing yield and gain, and the chemical fertilizer farming technology can enable the roots of crops to be developed, the stems to grow robustly and the fruits to be full, so that the crops and the soybeans benefit, and farmers benefit more.
Although the existing chemical fertilizer farming technology has great achievements in the aspect of agriculture in China, the technology is also step by step difficult in the aspect of sustainable development. The fertilizer is used in a large amount, and although the yield is increased in the early stage, the fertilizer is applied for a long time, so that the yield of the soybeans is necessarily reduced, and the soybean yield is reduced as follows: causing lodging or weak seedlings of the crops: the long-term application of chemical fertilizer can cause concentration difference, the root of crops is permeated, root cells not only do not absorb the moisture of soil, but also pour the moisture in cytoplasm into the soil, thereby causing crop damage, namely 'seedling burning'. Thereby causing the root and stem to be weak and leading to lodging of crops. The plant root cells breathe poorly, while the nitrogen and other nutrients exist in ionic state, so that the energy produced by cell metabolism is consumed during absorption, and the respiration is weakened, so that the energy supply is insufficient and the nutrient absorption is affected.
Nitrogen, phosphorus and potassium are three major elements of nutrition required by crops. The nitrogen in the atmosphere is in a nitrogen form which is unavailable to most crops, and the nitrogen must be converted into other forms which can be utilized by crops as fertilizer, and a great deal of energy is consumed in the conversion process to convert the nitrogen form into ammonia active nitrogen. In the past, the main energy sources for producing nitrogen fertilizer are petroleum, natural gas and coal are mainly used as raw materials, and the energy sources are short resources, and the shortage leads to high production cost of the fertilizer, so that the cost of applying the fertilizer by farmers is higher and higher, and the quality of produced agricultural products is lower and worse.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide the microbial mixed microbial agent for promoting the growth of soybeans, which can stimulate plant cells to secrete various growth hormones, strongly promote root growth, rooting, root strengthening and capillary root growth, increase the length or width of the microbial mixed microbial agent, enhance the water and fertilizer absorption capacity of crops, enable the crops to have the effects of growing and thickening stems, resisting lodging and the like, further increase the yield of the crops, and solve the problem of abusing or long-term use of fertilizers by farmers.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
the first object of the invention is to provide a microbial mixed inoculant, which comprises fermentation liquor of azospirillum brasilense and hydrogenophila praecox, wherein the concentration of azospirillum brasilense is 1 multiplied by 10 6 cfu/ml～1×10 10 cfu/ml; the concentration of the Paraffin hydrogen bacteria is 1 multiplied by 10 6 cfu/ml～1×10 10 cfu/ml。
The azospirillum brasilense (Azospirillum brasilense) is a spiral gram-negative bacterium, is facultative anaerobic, has polar flagella which can move to help the flagella to adhere to root systems, can be metabolized by various carbon sources and nitrogen sources, can form conceptual cytokinins under bad environments, can secrete a large amount of polysaccharide, and can form poly-B-hydroxybutyric acid in bodies as energy sources to spend the bad environments. The growth temperature and PH conditions are wide, and the nitrogen fixation enzyme has high activity under the proper conditions, and simultaneously has the function of generating H 2 Secretion of plant hormone and autonomous nitrogen fixation to stimulate crop growth.
The hydrogen philic pareto (Hydrogenophaga palleronii) is a phylum of the Paramyxotropy, and can decompose organic matters into micromolecular organic matters or inorganic matters, thereby helping crops absorb nutrients and enabling the crops to grow vigorously.
The Bacillusazotobacter can treat N in air 2 Absorbing and then forming ammonium ions (NH) by the nitroxidase in the azoospira bazedox 4 And salt ions are combined to form ammonium salt. This product provides the nitrogen element required by the crop, thereby forming a biofertilizer providing the inorganic matter required by the crop. So that the crop can absorb the biological fertilizer to flourish and promote the growth of rootstalk and leaf, thereby achieving mass production. The inventor finds that the combination of the azospirillum bassinense and the hydrogenophila paspaliana has good effect of promoting the growth of rhizomes and leaves of the soybeans and increases the yield of the soybeansAnd the quality of the soybean grains is enhanced, the loss of non-renewable resources such as natural gas, coal, petroleum and the like is reduced, and the concept of sustainable development is met.
As a preferred embodiment of the microbial mixed bacteria of the present invention, the concentration of the azospirillum brasilense is 1.9X10 8 cfu/ml; the concentration of the Paraffin hydrogen bacteria is 6.8X10 7 cfu/ml。
The inventor found by implementation that when the concentration of azospirillum brasilense is 1×10 6 cfu/ml～1×10 10 cfu/ml; the concentration of the hydrogen philic Paraffin is 1×10 6 cfu/ml～1×10 10 The effect of promoting soybean growth is better when cfu/ml is reached, and the concentration of azospirillum brasilense is 1.9X10 8 cfu/ml; the concentration of the hydrogen philic Paraffin is 6.8X10 7 The effect is best when cfu/ml.
As a preferred embodiment of the microbial mixed microbial inoculum, the volume ratio of the fermentation liquor of the azospirillum brasilense to the fermentation liquor of the hydrogenophila praecox is (10-13) (7-10).
As a preferred embodiment of the microbial mixed microbial agent, the volume ratio of the fermentation liquor of the azospirillum brasilense to the fermentation liquor of the hydrogenophila praecox is 13:7.
As a preferred implementation mode of the microbial mixed bacterial agent, the azoospermia Brazilian is derived from China general microbiological culture collection center, and the preservation number is CGMCC 1.10379.
As a preferred implementation mode of the microbial mixed bacterial agent, the hydrogen philic pareto comes from China general microbiological culture collection center with the preservation number of CGMCC 1.12500.
A second object of the present invention is to provide a method for soybean breeding comprising the steps of: the microbial mixed bacterial agent is sprayed on the root of soybean in seedling period.
The third object of the invention is to provide the application of the microbial mixed bacteria agent in preparation of a preparation for promoting crop growth.
As a preferred embodiment of the use according to the invention, the crop comprises soybean. In the present invention, the crop is not limited to soybean, and the crop may be other crops such as wheat, rice, etc.
The fourth object of the invention is to provide a culture medium which comprises the microbial mixed bacterial agent.
Compared with the prior art, the invention has the following beneficial effects:
the microbial mixed microbial inoculum can stimulate plant cells to secrete various growth hormones, strongly promote root growth, rooting, root strengthening and capillary root growth, increase the length or width of the microbial mixed microbial inoculum, enhance the water and fertilizer absorption capacity of crops, enable the crops to have the effects of growing and thickening stems, resisting lodging and the like, and further increase the yield of the crops.
Drawings
FIG. 1 is an optical microscopy image of Azospirillum brasilense (Azospirillum brasilense);
FIG. 2 is a plate coating of Azospirillum brasilense (Azospirillum brasilense);
FIG. 3 is an optical microscopy image of H.Paraffin (Hydrogenophaga palleronii);
FIG. 4 is a plate coating of H.Paraffin (Hydrogenophaga palleronii);
FIG. 5 is an optical microscopic view of the microbial blend of the present invention;
FIG. 6 is a plate coating pattern of the microbial blend of the present invention;
FIG. 7 is a graph showing comparison of soybean seedling stage growth in a control group and a microbial mixed inoculant of the present invention;
FIG. 8 is a graph comparing soybean pod-bearing drum-phase growth in a control group and a microbial mixed inoculant of the present invention;
FIG. 9 is a graph showing comparison of soybean maturity results for a control blank and a microbial mixed inoculant of the present invention;
FIG. 10 is a view of soybean stem paraffin sections from a control blank and a microbial mixed inoculant of the present invention;
FIG. 11 is a view showing paraffin sections of soybean leaves from a blank group and an experimental group of the microbial mixed inoculum of the invention.
Detailed Description
For a better description of the objects, technical solutions and advantages of the present invention, the present invention will be further described with reference to the accompanying drawings and specific embodiments. The various microorganisms in the invention can be purchased commercially or can be obtained from China general microbiological culture Collection center, and the preparation method of the various microorganism culture broths adopts corresponding standard culture media and culture broths according to conventional culture methods.
In the following examples, the azoospermia barceliac (Azospirillum brasilense) is derived from China general microbiological culture collection center (CGMCC) 1.10379. The hydrogen philic parvobacteria (Hydrogenophaga palleronii) is derived from China general microbiological culture collection center, and the preservation number is CGMCC 1.12500.
The formulation of the nutrient broth medium of China center for type culture Collection (0002) is shown in Table 1.
TABLE 1
| Peptone | 10.0g | Beef extract | 3.0g |
| NaCl | 5.0g | Distilled water | 1L |
EXAMPLE 1 Azospirillum bazeri (Azospirillum brasilense), H.praecox
Monoclonal culture of (Hydrogenophaga palleronii)
The obtained azotobacter bazedox (Azospirillum brasilense) is dyed by a gram-dyeing method, and the azotobacter bazedox (Azospirillum brasilense) is inspected to be gram-negative and has a spiral rod-shaped structure by an optical lens. The colonies were visually observed to be morphologically circular, low-bulge, rounded complete edges, and pale red in color (see fig. 1 and 2).
Taking a nutrient gravy culture medium with the specification of 1L of beaker dissolution culture medium, regulating the pH value to between 6.8 and 7.2 before high-temperature sterilization of the culture medium, pouring 200ml of prepared culture medium into a triangular flask with the capacity of 500ml, sealing a high-temperature-resistant sealing film on the bottle mouth of the triangular flask, placing the triangular flask for high-temperature sterilization, sterilizing at 121 ℃ for 30 minutes, adding the azoospermia bazeylanica (Azospirillum brasilense) after cooling to normal temperature, setting the culture temperature to between 25 and 30 ℃, and placing the triangular flask at constant temperature for shake culture for 24 hours to obtain the fermentation liquor containing the azoospermia bazemakinsonii.
The obtained hydrogen philic Paraffin (Hydrogenophaga palleronii) was stained by a gram staining method, and examined by an optical microscope for the hydrogen philic Paraffin (Hydrogenophaga palleronii) as a gram negative, spiral structure. The colonies were visually observed to be morphologically circular, low-bulge, rounded complete edges, and white in color (see fig. 3 and 4).
Taking a nutrient gravy culture medium with the specification of 1L and the number (0002) of China center for culture collection, taking a beaker with the specification to dissolve the culture medium to a constant volume of 1L, adjusting the pH value to between 6.8 and 7.2 before high-temperature sterilization of the culture medium, pouring 200ml of prepared culture medium into a triangular flask with the volume of 500ml, sealing a high-temperature resistant sealing film on the bottle mouth of the triangular flask, placing the triangular flask for high-temperature sterilization, sterilizing at 121 ℃ for 30 minutes, adding the hydrogen-philic Paraffin (Hydrogenophaga palleronii) after cooling to normal temperature, setting the culture temperature to between 25 and 30 ℃, placing the culture medium at constant temperature, and shaking and culturing for 24 hours to obtain the fermentation broth containing the hydrogen-philic Paraffin.
EXAMPLE 2 preparation of the microbial Mixed inoculant of the present invention
The fermentation broth containing azospirillum brasilense and the fermentation broth containing hydrogenophila praecox cultured in example 1 are taken as volumeMixing at a ratio of 13:7 to obtain microorganism mixed microbial inoculum (1L) (see figure 5), wherein the optimal concentration of azospirillum brasilense is 1.9X10 8 cfu/ml; the optimal concentration of the hydrogen philic Paraffin is 6.8X10 7 cfu/ml, total thallus concentration is not less than 1×10 8 cfu/mL。
200ml of the modified AGAR culture medium (the formula is shown in table 2) or the BPY culture medium (the formula is shown in table 3) is poured into a triangular flask with the capacity of 500ml, a high-temperature-resistant sealing film is sealed on the opening of the triangular flask, the triangular flask is placed for high-temperature sterilization and then is sterilized at 121 ℃ for 30 minutes, the microbial mixed bacteria can be added after the mixture is cooled to normal temperature, the culture temperature is set between 25 ℃ and 30 ℃, and the triangular flask is placed for constant-temperature shake culture for 24 hours.
TABLE 2 formulation of improved AGAR Medium
| Yeast extract | 15g | KH 2 PO 4 | 0.1g |
| Prion protein | 0.25g | MgSO 4 ·7H 2 O | 0.1g |
| Casein amino acids | 0.25g | FeCl 3 | 0.05g |
| Glucose | 15g | NaMoO 4 ·2H 2 | 0.05g |
| Peptone | 5g | Soluble starch | 0.25g |
| K 2 HPO 4 | 0.6g | Pyruvic acid sodium salt | 0.15g |
| CaCl 2 | 0.05g | Distilled water | 1L |
TABLE 3 formulation of BPY Medium
| Beef extract | 5g | Yeast extract | 5g |
| Peptone | 5g | Glucose | 5g |
| NaCl | 5g | Distilled water | 1L |
Example 3 plate calculation of colony count of Azospirillum bassinense and Hydrogen-philia Paramycolatopsis in the microbial Mixed inoculant of the present invention
Measuring amount
1ml of the microbial mixed bacteria prepared in example 2 is taken and subjected to gradient dilution to 10 5 The diluted bacterial liquid (25 ul) is used for coating (NA) nutrition agar plate, and the plate is inverted and cultured for 48 hours at a biochemical constant temperature, so that the following results are obtained (see FIG. 6):
white punctuation is azospirillum brasilense (Azospirillum brasilense), black punctuation is hydrogenophila praecox (Hydrogenophaga palleronii);
counting to obtain 14 black punctuations in the graph 6, 20 black punctuations in the succession of the graph 6, and 17 arithmetic average;
counting to obtain 50 white punctuations in the graph 6, 45 white punctuations in the graph 6, and 47.5 arithmetic average;
the total number of marks in fig. 6 is 64, the number of marks in the subsequent total number of marks in fig. 6 is 65, and the arithmetic average is 64.5; the average colony count x dilution x (1000/drop spread) at uniform dilution was calculated by plate counting according to the formula (cfu/ml) (see table 4):
TABLE 4 Table 4
| Name of single strain | Concentration (cfu/ml) |
| Azospirillum brasilense (Azospirillum brasilense) cfu: | 1.9×10 8 cfu/ml |
| hydrogen pervophila (Hydrogenophaga palleronii) cfu: | 6.8×10 7 cfu/ml |
| total bacterial concentration (cfu/ml): | 2.6×10 8 cfu/ml |
through the test, the experimental data all reach the standard.
Example 4 determination of Nitrogen fixation Activity of microbial Mixed inoculant of the present invention by acetylene reduction method
Methods for determining nitrogen fixation activity are described in Williams et al (1987) and Wilson et al (2012). The acetylene reducer was sealed, 5% of the air was first withdrawn, and then an equal volume of acetylene was injected to give an acetylene concentration of 5% in the reactor. Acetylene gas is generated in the gas storage bag through the reaction of calcium carbide and pure water by a self-made acetylene reactor. After the reactor is kept stand for reaction for 8 hours at 25 ℃, gas in the reactor is collected and injected into a gas collection bottle, and the yield of ethylene is analyzed by a gas chromatograph.
The amount of acetylene produced was measured by a gas chromatograph (Shanghai electric analysis). The chromatographic column is GDX-50260/80mesh 3mm diameter packed column. The concentration of acetylene standard gas is 100mg L-1. 1ml of each of a microorganism mixed microbial inoculum sample, a single strain of cultivated azoospira Brazilian (Azospirillum brasilense), a hydrogen philic Paraffin (Hydrogenophaga palleronii) and a blank (distilled water) cultured at 28 ℃ for 48 hours in a constant temperature shaker is sampled to measure acetylene reduction activity, a fresh root nodule sample is cleaned, superfluous water is removed, the fresh root nodule sample is placed in a serum bottle with a plug, a certain amount of acetylene gas is injected, and the produced ethylene amount is measured by shaking for more than half an hour at 28 ℃. After the determination, the root nodule is put into an oven at 80 ℃ and baked for 16 hours to weigh the dried nodule.
According to the formula, the activity of the azotase (nmol/mg.h) =C 2 H 4 nmol/[ amount of mycoprotein (mg) X reaction time (h)]Wherein (C) 2 H 4 nmol＝1000X C 2 H 4 Volume (. Mu.L) X273X P/[22.4X (273+t ℃ C.) X760]Where P is the gas pressure (mm Hg) and t is the reaction temperature.
Calculated according to a standard curve of the corresponding relation between the concentration of ethylene and the chromatographic peak area and an acetylene reduction method chromatogram calculation formula (see table 5):
TABLE 5
| Microbiome | (nmol C 2 H 4 *g -1 *h -1 ) |
| Microbial mixed microbial agent | 38.953 |
| Azospirillum brasilense (Azospirillum brasilense) | 37.471 |
| Hydrogen parer (Hydrogenophaga palleronii) | 1.482 |
| Blank distilled water | 0 |
According to the experiment, the data table shows that the azotobacter bazedoxorum (Azospirillum brasilense) and the hydrogenophilic paspalum (Hydrogenophaga palleronii) have nitrogen fixation function, but mainly the azotobacter bazedoxorum (Azospirillum brasilense) has nitrogen fixation function values.
Example 5 determination of Hydrogen utilization ability of the microbial Mixed inoculant of the present invention by gas chromatography
Hydrogen Paraffin (Hydrogenophaga palleronii) is capable of assimilating CO with H2 in the presence of hydrogenase 2 And H 2 Synthesizing the substance, and performing chemoautotrophy. In the invention, the strain absorption H is measured by utilizing a gas chromatography 2 Capability. Adding 5mL NA agar into two test tubes to obtain slant culture medium, respectively dipping individual strain of bacteria Paraffin hydrogen bacteria (Hydrogenophaga palleronii) and blank control distilled water, sealing and culturing with soft rubber plug, standing at 28deg.C for 48 hr until Paraffin hydrogen bacteria (Hydrogenophaga palleronii) grow out of bacterial plaque, simultaneously pumping 10% gas into two test tubes, adding equal volume of H 2 After 48 hours of culture, 100. Mu.L of the gas in the test tube was extracted, and the hydrogen consumption was measured by a gas chromatograph, without stopping any operation in the middle, waiting for the peak value, and ensuring the precision of quantitative analysis.
According to the curve run, the peak time, the analysis results were as follows (see table 6):
TABLE 6
Absorption H by H.Paraffin (Hydrogenophaga palleronii) was determined 2 The capability is taken as the dominant effect of the microbial mixed bacterial agent of the invention, and can be used as functional bacteria for hydrogen oxidation.
Example 6 method for promoting Soybean growth Using the microbial Mixed inoculant of the present invention
And (3) carrying out field planting on the nine vines at 3 months and 15 days in 2020, adding water to dilute the microbial mixed microbial inoculum by 1:30 from the seedling stage, irrigating roots, and irrigating the microbial mixed microbial inoculum for 1 time every 2 weeks, wherein the irrigation is carried out continuously until 30 days in 2020, obtaining soybean data, wherein no chemical fertilizer is applied in the period, and only the microbial mixed microbial inoculum is applied. The results are shown in FIGS. 7-9 (the left side of the figure represents the blank control group; the right side represents the experimental group).
Referring to fig. 7, the soybean seedlings to which the microbial mixed inoculant of the present invention was applied were grown to a higher level than those of the control group, and the soybean seedlings of the experimental group were grown to a higher level than those of the control group.
Referring to fig. 8, experimental data shows a 4-fold increase in root nodule count for soybean with more soybean pod drums in the soybean pod stage experimental group than in the blank control group.
Referring to fig. 9, the blank group soybean has a hundred grain weight of 8.646g, the experimental group soybean has a hundred grain weight of 19.196g, and the yield is improved by more than 45%, and the data are from nine field planting of vinca.
EXAMPLE 7 Paraffin section of the stems and leaves of Soybean
The stem of the experimental group using the microbial mixed inoculant of the present invention is tough and strong, and compared with the control group, the stem of the experimental group has full tissue, high strength and lodging resistance. The number of vascular bundles in the experimental group is large, the number of cell layers is large, the cell arrangement is compact, and the storage materials are large. The microbial mixed microbial agent can increase the strength and toughness of the stems of soybeans and increase the lodging resistance.
The main leaf vein and leaf vein of soybean in the experimental group are compared and thickened, and the corresponding leaf vein (containing bundle sheath cells) and mesophyll tissue (containing mesophyll cells) are more and stronger than those in the control group, so that the cells are arranged more and more tightly, and the stored substances are also more.
The experimental data show that the microbial mixed microbial inoculum can strongly promote root growth, rooting, root strengthening and capillary root growth, the length or width of the microbial mixed microbial inoculum is increased, the water and fertilizer absorption capacity of crops is enhanced, the soybeans have the properties of thick and long stems, lodging resistance and the like, and the yield of the crops is further increased.
Finally, it should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted equally without departing from the spirit and scope of the technical solution of the present invention.
Claims (7)
1. A microbial mixed bacterial agent is characterized by comprising fermentation liquor of azospirillum brasilense (Azospirillum brasilense) and hydrogenophila praecox (Hydrogenophaga palleronii), wherein the concentration of azospirillum brasilense is 1 multiplied by 10 6 cfu/ml～1×10 10 cfu/ml; the concentration of the Paraffin hydrogen bacteria is 1 multiplied by 10 6 cfu/ml～1×10 10 cfu/ml; the azoospermia Brazilian is derived from China general microbiological culture collection center, and the preservation number is CGMCC 1.10379; the hydrogen philic parliament is from China general microbiological culture collection center, and the preservation number is CGMCC 1.12500.
2. The microbial mixed inoculant according to claim 1, wherein the azospirillum brasilense concentration is 1.9x10 8 cfu/ml; the concentration of the Paraffin hydrogen bacteria is 6.8X10 7 cfu/ml。
3. The microbial mixed inoculant according to claim 1, wherein the volume ratio of the fermentation broth of azospirillum brasilense to the fermentation broth of hydrogenophila paspaliana is (10-13): 7-10.
4. A microbial mixed inoculant according to claim 3, wherein the volume ratio of the fermentation broth of azospirillum brasilense to the fermentation broth of hydrogenophila paspaliana is 13:7.
5. A method of soybean breeding comprising the steps of: the microbial mixed inoculant according to any one of claims 1 to 4 is sprayed on the roots of soybeans during seedling stage.
6. Use of the microbial mixed inoculant according to any one of claims 1-4 for the preparation of a soy bean growth promoting formulation.
7. A culture medium comprising the microbial mixed inoculum according to any one of claims 1-4.
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Effective date of registration: 20240929 Address after: No. 202, 2nd Floor, Building A3, No. 6 Chuangye 1st Road, Qingyuan High tech Industrial Development Zone, Qingyuan City, Guangdong Province, China 511500 Patentee after: Guangdong Lihao Biological Agriculture Co.,Ltd. Country or region after: China Address before: 137400 8-129 - (24), No.5 building, phase I, Anbang commercial and residential group, south of hanshangdong street, east of Yunxi Road, west of Gongye Road, Wulanhaote City, Xing'an League, Inner Mongolia Autonomous Region Patentee before: Xing'an League Laishen Biological Agriculture Co.,Ltd. Country or region before: China |
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