CN103396971B - Burkholderia cepacia and application thereof - Google Patents
Burkholderia cepacia and application thereof Download PDFInfo
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- CN103396971B CN103396971B CN201310368528.XA CN201310368528A CN103396971B CN 103396971 B CN103396971 B CN 103396971B CN 201310368528 A CN201310368528 A CN 201310368528A CN 103396971 B CN103396971 B CN 103396971B
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Abstract
The invention discloses Burkholderia cepacia and an application thereof. A bacterial strain is Burkholderia cepacia pt6, and a preservation number of the bacterial strain is CGMCC No. 7598 (China General Microbiological Culture Collection Center Number 7598). The invention further discloses a microorganism bacterium agent containing the bacterial strain and a preparation method. An experiment indicates that fermentation liquor of the Burkholderia cepacia pt6 contains a large number of phosphate solubilizing bacteria; the phosphate solubilizing bacteria have definite positive effects on soil improvement; and phosphate solubilizing functions of the phosphate solubilizing bacteria have important significance in increasing a use ratio of a phosphate fertilizer, increasing a yield of crop and improving the quality of the crop.
Description
Technical field
The invention belongs to phosphate solubilizing microorganism fertilizer field.Particularly, the present invention relates to a strain can efficient phosphate-solubilizing bacterium Burkholderia cepacia and uses thereof, and the biologic phosphorus fertilizer that adopts this Burkholderia cepacia to prepare.
Background technology
Phosphorus is the basic comprising element of hereditary material DNA and energy matter ATP in vegetable cell, in the whole vital process of plant, play an important role, phosphorus is one of essential element of growth and development of plants, high-energy phosphate bond is the carrier of energy, and the biological process in photosynthesis and the body of plant all must have phosphorus to participate in.Phosphorus is the important substance guarantee of agriculture production, is again non-renewable mineral resource.In China's arable soil, the massfraction of full phosphorus is generally at 0.2～1.l g/kg(soil) between variation, many soil phosphoruses are under-supply, and therefore utilizing phosphate solubilizing microorganism directionally to regulate Phosphorus Status of The Soils and improve phosphate fertilizer utilization efficiency is to reach one of high crop yield, colory important channel.
Burkholderia is a kind of gram negative bacterium that is extensively present in occurring in nature, this bacterium has the several functions such as biological control, biological degradation and Promoting plant growth in agriculture field, show its application prospect good in agriculture production simultaneously, become a study hotspot in microbiology field.Burkholderia can produce the multiple secondary metabolites such as pyrrolnitrin, azophenlyene, phenylpyrrole and bring into play Biocontrol Effect.From the eighties in 20th century, Burkholderia starts as biotic pesticide and biological bactericide successively at U.S.'s official registration, as Type wisconsin, Deny, Blue circle etc.; This bacterium is also being used mainly as biological pesticide in China, has registered similar biological product in 1996---and " sub-precious ".The molten phosphorus mechanism of bacterium is very complicated, the molten phosphorus of some bacterium is mainly that proton works, and some is mainly that organic acid works, and some bacterial strain two kinds of mechanism all exists, also some bacterial strain is owing to producing other chelating material in process of growth, thereby causes the dissolving of ground phosphate rock.Make full use of this characteristic of phosphate solubilizing microorganism, not only can utilize the phosphorus of stationary state, ADSORPTION STATE, effectively alleviate the present situation that lacks phosphorus in soil, and to improving phosphate fertilizer utilization efficiency, the environmental pollution that the use of minimizing phosphate fertilizer causes, even, to improving crop yield and quality thereof, all have important practical significance.
Summary of the invention
The object of this invention is to provide a kind of for efficient phosphate-solubilizing bacterium---Burkholderia cepacia bacterial strain pt6.Burkholderia cepacia bacterial strain of the present invention has that fast growth, phosphorus decomposing ability are strong, strong stress resistance, the feature such as can rapid, high volume surely grows at plant rhizosphere, therefore has a good application prospect.
Another object of the present invention be to provide a kind of adopt prepared by this new Burkholderia cepacia can efficient phosphate-solubilizing microbiobacterial agent.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
Bacterial strain provided by the present invention is Burkholderia cepacia (Burkholderia cepacia) pt6, from the soil of Honghu City, Hubei Province, separate and obtain, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on May 15th, 2013 and (be called for short CGMCC, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica), preserving number is CGMCC No.7598.It has following biological characteristics: on NA substratum, in the time cultivating for 30 DEG C, bacterial strain bacterium colony is faint yellow, is canescence, circle, smooth surface, protuberance, neat in edge, diameter 2～4mm on PDA flat board.Thalline direct rod shape, size is (0.5～0.7) μ m × (1.5～1.8) μ m, the tool flagellum of growing thickly, Gram-negative, without gemma, capsule stain negative (without pod membrane), can utilize malonate, Citrate trianion, the catalase positive, the gelatine liquefication positive, can not hydrolyzed starch.
Cultural method or the propagation method of Burkholderia cepacia of the present invention (Burkholderia cepaciais) bacterial strain pt6 comprise:
(1) common cultivation is preserved and is adopted NA substratum (Nutrient agar), formula: 0.5% peptone, and 0.3% extractum carnis, 0.5%NaCl, 1.5% agar, distilled water, pH is adjusted to 7.0.
(2) laboratory fluids is cultivated and is adopted NA substratum: 0.5% peptone, and 0.3% extractum carnis, 0.5%NaCl, distilled water, pH is adjusted to 7.0.
(3) seeding tank liquid fermentation medium, formula: Semen Maydis powder 15g, glucose 4g, soybean cake powder 21g, fish meal 4g, CaCO
39g, (NH
4)
2sO
41.5g, K
2hPO
40.5g, MgSO
47H
2o0.1g, NaCl0.1g, water 1000mL, pH is adjusted to 7.5.
(4) bulk fermentation substratum is with seeding tank liquid fermentation medium.
The present invention also provides a kind of microbiobacterial agent that can efficient phosphate-solubilizing, and described microbiobacterial agent contains described Burkholderia cepacia (Burkholderia cepacia) bacterial strain pt6.
The present invention also provides above-mentioned Burkholderia cepacia (Burkholderia cepacia) bacterial strain pt6 or microbiobacterial agent in the utilization ratio that improves phosphate fertilizer, improves Soil structure, improves the output of crop and the application of quality aspect.
The preparation method of mentioned microorganism microbial inoculum, comprising:
(1) seed shaking flask: the phosphate-solubilizing bacteria Burkholderia cepacia bacterial strain pt6 having activated is inoculated in the triangular flask that NA liquid nutrient medium is housed to 30 DEG C of 200r/min shaking culture 12h;
(2) seeding tank fermentation: the shake-flask seed liquid that step (1) is prepared is by 1%(v/v) inoculum size be seeded in the seeding tank that seeding tank liquid fermentation medium is housed, to ferment to thalline and enter logarithmic phase; Temperature is 30～32 DEG C, and tank pressure is 0.5kg/cm
2, ventilation 0.5～0.8:1;
Described seeding tank liquid fermentation medium is:: Semen Maydis powder 15g, glucose 4g, soybean cake powder 21g, fish meal 4g, CaCO
39g, (NH
4)
2sO
41.5g, K
2hPO
40.5g, MgSO
47H
2o0.1g, NaCl0.1g, water 1000mL, pH is adjusted to 7.5.
(3) fermentor tank: by the seed liquor of step (2) seeding tank by 10%(v/v) inoculum size be seeded to fermentor tank and ferment, after fermentation 20～30h, biomass reaches 50～80 × 10
8cfu/mL, can go out tank.The same step of fermentation condition and medium component (2).
Experiment shows: in Burkholderia cepacia bacterial strain of the present invention (Burkholderia cepacia) pt6 fermented liquid, contain a large amount of phosphate solubilizing bacterias, soil improvement is had to certain active effect, the molten phosphorus function that it has, in the utilization ratio that improves phosphate fertilizer, the output and the quality aspect that improve crop have great importance.
Brief description of the drawings
Fig. 1 is phosphorus typical curve.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage and disadvantage of the present invention will be more clear along with description.But these embodiment are only exemplary, scope of the present invention are not formed to any restriction.It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can the details of technical solution of the present invention and form be modified or be replaced, but these amendments and replacement all fall within the scope of protection of the present invention.
Embodiment 1
1, the isolation and purification of efficient phosphate-solubilizing bacterium Burkholderia cepacia (B.cepacia) pt6
Burkholderia cepacia of the present invention (B.cepacia) pt6 adopts dilution-plate method to separate and obtain with plate streak from soil, and separation method is: adopt Meng Jinna inorganic phosphorus solid medium to screen.The collection of soil sample, sample collecting place is Honghu City, Hubei Province, and the sampling time is on February 13rd, 2012, and employing 5 point samplings, gather respectively plot surrounding and the soil of the central degree of depth within the scope of 10～20cm is appropriate, and equivalent mixes.Indicate collecting location, time and collection people.Take 1g soil sample in 100mL sterilized water, be placed in 30 DEG C of shaking table 150r/m concussion 10min, get 100 μ L10
-2, 10
-3, 10
-4diluent is coated on inorganic phosphorus solid medium flat board, three of each gradient coatings are parallel, cultivate after 7d at 30 DEG C, observation has or not molten phosphorus circle, and according to the size of molten phosphorus loop diameter (D) and colony diameter (d) ratio tentatively determine phosphorus decomposing ability by the bacterial strain that has molten phosphorus effect perform record for subsequent use.
The inventor obtains by a large amount of screening operations Burkholderia cepacia (B.cepacia) pt6 that a strain can efficient phosphate-solubilizing.Experiment showed, there is fast growth, phosphorus decomposing ability is strong, strong stress resistance, the feature such as can rapid, high volume surely grows at plant rhizosphere, therefore have a good application prospect.
Phosphate solubilizing bacteria screening adopts Meng Jinna inorganic phosphorus solid medium: glucose 10g, (NH
4)
2sO
40.5g, NaCl0.3g, KCl0.3g, FeSO
4.7H
2o0.03g, MgSO
4.7H
2o0.3g, MnSO
4.4H
2o0.03g, Ca
3(PO
3)
210g, yeast extractive substance 0.4g, agar 20g, deionized water 1000mL, pH is adjusted to 7.0.
The identification of strains of embodiment 2 efficient phosphate-solubilizing bacterium Burkholderia cepacia (B.cepacia) pt6
(1) Microbiological Characteristics: on NA substratum, bacterial strain bacterium colony is faint yellow in the time cultivating for 30 DEG C, is canescence, circle, smooth surface, protuberance, neat in edge, diameter 2～4mm on PDA flat board.Thalline direct rod shape, size is (0.5～0.7) μ m × (1.5～1.8) μ m, the tool flagellum of growing thickly, Gram-negative, without gemma, capsule stain negative (without pod membrane), can utilize malonate, Citrate trianion, the catalase positive, the gelatine liquefication positive, can not hydrolyzed starch.
(2) molecular biological characteristic (classification qualification)
The 16s rDNA gene sequencing result following (SEQ No.1) of this bacterial strain:
TGCAAGTCGAACGGCAGCACGGGTGCTTGCACCTGGTGGCGAGTGGCGAACGGGT?GAGTAATACATCGGAACATGTCCTGTAGTGGGGGATAGCCCGGCGAAAGCCGGATTAAT?ACCGCATACGATCTACGGATGAAAGCGGGGGACCTTCGGGCCTCGCGCTATAGGGTTGG?CCGATGGCTGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCAGTAGCT?GGTCTGAGAGGACGACCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGA?GGCAGCAGTGGGGAATTTTGGACAATGGGCGAAAGCCTGATCCAGCAATGCCGCGTGT?GTGAAGAAGGCCTTCGGGTTGTAAAGCACTTTTGTCCGGAAAGAAATCCTTGGTTCTAA?TATAGCCGGGGGATGACGGTACCGGAAGAATAAGCACCGGCTAACTACGTGCCAGCAGC?CGCGGTAATACGTAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAG?GCGGTTTGTTAAGACCGATGTGAAATCCCCGGGCTCAACCTGGGAACTGCATTGGTGAC?TGGCAAGCTAGAGTATGGCAGAGGGGGGTAGAATTCCACGTGTAGCAGTGAAATGCGTA?GAGATGTGGAGGAATACCGATGGCGAAGGCAGCCCCCTGGGCCAATACTGACGCTCATG?CACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGAT?GTCAACTAGTTGTTGGGGATTCATTTCCTTAGTAACGTAGCTAACGCGTGAAGTTGACCG?CCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGACCCGCACAAG?CGGTGGATGATGTGGATTAATTCGATGCAACGCGAAAAACCTTACCTACCCTTGACATGG?TCGGAATCCCGCTGAGAGGTGGGAGTGCTCGAAAGAGAACCGGCGCACAGGTGCTGCA?TGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCC?TTGTCCTTAGTTGCTACGCAAGAGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGG?AAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGGGTAGGGCTTCACACGTCATAC?AATGGTCGGAACAGAGGGTTGCCAACCCGCGAGGGGGAGCTAATCCCAGAAAACCGAT?CGTAGTCCGGATTGCACTCTGCAACTCGAGTGCATGAAGCTGGAATCGCTAGTAATCGC?GGATCAGCATGCCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCA?TGGGAGTGGGTTTTACCAGAAGTGGCTAGTCTAACCGCAAGGAGGACGTCACCACG
The mensuration of embodiment 3 efficient phosphate-solubilizing bacterium Burkholderia cepacia (B.cepacia) pt6 phosphorus decomposing abilities
Adopt the phosphorus decomposing ability of Meng Jinna inorganic phosphorus liquid culture based assays phosphate solubilizing bacteria, first the Burkholderia cepacia of preserving on inclined-plane (B.cepacia) pt6 is inoculated in NB liquid nutrient medium as seed, in 30 DEG C, shaking culture 12h in 180r/min constant-temperature table, by 1% inoculum size (v/v), cultured bacterium liquid is inoculated into 250mL containing in 100mL Meng Jinna inorganic phosphorus liquid nutrient medium triangular flask subsequently, to inoculate 1% aseptic NB liquid nutrient medium to Meng Jinna inorganic phosphorus liquid nutrient medium as blank, in 30 DEG C, shaking culture 7d in 180r/min constant-temperature table.
Adopt molybdenum antimony resistance colorimetric method to measure the available phosphorus content in fermented liquid, phosphate solubilizing bacteria fermented liquid available phosphorus content is higher shows that this phosphate solubilizing bacteria phosphorus decomposing ability is stronger.Fermented liquid is shaken up, get 20mL in centrifuge tube, 8000r/min, centrifugal 10min; Draw 0.5～2mL supernatant liquor in the volumetric flask of 50mL, add distilled water to volume to be about 30mL, add 2 2.6-dinitrophenol indicator, dropwise add NaOH solution to the liquid of 4mol/L to present yellow, then add the H of 1 1mol/L
2sO
4just decorporate to solution yellow, then add the anti-reagent of molybdenum antimony of 5mL, water is settled to scale, and then the 30min that develops the color at 25 DEG C measures absorbance on 721 type ultraviolet-visible pectrophotometers, and wavelength set is 700nm.Calculate corresponding available phosphorus content according to typical curve.
The drafting of phosphorus typical curve: by 10 times of the phosphorus mark liquid dilutions of 50mg/L, for 5mg/L, draw respectively in the phosphorus mark liquid 0,2,4,6,8,10mL to 50mL volumetric flask of 5mg/L, add distilled water to volume to be about 30mL, add 2 2.6-dinitrophenol indicator, the NaOH solution that dropwise adds 4mol/L to liquid in volumetric flask presents micro-yellow, then adds the H of 1 1mol/L
2sO
4to just cancellation of yellow, then add the anti-reagent of 5mL molybdenum antimony, and be settled to scale with distilled water.The 30min that develops the color at 25 DEG C, then coexists on 721 type ultraviolet-visible pectrophotometers and measures absorbance with the nitrite ion of above-mentioned sample, and wavelength set is 700nm.Taking phosphorus concentration as X-coordinate, taking absorbance as ordinate zou, draw phosphorus typical curve, as shown in Figure 1.
The available phosphate concentration not connecing in the blank solution of bacterium is 83.6mg/L, in the fermented liquid of phosphate-solubilizing bacteria Burkholderia cepacia (B.cepacia) pt6, available phosphorus content is 158.7mg/L, shows that phosphate-solubilizing bacteria Burkholderia cepacia (B.cepacia) pt6 is a plant height effect phosphate-solubilizing bacteria.
NB substratum, formula: 0.5% peptone, 0.3% extractum carnis, 0.5%NaCl, all the other are distilled water, pH is adjusted to 7.0.
Meng Jinna inorganic phosphorus liquid nutrient medium: glucose 10g, (NH
4)
2sO
40.5g, NaCl0.3g, KCl0.3g, FeSO
4.7H
2o0.03g, MgSO
4.7H
2o0.3g, MnSO
4.4H
2o0.03g, Ca
3(PO
3)
210g, deionized water 1000mL, pH is adjusted to 7.0, for the phosphorus decomposing ability of quantitative assay phosphate solubilizing microorganism.
A large amount of liquid fermentings of embodiment 4 efficient phosphate-solubilizing bacterium Burkholderia cepacia (B.cepacia) pt6
1. inclined-plane seed: phosphate-solubilizing bacteria bacterial classification Burkholderia cepacia (B.cepacia) pt6 is kept on NA substratum, is stored in 4 DEG C of thermostat containers, before using, takes out and activates twice on NA substratum.
2. seed shaking flask: it is that 200mL contains in the 500mL triangular flask of NA liquid nutrient medium that phosphate-solubilizing bacteria Burkholderia cepacia (B.cepacia) pt6 having activated is inoculated into liquid amount, 30 DEG C of 200r/min shaking culture 12h.
3. seeding tank fermentation: by the above-mentioned shake-flask seed liquid preparing by 1%(v/v) inoculum size be seeded in the seeding tank that seeding tank liquid fermentation medium is housed and ferment;
Seeding tank liquid fermentation medium: Semen Maydis powder 15g, glucose 4g, soybean cake powder 21g, fish meal 4g, CaCO
39g, (NH
4)
2sO
41.5g, K
2hPO
40.5g, MgSO
47H
2o0.1g, NaCl0.1g, water 1000mL, pH is adjusted to 7.5.
Fermentation condition: temperature is 30～32 DEG C, tank pressure is 0.5kg/cm
2, ventilation 0.5～0.8:1; After inoculation, at interval of sampling in 2 hours once, with after 10～100 times of aqua sterilisa dilutions, count somatic cells with Hematocyte Counter under the microscope, and adopt crystal violet staining assay, whether normal with oily sem observation Growth of Cells, whether pollute etc.If thalline size is basically identical, and when quantity sudden fast rush, enter logarithmic phase (approximately needing 12h), now can be to producing tank inoculation.
4. fermentor tank: by the seed liquor of above-mentioned seeding tank by 10%(v/v) inoculum size be seeded to fermentor tank and ferment, the same seeding tank of fermentation condition and substratum.Determining not pollution, under normal circumstances, after fermentation 20～30h, biomass should reach 50～80 × 10 in fermentation
8cfu/mL.
Embodiment 5 efficient phosphate-solubilizing bacterium Burkholderia cepacia (B.cepacia) pt6 plot experiments
Test materials: the fermented liquid of efficient phosphate-solubilizing bacterium Burkholderia cepacia (B.cepacia) pt6, there is effect of molten phosphorus, living bacteria count is greater than 50 × 10
8cfu/mL.
Test period: in October, 2012～2013 year May
Test site: Shandong Forest Science Academy's pilot scale base
Experimental cultivar: wheat (mountain agriculture 21)
Test soil: vegetable garden soil
Test design: dress seed in contrast with matrix substratum, dress seed as demonstration using the fermented liquid of different extent of dilution (diluting 10 times, 20 times, 30 times), three of each processing are parallel, community area 20m
2, stochastic distribution.
Rate of fertilizer application and fertilizing method: before sowing, adopt the fermented liquid seed dressing of different extent of dilution (diluting 10 times, 20 times, 30 times), dress seed in contrast with matrix substratum, other measures are same local, the field management measures such as unification is watered, weeding.
Economical character and yield and quality investigation: measure plant height, tiller number, effective fringe, the percentage of earbearing tiller, grain number per spike, thousand seed weight the day before yesterday in gathering in the crops.
Table 1 Agronomic Characters in Wheat questionnaire
Note: pt6 refers to efficient phosphate-solubilizing bacterium Burkholderia cepacia (B.cepacia) bacterial strain
As can be seen from Table 1, adopt the wheat of the diluent seed dressing of phosphate-solubilizing bacteria Burkholderia cepacia (B.cepacia) pt6 fermented liquid, its ratio aspect economical character has advantage clearly with the wheat of matrix substratum seed dressing.
After results wheat, measure the output of each community, completely count Different treatments, the difference of wheat yield simultaneously.As table 2.
Table 2 wheat yield questionnaire
As can be seen from Table 2, adopt the wheat of the diluent seed dressing of phosphate-solubilizing bacteria Burkholderia cepacia (B.cepacia) pt6 fermented liquid, its output is than high with the contrast of matrix substratum seed dressing, the wherein diluent seed dressing of 10 times, 20 times of phosphate-solubilizing bacteria Burkholderia cepacia (B.cepacia) pt6 fermented liquids and 30 times, the output of wheat increases by 16.7%, 13.2% and 8.7% than contrast respectively.The wheat of dressing seed with 10 times of diluents of phosphate-solubilizing bacteria Burkholderia cepacia (B.cepacia) pt6 fermented liquid, its output utmost point is significantly higher than contrast, and effect of increasing production is fairly obvious.
Because containing a large amount of phosphate solubilizing bacterias in phosphate-solubilizing bacteria Burkholderia cepacia (B.cepacia) pt6 fermented liquid, soil improvement is had to certain active effect, the molten phosphorus function that it has, to improving Soil structure, the specific absorption of enhancing crop on fertilizer etc. has very large positive impact.
Use phosphate-solubilizing bacteria Burkholderia cepacia (B.cepacia) pt6 fermented liquid, the improvement to soil and the growth of crop all have very large active effect.
Claims (7)
1. Burkholderia cepacia (Burkholderia cepacia) bacterial strain pt6, the deposit number of described bacterial strain is CGMCC No.7598.
2. Burkholderia cepacia claimed in claim 1 (Burkholderia cepacia) bacterial strain pt6 is improving the utilization ratio of phosphate fertilizer, is improving the purposes aspect Soil structure.
3. the purposes of Burkholderia cepacia claimed in claim 1 (Burkholderia cepacia) bacterial strain pt6 aspect output and the quality of raising crop.
4. a microbiobacterial agent, contains Burkholderia cepacia claimed in claim 1 (Burkholderia cepacia) bacterial strain pt6.
5. a method of preparing microbiobacterial agent claimed in claim 4, is characterized in that, comprises the following steps:
(1) seed shaking flask: the phosphate-solubilizing bacteria Burkholderia cepacia bacterial strain pt6 having activated is inoculated in the triangular flask that NA liquid nutrient medium is housed to 30 DEG C of 200r/min shaking culture 12h; Described NA liquid nutrient medium is: 0.5% peptone, and 0.3% extractum carnis, 0.5%NaCl, distilled water, pH is adjusted to 7.0;
(2) seeding tank fermentation: the shake-flask seed liquid that step (1) is prepared by volume 1% inoculum size is seeded to ferment in the seeding tank that seeding tank liquid fermentation medium is housed to thalline and enters logarithmic phase; Leavening temperature is 30～32 DEG C, and ventilating ratio is 0.5～0.8:1;
Described seeding tank liquid fermentation medium is: Semen Maydis powder 15g, glucose 4g, soybean cake powder 21g, fish meal 4g, CaCO
39g, (NH
4)
2sO
41.5g, K
2hPO
40.5g, MgSO
47H
2o0.1g, NaCl0.1g, water 1000mL, pH is adjusted to 7.5;
(3) fermentor tank: by the seed liquor of step (2) seeding tank by volume 10% inoculum size be seeded to fermentor tank and ferment, fermentation 20～30h; The same step of fermentation condition and medium component (2).
6. microbiobacterial agent claimed in claim 4 is improving the utilization ratio of phosphate fertilizer, is improving the purposes aspect Soil structure.
7. the purposes of microbiobacterial agent claimed in claim 4 aspect output and the quality of raising crop.
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| CN107988100B (en) * | 2017-12-05 | 2021-06-22 | 湖南豫园生物科技股份有限公司 | Inorganic phosphate solubilizing bacteria, microbial fertilizer and application |
| CN108265015B (en) * | 2018-01-09 | 2021-04-06 | 华中农业大学 | Multifunctional potassium-decomposing bacterium and application thereof |
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| CN109913393B (en) * | 2019-04-03 | 2022-05-17 | 贵州大学 | Burkholderia cepacia P10 and application thereof |
| CN112279710B (en) * | 2020-07-10 | 2022-05-24 | 江西农业大学 | Phosphorus-dissolving bacterial fertilizer and application thereof |
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| CN116218740B (en) * | 2023-04-03 | 2023-09-01 | 上海交通大学 | Burkholderia cepacia and application thereof |
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