CN107586743A - One plant of forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium and application - Google Patents
One plant of forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium and application Download PDFInfo
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- 238000004090 dissolution Methods 0.000 title claims abstract description 16
- 239000002689 soil Substances 0.000 claims abstract description 45
- 241000894006 Bacteria Species 0.000 claims abstract description 25
- 210000003608 fece Anatomy 0.000 claims abstract description 8
- 230000012010 growth Effects 0.000 claims abstract description 8
- 239000010871 livestock manure Substances 0.000 claims abstract description 8
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 6
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 6
- 230000001737 promoting effect Effects 0.000 claims abstract description 5
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 7
- 239000001506 calcium phosphate Substances 0.000 claims description 5
- 229910000389 calcium phosphate Inorganic materials 0.000 claims description 5
- 235000011010 calcium phosphates Nutrition 0.000 claims description 5
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims description 5
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- 229910052760 oxygen Inorganic materials 0.000 claims description 4
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- 229910052698 phosphorus Inorganic materials 0.000 abstract description 31
- 239000011574 phosphorus Substances 0.000 abstract description 31
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- 229910019142 PO4 Inorganic materials 0.000 description 10
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Abstract
The invention discloses one plant of forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium, deposit number is CCTCC NO.M 2017454, and Classification And Nomenclature is bacillus megaterium (Bacillus megaterium) L205.Experiment shows that the amount of phosphorus dissolved of the bacillus megaterium reaches 203.04mg/L, and its liquid bacterium solution can effectively dissolve slightly solubility Phos, increases the available phosphorus content in red-yellow soil soil, and promote the growth of Forest Seedlings.The present invention enriches the population storehouse of the bacillus megaterium of DSMZ, and the microbial population resource for preferably utilization bacillus megaterium provides conveniently.Simultaneously, Bacillus megatherium bacterial strain L205 can secrete organic acid, dissolved metals, improve plant and phosphorus element is absorbed, especially suitable for the forest bio-bacterial manure of south China Red-yellow and the production of forest growth promoting bacteria agent, composite microbial manure and microbial inoculum can be further developed.
Description
Technical field
The invention belongs to microbial technology field, more particularly to one plant of forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium and should
With.
Background technology
Rhizosphere is that soil-root system-microorganism three combines closely interactional place, and soil micro-ecosystem system
Important component.Sperber etc. has found that most of Soluble phosphorus microorganism appeared in plant rhizosphere soil, rhizosphere phosphorus-solubilizing bacteria with
Root system has the function that to mutually promote.Most of China soil is in the phosphorus state of lacking, more than 95% phosphorus in southern china soil
All it is that indissoluble state phosphorus thus can not directly be absorbed by plant, phosphorus element turns into the Main Factors for influenceing plant growth.Meanwhile south
It is Artificial Pure that fast-growing, high-yield woods is most of, and bio-diversity is simple, and the ecosystem is weak, and a large amount of irrational chemical fertilizer are applied
With causing eco-environmental quality to decline, getting worse the problems such as water and soil pollution and soil hardening.
Dry branches and fallen leaves is more in timbered soil, organic matter enriches, and edaphon is relative to agricultural soil more horn of plenty, Yin Rui
Age research is found, in the Soluble phosphorus microorganism of Chinese dryland soil, bacterial number proportion is maximum.The phosphorus-solubilizing bacteria reported at present
Species mainly has：Bacillus (Bacillus), Pseudomonas alba (Pseudomonas), salmonella (Salmonella), production
Alkali bacterium (Alcaligenes) etc., wherein bacillus megaterium (Bacillus megaterium) are to find earliest, Soluble phosphorus phosphorus effect
Fruit is best and is agriculturally applying the most commonly used bacterial strain of area.
Different soil environment climate characteristics, different plants its rhizosphere situations also difference, therefore special soil environment
It may result in the drawbacks such as poor, the strain mortality height of non-indigenous bacterial strain colonization ability.Guangxi soil is mainly red-yellow soil, red-yellow soil
And the typical soil types of south China.Existing report on China red-yellow soil area forest rhizosphere phosphorus-solubilizing bacteria is simultaneously few.
Due to its special soil-forming process, clay, iron oxide and aluminum oxide are higher in soil, and available phosphorus lacks, and governs this area woods
The raising of wooden accumulation.The isolated plant height effect phosphorus-solubilizing bacteria Y5 from the Yingtan rice soil of Jiangxi such as Dai Shenyan, it is to calcium phosphate
Amount of phosphorus dissolved be 159.1mg/L.At present, the domestic research for forest land red-yellow soil forest rhizosphere phosphorus-solubilizing bacteria is also fewer, screening
What is gone out derives from crops soil of ploughing, and the application effect in forest production is limited.
The content of the invention
The technical problem to be solved in the present invention is to provide one plant of effect of solubilizing phosphate is good, colonization ability is strong, have a wide range of application, produce
The low forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium of cost and application, the bacterial strain are given birth to especially suitable for south China Red-yellow forest
The production of thing bacterial manure and forest growth promoting bacteria agent.
In order to solve the above technical problems, the present invention uses following technical scheme：
One plant of forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium, deposit number are CCTCC N0.M 2017454, classification life
Entitled bacillus megaterium (Bacillus megaterium) L205.
Application of the above-mentioned forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium in slightly solubility Phos is dissolved.
Slightly solubility Phos comes from red-yellow soil soil.
Slightly solubility Phos is calcium phosphate.
In above-mentioned application, the numerous condition of expansion of Bacillus megatherium is 15~35 DEG C, pH 6.0~9.0 of temperature, dissolved oxygen 0.2
~2.0mg/L, carbon-nitrogen mass ratio 2: 1~5: 1.
Application of the above-mentioned forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium in Forest Seedlings growth is promoted.
Forest Seedlings are grown on red-yellow soil soil.
In above-mentioned application, the numerous condition of expansion of Bacillus megatherium is 15~35 DEG C, pH 6.0~9.0 of temperature, dissolved oxygen 0.2
~2.0mg/L, carbon-nitrogen mass ratio 2: 1~5: 1.
Above-mentioned forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium is used to produce forest bio-bacterial manure or forest growth promoting bacteria agent.
Bacillus megaterium L205 or Mixed Microbes sum are no less than 0.2*108cfu/mL(g)。
Inventor's isolation and selection from the forest rhizosphere soil of Guangxi Red-yellow obtains one plant of effective Soluble phosphorus bacterial strain, passes through
Sequence progress Blast in 16SrDNA and GenBank, which is compared, is accredited as bacillus megaterium.The forest rhizosphere efficient phosphorus-dissolution is huge
Big bacillus, deposit number are CCTCC N0.M 2017454, and Classification And Nomenclature is bacillus megaterium (Bacillus
megaterium)L205.In the research of bacterial strain application, inventor is cultivated by purebred expansion, liquid fermentation process optimization, is obtained
Obtain the strain and expand numerous technology.Experiment shows that the amount of phosphorus dissolved of the bacillus megaterium reaches 203.04mg/L, its liquid bacteria
Liquid can effectively dissolve slightly solubility Phos, increase the available phosphorus content in red-yellow soil soil, and promote the growth of Forest Seedlings.
Soluble phosphorus bacterial strain of the present invention enriches the population storehouse of the bacillus megaterium of DSMZ, preferably to develop huge bud
The microbial population resource of spore bacillus provides conveniently.Meanwhile Bacillus megatherium bacterial strain L205 can secrete organic acid, nothing is dissolved
Machine phosphorus, improve plant and phosphorus element is absorbed, especially suitable for the forest bio-bacterial manure of south China Red-yellow and forest Promoting bacteria
The production of agent, it can further develop composite microbial manure and microbial inoculum.In a word, forest rhizosphere efficient phosphorus-dissolution of the invention is huge
Big bacillus effect of solubilizing phosphate is good, colonization ability is strong, have a wide range of application, production cost is low, has larger market application potential.
Brief description of the drawings
Fig. 1 is bacillus megaterium L205 of the present invention phylogenetic tree.
Fig. 2 is bacillus megaterium L205 of the present invention Soluble phosphorus result figure, in figure：A flat board front elevations, B flat board back views.
Preservation information explanation
Bacillus megaterium (Bacillus megaterium) L205, deposit number are CCTCC NO.M 2017454, are protected
Hide the date：On August 28th, 2017, preservation address are：Wuhan City, Hubei Province Wuchang District Bayi Road Luo Jia Shan, postcode 430072, protect
Hide unit：China typical culture collection center.
Embodiment
With reference to embodiments and accompanying drawing, the present invention is described in further detail.
First, the bacterium source of bacillus megaterium, seed selection, gene sequencing
Bacterium source：Bacillus megaterium (Bacillus megaterium) L205 of the present invention is from Guangxi red-yellow soil
In area's forest rhizosphere soil obtained by isolation and selection, preservation information is same as above.
Bacillus megaterium L205 morphological feature is：Colony colour is white, and thalline is in shaft-like, there is gemma.
Bacillus megaterium L205 physiological and biochemical property is：Gram's staining is positive, and contacts enzyme positive, oxidizing ferment sun
Property.
Bacillus megaterium L205 is obtained by following specific separating step：
A soil collections：Adopt the forest Rhizosphere Soil that the ground such as Guangxi Red-yellow Wuzhou, Ningming, Luzhai, Tianlin County grow fine
Earth, 0~20cm rhizospheres non-rhizosphere soils and root system sample are taken, be fitted into the freshness protection package without opening, labelled, sealing,
It is placed in ice bag and takes back laboratory progress Soluble phosphorus strain isolation at once.
The indices measurement result of pedotheque is shown in Table 1.As shown in Table 1：Gather from Guangxi Wuzhou, Ningming, Luzhai, field
The content of organic matter of woods soil sample is both greater than 36.75mg/kg, and the content of organic matter of four pedotheques is in 36.75-56.51mg/kg
Between, the minimum soil sample of the content of organic matter picks up from Luzhai.The soil total phosphorus content highest in Wuzhou is picked up from, is 1.48g/kg, picks up from
The quick-acting contents of samples-soil of Tianlin County are that highest is 16.53mg/kg.The pH value of each soil sample slant acidity between 4.15-5.02.
Table 1 picks up from the soil sample analysis result of four, Guangxi different zones
B strain isolations：The eucalyptus rhizosphere soil of different regions is weighed into sample 5g, is placed in and has sterilized equipped with 45mL
In the 150mL triangular flasks of 0.85%NaCl solution, 30min is vibrated, suspension is made.1mL is therefrom drawn with 1mL micro sample adding appliances
Suspension is injected in the test tube for filling 0.85% sterile NaCl solutions of 9mL, is fully mixed, then draws 1mL from this test tube, is injected
In another test tube for filling 0.85% sterile NaCl solutions of 9mL, by that analogy, content (g) and NaCl solution (mL) ratio is made
It is followed successively by 10-3、10-4、10-53 kinds of dilution gradients Soil Slurry.50 μ L coatings are drawn, each gradient repeats 3, and makees
Good mark.28 DEG C of constant temperature, 2~5d is cultivated, screening on the culture medium of phosphorus-solubilizing bacteria has the bacterial strain of Soluble phosphorus circle according to Soluble phosphorus loop diameter
(D) and colony diameter (d) ratio size be HE values determine Soluble phosphorus power, by the stronger bacterial strain of phosphate solubilization, be inoculated into LB respectively
Purified, be then saved on LB slant mediums on flat board, with standby in 4 DEG C of refrigerator.
C phosphate solubilizations determine：100mL liquid PKO culture mediums are loaded into 250mL triangular flasks, 121 DEG C of sterilizing 20min, cooling
Afterwards, by each strain to be tested bacteria suspensions (10 of 1mL8Cfu/mL) it is seeded in triangular flask.Each bacterial strain sets 3 repetitions, control
Not connecing bacterium adds 1ml sterilized waters to replace.Above-mentioned triangular flask is placed in 28 DEG C, 160r/min shaking table culture 3d, is measured and cultivated with pH meter
Liquid pH value.And 10min is centrifuged in 4 DEG C of 100 00r/min, supernatant 5mL molybdenum antimony scandium colorimetric methods are taken, are surveyed under wavelength 700nm
Determine available phosphorus content.
D Liquid Cultures：From the ring bacillus megaterium of single bacterium colony test tube slant picking one, it is inoculated in fluid nutrient medium,
On 150-220rpm rotating speed shaking tables, 32 DEG C are cultivated 24 hours, and viable count reaches 6 × 108Cfu/mL or so.
E bacterial strains preserve：Aforesaid liquid is fermented to obtain the addition of bacillus megaterium bacterium solution by the glycerine of high-temperature sterilization, made
Final concentration of 20% (V/V) of the glycerine in seed liquor is obtained, bacillus megaterium Freezing Glycerine pipe is made, is stored in -80 DEG C of ice
Case preserves, or is stored in -196 DEG C of liquid nitrogen containers and carries out cryopreservation.
F gene sequencings：Hai Meiji biotech companies are served into bacillus megaterium test tube slant and carry out 16sRNA
Gene sequencing, its 16s rRNA gene order such as sequence table SEQ .ID.No.1 base sequence.Its phylogenetic tree such as Fig. 1.
2nd, Soluble phosphorus is tested
PKO (Pikovskaya ' s Medium) culture medium：Glucose 10g, calcium phosphate 3g, ammonium sulfate 0.5g, sodium chloride
0.1g, epsom salt 0.1g, potassium chloride 0.2g, manganese sulfate 0.004g, ferrous sulfate 0.002g, yeast extract 0.5g, water
1000ml, pH7.2.
Bacillus megaterium CCTCC M 2017454 are inoculated on PK0 culture mediums, 30 DEG C of culture 5d, determine its Soluble phosphorus
Circle is that the ratio of transparent loop diameter (D)/bacterium colony size (d) is 2.67.Show that the bacillus megaterium has good Soluble phosphorus effect
Fruit.3rd, amount of phosphorus dissolved determines
PKO (Pikovskaya ' s Medium) culture medium：Glucose 10g, calcium phosphate 3g, ammonium sulfate 0.5g, sodium chloride
0.1g, epsom salt 0.1g, potassium chloride 0.2g, manganese sulfate 0.004g, ferrous sulfate 0.002g, yeast extract 0.5g, water
1000ml, pH7.2.
100mL liquid PK0 culture mediums are loaded into 250mL triangular flasks, 121 DEG C of sterilizing 20min, after cooling, by the huge buds of 1mL
Spore bacillus CCTCC M 2017454 (108Cfu/mL) it is seeded in triangular flask.Each bacterial strain sets 3 repetitions, and control does not connect
Bacterium adds 1ml sterilized waters to replace.Above-mentioned triangular flask is placed in 28 DEG C, 160r/min shaking table culture 3d, nutrient solution pH is measured with pH meter
Value.And 10min is centrifuged in 4 DEG C of 100 00r/min, supernatant 5mL molybdenum antimony scandium colorimetric methods are taken, being determined under wavelength 700nm has
Imitate phosphorus content.
Pass through L205 phosphate solubilizations under the conditions of liquid submerged culture, the results showed that the available phosphorus of the bacterial strain dissolved metals bacterium
Increment is for 203.04mg/L.
4th, the preparation of forest phosphate solubilizing microorganism bacterium solution
(1) prepared by seed
Seed culture medium：Tryptone 10g, yeast extract powder 5g, NaCl 10g, pH value 7.0, distilled water 1000mL.Will
Bacillus megaterium CCTCC M 2017454 are inoculated with 1 ring in the 250mL triangular flasks equipped with the above-mentioned seed culture mediums of 100mL,
Under rotary shaker 180rpm rotating speeds, 30 DEG C of culture 24h, that is, the seed suitable for inoculation is obtained.
(2) liquid fermentation and culture
Fermentation medium：Tryptone 10g, yeast extract powder 5g, NaCl 10g, pH value 7.0, distilled water 1000mL.Will
(1) the bacillus megaterium seed obtained in is inoculated in liquid fermentation medium according to the 5% of fermentation medium volume, is being revolved
Under rotatable shaking table 180rpm rotating speeds, 30 DEG C of culture 48h.
(3) pH value of zymotic fluid is adjusted after fermentation ends between 5.0-6.0, you can obtain liquid bacterial agent.
5th, the preparation of forest phosphate solubilizing microorganism microbial inoculum
(1) prepared by seed
Seed culture medium：Tryptone 10g, yeast extract powder 5g, NaCl 10g, pH value 7.0, distilled water 1000mL.Will
Bacillus megaterium CCTCC M 2017454 are inoculated with 1 ring in the 250mL triangular flasks equipped with the above-mentioned seed culture mediums of 100mL,
Under rotary shaker 180rpm rotating speeds, 30 DEG C of culture 24h, that is, the seed suitable for inoculation is obtained.
(2) liquid fermentation and culture
Fermentation medium：Tryptone 10g, yeast extract powder 5g, NaCl 10g, pH value 7.0, distilled water 1000mL.Will
(1) the bacillus megaterium seed obtained in is inoculated in liquid fermentation medium according to the 5% of fermentation medium volume, is being revolved
Under rotatable shaking table 180rpm rotating speeds, 30 DEG C of culture 48h.
(3) after fermentation ends, take liquid fermentation medium to be placed in supercentrifuge and centrifuge 5 minutes, remove supernatant, add
Enter 1% Tween 80, be freeze-dried, that is, obtain forest phosphate solubilizing microorganism microbial inoculum.
6th, application of the forest phosphate solubilizing microorganism bacterium solution in the cultivation of forest cup seedling
Using pouring root inocalation method, bacterium solution obtained by step 4 is diluted 10 times with sterilized water.Take pine tree, the China fir of growing way identical
Wooden seedling, the bacterium solution poured after 10mL dilutions, sterilized water 50mL is poured every 2d, is affixed by the Glan nutrition suddenly of scarce phosphorus every 10d
Liquid.Each concentration handles 25 plants, and using clear water as control, test temperature is 15~30 DEG C.Each processing nursery stock is calculated after inoculation 30d to deposit
Work number, plant average height of seedling, ground diameter.
Measurement result shows：The pine tree cup seedling of phosphate solubilizing microorganism microbial inoculum is added 12.3% than control group height of seedling gain, ground
Footpath gain 10.6%, China fir cup seedling is than control group height of seedling gain 9.5%, ground diameter gain 12.8%.Illustrate the present invention's
Microorganism formulation has relatively good growth-promoting effect to forest.
Sequence table
<110>Inst. of Forestry Science, Guangxi Zhuang Autonomous Region
<120>One plant of forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium and application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1398
<212> DNA
<213>Bacillus megaterium L205 (Bacillus megaterium L205)
<400> 1
gcagtcgagc gaactgatta gaagcttgct tctatgacgt tagcggcgga cgggtgagta 60
acacgtgggc aacctgcctg taagactggg ataacttcgg gaaaccgaag ctaataccgg 120
ataggatctt ctccttcatg ggagatgatt gaaagatggt ttcggctatc acttacagat 180
gggcccgcgg tgcattagct agttggtgag gtaacggctc accaaggcaa cgatgcatag 240
ccgacctgag agggtgatcg gccacactgg gactgagaca cggcccagac tcctacggga 300
ggcagcagta gggaatcttc cgcaatggac gaaagtctga cggagcaacg ccgcgtgagt 360
gatgaaggct ttcgggtcgt aaaactctgt tgttagggaa gaacaagtac gagagtaact 420
gcttgtacct tgacggtacc taaccagaaa gccacggcta actacgtgcc agcagccgcg 480
gtaatacgta ggtggcaagc gttatccgga attattgggc gtaaagcgcg cgcaggcggt 540
ttcttaagtc tgatgtgaaa gcccacggct caaccgtgga gggtcattgg aaactgggga 600
acttgagtgc agaagagaaa agcggaattc cacgtgtagc ggtgaaatgc gtagagatgt 660
ggaggaacac cagtggcgaa ggcggctttt tggtctgtaa ctgacgctga ggcgcgaaag 720
cgtggggagc aaacaggatt agataccctg gtagtccacg ccgtaaacga tgagtgctaa 780
gtgttagagg gtttccgccc tttagtgctg cagctaacgc attaagcact ccgcctgggg 840
agtacggtcg caagactgaa actcaaagga attgacgggg gcccgcacaa gcggtggagc 900
atgtggttta attcgaagca acgcgaagaa ccttaccagg tcttgacatc ctctgacaac 960
tctagagata gagcgttccc cttcggggga cagagtgaca ggtggtgcat ggttgtcgtc 1020
agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt gatcttagtt 1080
gccagcattt agttgggcac tctaaggtga ctgccggtga caaaccggag gaaggtgggg 1140
atgacgtcaa atcatcatgc cccttatgac ctgggctaca cacgtgctac aatggatggt 1200
acaaagggct gcaagaccgc gaggtcaagc caatcccata aaaccattct cagttcggat 1260
tgtaggctgc aactcgccta catgaagctg gaatcgctag taatcgcgga tcagcatgcc 1320
gcggtgaata cgttcccggg ccttgtacac accgcccgtc acaccacgag agtttgtaac 1380
acccgaagtc ggtggagt 1444
Claims (10)
1. one plant of forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium, it is characterised in that deposit number is CCTCC NO.M
2017454, Classification And Nomenclature is bacillus megaterium (Bacillus megaterium) L205.
2. application of the forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium described in claim 1 in slightly solubility Phos is dissolved.
3. application according to claim 2, it is characterised in that：The slightly solubility Phos comes from red-yellow soil soil.
4. application according to claim 2, it is characterised in that：The slightly solubility Phos is calcium phosphate.
5. according to any described application of claim 2 to 4, it is characterised in that：The numerous condition of expansion of the Bacillus megatherium is
15~35 DEG C, pH 6.0~9.0,0.2~2.0mg/L of dissolved oxygen of temperature, carbon-nitrogen mass ratio 2: 1~5: 1.
6. application of the forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium described in claim 1 in Forest Seedlings growth is promoted.
7. application according to claim 6, it is characterised in that：The Forest Seedlings are grown on red-yellow soil soil.
8. according to any described application of claim 6 to 7, it is characterised in that：The numerous condition of expansion of the Bacillus megatherium is
15~35 DEG C, pH 6.0~9.0,0.2~2.0mg/L of dissolved oxygen of temperature, carbon-nitrogen mass ratio 2: 1~5: 1.
9. forest rhizosphere efficient phosphorus-dissolution Bacillus megatherium described in claim 1 is used to produce forest bio-bacterial manure or forest growth-promoting
Microbial inoculum.
10. forest bio-bacterial manure according to claim 9 or forest growth promoting bacteria agent, it is characterised in that：The huge gemma
Bacillus L205 or Mixed Microbes sum are no less than 0.2*108cfu/mL(g)。
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108676744A (en) * | 2018-05-16 | 2018-10-19 | 南京农业大学 | A kind of bacillus megaterium bacterial strain ZT-P and its application |
| CN110684696A (en) * | 2019-11-11 | 2020-01-14 | 乔志伟 | Bacillus megaterium QZY-3 and application thereof |
| CN111676164A (en) * | 2020-06-19 | 2020-09-18 | 中国林业科学研究院林业研究所 | Phosphorus-solubilizing bacterium RP2, fermentation product, microbial inoculum and application thereof |
| CN112852687A (en) * | 2021-04-15 | 2021-05-28 | 河南科技学院 | Peony rhizosphere bacillus megatherium FD3-13 for three-year-old oil and application |
| CN114574395A (en) * | 2022-03-17 | 2022-06-03 | 广西壮族自治区林业科学研究院 | Bacillus megaterium and biofertilizer capable of antagonizing colletotrichum gloeosporioides |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103320360A (en) * | 2013-06-25 | 2013-09-25 | 云南省烟草公司玉溪市公司 | Bacillus megatherium SJ-7 strain and application thereof |
| CN103992963A (en) * | 2014-03-19 | 2014-08-20 | 南京博农生物科技有限公司 | Bacillus megaterium and application thereof |
| CN104560787A (en) * | 2014-12-13 | 2015-04-29 | 郑州市污水净化有限公司 | Peanut rhizosphere growth-promoting bacteria HS11 and application thereof |
-
2017
- 2017-10-20 CN CN201710989469.6A patent/CN107586743B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103320360A (en) * | 2013-06-25 | 2013-09-25 | 云南省烟草公司玉溪市公司 | Bacillus megatherium SJ-7 strain and application thereof |
| CN103992963A (en) * | 2014-03-19 | 2014-08-20 | 南京博农生物科技有限公司 | Bacillus megaterium and application thereof |
| CN104560787A (en) * | 2014-12-13 | 2015-04-29 | 郑州市污水净化有限公司 | Peanut rhizosphere growth-promoting bacteria HS11 and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| ZHONG CHUAN-QING等: "Dissolving mechanism of strain P17 on insoluble phosphorus of yellow-brown soil", 《BRAZILIAN JOURNAL OF MICROBIOLOGY》 * |
| 张维娜等: "巨大芽孢杆菌JD-2的解磷效果及对土壤有效磷化的研究", 《吉林农业科学》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108676744A (en) * | 2018-05-16 | 2018-10-19 | 南京农业大学 | A kind of bacillus megaterium bacterial strain ZT-P and its application |
| CN108676744B (en) * | 2018-05-16 | 2020-07-28 | 南京农业大学 | Bacillus megaterium strain ZT-P and application thereof |
| CN110684696A (en) * | 2019-11-11 | 2020-01-14 | 乔志伟 | Bacillus megaterium QZY-3 and application thereof |
| CN110684696B (en) * | 2019-11-11 | 2022-04-19 | 安顺学院 | Bacillus megaterium QZY-3 and application thereof |
| CN111676164A (en) * | 2020-06-19 | 2020-09-18 | 中国林业科学研究院林业研究所 | Phosphorus-solubilizing bacterium RP2, fermentation product, microbial inoculum and application thereof |
| CN111676164B (en) * | 2020-06-19 | 2022-05-20 | 中国林业科学研究院林业研究所 | Phosphate solubilizing bacterium RP2, and fermentation product, microbial inoculum and application thereof |
| CN112852687A (en) * | 2021-04-15 | 2021-05-28 | 河南科技学院 | Peony rhizosphere bacillus megatherium FD3-13 for three-year-old oil and application |
| CN114574395A (en) * | 2022-03-17 | 2022-06-03 | 广西壮族自治区林业科学研究院 | Bacillus megaterium and biofertilizer capable of antagonizing colletotrichum gloeosporioides |
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