CN111484951A - Bacillus for dissolving phosphorus and fixing nitrogen and application thereof in growth promotion - Google Patents
Bacillus for dissolving phosphorus and fixing nitrogen and application thereof in growth promotion Download PDFInfo
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- CN111484951A CN111484951A CN201910082120.3A CN201910082120A CN111484951A CN 111484951 A CN111484951 A CN 111484951A CN 201910082120 A CN201910082120 A CN 201910082120A CN 111484951 A CN111484951 A CN 111484951A
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- bacillus
- phosphorus
- growth
- nitrogen
- promoting
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 title claims abstract description 90
- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 66
- 229910052698 phosphorus Inorganic materials 0.000 title claims abstract description 59
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 title claims abstract description 57
- 239000011574 phosphorus Substances 0.000 title claims abstract description 57
- 229910052757 nitrogen Inorganic materials 0.000 title claims abstract description 47
- 230000012010 growth Effects 0.000 title claims abstract description 28
- 230000008635 plant growth Effects 0.000 claims abstract description 12
- 238000004321 preservation Methods 0.000 claims abstract description 10
- 244000005700 microbiome Species 0.000 claims abstract description 6
- 238000000855 fermentation Methods 0.000 claims description 25
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- 239000007788 liquid Substances 0.000 claims description 24
- 241000196324 Embryophyta Species 0.000 claims description 22
- 230000001737 promoting effect Effects 0.000 claims description 22
- 230000003381 solubilizing effect Effects 0.000 claims description 19
- 229910019142 PO4 Inorganic materials 0.000 claims description 17
- 230000035784 germination Effects 0.000 claims description 16
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 15
- 239000010452 phosphate Substances 0.000 claims description 15
- 239000002068 microbial inoculum Substances 0.000 claims description 6
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- 229910052921 ammonium sulfate Inorganic materials 0.000 description 3
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- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
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- 239000011609 ammonium molybdate Substances 0.000 description 2
- 235000018660 ammonium molybdate Nutrition 0.000 description 2
- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical compound [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 description 2
- 229940010552 ammonium molybdate Drugs 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- RJGDLRCDCYRQOQ-UHFFFAOYSA-N anthrone Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 RJGDLRCDCYRQOQ-UHFFFAOYSA-N 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
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- 235000013311 vegetables Nutrition 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- UFBJCMHMOXMLKC-UHFFFAOYSA-N 2,4-dinitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1[N+]([O-])=O UFBJCMHMOXMLKC-UHFFFAOYSA-N 0.000 description 1
- JCRIDWXIBSEOEG-UHFFFAOYSA-N 2,6-dinitrophenol Chemical compound OC1=C([N+]([O-])=O)C=CC=C1[N+]([O-])=O JCRIDWXIBSEOEG-UHFFFAOYSA-N 0.000 description 1
- AGJBKFAPBKOEGA-UHFFFAOYSA-M 2-methoxyethylmercury(1+);acetate Chemical compound COCC[Hg]OC(C)=O AGJBKFAPBKOEGA-UHFFFAOYSA-M 0.000 description 1
- -1 Anthracene ketone Chemical class 0.000 description 1
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Natural products OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- JJLJMEJHUUYSSY-UHFFFAOYSA-L Copper hydroxide Chemical compound [OH-].[OH-].[Cu+2] JJLJMEJHUUYSSY-UHFFFAOYSA-L 0.000 description 1
- 239000005750 Copper hydroxide Substances 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 241000589157 Rhizobiales Species 0.000 description 1
- 240000004922 Vigna radiata Species 0.000 description 1
- 235000010721 Vigna radiata var radiata Nutrition 0.000 description 1
- 235000011469 Vigna radiata var sublobata Nutrition 0.000 description 1
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- MWPLVEDNUUSJAV-UHFFFAOYSA-N anthracene Natural products C1=CC=CC2=CC3=CC=CC=C3C=C21 MWPLVEDNUUSJAV-UHFFFAOYSA-N 0.000 description 1
- 229940026189 antimony potassium tartrate Drugs 0.000 description 1
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- 229960005070 ascorbic acid Drugs 0.000 description 1
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- WBTCZEPSIIFINA-MSFWTACDSA-J dipotassium;antimony(3+);(2r,3r)-2,3-dioxidobutanedioate;trihydrate Chemical compound O.O.O.[K+].[K+].[Sb+3].[Sb+3].[O-]C(=O)[C@H]([O-])[C@@H]([O-])C([O-])=O.[O-]C(=O)[C@H]([O-])[C@@H]([O-])C([O-])=O WBTCZEPSIIFINA-MSFWTACDSA-J 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
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- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- TYQCGQRIZGCHNB-JLAZNSOCSA-N l-ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(O)=C(O)C1=O TYQCGQRIZGCHNB-JLAZNSOCSA-N 0.000 description 1
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- 230000007774 longterm Effects 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- VTHJTEIRLNZDEV-UHFFFAOYSA-L magnesium dihydroxide Chemical compound [OH-].[OH-].[Mg+2] VTHJTEIRLNZDEV-UHFFFAOYSA-L 0.000 description 1
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- 229910001862 magnesium hydroxide Inorganic materials 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 1
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- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
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- 238000011392 neighbor-joining method Methods 0.000 description 1
- 150000007523 nucleic acids Chemical group 0.000 description 1
- VSIIXMUUUJUKCM-UHFFFAOYSA-D pentacalcium;fluoride;triphosphate Chemical compound [F-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O VSIIXMUUUJUKCM-UHFFFAOYSA-D 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 230000000413 phospholytic effect Effects 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- LJCNRYVRMXRIQR-OLXYHTOASA-L potassium sodium L-tartrate Chemical compound [Na+].[K+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O LJCNRYVRMXRIQR-OLXYHTOASA-L 0.000 description 1
- 229940074439 potassium sodium tartrate Drugs 0.000 description 1
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- 230000035040 seed growth Effects 0.000 description 1
- HELHAJAZNSDZJO-OLXYHTOASA-L sodium L-tartrate Chemical compound [Na+].[Na+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O HELHAJAZNSDZJO-OLXYHTOASA-L 0.000 description 1
- 235000011006 sodium potassium tartrate Nutrition 0.000 description 1
- 239000001433 sodium tartrate Substances 0.000 description 1
- 229960002167 sodium tartrate Drugs 0.000 description 1
- 235000011004 sodium tartrates Nutrition 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
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- 239000011550 stock solution Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 229910000391 tricalcium phosphate Inorganic materials 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
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- 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
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- 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
- C12R2001/07—Bacillus
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- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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- C12N1/20—Bacteria; Culture media therefor
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
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Abstract
The invention provides a phosphate-solubilizing nitrogen-fixing Bacillus and application thereof in growth promotion, wherein the strain is Siamese Bacillus FJAT-47169 with the scientific name of Bacillus siemensis FJAT-47169 which is preserved in the common microorganism center of China Committee for culture Collection of microorganisms with the preservation number of CGMCC NO. 16250. The bacillus of the invention can not only dissolve phosphorus but also fix nitrogen, can improve soil fertility, provide sufficient nutrition for plant growth, can also obviously promote the growth of crops, develop root systems of the crops and enhance the stress resistance of the crops.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a bacillus strain for dissolving phosphorus and fixing nitrogen and application thereof in promoting plant growth.
Background
The phosphorus supply level of soil is one of the key factors influencing the growth of plants, 95 percent of phosphorus in the soil is in an invalid form, and the plants are difficult to directly absorb and utilize, so that the phosphorus deficiency phenomenon exists in 74 percent of cultivated land soil in China.
In a crop-microorganism interaction system, Plant growth-promoting rhizobacteria (PGPR) are colonized in the rhizosphere soil of crops, and can effectively decompose insoluble and fixed elements (phosphorus, potassium and the like) in the soil, promote the absorption of the crops on fertilizers and elements in the soil, and further promote the growth, yield increase, disease resistance and the like of the crops. Therefore, the microbial fertilizer with the efficient growth promoting function is screened and developed and applied to agricultural production, potential element resources of soil are fully utilized, and the microbial fertilizer has important significance for improving element shortage of soil such as phosphorus and potassium and the like, reducing environmental pollution and promoting agricultural sustainable development.
Disclosure of Invention
Therefore, a strain for promoting growth, dissolving phosphorus and fixing nitrogen is needed to be provided, and the problem that elements such as phosphorus, potassium and the like in soil cannot be absorbed and utilized by plants is solved.
In order to achieve the purpose, the inventor provides the following technical scheme:
a bacillus for dissolving phosphorus and fixing nitrogen is characterized in that: the Bacillus is Siamese Bacillus FJAT-47169 with the scientific name of Bacillus siemensis FJAT-47169, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, has the preservation number of CGMCC NO.16250, the preservation date of 2018, month 13 and the preservation address of the institute of microorganisms of China national academy of sciences in Beijing, China.
The colony morphology of the bacillus FJAT-47169 is as follows: round, light yellow, opaque, well-defined, protruding, moist microcolonies.
Further, application of the bacillus for dissolving phosphorus and fixing nitrogen in degradation of insoluble phosphate
Further, the application of the bacillus for solubilizing phosphorus and fixing nitrogen in promoting plant seed germination is realized by the specific method that the bacillus fermentation liquor is prepared into the bacillus fermentation liquor with the concentration of 1 × 105-3×105cfu/m L, soaking the strain for 2-3 days, and placing at 25-30 deg.C under illumination for 16-20 h/day.
Further, the application of the bacillus for dissolving phosphorus and fixing nitrogen in promoting the growth of plant seedlings is realized by the specific method that the bacillus fermentation liquor is prepared into the fermentation liquor with the concentration of 0.8 × 106-1.2×106cfu/m L, planting the young plant in planting medium for 6-8 days, irrigating root with bacterial suspension once every 5-7 days, irrigating once every 2-3 days, and irrigating plant nutrient solution (such as Hoagland solution) once every 10-14 days to supplement nutrient elements.
The planting matrix comprises N, P, K and organic matters, wherein N, P, K mass percent is not less than 3%, 3545% of the organic matters, and the pH value of the matrix is 5.5-6.5. The planting substrate comprises a vegetable planting substrate.
Further, the application of the bacillus for dissolving phosphorus and fixing nitrogen in plant nitrogen fixation.
Further, the bacillus for solubilizing phosphorus and fixing nitrogen is applied to preparation of the composite microbial inoculum for solubilizing phosphorus and fixing nitrogen.
A plant growth promoting microbial inoculum comprises the bacillus.
The invention has the beneficial effects that:
(1) the bacillus of the invention can effectively degrade inorganic phosphorus, promote insoluble phosphate to release phosphorus, and improve the content of soluble phosphorus in soil, thereby obviously promoting the growth of crops, enabling the roots of the crops to be developed, and enhancing the stress resistance of the crops.
(2) The bacillus of the invention can improve the activity of plant seeds, promote the seeds to take root and sprout, and effectively shorten the growth cycle of plants.
(3) The bacillus of the invention has double effects of dissolving phosphorus and fixing nitrogen, and can be used as a microbial organic fertilizer for improving soil fertility. Nitrogen and phosphorus are essential elements for plant growth, and the bacteria capable of fixing nitrogen and dissolving phosphorus can provide sufficient nutrition for plants in soil deficient in phosphorus and less in nitrogen and improve the soil nutrition structure. If different azotobacter and phosphate-solubilizing bacteria are prepared into bacterial manure, competition between two or more bacteria may exist, and the effects of bacterial strain colonization and phosphate-solubilizing and nitrogen-solubilizing are influenced. The bacterial manure prepared from the bacterial strain with the functions of dissolving phosphorus and fixing nitrogen can avoid the competition effect and better provide nutrition for the growth of plants.
Drawings
FIG. 1 shows colony morphology of Bacillus strain FJAT-47169, wherein the left image shows colony growth state of FJAT-47169 strain on a whole plate, and the right image shows a partial enlarged view of the colony on the left image.
FIG. 2 is a tree showing the results of identifying the 16S rRNA sequence of Bacillus FJAT-47169 in accordance with an embodiment.
FIG. 3 shows the effect of different concentrations of Bacillus FJAT-47169 on the growth of tomato seeds.
FIG. 4 shows the effect of different concentrations of Bacillus FJAT-47169 on germination index and germination rate of tomato seeds.
FIG. 5 shows the effect of different concentrations of Bacillus FJAT-47169 on the root length, stem length and vigor of tomato seeds.
FIG. 6 shows the growth of tomato seedlings according to the embodiment, in which the left side of the diagram is CK-W group and the right side is B-1 group.
FIG. 7 shows the effect of Bacillus FJAT-47169 on the plant height, root length and stem thickness of tomato seedlings.
FIG. 8 shows the effect of Bacillus FJAT-47169 on the fresh weight of tomato plants according to an embodiment.
FIG. 9 is a graph showing the effect of Bacillus FJAT-47169 on the dry weight of tomato plants according to an embodiment.
Detailed Description
To explain technical contents, achieved objects and effects of the technical solutions in detail, the following detailed description is given with reference to the accompanying drawings in combination with specific embodiments.
EXAMPLE 1 phosphate solubilization of Bacillus
1. Test materials
1.1 test strains
Test strains: the bacillus FJAT-47169 is separated from soil samples near Mema day slaughtered in double lakes of Tibet, is frozen and preserved in glycerol at-80 ℃ in the China general microbiological culture Collection center with the preservation number of CGMCC No. 16250.
1.2 culture Medium
L B solid culture medium (purchased from the manufacturer) formula, i.e. 10g tryptone, 10g sodium chloride, 5g yeast powder, 15g agar, 1000m L water, pH 7.0, (2) L B liquid culture medium (purchased from the manufacturer) formula, i.e. 10g tryptone, 10g sodium chloride, 5g yeast powder, 1000m L water, pH 7.0.
Inorganic phosphorus medium (NBRIP medium): glucose 10g, Ca3(PO4)25g，MgCl2·6H2O 5g，KCl0.2g，MgSO4·7H2O 0.25g，(NH4)2SO40.1g, distilled water 1000m L, natural pH.
The organic phosphorus growth medium comprises 10g of glucose, 0.5g of ammonium sulfate, 0.5g of yeast extract powder, 0.3g of sodium chloride, 0.3g of potassium chloride, 0.3g of magnesium sulfate, 0.03g of ferrous sulfate, 0.03g of manganese sulfate, 0.2g of lecithin, 1.0g of calcium carbonate, 1000m of distilled water L, 15g of agar and pH 7.0-7.5.
1.3 preparation of test reagents
153m L concentrated sulfuric acid (analytically pure, density 1.84g/m L) is measured, slowly added into 400m L distilled water, continuously stirred and cooled, 10g of ground ammonium molybdate is also weighed and poured into the solution, stirred and dissolved, 0.5% (5 g/L) of antimony potassium tartrate solution 100m L is added, after cooling, water is added to dilute the solution to 1000m L, the solution is shaken evenly and stored in a brown reagent bottle, and the stock solution contains 1% of ammonium molybdate and 2.75mo L/L of sulfuric acid.
The molybdenum-antimony color-developing agent is prepared by weighing 1.50g ascorbic acid and dissolving in 100m L molybdenum-antimony storage solution, wherein the solution has short effective period and is suitable for use.
5 mg/L phosphorus Standard solution 0.4394g of potassium dihydrogen phosphate (KH) dried at 50 deg.C2PO4Analytically pure), 100m L water, 5m L concentrated sulfuric acid (preservation), water to make the volume 1L, the concentration of phosphorus 100 mg/L, the solution can be preserved for a long time, the solution 5m L is absorbed into a 100m L volumetric flask, water to make the volume 5 mg/L phosphorus standard solution, the solution is not suitable for long-term preservation.
2. Test method
2.1 determination of phosphate solubilizing ability
2.1.1 activation of the test strains
Taking out the test strain from a refrigerator at the temperature of minus 80 ℃, after the test strain is warmed to room temperature, streaking the test strain in an ultra-clean bench into a L B solid agar medium plate, inversely placing the test strain in a biological incubator for 2 d.2d at the temperature of 30 ℃, observing the colony morphology, selecting a single colony for secondary streaking culture, ensuring that the activated colony morphology is single, selecting a proper amount of the single colony in a L B liquid medium, and performing shake culture at the temperature of 30 ℃ for 2d to obtain a seed solution.
2.1.2 liquid Shake flask fermentation
Diluting the seed liquid by 2 times, and detecting OD by an enzyme-labeling instrument600nmCombining with bacterial count under microscope, diluting properly, and adjusting bacterial density to 108cfu/m L (bacterial liquid OD diluted 2 times)600nmBetween 0.3 and 0.5), 200 μ L was inoculated into 50m L centrifuge tubes containing 10m L liquid culture medium of organophosphorus and inorganic phosphorus, respectively, and shake-cultured at 230rpm and 30 ℃ for 6d, and two replicates of each test bacterium were inoculated with 200 μ L sterile water as a control.
2.1.3 detection of effective phosphorus content in supernatant by MoSb antibody method
a. Preparation of supernatant
And centrifuging the fermentation liquor cultured for 6d at 1200rpm for 30min, taking supernatant, and discarding the precipitate.
b. Drawing of standard curve
Respectively and accurately sucking 0, 2, 4, 6, 8 and 10m L of 5 mg/L phosphorus standard solution into a 50m L volumetric flask, diluting the solution to a position with water with the total volume of about 3/5, adding 2 drops of 2, 6-dinitrophenol as an indicator, adjusting 50m L/L dilute sulfuric acid (or hydrochloric acid) and 10 percent sodium hydroxide until the solution is just yellowish, accurately adding 5m L molybdenum-antimony anti-color developing agent, shaking up, adding water to fix the volume to obtain a standard solution series with phosphorus contents of 0.0, 0.2, 0.4, 0.8 and 1.0 mg/L, respectively, shaking up, standing at room temperature of more than 15 ℃, measuring the absorbance at a wavelength of 700nm, taking the absorbance as a vertical coordinate and the phosphorus concentration (mg/L) as a horizontal coordinate, and drawing a standard curve.
c. Determination of available phosphorus content in supernatant
Transferring appropriate amount of supernatant into 50m L volumetric flask, diluting with water to total volume of about 3/5, adding 1-2 drops of dinitrophenol indicator, accurately adding 5m L molybdenum antimony anti-color developing agent, shaking, adding water to desired volume, standing at room temperature above 15 deg.C for 30min, and reading absorbance OD700nmThen, the corresponding phosphorus content is checked from the standard curve.
d. Calculating the effective dissolved phosphorus content in the supernatant
The effective phosphorus content p (mg/L) of the supernatant is × colorimetric volume of the supernatant, × times of division/total volume of fermentation liquor
Wherein, the concentration of the supernatant fluid is that the concentration of phosphorus mg/L is found from a standard curve;
the colorimetric volume is 50m L of constant volume;
dividing times are the total volume of the fermentation liquor/sampling volume.
Effective phosphorus-dissolving amount P (mg/L) ═ effective phosphorus content of strain supernatant-control supernatant phosphorus content
2.2 phospholytic bacteria morphology and 16s rRNA identification
And (3) the morphology of the phosphate solubilizing bacteria, namely inoculating the purified bacillus onto an L B plate by streaking, culturing at the constant temperature of 30 ℃ for 48h, and observing the characteristics of the size, the color, the edge uniformity, the wettability and the like of a bacterial colony after the bacterial colony grows out.
The molecular identification is that pure strains are inoculated to L B liquid culture medium, the shaking table is placed at 30 ℃, after the strains are cultured to a logarithmic phase, the genome DNA of the strain FJAT-47169 is extracted by adopting a Tris-saturated phenol method, 16S rRNA gene universal primers 27F and 1492R are adopted for PCR amplification, the PCR reaction program refers to the literature of Zhengxuefang and the like (Zhengxuefang, Liubo, Zhuyanqing, and the like, the screening and identification of the tomato bacterial wilt biocontrol bacillus is [ J ]. Chinese biological prevention and control institute, 2016,32(5):657 and 665.), the PCR products are sent to Shanghai Boshang biotechnology Limited for Sanger sequencing, EZBIOCou is used for completing sequence homology comparison, and the sequence is analyzed by MEGA 6.0.6 software and the development tree is constructed.
3. Test results
3.1 determination of phosphate solubilizing ability
The phosphate solubilizing ability of Bacillus FJAT-47169 on organic phosphorus and inorganic phosphorus is shown in Table 1. Experimental results show that the bacillus FJAT-47169 has a relatively obvious phosphate solubilizing effect on inorganic phosphorus and a relatively poor phosphate solubilizing effect on organic phosphorus.
TABLE 1 phosphate solubilizing ability of Bacillus FJAT-47169
It can be seen that Bacillus FJAT-47169 can promote apatite Ca as a phosphate solubilizing (inorganic phosphate degrading) microorganism3(PO4)2The insoluble phosphate releases phosphorus, and the content of soluble phosphorus in the soil is improved, so that the growth of crops can be obviously promoted, the root system of the crops is developed, and the stress resistance of the crops is enhanced.
3.2 identification of the Strain
3.2.1 Strain morphology
The colony morphology of FJAT-47169 is: round, light yellow, opaque, well-defined, protruding, moist microcolonies. The colony morphology is shown in FIG. 1.
Identification of 3.2.216S rRNA Gene
The nucleic acid sequence of the 16S rRNA gene of the strain FJAT-47169 is as follows:
TGCAGTCGAGCGGACAGATGGGAGCTTGCTCCCTGATGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAAACCGGGGCTAATACCGGATGGTTGTTTGAACCGCATGGTTCAGACATAAAAGGTGGCTTCGGCTACCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTTGTTAGGGAAGAACAAGTGCCGTTCAAATAGGGCGGCACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCTCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACAATCCTAGAGATAGGACGTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACAGAACAAAGGGCAGCGAAACCGCGAGGTTAAGCCAATCCCACAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTAGGAGCCAGCCGC(SEQ ID NO:1)
comparing the 16S rRNA gene sequence SEQ ID NO 1 of the strain FJAT-47169 with an EZBIOCloud gene database, wherein the genetic relationship between the strain FJAT-47169 and Bacillus siamensis is nearest, the homology of the 16S rRNA gene is 99.93 percent, so the strain FJAT-47169 belongs to Bacillus siamensis Siamese Bacillus. Downloading 16S rRNA gene sequences of strains with higher homology, carrying out comparative analysis, constructing a phylogenetic tree, and forming the phylogenetic tree as shown in figure 2 when a neighbor-Joining method is adopted and the Bootstrap value is 1000 times. In the constructed phylogenetic tree, the strain FJAT-47169 and Bacillus siamensis are gathered in the same branch.
EXAMPLE 2 growth promoting action of Bacillus
1. Tomato seed growth promotion test method
Selecting single colony of test strain, inoculating into 250m L conical flask containing 100m L L B liquid culture, and culturing at 30 deg.C for 48 hr (counting bacteria under microscope, bacterial density is 10%8cfu/m L or more), diluting the test bacteria fermentation broth according to a gradient, and taking 1400 times, 1200 times, 1000 times, 800 times and 600 times of bacteria liquid and clear water as a control (ck).
Selecting tomato seeds with relatively consistent growth vigor, placing the tomato seeds in a transparent culture box of 9cm with 2-3 layers of filter paper laid at the bottom, placing 15 mung bean seeds in a constant-temperature artificial climate box at 27 ℃, and illuminating for 16h and dark for 8 h. Tracking and observing the tomato germination condition, recording the germination number until no new germination grains appear in 3d continuously, measuring the germ length of the tomato germination radicle machine, and analyzing the promoting effect of the test bacteria on tomato seed germination.
The germination percentage (number of germinated seeds on specified days/number of test seeds) is × 100%
The germination index is ∑ (Gt/Dt), wherein Gt is the number of seeds to be germinated at t d, and Dt is the corresponding number of days to be germinated.
Vigor index (germination index × embryo root length (cm)
The test data adopts DPS software and a new double-polarization method (Duncan) to carry out the significance test of the data difference among treatments.
2. Tomato seedling growth promotion test method
Inoculating the test strain into L B liquid culture medium, culturing at 30 deg.C under 170rpm for 48 hr until the density is about 0.8 × 108-1.2×108CFU/mL(OD600nmAbout 0.8), diluting the bacterial liquid by 100 times, and placing the diluted bacterial liquid in a refrigerator at 4 ℃ for later use.
Tomato seedlings with similar growth vigor are selected and transplanted into flowerpots, 4 tomato seedlings per pot are set with a test group B-1 and a control group CK-W, 20 tomato seedlings (5 pots) in each treatment group are planted indoors for 7 days, then the roots of the tomato seedlings are irrigated with fermentation liquor, the fermentation liquor is diluted by 100 times, 200m L is irrigated in each pot every week, the tomato seedlings are irrigated once every 2 days in the plant growth period, and 100m L per pot of Hoagland liquid is irrigated every 2 weeks.
The experiments were performed in a smart greenhouse. The temperature of day and night is respectively 25 deg.C/20 deg.C, the sunshine time is 14h, and the diameter of the basin is 14 cm. 1kg of vegetable planting substrate sterilized at 121 ℃ is filled in each pot.
After 35d, the plant height, stem thickness (stem diameter), root length, fresh weight, dry weight of root, fresh weight of aerial parts, dry weight of aerial parts of each group of tomato plants were measured.
3. Garlic growth promotion test method
Inoculating a test strain into L B liquid culture medium, culturing for 48h in a shaking table at 30 ℃ and 170rpm to obtain fermentation liquor, diluting the fermentation liquor by 1000 times with clear water, then putting the diluted fermentation liquor into a 150m L conical flask, putting garlic on the mouth of the conical flask to ensure that the bottom of the garlic is soaked by bacterial liquid, taking the clear water as a Control (CK), replenishing water at proper time, and measuring the root length after culturing for 14d at 25 ℃ in a greenhouse.
4. Test results
4.1 growth-promoting results of tomato seeds
The experimental results show that the early germination condition is better 1400 times bacterial liquid dilution, the later growth condition is better 800 times bacterial liquid dilution, and the overall experimental group has the best growth promotion effect of 800 times bacterial liquid dilution, namely the bacterial concentration of the bacillus FJAT-47169 is about 1.25 × 105cfu/m L, the germination rate, germination index, root length, stem length and seed vigor index of the tomato seed are 108.34%, 107.54%, 116.23%, 114.81% and 124.99% of ck control, so that the tomato seed is soaked in the fermentation liquid of the bacillus FJAT-47169 to promote germination and growth.
TABLE 2 growth promoting ability of Bacillus FJAT-47169 on tomato seeds
Note: in the above table, abcd represents significant difference (P < 0.05).
4.2 growth-promoting results of tomato seedlings
The growth promoting experiment results of the bacillus on tomato seedlings are shown in table 3. Experimental results show that after the bacillus FJAT-47169 bacterial solution is applied, the root weight, stem thickness, root length, root fresh weight, dry weight, overground part dry weight and fresh weight of tomato seedlings are all improved compared with those of a control group.
Compared with a clear water control group (CK-W), the tomato plant height, the fresh weight and the dry weight of the overground part of the tomato of the treatment group (B-1) applied with the bacterial liquid are all obviously improved (P is less than 0.05), and are respectively improved by 59.49%, 114.72% and 102.80%. The bacillus FJAT-47169 fermentation liquor has the functions of promoting tomato seedling plant height increase, tomato seedling growth and tomato seedling leaf growth and promoting root system development.
TABLE 3 growth promoting effect of Bacillus FJAT-47169 on tomato seedlings
Note: different lower case letters after the same column of data in the table above indicate that the difference is up to a significance level (P < 0.05).
4.3 growth-promoting results of Garlic
The experimental result shows that the root length of the garlic in the CK group is 70.8mm, and the root length of the garlic in the FJAT-47169 bacterial liquid soaking group is 148.2 mm. Compared with CK group, the root length of garlic in the bacteria solution soaking group is increased by 109.32%. Experimental results show that the FJAT-47169 bacterial liquid has an obvious effect of promoting plant rooting.
Example 3 Nitrogen fixation of Bacillus
1. Nitrogen fixation efficiency determination method (national standard NY411-2000)
Adding 100m L nitrogen-free medium (formula: 0.2g of monopotassium phosphate, 0.2g of magnesium sulfate heptahydrate, 0.2g of sodium chloride, 5g of calcium sulfate, 10g of mannitol, 0.1g of calcium sulfate dihydrate, 1000m L of water and pH 7.0) into a 500m L triangular flask, sterilizing at 121 ℃ for 30min, performing aseptic operation, adding two rings of strains to be detected (or fermentation liquor obtained by culturing 1m L for 3 d) into each flask, placing the flasks on a shaking bed, performing shaking culture at 30 ℃ for 5-7d (120r/min), taking out the solution, determining sugar and nitrogen, determining sugar by adopting an anthrone photoelectric colorimetry method, and determining nitrogen by adopting a micro photoelectric colorimetry.
1.1 Anthracene ketone photoelectric colorimetry
1.1.1 test treatment, namely taking 1-4m L (depending on the sugar content) of fermentation culture liquid, diluting to 100m L, taking 1.00m L of the diluted liquid in a colorimetric tube, adding water to 2m L, adding 4m L of anthrone in each tube, shaking up, boiling and heating for 15min, cooling, carrying out colorimetric determination at 620nm, recording absorbance, and simultaneously carrying out blank test.
1.1.2 Standard Curve is drawn, 1.00, 2.00, 4.00, 6.00, 8.00, 10.00 and 20.00m L of 1.000mg/m L glucose standard solution is sucked into a 100m L volumetric flask, water is added to the volumetric flask to the scale, the solution contains 10, 20, 40, 60, 80, 100 and 200 mug/m L of sugar respectively, 1.00m L is sucked into a colorimetric tube, water is added to 2.00m L, the absorbance is recorded according to the method of 1.1.1, the abscissa is defined by the sugar content of the standard system, and the standard curve is drawn by the ordinate.
1.1.3 calculation of analytical results
The sugar content (X) in the 100m L solution was calculated according to the formula (C1):
in the formula: m is1- -the test sugar content, μ g/m L, found on the standard curve;
m2- -the sugar content of the blank examined on the standard curve,. mu.g/m L;
m-suction fermentation volume, m L.
Tolerance difference: a) taking the arithmetic mean value of the parallel measurement as a measurement result;
b) the permissible difference of the results of the parallel measurement is not more than 0.005g/100m L.
1.2 azotobacter liquid total nitrogen colorimetric determination method
1.2.1 preparation and testing of samples, absorbing 1.00m L of azotobacteria liquid into a 30m L digestion tube, adding 3m L of sulfuric acid, adding 0.1g of catalyst and 5 drops of hydrogen peroxide for digestion until the solution is clear, adding 1-2 drops of hydrogen peroxide after cooling if the reaction solution is not well cooked, continuing to cook until the solution is colorless and transparent, taking down and cooling, adding a little distilled water, shaking up, adding 40% (m/V) of sodium hydroxide solution until copper hydroxide precipitation (about 11-12m L) appears, adding 20 drops of 50% (m/V) of potassium sodium tartrate solution to remove the calcium and magnesium hydroxide precipitate, transferring the test tube liquid into a 100m L volumetric flask, pouring the digestion tube into the volumetric flask together with the digestion solution, diluting to the scale, shaking up, filtering, placing the filtrate 10.00m 2 into a 493 tube, adding 2 mol/1.00 m L of sodium hydroxide, adding 1.00m 5 of sodium tartrate, adding 7.1g of Neishi reagent, 10.00m 2 into a 493 2, adding water, stirring and then pouring into a 5m of sodium hydroxide solution into a yellow bottle, stirring, standing still, adding a small amount of sodium hydroxide solution, dissolving the solution into a yellow wine bottle, adding water, standing and cooling, adding water, dissolving the solution before adding water, dissolving the solution into a yellow wine, standing until the solution with absorbance measuring, measuring the solution, adding water, stirring, standing and measuring the solution, adding water, stirring.
1.2.2 drawing of Standard Curve
Accurately weighing 0.4716g of high-grade reagent ammonium sulfate which is baked at 105 +/-5 ℃ for 1h till constant weight, dissolving in water, diluting to 100m L, wherein the solution contains nitrogen of 1mg/m L, taking 0.05, 0.10, 0.25, 0.50, 1.00 and 2.00m L of the solution, putting the solution into a 100m L volumetric flask, diluting to scale with water, wherein the nitrogen content is respectively 0.50, 1.00, 2.50, 5.00, 10.00 and 20.00 mu g/m L, taking 1m L of each standard solution, putting the standard solution into a colorimetric tube, adding water to 2m L, measuring according to a 1.2.1 method, and recording the absorbance.
And drawing a working curve by using the nitrogen content of the standard solution as an abscissa and the corresponding absorbance as an ordinate.
1.2.3 calculation of analytical results
The nitrogen (X) content, expressed in g/100m L, is calculated according to formula (C2):
in the formula: m is1- -the test nitrogen content found on the standard curve,. mu.g/m L;
m2- -the nitrogen content of the blank, μ g/m L, found on the standard curve;
m-suction fermentation volume, m L.
Tolerance difference: a) taking the arithmetic mean value of the parallel measurement as a measurement result;
b) the permissible difference of the results of the parallel measurement is not more than 0.005 g.
1.3 Nitrogen fixation efficacy calculation
Nitrogen-fixing bacteria take up milligrams of nitrogen from the air per 1g of carbohydrate (sugar) consumed, and nitrogen-fixing potency is expressed as mg nitrogen/g sugar.
2. Test results
The experiment shows that the sugar consumption of the bacillus FJAT-47169 is 151.08 mg/L, the nitrogen fixation amount is 1.78 mg/L, and the nitrogen fixation efficiency is 11.80 mg/g.
In the practical application process, fermentation liquor diluted by 800 times, namely the bacterial concentration 1 × 10 can be used5-2×105cfu/m L for soaking plant seed to promote germination and growth, or diluting 100 times fermentation liquid to about 10 times concentration6The cfu/m L is used to treat plant seedlings by irrigating roots to promote the enrichment of nutrients in plant roots, so promoting the vigorous growth of plant seedlings, and the fermentation liquid diluted by 1000 times, i.e. bacterial concentration 1 × 105-2×105On the other hand, the bacillus FJAT-47169 has better phosphate solubilizing effect and nitrogen fixation efficiency, can be directly applied as a microbial organic fertilizer to improve soil fertility and better provide nutrition for plant growth, or can be freeze-dried to prepare a solid microbial organic fertilizer for convenient storage and transportation and used after being dissolved in water, and can also be mixed with other functional strains to prepare a composite microbial inoculum so as to further improve the phosphate solubilizing and nitrogen fixation effect.
It should be noted that, although the above embodiments have been described herein, the invention is not limited thereto. Therefore, based on the innovative concepts of the present invention, the technical solutions of the present invention can be directly or indirectly applied to other related technical fields by making changes and modifications to the embodiments described herein, or by using equivalent structures or equivalent processes performed in the content of the present specification and the attached drawings, which are included in the scope of the present invention.
Sequence listing
<110> institute of agricultural biological resources of academy of agricultural sciences of Fujian province
<120> bacillus for dissolving phosphorus and fixing nitrogen and application thereof in growth promotion
<130>47169
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>1416
<212>DNA
<213> Bacillus siamensis
<400>1
tgcagtcgag cggacagatg ggagcttgct ccctgatgtt agcggcggac gggtgagtaa 60
cacgtgggta acctgcctgt aagactggga taactccggg aaaccggggc taataccgga 120
tggttgtttg aaccgcatgg ttcagacata aaaggtggct tcggctacca cttacagatg 180
gacccgcggc gcattagcta gttggtgagg taacggctca ccaaggcgac gatgcgtagc 240
cgacctgaga gggtgatcgg ccacactggg actgagacac ggcccagact cctacgggag 300
gcagcagtag ggaatcttcc gcaatggacg aaagtctgac ggagcaacgc cgcgtgagtg 360
atgaaggttt tcggatcgta aagctctgtt gttagggaag aacaagtgcc gttcaaatag 420
ggcggcacct tgacggtacc taaccagaaa gccacggcta actacgtgcc agcagccgcg 480
gtaatacgta ggtggcaagc gttgtccgga attattgggc gtaaagggct cgcaggcggt 540
ttcttaagtc tgatgtgaaa gcccccggct caaccgggga gggtcattgg aaactgggga 600
acttgagtgc agaagaggag agtggaattc cacgtgtagc ggtgaaatgc gtagagatgt 660
ggaggaacac cagtggcgaa ggcgactctc tggtctgtaa ctgacgctga ggagcgaaag 720
cgtggggagc gaacaggatt agataccctg gtagtccacg ccgtaaacga tgagtgctaa 780
gtgttagggg gtttccgccc cttagtgctg cagctaacgc attaagcact ccgcctgggg 840
agtacggtcg caagactgaa actcaaagga attgacgggg gcccgcacaa gcggtggagc 900
atgtggttta attcgaagca acgcgaagaa ccttaccagg tcttgacatc ctctgacaat 960
cctagagata ggacgtcccc ttcgggggca gagtgacagg tggtgcatgg ttgtcgtcag 1020
ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg caacccttga tcttagttgc 1080
cagcattcag ttgggcactc taaggtgact gccggtgaca aaccggagga aggtggggat 1140
gacgtcaaat catcatgccc cttatgacct gggctacaca cgtgctacaa tggacagaac 1200
aaagggcagc gaaaccgcga ggttaagcca atcccacaaa tctgttctca gttcggatcg 1260
cagtctgcaa ctcgactgcg tgaagctgga atcgctagta atcgcggatc agcatgccgc 1320
ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcac accacgagag tttgtaacac 1380
ccgaagtcgg tgaggtaacc tttaggagcc agccgc 1416
Claims (9)
1. A bacillus for dissolving phosphorus and fixing nitrogen is characterized in that: the Bacillus is Siamese Bacillus FJAT-47169 with the scientific name of Bacillus siemensis FJAT-47169, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, has the preservation number of CGMCC NO.16250, the preservation date of 2018, month 13 and the preservation address of the institute of microorganisms of China national academy of sciences in Beijing, China.
2. Use of the bacillus of claim 1 for degrading insoluble phosphate.
3. Use of the bacillus for solubilizing phosphorus and fixing nitrogen as defined in claim 1 for promoting germination of plant seeds.
4. The application of the Bacillus subtilis for solubilizing phosphorus and fixing nitrogen in promoting the germination of plant seeds as claimed in claim 3, characterized in that the Bacillus fermentation liquid is prepared to have a concentration of 1 × 105-3×105cfu/m L, soaking the strain for 2-3 days, and placing at 25-30 deg.C under illumination for 16-20 h/day.
5. Use of the bacillus of claim 1 for promoting growth of young plants.
6. The use of the Bacillus bacteria for solubilizing phosphorus and fixing nitrogen in promoting the growth of plant seedlings as claimed in claim 5, wherein the Bacillus fermentation broth is prepared to a concentration of 0.8 × 106-1.2×106cfu/m L, planting the young plant in planting medium for 6-8 days, and irrigating the roots once every 5-7 days.
7. The application of the bacillus for solubilizing phosphorus and fixing nitrogen as claimed in claim 1 in plant nitrogen fixation.
8. The application of the bacillus for solubilizing phosphorus and fixing nitrogen as claimed in claim 1 in preparing a composite microbial inoculum for solubilizing phosphorus and fixing nitrogen.
9. A plant growth promoting microbial inoculum is characterized in that: the microbial inoculum comprises the bacillus of claim 1.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113549578A (en) * | 2021-08-03 | 2021-10-26 | 华南农业大学 | Siamese bacillus BsNlG13 for inhibiting rice blast germs and promoting seed germination and application thereof |
| CN114591865A (en) * | 2022-03-24 | 2022-06-07 | 江西益地生物科技有限公司 | Siamese bacillus KY758 capable of decomposing potassium and resisting diseases and application thereof |
| CN116656570A (en) * | 2023-06-27 | 2023-08-29 | 东北农业大学 | Siamese bacillus CM-19 and culture method, microbial inoculum and bacterial fertilizer thereof, and preparation method and application of microbial inoculum and bacterial fertilizer |
| CN116814479A (en) * | 2023-06-08 | 2023-09-29 | 贵州大学 | Bacillus GM-12-PT and method for promoting germination of Pterocarpus marsupium seeds by using same |
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| CN106635922A (en) * | 2017-01-22 | 2017-05-10 | 浙江农林大学 | Salt-tolerant biocontrol bacterium B268 for bacterial wilt of horsetail beefwood and application of salt-tolerant biocontrol bacterium B268 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113549578A (en) * | 2021-08-03 | 2021-10-26 | 华南农业大学 | Siamese bacillus BsNlG13 for inhibiting rice blast germs and promoting seed germination and application thereof |
| CN114591865A (en) * | 2022-03-24 | 2022-06-07 | 江西益地生物科技有限公司 | Siamese bacillus KY758 capable of decomposing potassium and resisting diseases and application thereof |
| CN114591865B (en) * | 2022-03-24 | 2023-08-22 | 江西益地生物科技有限公司 | Siamese bacillus KY758 for potassium decomposition and disease resistance and application thereof |
| CN116814479A (en) * | 2023-06-08 | 2023-09-29 | 贵州大学 | Bacillus GM-12-PT and method for promoting germination of Pterocarpus marsupium seeds by using same |
| CN116814479B (en) * | 2023-06-08 | 2024-05-07 | 贵州大学 | Bacillus GM-12-PT and method for promoting germination of Pterocarpus marsupium seeds by using same |
| CN116656570A (en) * | 2023-06-27 | 2023-08-29 | 东北农业大学 | Siamese bacillus CM-19 and culture method, microbial inoculum and bacterial fertilizer thereof, and preparation method and application of microbial inoculum and bacterial fertilizer |
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| CN111484951B (en) | 2022-07-08 |
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Effective date of registration: 20231214 Address after: Comprehensive Experimental Building (Crop Institute), No. 104 Pudang Road, Xindian Town, Jin'an District, Fuzhou City, Fujian Province, 350013 Patentee after: Crop Research Institute of Fujian Academy of Agricultural Sciences (Fujian Provincial Germplasm Resources Center) Address before: Room 1309, high tech building, Academy of Agricultural Sciences, 247 Wusi Road, Gulou District, Fuzhou, Fujian 350003 Patentee before: AGRICULTURAL BIORESOURCES INSTITUTE OF FUJIAN ACADEMY OF AGRICULTURAL SCIENCES |
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