CN116121134B - Bacillus terreus in phyllostachys pubescens and application thereof - Google Patents
Bacillus terreus in phyllostachys pubescens and application thereof Download PDFInfo
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- CN116121134B CN116121134B CN202211709044.2A CN202211709044A CN116121134B CN 116121134 B CN116121134 B CN 116121134B CN 202211709044 A CN202211709044 A CN 202211709044A CN 116121134 B CN116121134 B CN 116121134B
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- 235000003570 Phyllostachys pubescens Nutrition 0.000 title claims abstract description 20
- 241001520913 Phyllostachys edulis Species 0.000 title claims abstract 6
- 241000193830 Bacillus <bacterium> Species 0.000 title claims description 7
- 241000626621 Geobacillus Species 0.000 claims abstract description 16
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 12
- 241000530609 Terribacillus goriensis Species 0.000 claims abstract description 10
- 108091005804 Peptidases Proteins 0.000 claims abstract description 6
- 239000004365 Protease Substances 0.000 claims abstract description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 6
- 238000009629 microbiological culture Methods 0.000 claims abstract description 3
- 239000002689 soil Substances 0.000 claims description 11
- 244000005700 microbiome Species 0.000 claims description 6
- 230000008635 plant growth Effects 0.000 claims description 3
- 238000000354 decomposition reaction Methods 0.000 claims description 2
- 102000035195 Peptidases Human genes 0.000 claims 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 abstract description 9
- 239000011574 phosphorus Substances 0.000 abstract description 9
- 229910052698 phosphorus Inorganic materials 0.000 abstract description 9
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract description 5
- 230000012010 growth Effects 0.000 abstract description 3
- -1 pplication Species 0.000 abstract description 3
- 239000007788 liquid Substances 0.000 description 16
- 244000302661 Phyllostachys pubescens Species 0.000 description 14
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 11
- 235000017491 Bambusa tulda Nutrition 0.000 description 11
- 241001330002 Bambuseae Species 0.000 description 11
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 11
- 230000001580 bacterial effect Effects 0.000 description 11
- 239000011425 bamboo Substances 0.000 description 11
- 238000012258 culturing Methods 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 9
- 239000002504 physiological saline solution Substances 0.000 description 8
- 229920001817 Agar Polymers 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000012880 LB liquid culture medium Substances 0.000 description 5
- 230000003213 activating effect Effects 0.000 description 5
- 230000004913 activation Effects 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 239000002028 Biomass Substances 0.000 description 3
- 238000012300 Sequence Analysis Methods 0.000 description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 3
- 235000019341 magnesium sulphate Nutrition 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 235000020183 skimmed milk Nutrition 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000011790 ferrous sulphate Substances 0.000 description 2
- 235000003891 ferrous sulphate Nutrition 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 2
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 229940099596 manganese sulfate Drugs 0.000 description 2
- 239000011702 manganese sulphate Substances 0.000 description 2
- 235000007079 manganese sulphate Nutrition 0.000 description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 239000006916 nutrient agar Substances 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 description 2
- 229910052939 potassium sulfate Inorganic materials 0.000 description 2
- 235000011151 potassium sulphates Nutrition 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000012138 yeast extract Substances 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
- 241000894006 Bacteria Species 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
- 241000282596 Hylobatidae Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 108091036078 conserved sequence Proteins 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229910052816 inorganic phosphate Inorganic materials 0.000 description 1
- 229910001710 laterite Inorganic materials 0.000 description 1
- 239000011504 laterite Substances 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate 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
- 238000012257 pre-denaturation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000005067 remediation Methods 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 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 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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Abstract
The invention discloses a phyllostachys pubescens geobacillus and application thereof, wherein the phyllostachys pubescens geobacillus is classified and named Terribacillus goriensis CS, and is preserved in China general microbiological culture collection center (CGMCC) No. 26119 in the 11 th month 11 of 2022. The phyllostachys pubescens geobacillus has the capabilities of fixing nitrogen, decomposing organic phosphorus and inorganic phosphorus, and can also produce protease; and the culture is convenient, and the growth speed is high.
Description
Technical Field
The invention relates to a phyllostachys pubescens geobacillus and application thereof, and belongs to the technical field of microorganisms and ecological restoration.
Background
How to solve the environmental problems existing in the management and industry development of the bamboo forests, improve the utilization efficiency of bamboo resources, reduce the investment of the management of the bamboo forests, and realize the high-quality development of the bamboo industry is one of the most important problems in the current research of the bamboo industry.
Soil microorganisms are important components for maintaining underground biological activity, and regulate processes such as decomposition of soil animal and plant residues and soil organic matters and other harmful compounds, biochemical circulation, formation of soil structures, and the like. The microorganisms are sensitive to external interference, and the community structure and activity change of the microorganisms can sensitively reflect soil quality and health conditions. The area, the types and the reserves of the bamboo resources in China all stay in the front of the world, wherein the area of the bamboo forest is 641.16 ten thousand hm 2 The ecological agent occupies about 2.94% of the area of forests in the whole country, contains abundant biological resources and huge biomass, fully exerts the advantages of the biological resources of the bamboo forests, and digs various beneficial microorganism resources in the ecological system of the bamboo forests, and has huge potential and broad prospect.
Disclosure of Invention
The invention provides a phyllostachys pubescens geobacillus and application thereof.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
the Geobacillus mioga is classified and named Terribacillus goriensis CS, and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of 26119 in the 11 th year 2022.
The bacillus terreus in the phyllostachys pubescens is obtained by self-screening of the inventor, and the inventor discovers that the bacillus terreus Terribacillus goriensis CS in the phyllostachys pubescens can be used for soil restoration and plant growth promotion through research and accidents.
The phyllostachys pubescens geobacillus Terribacillus goriensis CS3 can be used for nitrogen fixation.
The phyllostachys pubescens geobacillus Terribacillus goriensis CS3 can be used for decomposing inorganic phosphorus and organic phosphorus.
The phyllostachys pubescens geobacillus Terribacillus goriensis CS3 can be used for producing protease.
The screening method of the phyllostachys pubescens geobacillus Terribacillus goriensis CS3 comprises the following steps:
1) Preparing a diluent: fully and evenly shaking and mixing the phyllostachys pubescens soil and sterile physiological saline, standing for 3-10 minutes, and diluting the upper bacterial liquid by using the sterile physiological saline;
2) Culturing: culturing the diluted bacterial liquid obtained in the step 1) in an LB solid culture medium at 30+/-2 ℃ for 48-72 h;
3) Isolation and purification of strains: after single colony is grown, single colony is selected for streak culture, and pure bacterial strain is obtained through repeated plate streaking (6-8 times).
In the step 1), the mass ratio of the phyllostachys pubescens soil to the sterile normal saline is 1: (8-10), the concentration of the sterile physiological saline is 0.90%.
The dilution factor in step 1) was 10 4 ～10 6 。
The bacillus laterite Terribacillus goriensis CS in the bamboo forest has the colony diameter of 0.5-1.0 mm on a nutrient agar plate, and the bacteria are irregularly round, rough, opaque and white. Culturing in LB culture solution, and the number of strains can reach 1×10 8 cfu/ml。
The technology not mentioned in the present invention refers to the prior art.
The phyllostachys pubescens geobacillus can be used for soil remediation and plant growth promotion, has the capabilities of fixing nitrogen, decomposing organic phosphorus and inorganic phosphorus, and can also produce protease; and the culture is convenient, and the growth speed is high.
Drawings
FIG. 1 shows that the Geobacillus macerans CS3 of the present invention is grown on a plate of organophosphorus-degrading medium;
FIG. 2 shows that the Geobacillus macerans CS3 of the present invention is grown on a plate of inorganic phosphate solubilizing medium;
FIG. 3 shows the growth of Geobacillus Moso CS3 of the present invention on a plate of skimmed milk powder medium.
Detailed Description
For a better understanding of the present invention, the following examples are further illustrated, but are not limited to the following examples.
Example one, acquisition of Bacillus Terribacillus goriensis CS3
1) Preparing a diluent:
10g of phyllostachys pubescens soil from Anji county of Zhejiang province is weighed and placed in 90mL of 0.90% sterile physiological saline, fully vibrated and mixed uniformly, and kept still for 5 minutes, and then the bacterial liquid is diluted by the gradient of the sterile physiological saline.
2) Culturing:
selection 10 -4 、10 -5 、10 -6 Three dilution gradients were each aspirated 100. Mu.L of bacterial liquid in LB solid medium (peptone 10g/L, naCl 10g/L, yeast extract 5g/L, agar 18g/L, balance water, pH7.0, sterilization at 121℃for 20 min), smeared with sterile smears, and finally placed upside down in a thermostated incubator at 37 ℃.
3) Isolation and purification of strains:
culturing in LB solid culture medium at 37deg.C for 48 hr, and the number of strains reaches 10 8 ～10 9 cfu/ml. Single colonies were picked with an inoculating needle, streaked on the surface of an agar medium, and pure strains were obtained by repeating the streaking of the plate 6 times.
By glycerol preservation
Adding 0.7-0.8 ml of CS3 culture solution and 0.2-0.3 ml of glycerol into a sterile strain preservation tube, fully mixing and preserving in a refrigerator at-80 ℃.
Identification of species
(1) Morphological features
CS3 was homogeneously turbid with trace amounts of precipitate in LB liquid medium (peptone 10g/L, naCl 10g/L, yeast extract 5g/L, balance water, pH7.0, 121℃for 20 minutes), and the precipitate was dispersed by gentle shaking. The colony diameter on the nutrient agar plate is 0.5-1.0 mm, the shape of the platform is saw-tooth, the edge is opaque, white and the texture is softer.
(2) 16S rRNA sequence analysis
Further 16S rRNA sequence analysis and alignment were performed on isolated strain CS 3. A pair of primers was designed based on the conserved sequence of bacterial 16SrDNA,
upstream primer (P1): 5'-AGAGTTTGATCCTGGTCAGAACGAACGCT-3'
Downstream primer (P6): 5'-TACGGCTACCTTGTTACGACTTCACCCC-3'
PCR amplification is carried out by taking the strains obtained by screening as templates. Reaction conditions: pre-denaturation at 95℃for 5min, denaturation at 94℃for 50s, annealing at 52℃for 1min, extension at 72℃for 1min 30s, 30 cycles of reaction.
Cloning and sequencing the target fragment according to a conventional method (Sambrook et al, 2001), and comparing the result with the 16S rRNA sequences registered in GenBank (accession numbers: NZ CP008876.1, NZ FMZB01000020.1, etc.), and analyzing the result to achieve the gene homology of more than 99%. Strain CS3 was identified as Geobacillus terrestris by morphological characterization and 16S rRNA sequence analysis according to Bergey' S Mannual of Determinative Bacteriology (Holt, J.G., gibbons, N.E., 1994) and the handbook of common bacterial System identification (Dongxiu bead and Cai Miaoying et al, 2001).
Example two, determination of Nitrogen fixation Capacity of Strain
Activating the strain stored on the inclined surface of the test tube, inoculating the strain to an LB liquid culture medium for activation, culturing for 48 hours at 37 ℃ and then taking the strain as seed liquid, inoculating the strain to an Abbe's shell culture medium (0.2 g/L of monopotassium phosphate, 0.2g/L of sodium chloride, 0.2g/L of magnesium sulfate, 5.0g/L of calcium carbonate, 0.1g/L of calcium sulfate, 10g/L of mannitol, 15.0g/L of agar and high-pressure sterilization for 30 minutes at the pH value of 7.0-7.5,115 ℃), and continuously transferring for 5 times to ensure that the strain has the nitrogen fixation capability.
Example three determination of the organophosphorus-degrading Capacity of Strain
Activating the strain stored on the inclined surface of the test tube, inoculating the strain to an LB liquid culture medium for activation, culturing for 48 hours at 37 ℃ to obtain seed liquid, inoculating 10ul of the seed liquid to an organophosphorus culture medium plate (10.0 g/L of glucose, 0.5g/L of ammonium sulfate, 0.3g/L of sodium chloride, 0.3g/L of magnesium sulfate, 0.03g/L of manganese sulfate, 0.3g/L of potassium sulfate, 0.03g/L of ferrous sulfate, 5.0g/L of lecithin, 15.0g/L of agar and autoclaving for 30min at the pH value of 7.0-7.5,115 ℃), and repeating the culture for 2 planting points per dish three times. The strain was incubated at 37℃for 5 days, and as shown in FIG. 1, a transparent ring was observed around the strain, indicating that the strain had the ability to decompose organic phosphorus.
Example four determination of inorganic phosphorus-degrading Capacity of Strain
Activating the strain stored on the inclined surface of the test tube, inoculating the strain to an LB liquid culture medium for activation, culturing for 48 hours at 37 ℃ to obtain seed liquid, inoculating 10ul of the seed liquid to an inorganic phosphorus culture medium flat plate (10.0 g/L of glucose, 0.5g/L of ammonium sulfate, 0.3g/L of sodium chloride, 0.3g/L of magnesium sulfate, 0.03g/L of manganese sulfate, 0.3g/L of potassium sulfate, 0.03g/L of ferrous sulfate, 5.0g/L of calcium phosphate, 15.0g/L of agar and sterilizing at the pH value of 7.0-7.5,115 ℃ under high pressure for 30 minutes), and repeating the culture for three times every 2 planting points. The strain was incubated at 37℃for 5-7 days, as shown in FIG. 2, and a transparent ring was observed around the strain, indicating that the strain had no ability to hydrolyze organic phosphorus.
Example five, measurement of protease production Capacity of Strain
Activating the strain stored on the inclined surface of the test tube, inoculating the strain on an LB liquid culture medium for activation, culturing for 48 hours at 37 ℃ to obtain seed liquid, inoculating 10ul of the seed liquid on a skim milk powder culture medium plate (15 g of skim milk powder, 16g of agar powder, 1000mL of water, and sterilizing for 15min at the pH value of 7.0-7.2,108 ℃), and repeating the steps for 2 planting points on each dish for three times. Incubation at 37℃for 5d, as shown in FIG. 3, a transparent ring around the strain was observed, indicating that the strain can produce protease.
Example six potted plant experiment of Strain
Activating the strain stored in the inclined surface of the test tube, inoculating the strain on an LB liquid culture medium for activation, and culturing at 37 ℃ for 48 hours to obtain seed liquid. The bacterial liquid after fermentation is centrifuged for 10min at 8000rpm at 4 ℃, the supernatant is discarded, and the bacterial liquid is resuspended in 0.90% sterile physiological saline, and then centrifuged for 10min at 8000rpm at 4 ℃, and the bacterial liquid is collected repeatedly three times. The collected cells were adjusted to 10 with 0.90% sterile physiological saline 8 cfu/ml was used as an experimental bacterial agent. 8ml of the microbial inoculum is inoculated to the rhizosphere of 1-year-old phyllostachys pubescens seedlings, and 6 replicates of each treatment are treated by taking the same amount of 0.90% sterile physiological saline as a control. The potted bamboo seedlings are subjected to natural illumination and unified moisture management. After 90d inoculation, the overground biomass and the total biomass of the strain CS3 are obviously improved by 30.12 to 40.31 percent and 35.67 to 43.88 percent respectively compared with the control。
Claims (5)
1. The bacillus terrestris in phyllostachys pubescens is characterized in that: the classification is Terribacillus goriensis CS, and the collection number is CGMCC NO 26119 which is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms at 11 and 11 of 2022.
2. The use of the geobacillus for phyllostachys pubescens as claimed in claim 1, characterized in that: is used for soil restoration and plant growth promotion.
3. The use according to claim 2, wherein: for nitrogen fixation.
4. The use according to claim 2, wherein: for the decomposition of inorganic and organic phosphors.
5. The use according to claim 2, wherein: use for producing proteases.
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| ES2510765A1 (en) * | 2013-01-02 | 2014-10-21 | Universidad De Sevilla | Microbial strain terribacillus sp-ae2b 122 with capacity to carry out transesterification reactions and uses thereof (Machine-translation by Google Translate, not legally binding) |
| CN107548280A (en) * | 2014-07-28 | 2018-01-05 | 氮技术有限公司 | Agricultural methods |
| CN111718868A (en) * | 2020-06-03 | 2020-09-29 | 青岛北方茶仓茶文化有限公司 | Edinglake terribacillus LBX capable of improving free radical scavenging capacity and fermentation product and application thereof |
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| ES2510765A1 (en) * | 2013-01-02 | 2014-10-21 | Universidad De Sevilla | Microbial strain terribacillus sp-ae2b 122 with capacity to carry out transesterification reactions and uses thereof (Machine-translation by Google Translate, not legally binding) |
| CN107548280A (en) * | 2014-07-28 | 2018-01-05 | 氮技术有限公司 | Agricultural methods |
| CN111718868A (en) * | 2020-06-03 | 2020-09-29 | 青岛北方茶仓茶文化有限公司 | Edinglake terribacillus LBX capable of improving free radical scavenging capacity and fermentation product and application thereof |
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