CN114292778B - Efficient potassium-decomposing phosphate-decomposing bacteria and application thereof - Google Patents
Efficient potassium-decomposing phosphate-decomposing bacteria and application thereof Download PDFInfo
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- 241000894006 Bacteria Species 0.000 title claims abstract description 27
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 35
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 35
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims abstract description 35
- 239000011591 potassium Substances 0.000 claims abstract description 35
- 229910052700 potassium Inorganic materials 0.000 claims abstract description 35
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims abstract description 32
- 239000011574 phosphorus Substances 0.000 claims abstract description 32
- 229910052698 phosphorus Inorganic materials 0.000 claims abstract description 32
- 239000002689 soil Substances 0.000 claims abstract description 28
- 239000003337 fertilizer Substances 0.000 claims abstract description 15
- 230000000813 microbial effect Effects 0.000 claims abstract description 5
- 238000009629 microbiological culture Methods 0.000 claims abstract description 5
- 238000004321 preservation Methods 0.000 claims abstract description 4
- 238000000855 fermentation Methods 0.000 claims description 36
- 230000004151 fermentation Effects 0.000 claims description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 239000000843 powder Substances 0.000 claims description 15
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 14
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 10
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- 230000003213 activating effect Effects 0.000 claims description 9
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- 229910000323 aluminium silicate Inorganic materials 0.000 abstract description 2
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 abstract description 2
- 239000011777 magnesium Substances 0.000 abstract description 2
- 229910052749 magnesium Inorganic materials 0.000 abstract description 2
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- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 6
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- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
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- 229910019142 PO4 Inorganic materials 0.000 description 4
- 235000010755 mineral Nutrition 0.000 description 4
- 239000011707 mineral Substances 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 4
- 239000010452 phosphate Substances 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 3
- 229910002651 NO3 Inorganic materials 0.000 description 3
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 3
- 229930003268 Vitamin C Natural products 0.000 description 3
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- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
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- NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 description 2
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- 230000002950 deficient Effects 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 239000011790 ferrous sulphate Substances 0.000 description 2
- 235000003891 ferrous sulphate Nutrition 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
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 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
- 230000000877 morphologic effect Effects 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- RKORKXFKXYYHAQ-UHFFFAOYSA-N 2-(4-iodophenoxy)acetic acid Chemical compound OC(=O)COC1=CC=C(I)C=C1 RKORKXFKXYYHAQ-UHFFFAOYSA-N 0.000 description 1
- 102000011931 Nucleoproteins Human genes 0.000 description 1
- 108010061100 Nucleoproteins Proteins 0.000 description 1
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- 229920002472 Starch Polymers 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- WYWFMUBFNXLFJK-UHFFFAOYSA-N [Mo].[Sb] Chemical compound [Mo].[Sb] WYWFMUBFNXLFJK-UHFFFAOYSA-N 0.000 description 1
- 230000009102 absorption Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- BIGPRXCJEDHCLP-UHFFFAOYSA-N ammonium bisulfate Chemical compound [NH4+].OS([O-])(=O)=O BIGPRXCJEDHCLP-UHFFFAOYSA-N 0.000 description 1
- 239000011609 ammonium molybdate Substances 0.000 description 1
- 235000018660 ammonium molybdate Nutrition 0.000 description 1
- APUPEJJSWDHEBO-UHFFFAOYSA-P ammonium molybdate Chemical compound [NH4+].[NH4+].[O-][Mo]([O-])(=O)=O APUPEJJSWDHEBO-UHFFFAOYSA-P 0.000 description 1
- 229940010552 ammonium molybdate Drugs 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- FFBHFFJDDLITSX-UHFFFAOYSA-N benzyl N-[2-hydroxy-4-(3-oxomorpholin-4-yl)phenyl]carbamate Chemical compound OC1=C(NC(=O)OCC2=CC=CC=C2)C=CC(=C1)N1CCOCC1=O FFBHFFJDDLITSX-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 229910021538 borax Inorganic materials 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000010433 feldspar Substances 0.000 description 1
- -1 feldspar and mica Chemical compound 0.000 description 1
- 229960002089 ferrous chloride Drugs 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000009616 inductively coupled plasma Methods 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- NMCUIPGRVMDVDB-UHFFFAOYSA-L iron dichloride Chemical compound Cl[Fe]Cl NMCUIPGRVMDVDB-UHFFFAOYSA-L 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000010445 mica Substances 0.000 description 1
- 229910052618 mica group Inorganic materials 0.000 description 1
- 239000002366 mineral element Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 150000003112 potassium compounds Chemical class 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 239000004323 potassium nitrate Substances 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
- 238000001243 protein synthesis Methods 0.000 description 1
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- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
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- 229910052604 silicate mineral Inorganic materials 0.000 description 1
- 239000004328 sodium tetraborate Substances 0.000 description 1
- 235000010339 sodium tetraborate Nutrition 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
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- 238000004611 spectroscopical analysis Methods 0.000 description 1
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- 235000000346 sugar Nutrition 0.000 description 1
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- 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
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- 235000019731 tricalcium phosphate Nutrition 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 229960001763 zinc sulfate Drugs 0.000 description 1
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fertilizers (AREA)
Abstract
The invention provides a high-efficiency potassium-decomposing phosphate-dissolving bacterium and application thereof, belonging to the technical field of microorganisms. The taxonomy of the high-efficiency potassium-decomposing phosphorus-decomposing bacteria provided by the invention is named as bacillus subtilis, the strain number is FTB-K1, and the bacillus subtilis is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No.23346. The bacillus subtilis FTB-K1 separated from cow dung has higher potassium and phosphorus decomposing capacity, can increase the content of effective potassium and phosphorus in soil, and improves the utilization efficiency of insoluble potassium and phosphorus in soil; can decompose insoluble aluminosilicate inorganic mineral substances such as potassium feldspar and the like, promote the conversion of nutrient elements such as insoluble potassium, silicon, magnesium and the like into soluble nutrients, can be used as a microbial fertilizer, increase the content of available nutrients in soil, promote the growth and development of crops and improve the yield. Has higher survival rate in soil and can be applied to actual agricultural production.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a high-efficiency potassium-decomposing phosphorus-decomposing bacterium and application thereof.
Background
Phosphorus is one of indispensable elements in the growth and development process of plants, phosphorus is a main component forming phospholipid and nucleoprotein, is an important raw material for constructing plant cells, and has the participation of phosphorus in many vital activities of plants such as photosynthesis, respiration, absorption of moisture and nutrients by plant roots, synthesis, conversion and the like of various substances such as sugar, starch, grease, protein and the like. The effective phosphorus in the soil can be absorbed and utilized by plants only accounting for 2% -3% of the total phosphorus, but most of the phosphorus cannot be directly absorbed by plants, and can be finally converted into substances which can be utilized by plants through a series of physical, chemical and biological reactions. Phosphorus in soil is a non-renewable mineral resource, 1.3 hundred million hectares of farmland exist in China, 74% of farmland have high phosphorus content, but the content of effective phosphorus is less than 5%, and the rest of farmland have serious phosphorus deficiency.
Potassium is not an essential element for constructing plants, but potassium is an extremely important substance in plant life activities. Potassium is mainly concentrated in organs or tissues where plant vital activities are most active, such as root tips, stem tips, young leaves, cambium, etc. Potassium can promote carbohydrate synthesis and transport and protein synthesis. The content of potassium in the plant body is generally 0.2% -4.1% by weight of the dry matter, next to nitrogen. However, the total nitrogen content of most cultivated lands in China is below 0.2%, the nitrogen-deficient soil is nitrogen-deficient soil, the content of potassium directly available in the soil is only 2% -10% of total potassium, and the rest more than 90% of potassium exists in the form of mineral potassium, such as silicate minerals like feldspar and mica, and the mineral has stable chemical properties and can be absorbed and utilized by plants only when released from the minerals.
In view of the above reality, most of the fields adopt a mode of applying fertilizer rich in phosphorus and potassium to meet the requirements of crops on nutrient elements, but a large amount of fertilizer application can cause environmental problems such as environmental pollution, soil hardening, soil fertility decline, agricultural product quality reduction, reduction of the number of soil microorganisms, ecological deterioration, and unbalanced ecological system of cultivated land soil. Therefore, how to improve the utilization rate of nutrient elements potassium and phosphorus in the cultivated field soil is a technical problem which needs to be solved in the field. As reported in the prior literature, although related phosphate-dissolving strains and potassium-dissolving strains are reported, the effects of the strains are poor, and the strains with the functions of dissolving potassium and dissolving phosphorus are fewer, so that the survival rate in soil is low, and the strain can not be applied to actual agricultural production. Therefore, the screening of the wild strain which has the function of efficiently decomposing potassium and phosphorus and can be applied to actual agricultural production has great significance for the development of agriculture in China.
Disclosure of Invention
Therefore, the invention aims to provide a high-efficiency phosphate and potassium decomposing bacterium which can degrade indissolvable inorganic phosphorus and potassium in soil, can provide high-quality phosphorus and potassium elements for plants and can be directly absorbed, and can be applied to actual agricultural production.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a high-efficiency potassium-decomposing phosphorus-decomposing bacterium, which is named as bacillus subtilis in taxonomy, has a strain number of FTB-K1 and is preserved in China general microbiological culture collection center (CGMCC) No.23346.
The invention also provides application of the high-efficiency potassium-decomposing phosphorus-decomposing bacteria in preparation of potassium-decomposing and/or phosphorus-decomposing microbial fertilizers.
The invention also provides application of the efficient potassium-decomposing phosphorus-decomposing bacteria in soil potassium-decomposing and/or phosphorus-decomposing.
Preferably, the fermentation broth of the efficient potassium-decomposing phosphorus-decomposing bacteria is used for decomposing potassium and/or phosphorus in soil.
Preferably, the application amount of the fermentation liquor is 0.4-0.8L/m 2 。
Preferably, the preparation method of the fermentation broth comprises the following steps: activating the efficient potassium-decomposing phosphate-dissolving bacteria, inoculating the activated bacteria into a fermentation medium, and culturing for 23-30h at 29-33 ℃.
Preferably, the activation time is 24-28 hours.
Preferably, the activating culture medium for activation comprises the following components in parts by weight: 10-14 parts of peptone, 5-9 parts of beef extract powder, 5-9 parts of sodium chloride, 12-16 parts of agar, 1000-1400 parts of water and pH7.2-7.4.
Preferably, the fermentation medium comprises the following components in parts by weight: 12-18 parts of bean cake powder, 4-10 parts of fish meal, 3-8 parts of glucose, 2-4 parts of compound inorganic salt, 1000-2000 parts of water and 6.8-7.3 of pH value.
Preferably, in the composite inorganic salt (NH) 4 ) 2 SO 4 ：K 2 HPO 4 ：MnSO 4 ·H 2 O：MgSO 4 ·7H 2 The weight ratio of O is 8-12:1-4:1-3:1-3.
The invention has the beneficial effects that:
the bacillus subtilis FTB-K1 separated from cow dung has high potassium-dissolving capacity and good phosphorus-dissolving capacity, can increase the content of effective potassium and phosphorus in soil, and improves the utilization efficiency of insoluble potassium and phosphorus in soil; can decompose insoluble aluminosilicate inorganic mineral substances such as potassium feldspar and the like, promote the conversion of nutrient elements such as insoluble potassium, silicon, magnesium and the like into soluble nutrients, can be used as a microbial fertilizer, increase the content of available nutrients in soil, promote the growth and development of crops and improve the yield.
The bacillus subtilis FTB-K1 has higher survival rate in soil, can be applied to actual agricultural production, can quickly colonize beneficial microorganisms after being used, decompose and activate indissolvable mineral elements, can convert indissolvable inorganic phosphorus and potassium in soil into high-quality phosphorus and potassium compounds which can be directly absorbed and utilized by plants, and can greatly improve the bioavailability of indissolvable phosphorus and potassium in the soil and the nutrient utilization rate of phosphorus and potassium fertilizers.
Preservation description
The bacillus subtilis FTB-K1 (Bacillus subtilis FTB-K1) is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) for 2021, 9 months and 2 days, and has a preservation address of CGMCC No.23346, namely China academy of microorganisms of national academy of sciences No.1 and No.3 of the Korean area of Beijing.
Drawings
FIG. 1 is a colony morphology of FTB-K1;
FIG. 2 is a diagram of FTB-K1 cells under a microscope.
Detailed Description
The invention provides a high-efficiency potassium-decomposing phosphorus-decomposing bacterium, which is named as bacillus subtilis in taxonomy, has a strain number of FTB-K1 and is preserved in China general microbiological culture collection center (CGMCC) No.23346.
The bacillus subtilis FTB-K1 is obtained by screening cow dung in Hebei mountain city, yutian county and Hebei natural pasture. The FTB-K1 strain obtained by screening is determined to be bacillus subtilis (Bacillus subtilis) FTB-K1 by morphological identification and 16S rRNA sequencing analysis. The bacterial strain can grow in large quantity by aerobic culture for 18 hours at 30 ℃ on an agar nutrient medium, and the bacterial strain has a yellowish color, a rough and opaque surface, a saw-toothed edge and irregular shape as shown in a bacterial strain morphological result shown in figure 1. Microscopic observation shows that the thallus ellipse, single cell 0.7-0.8 μm×2-3 μm, no capsule, periphyton, and capable of movement, spore 0.6-0.9 μm×1.0-1.5 μm, is located in the center of thallus or slightly offset, as shown in fig. 2. The bacterial 16S rDNA universal primers are shown as SEQ ID NO.2 and SEQ ID NO. 3:
27F(5’-AGTTTGATCMTGGCTCAG-3’)
1492R (5'-GGTTACCTTGTTACGACTT-3'), wherein M represents degenerate base, M=A/C, namely M is A or C, the strain FTB-K1 is amplified and sequenced in forward direction, the length of the obtained sequence is 1390bp, and the specific sequence is shown as SEQ ID NO. 1.
And comparing the sequencing results in NCBI database, and finally identifying and screening to obtain a sample which is bacillus subtilis (Bacillus subtilis) FTB-K1.
The invention also provides application of the high-efficiency potassium-decomposing phosphorus-decomposing bacteria in preparation of potassium-decomposing and/or phosphorus-decomposing microbial fertilizers.
The invention also provides application of the efficient potassium-decomposing phosphorus-decomposing bacteria in soil potassium-decomposing and/or phosphorus-decomposing.
When the efficient potassium-decomposing and phosphorus-decomposing bacteria are used for decomposing potassium and/or phosphorus in soil, the fermentation liquor of the bacillus subtilis FTB-K1 is preferably used, and the application amount of the fermentation liquor is preferably 0.4-0.8L/m 2 More preferably 0.5 to 0.7L/m 2 。
In the invention, the preparation method of the bacillus subtilis FTB-K1 fermentation broth preferably comprises the following steps: after the efficient potassium-decomposing phosphate-dissolving bacteria are activated, the bacteria are inoculated into a fermentation culture medium and are cultured for 23-30 hours at 29-33 ℃.
In the present invention, the time for activating the Bacillus subtilis FTB-K1 is preferably 24 to 28 hours, more preferably 25 to 27 hours. The activation medium for activating the bacillus subtilis FTB-K1 strain preferably comprises the following components in parts by weight: 10-14 parts of peptone, 5-9 parts of beef extract powder, 5-9 parts of sodium chloride, 12-16 parts of agar, 1000-1400 parts of water and pH7.2-7.4, and more preferably comprises the following components in parts by weight: 12-13 parts of peptone, 7-8 parts of beef extract powder, 7-8 parts of sodium chloride, 13-15 parts of agar, 1100-1300 parts of water and pH 7.3. The sources of the above components are not particularly limited in the present invention, and conventional commercially available products in the art can be used.
In the invention, the fermentation medium preferably comprises the following components in parts by weight: 12-18 parts of bean cake powder, 4-10 parts of fish meal, 3-8 parts of glucose, 2-4 parts of compound inorganic salt, 1000-2000 parts of water and pH value of 6.8-7.3, and more preferably comprises the following components in parts by weight: 14-16 parts of bean cake powder, 6-8 parts of fish meal, 5-6 parts of glucose, 3 parts of composite inorganic salt, 1400-1600 parts of water and pH value of 7.1-7.2. The composite inorganic salt of the present invention is preferably prepared from (NH) 4 ) 2 SO 4 、K 2 HPO 4 、MnSO 4 ·H 2 O and MgSO 4 ·7H 2 O, and the weight ratio of the four is preferably 8-12:1-4:1-3:1-3, more preferably 9-11:2-3:2:2. the sources of the above components are not particularly limited in the present invention, and conventional commercially available products in the art can be used.
In the present invention, the temperature of the fermentation culture is preferably 30 to 32℃and the time of the fermentation culture is preferably 25 to 28 hours, more preferably 26 to 27 hours. The fermentation culture of the invention preferably adopts shaking culture, and the rotating speed of the shaking culture is preferably 90-130r/min, more preferably 100-120r/min.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Determination of potassium-decomposing ability of bacillus subtilis FTB-K1
Inoculating bacillus subtilis FTB-K1 into an activation culture medium of 10g of peptone, 5g of beef extract powder, 5g of sodium chloride, 12g of agar, 1000g of water and pH7.2, activating for 24 hours, and then inoculating into 12g of bean cake powder, 4g of fish meal, 3g of glucose and compound inorganic salt ((NH) 4 ) 2 SO 4 ：K 2 HPO 4 ：MnSO 4 ·H 2 O：MgSO 4 ·7H 2 The weight ratio of O) is 8:1:1: 1) 2g, 1000g of water and pH value of 6.8, and culturing for 25h at 29 ℃ and 90r/min to obtain bacterial suspension.
Preparing a potassium-dissolving culture medium (without water-soluble potassium ions): 5g of glucose, 0.5g of ammonium sulfate, 0.5g of yeast powder, 0.3g of magnesium sulfate, 2g of disodium hydrogen phosphate, 0.03g of ferrous sulfate, 0.03g of manganese sulfate, 2g of potassium feldspar (the potassium content is 11-13%), 1L of distilled water, uniformly stirring, adjusting the pH value to 7.2, and sterilizing at the high temperature of 121 ℃ for 15min.
500mL triangular flasks, each flask was filled with 100mL of potassium-solubilizing medium, 5mL of bacterial suspension was added, and each treatment was repeated 5 times with reference to the non-inoculated bacterial suspension. Shaking culture at 29 deg.C and 90r/min for 10d, centrifuging the culture solution for 20min at 4500r/min, collecting supernatant, adding 2mL of 6% H 2 O 2 Boiling for 1h, centrifuging, and collecting supernatant. And measuring the water-soluble potassium content in the supernatant by using an inductively coupled plasma spectrometry, comparing the water-soluble potassium content with control treatment, and calculating the potassium-decomposing amount of the potassium-decomposing bacteria.
Potassium-dissolving amount (mg/L) =k in bacterial suspension + content-K in control solution + The content is as follows.
The water-soluble potassium ion content of the experimental group is 150-170mg/L, the water-soluble potassium ion content of the control group approaches to 0, the potassium decomposition rate reaches more than 68%, and the potassium decomposition capacity is high.
Example 2
Determination of phosphate-solubilizing ability of bacillus subtilis FTB-K1
The preparation process of the bacillus subtilis FTB-K1 bacterial suspension is the same as that of example 1. 100ml of inorganic phosphorus liquid culture medium is respectively added into a 250ml triangular flask, and the culture medium comprises 10g of glucose, 0.5g of magnesium sulfate, 0.5g of ammonia sulfate, 0.1g of calcium carbonate, 0.005g of ferrous chloride, 5g of tricalcium phosphate and 1000ml of water. 2ml of the bacterial suspension was inoculated as an experimental group, the control group was not inoculated with the bacterial suspension, 2ml of water was inoculated, and all treatments were repeated 3 times. The culture solution was centrifuged at 12000r/min for 10min at 32℃and 140rpm for 5d, and the amount of phosphorus (water-soluble phosphorus content) was measured by molybdenum-antimony anti-colorimetry. The results are shown in Table 1.
TABLE 1 phosphate solubilizing amount of Bacillus subtilis FTB-K1
As can be seen from Table 1, the Bacillus subtilis FTB-K1 can be used for phosphate dissolution and has good phosphate dissolution effect.
Example 3
Inoculating bacillus subtilis FTB-K1 into an activation culture medium of 14g of peptone, 9g of beef extract powder, 9g of sodium chloride, 16g of agar, 1400g of water and pH 7.4, activating for 28h, and then inoculating 18g of bean cake powder, 10g of fish meal, 8g of glucose and a compound inorganic salt ((NH) 4 ) 2 SO 4 ：K 2 HPO 4 ：MnSO 4 ·H 2 O：MgSO 4 ·7H 2 The weight ratio of O) is 12:4:3: 3) 4g, 2000g of water and pH value of 7.3, and culturing for 28h at 33 ℃ and 130r/min to obtain fermentation liquor of the experimental group.
The preparation process of the fermentation liquor of the control group is the same as that of the fermentation liquor of the experimental group, and the difference is that the bacillus subtilis FTB-K1 is not added in the control group.
Selecting 100m of Yutian county in Tangshan city 2 The wheat planting field was divided into four test fields on average, 2 of which were set as test groups (according to 0.4L/m 2 To the experimental group broth) was applied, 2 pieces were set as a control group (according to 0.4L/m 2 The control group fermentation liquor is applied to the application amount of the test group), the test group and the control group are both fertilized in the tillering stage of the wheat, and the conditions of the field management and the like are the same except that the application amount of the fermentation liquor is different from that of the test group and the control group. The results of measuring various indexes such as wheat yield at the mature period of wheat are shown in Table 2.
TABLE 2 influence of Bacillus subtilis FTB-K1 on wheat yield
As can be seen from Table 2, the fermentation broth of the present invention can improve crop yield.
Example 4
Inoculating bacillus subtilis FTB-K1 into an activation culture medium of 13g of peptone, 8g of beef extract powder, 8g of sodium chloride, 15g of agar, 1300g of water and pH 7.3, activating for 25h, and then inoculating into 16g of bean cake powder, 8g of fish meal, 6g of glucose and a compound inorganic salt ((NH) 4 ) 2 SO 4 ：K 2 HPO 4 ：MnSO 4 ·H 2 O：MgSO 4 ·7H 2 The weight ratio of O) is 9:2:2: 2) 3g, 160 g of water and pH7.2, and culturing at 32 ℃ for 26 hours at 120r/min to obtain fermentation liquor of the experimental group.
The preparation process of the fermentation liquor of the control group is the same as that of the fermentation liquor of the experimental group, and the difference is that the bacillus subtilis FTB-K1 is not added in the control group.
Selecting 50m of Yutian county in Tangshan city 2 Melon planting fields were divided into 5 test fields on average, 3 of which were set as test groups (according to 0.7L/m 2 To the experimental group broth) was applied, 2 pieces were set as a control group (according to 0.7L/m 2 The control group fermentation liquor is applied to the application amount of the melon, the test group and the control group are fertilized after 3 days of fruit setting, and the conditions of the field management and the like are the same except that the fermentation liquor is different from the application amount of the test group and the control group. The melon vitamin C and nitrate content was measured at the melon maturity stage and the results are shown in table 3.
TABLE 3 influence of Bacillus subtilis FTB-K1 on melon yield and quality
| Treatment of | Vitamin C content (mg/100 g) | Nitrate content (mg/100 g) | Yield (kg/mu) |
| Experimental group | 9.2±0.4 | 13.2±0.3 | 4333.6±19 |
| Control group | 4.0±0.3 | 22.5±0.1 | 1800.1±22 |
As can be seen from Table 3, the fermentation broth of the invention can improve the crop yield and the crop quality, and compared with a control group without the microbial inoculum of the invention, the bacillus subtilis FTB-K1 of the invention can improve the vitamin C content of melons by more than one time, and can reduce the nitrate content by 41.3%.
Example 5
The preparation process of the fermentation broth of the experimental group is the same as that of the fermentation broth of the control group in example 3, except that the bacillus subtilis FTB-K1 is not added in the control group.
Selecting 100m of Yutian county in Tangshan city 2 The corn planting field is divided into 5 test fields on average, 5 treatments are set for the test, wherein the treatment 1 is not fertilized, namely blank control, and the treatment 2 is carried out according to the ratio of 0.9L/m 2 The fermentation liquor of the control group is applied to the application amount of the corn special fertilizer of 0.05kg/m 2 Treatment 4 application of a common corn-specific fertilizer in the field 0.05kg/m 2 And according to 0.9L/m 2 The fermentation liquor of the experimental group is applied, and the special fertilizer for corn in the field is applied in treatment 5 of 0.025kg/m 2 And according to 0.9L/m 2 The experimental group broth was applied at the amount of application. The special fertilizer for corn consists of 70kg of urea, 30kg of monopotassium phosphate, 60kg of potassium nitrate, 1kg of 2, 4-D1 kg of 4-iodophenoxyacetic acid, 1kg of borax, 1kg of ammonium molybdate, 1kg of zinc sulfate, 1kg of copper sulfate, 1kg of manganese sulfate and 1kg of ferrous sulfate. The 5 treatment groups are all fertilized once in spring sowing, except for different fertilizations, the other fields are all fertilizedConditions such as inter-management are the same. Corn yield was measured at corn maturity and the results are shown in table 4.
TABLE 4 influence of Bacillus subtilis FTB-K1 on corn yield
| Group of | Yield (kg/mu) |
| Process 1 | 533.4 |
| Process 2 | 733.4 |
| Process 3 | 1400.1 |
| Process 4 | 2133.4 |
| Process 5 | 1533.4 |
As shown in Table 4, the bacillus subtilis FTB-K1 microbial inoculum and the fertilizer are applied in a mixed mode, so that the corn yield can be improved, and when the special fertilizer for common corn is applied in a reduced amount of 50% by mass, the corn yield can still be improved, and the microbial inoculum can effectively reduce the application amount of the fertilizer.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Sequence listing
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Claims (10)
1. The efficient potassium-decomposing phosphate-decomposing bacteria is bacillus subtilis FTB-K1 and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of 2346.
2. The use of the high-efficiency potassium-decomposing phosphate-decomposing bacteria in claim 1 in the preparation of a potassium-decomposing and/or phosphate-decomposing microbial fertilizer.
3. The application of the high-efficiency potassium-decomposing phosphorus-decomposing bacteria in soil potassium and/or phosphorus decomposition.
4. The use according to claim 3, wherein the fermentation broth of the efficient potassium-decomposing and phosphorus-decomposing bacteria is used for decomposing potassium and/or phosphorus in soil, and the fermentation broth is the fermentation broth containing the efficient potassium-decomposing and phosphorus-decomposing bacteria.
5. The use according to claim 4, wherein the fermentation broth is applied in an amount of 0.4 to 0.8L/m 2 。
6. The use according to claim 4, wherein the method of preparing the fermentation broth comprises the steps of: activating the efficient potassium-decomposing phosphate-dissolving bacteria, inoculating the activated bacteria into a fermentation medium, and culturing for 23-30h at 29-33 ℃.
7. The use according to claim 6, wherein the activation time is 24-28h.
8. The use according to claim 6, wherein the activating medium for activation comprises the following components in parts by weight: 10-14 parts of peptone, 5-9 parts of beef extract powder, 5-9 parts of sodium chloride, 12-16 parts of agar, 1000-1400 parts of water and pH7.2-7.4.
9. The use according to claim 6, wherein the fermentation medium comprises the following components in parts by weight: 12-18 parts of bean cake powder, 4-10 parts of fish meal, 3-8 parts of glucose, 2-4 parts of compound inorganic salt, 1000-2000 parts of water and 6.8-7.3 of pH value.
10. The use according to claim 9, characterized in that in the complex inorganic salt (NH 4 ) 2 SO 4 ：K 2 HPO 4 ：MnSO 4 ·H 2 O：MgSO 4 ·7H 2 The weight ratio of O is 8-12:1-4:1-3:1-3.
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Effective date of registration: 20240205 Address after: 064100 Huxing Road, Houhu industrial cluster, Yutian County, Tangshan City, Hebei Province 1966 Patentee after: FENG TIAN BAO AGRICULTURE TECHNOLOGY CO.,LTD. Country or region after: China Patentee after: Hebei Fenglingguan Agricultural Technology Co.,Ltd. Address before: 064100 Huxing Road, Houhu industrial cluster, Yutian County, Tangshan City, Hebei Province 1966 Patentee before: FENG TIAN BAO AGRICULTURE TECHNOLOGY CO.,LTD. Country or region before: China |