CN115287227B - Bacillus for promoting nitrogen nutrition and growth of plants and application thereof - Google Patents
Bacillus for promoting nitrogen nutrition and growth of plants and application thereof Download PDFInfo
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- CN115287227B CN115287227B CN202210889156.4A CN202210889156A CN115287227B CN 115287227 B CN115287227 B CN 115287227B CN 202210889156 A CN202210889156 A CN 202210889156A CN 115287227 B CN115287227 B CN 115287227B
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 title claims abstract description 82
- 229910052757 nitrogen Inorganic materials 0.000 title claims abstract description 41
- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 35
- 235000016709 nutrition Nutrition 0.000 title claims abstract description 11
- 230000035764 nutrition Effects 0.000 title claims abstract description 11
- 230000001737 promoting effect Effects 0.000 title abstract description 9
- 230000008635 plant growth Effects 0.000 title abstract description 8
- 241000196324 Embryophyta Species 0.000 claims abstract description 23
- 244000105624 Arachis hypogaea Species 0.000 claims abstract description 20
- 235000020232 peanut Nutrition 0.000 claims abstract description 20
- 240000008042 Zea mays Species 0.000 claims abstract description 19
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 19
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 18
- 235000005822 corn Nutrition 0.000 claims abstract description 18
- 238000004321 preservation Methods 0.000 claims abstract description 9
- 230000029553 photosynthesis Effects 0.000 claims abstract description 7
- 238000010672 photosynthesis Methods 0.000 claims abstract description 7
- 239000002068 microbial inoculum Substances 0.000 claims description 8
- 230000000813 microbial effect Effects 0.000 claims description 5
- 230000002708 enhancing effect Effects 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 5
- 235000017060 Arachis glabrata Nutrition 0.000 abstract description 13
- 235000010777 Arachis hypogaea Nutrition 0.000 abstract description 13
- 235000018262 Arachis monticola Nutrition 0.000 abstract description 13
- 239000002689 soil Substances 0.000 abstract description 9
- 244000005700 microbiome Species 0.000 abstract description 6
- 238000010521 absorption reaction Methods 0.000 abstract description 3
- 238000009342 intercropping Methods 0.000 abstract description 2
- 235000015097 nutrients Nutrition 0.000 abstract description 2
- 241000192125 Firmicutes Species 0.000 abstract 1
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 24
- 230000001580 bacterial effect Effects 0.000 description 17
- 239000007788 liquid Substances 0.000 description 14
- 238000000034 method Methods 0.000 description 7
- 239000000618 nitrogen fertilizer Substances 0.000 description 7
- 230000000694 effects Effects 0.000 description 6
- 239000002609 medium Substances 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 229910052698 phosphorus Inorganic materials 0.000 description 4
- 239000011574 phosphorus Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 229930192334 Auxin Natural products 0.000 description 3
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 3
- 239000005977 Ethylene Substances 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 239000002363 auxin Substances 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- 208000003643 Callosities Diseases 0.000 description 2
- 206010020649 Hyperkeratosis Diseases 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000004178 biological nitrogen fixation Methods 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000000243 photosynthetic effect Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 238000004383 yellowing Methods 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- 239000012880 LB liquid culture medium Substances 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 1
- WYWFMUBFNXLFJK-UHFFFAOYSA-N [Mo].[Sb] Chemical compound [Mo].[Sb] WYWFMUBFNXLFJK-UHFFFAOYSA-N 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- HSFWRNGVRCDJHI-UHFFFAOYSA-N alpha-acetylene Natural products C#C HSFWRNGVRCDJHI-UHFFFAOYSA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 229910052786 argon Inorganic materials 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 229930002875 chlorophyll Natural products 0.000 description 1
- 235000019804 chlorophyll Nutrition 0.000 description 1
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 125000002534 ethynyl group Chemical group [H]C#C* 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 208000006278 hypochromic anemia Diseases 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 235000009973 maize Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- KKCBUQHMOMHUOY-UHFFFAOYSA-N sodium oxide Chemical compound [O-2].[Na+].[Na+] KKCBUQHMOMHUOY-UHFFFAOYSA-N 0.000 description 1
- 229910001948 sodium oxide Inorganic materials 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
Classifications
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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
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
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- 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
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- C—CHEMISTRY; METALLURGY
- 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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- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
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- Medicinal Chemistry (AREA)
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- Biomedical Technology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Soil Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to the technical field of microorganisms, in particular to bacillus for promoting nitrogen nutrition and growth of plants and application thereof. The bacillus is separated from peanut rhizosphere soil in the middle of a peanut and corn intercropping system, named 1603IPR-02 and subjected to biological preservation with the preservation number of CGMCC No.24753. The bacillus 1603IPR-02 provided by the invention is gram-positive bacteria, has strong nitrogen fixation capability, can convert nitrogen in air into nitrogen which can be utilized by plants, increases soil nutrients, promotes the absorption of nitrogen by the plants, can successfully colonize plant rhizosphere soil, improves plant photosynthesis and promotes plant growth, and has great application potential in the field of plant growth promotion.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to bacillus for promoting nitrogen nutrition and growth of plants and application thereof.
Background
Peanut is a common leguminous oil crop, has higher oil yield and wider distribution, and is widely applied to a plurality of fields in the prior art. Corn is a common food crop that is widely planted in tropical and temperate regions of the world. The corn has higher nutritive value, is widely applied to the fields of animal husbandry, breeding industry, aquaculture industry and the like, and has wide development and application prospects.
Nitrogen is called as a vital element, is an important component of biological macromolecules such as nucleic acid, protein, chlorophyll and the like, and participates in important vital processes such as photosynthesis, respiration and the like. The chemical nitrogen fertilizer is generally used excessively in the existing agricultural production, so that the fertilizer utilization efficiency is reduced, the symbiotic nitrogen fixation efficiency of peanuts is inhibited, the production cost is directly increased, and the economic benefit is reduced. Corn as a main grain crop has a great demand for nitrogen fertilizer and inevitably has a strong dependence on exogenous nitrogen fertilizer in the actual production process. However, a large amount of nitrogen fertilizer can cause irreversible damage to soil and ecological environment, so that the research of other green pollution-free ways to replace a large amount of nitrogen fertilizer application by laboratory means has very important practical significance.
The process of converting free nitrogen in air into compound nitrogen is called nitrogen fixation, wherein biological nitrogen fixation refers to the process that microorganisms catalyze N by nitrogen fixation enzymes 2 A process of reduction to ammonia. The 65% of nitrogen in the atmosphere is mainly immobilized in the form of biological nitrogen fixation, and the nitrogen can be utilized by microorganisms and plants. The global annual nitrogen content by biological fixation is approximately 200 ten thousand tons, accounting for 75% of the global plant nitrogen demand. Therefore, the reasonable utilization of nitrogen immobilized by microorganisms to reduce the application of chemical nitrogenous fertilizer is an energy-saving, green and healthy fertility supplementing mode in agriculture. The method for fixing nitrogen by utilizing microorganisms has important significance in reducing the application amount of nitrogen fertilizer, improving the yield of crops, reducing environmental pollution, promoting the sustainable development of agriculture and the like.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides bacillus for promoting nitrogen nutrition and growth of plants and application thereof.
The invention separates Bacillus from peanut rhizosphere soil in the middle of peanut corn intercropping system, and the Bacillus is named Bacillus 1603IPR-02, and the Bacillus colony is characterized in that: the bacterial colony is round, the center is concave, the color is white, the edge is rough, and the bacterial body is dry and semitransparent. The microscopic observation is characterized by rod-shaped, single cell, positive gram staining and spore.
The invention further carries out biological preservation on the strain, and the preservation information is as follows:
preservation number: CGMCC No.24753; classification naming: bacillus sp; preservation unit: china general microbiological culture Collection center (China Committee for culture Collection); preservation address: beijing, chaoyang area, north Chenxi way No. 1, no. 3, postal code 100101; preservation date: 2022, 4 and 22 days.
The invention further provides a fermentation product of Bacillus 1603 IPR-02.
Further, the culture medium adopted by the fermentation of the bacillus 1603IPR-02 is LB culture medium, and the composition of the culture medium is as follows: 5g of yeast extract; 10g of peptone; 10g of sodium oxide; distilled water is added to fix the volume to 1000mL; ph=7.0, sterilization at 121 ℃ for 20min; the fermentation conditions are as follows: the temperature is 30 ℃ and the rotating speed is 150-200rpm.
In a second aspect, the invention provides a microbial inoculant comprising said Bacillus sp 1603IPR-02 or a fermentation product thereof.
Further, the effective viable count of the Bacillus sp 1603IPR-02 is not less than 1×10 9 CFU/mL。
Further, the microbial inoculum is a plant photosynthesis improver or a plant nitrogen nutrition improver.
The invention further provides a biofertilizer comprising said Bacillus (Bacillus sp.) 1603IPR-02, or said microbial inoculum.
The invention further provides the use of said Bacillus (Bacillus sp.) 1603IPR-02, or said microbial inoculum, or said biofertilizer, for promoting nitrogen uptake in soil by plants.
The invention further provides the use of said Bacillus (Bacillus sp.) 1603IPR-02, or said microbial inoculum, or said biofertilizer for enhancing photosynthesis in plants.
Further, the plant comprises: leguminous crops and/or gramineous crops.
The invention has the following beneficial effects:
the Bacillus 1603IPR-02 is separated from plant rhizosphere soil, has high-efficiency nitrogen fixation capability, converts nitrogen in air into nitrogen which can be utilized by plants, increases soil nutrients, and can promote the absorption of peanuts and corns to soil nitrogen, improve plant nitrogen nutrition and improve plant photosynthesis. The bacillus 1603IPR-02 provided by the invention has no pollution, no residue and biological environmental protection in the application process, and is a growth promoting strain with good application prospect in the field of plant growth promotion.
Drawings
FIG. 1 is a photograph of the effect of Bacillus sp 1603IPR-02 on peanuts provided in example 2 of the present invention; wherein the left is control group and the right is Bacillus sp 1603IPR-02 treated peanut.
FIG. 2 is a statistical result of SPAD values of control group and Bacillus sp 1603IPR-02 treated peanut leaves provided in example 2.
FIG. 3 is a photograph of the effect of Bacillus sp 1603IPR-02 on corn provided in example 2 of the present invention; wherein the left is control group and the right is Bacillus sp 1603IPR-02 treated corn.
FIG. 4 is a graph showing statistics of SPAD values of control and maize leaves subjected to Bacillus sp 1603IPR-02 hammer, provided in example 2 of the present invention.
FIG. 5 is a graph showing nitrogen concentration statistics of control and Bacillus sp 1603IPR-02 treated corn leaves provided in example 2 of the present invention.
Detailed Description
The following examples are illustrative of the invention and are not intended to limit the scope of the invention.
The media used in the examples below were formulated as follows unless otherwise specified:
LB medium: 10g of tryptone, 5g of yeast extract and 10g of sodium chloride, the volume is fixed to 1L by distilled water, and the pH is adjusted to 7.0.
Example 1
The present example was validated for the growth promoting function of Bacillus sp 1603IPR-02, and the specific steps are as follows:
1. identification of nitrogen fixation ability of Bacillus (Bacillus sp.) 1603 IPR-02:
and inoculating the strain to be detected in the logarithmic growth phase into an LB liquid culture medium, and shake culturing at 30 ℃ for overnight. After the OD600 value of the strain was measured, the cells were collected by centrifugation, washed 3 times with physiological saline, and suspended in a nitrogen-limited medium, and the OD 600=0.2 was adjusted. Each anaerobic tube is connected with 1mL of bacterial liquid, the anaerobic tube is in a sealed state, air is removed, 2.5mL of fresh acetylene is added by injecting high-purity argon, and the anaerobic tube is subjected to shaking culture at 30 ℃ for 72h. 100 mu L of gas is taken from each anaerobic tube by a microsyringe, and the gas chromatograph is used for measuring the ethylene content.
The enzyme activity calculation formula is as follows:
nitrogen fixation enzyme activity = ethylene peak area on recorder x (test tube gas volume/sample injection amount)/(1 nmol standard ethylene peak area x reaction time)
As shown by the determination result of the bacterial strain nitrogen fixation enzyme activity, the Bacillus (Bacillus sp.) 1603IPR-02 has stronger nitrogen fixation capacity, and reaches 1.27 mu mol/(h.ml).
2. Identification of Bacillus (Bacillus sp.) 1603IPR-02 phosphate solubilizing ability:
single colonies were inoculated into Meng Jinna inorganic phosphorus medium and shake-cultured at 28℃and 180rmp for two days. The bacterial liquid is centrifuged for 10min at 10000rpm, 5mL of bacterial liquid supernatant is taken, and the bacterial liquid is filtered by a filter membrane with a filter hole of 0.22 mu m and diluted. And 5mL of diluent is taken and added into a 50mL volumetric flask, then 5mL of molybdenum-antimony colorimetric solution is added, and deionized water is added to the scale mark. After standing for 30min, the absorbance OD880 at 880nm was measured. And drawing a phosphorus concentration standard curve, and calculating the phosphorus content dissolved out of the bacterial liquid according to the phosphorus standard curve and the OD880 value of the bacterial liquid.
As shown by the determination result of the phosphate-dissolving capability of the strain, the Bacillus (Bacillus sp.) 1603IPR-02 has stronger phosphate-dissolving capability and reaches 4.11 mug/mL.
3. Identification of the ability of Bacillus (Bacillus sp.) 1603IPR-02 to produce auxin:
tryptophan was sterilized separately and added to LB liquid medium to a final concentration of 100mg/L. Shaking culture was performed for 1 day after single colonies were inoculated, at 28℃and shaking speed of 180rpm. 1mL of the bacterial liquid is centrifuged for 10min at 10000rpm, 100 mu L of supernatant is dripped on a white drip plate, a blank culture medium and 50mg/L IAA solution are respectively used as negative and positive controls, an equivalent amount of Salkowski chromogenic liquid is added, and the mixture is placed for 30min at room temperature in a dark place. 1mL of supernatant is taken and evenly mixed with an equal volume of Salkowski chromogenic solution, the chromogenic solution is placed in a water bath at 40 ℃ to react for 30min in a dark place, absorbance at 530nm wavelength is measured by a colorimetric method, and OD600 value of bacterial suspension is measured. And calculating the IAA amount generated by the bacterial liquid per unit volume when the bacterial liquid concentration OD600 = 1 by combining with an IAA concentration standard curve.
As can be seen from the results of the auxin production capacity measurement of the strain, the Bacillus sp 1603IPR-02 has a strong auxin secretion capacity, reaching 3.21 mug/mL.
TABLE 1 growth-promoting Capacity of Bacillus 1603IPR-02
Example 2
The present example further demonstrates the effect of Bacillus (Bacillus sp.) 1603IPR-02 on SPAD values and nitrogen nutrition of plants, as follows:
the strain 1603IPR-02 of the present invention was cultured in LB liquid medium at 30℃to 10 9 Centrifuging CFU/ml in a high-speed centrifuge at 5000rpm for 10min, pouring out supernatant, adding sterilized water with equal amount to obtain bacterial suspension, and standing.
Transplanting peanut and corn, wherein the peanut variety adopts Luhua 14, and the corn variety adopts Zhengdan 958. Bacterial liquid is added at a rate of 50 ml/strain every week after the first week of thinning, and the bacterial liquid concentration in the bacterial adding treatment: 1X10 9 CFU/ml, the control group was added with an equal amount of an equal concentration of sterile saline solution. Two sets of treatments each treated 4 pots, 6 biological replicates per pot of peanuts, 3 biological replicates per pot of corn. And (5) collecting samples three months after transplanting the peanuts and the corns. And measuring peanut SPAD value by using a SPAD instrument one week before sample collection to characterize photosynthetic capacity of the peanut SPAD value, and measuring nitrogen concentration of plants by adopting a Kjeldahl nitrogen determination method after sample collection of dried plant samples.
The results are shown in fig. 1-5, and the results in fig. 1 and 2 show that the nitrogen-deficiency yellowing phenomenon of the peanut is obviously relieved after the Bacillus (Bacillus sp.) 1603IPR-02 is applied, and the nitrogen-deficiency yellowing phenomenon is mainly reflected in that the SPAD value of the peanut leaf is obviously improved, and the amplitude reaches 3.4%. The results in figures 3-5 demonstrate that Bacillus 1603IPR-02 significantly improves corn nitrogen nutrition and remedies chlorosis symptoms after administration. The main expression is that after the Bacillus sp 1603IPR-02 is applied, the SPAD value of the corn leaf is obviously improved, and the amplification is 60.1%. After application of Bacillus 1603IPR-02, the nitrogen concentration of the corn leaf was significantly increased, with an amplitude of 15.0%. From the above results, it can be concluded that application of Bacillus (Bacillus sp.) 1603IPR-02 to plant rhizosphere can effectively promote absorption and utilization of nitrogen nutrition by plants and improve photosynthetic capacity of plants.
While the invention has been described in detail in the foregoing general description and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (7)
1. Bacillus cellBacillussp.) 1603IPR-02, wherein the preservation number is CGMCC No.24753.
2. A microbial inoculum, characterized by comprising the bacillus of claim 1Bacillus sp.) 1603IPR-02。
3. The microbial agent of claim 2, wherein the agent isIn the microbial inoculum, the bacillus is [ ]Bacillussp.) 1603IPR-02 is not less than 1×10 9 CFU/mL。
4. A microbial agent according to claim 2 or 3, wherein the microbial agent is a plant photosynthesis improver or a plant nitrogen nutrition improver.
5. A biofertilizer comprising the bacillus of claim 1Bacillussp.) 1603IPR-02, or the microbial agent of any one of claims 2-4.
6. The bacillus strain of claim 1Bacillussp.) 1603IPR-02, or the microbial inoculum of any one of claims 2-4, or the use of the biofertilizer of claim 5 for increasing nitrogen fixation capacity of peanuts or corn.
7. The bacillus strain of claim 1Bacillussp.) 1603IPR-02, or the microbial inoculum of any one of claims 2-4, or the biofertilizer of claim 5 for use in enhancing photosynthesis of peanuts or corn.
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| CN202210889156.4A CN115287227B (en) | 2022-07-26 | 2022-07-26 | Bacillus for promoting nitrogen nutrition and growth of plants and application thereof |
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