CN106754463A - One plant of tool dissolving P capacity Burkholderia bacterium NJAU B8 and its microbial manure of development - Google Patents

One plant of tool dissolving P capacity Burkholderia bacterium NJAU B8 and its microbial manure of development Download PDF

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CN106754463A
CN106754463A CN201611001344.XA CN201611001344A CN106754463A CN 106754463 A CN106754463 A CN 106754463A CN 201611001344 A CN201611001344 A CN 201611001344A CN 106754463 A CN106754463 A CN 106754463A
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njau
fertilizer
plant
burkholderia
nutrient
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CN106754463B (en
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沈其荣
乔策策
李�荣
张瑞福
王若斐
寻佳佳
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Meishan Huiyu Agricultural Development Co.,Ltd.
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Nanjing Agricultural University
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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
    • 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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05B—PHOSPHATIC FERTILISERS
    • C05B1/00—Superphosphates, i.e. fertilisers produced by reacting rock or bone phosphates with sulfuric or phosphoric acid in such amounts and concentrations as to yield solid products directly
    • C05B1/02—Superphosphates
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • 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
    • 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
    • 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

Abstract

The invention discloses one plant of tool dissolving P capacity Burkholderia bacterium NJAU B8 and its microbial manure of development, the strain isolation is in field corn rhizosphere soil, it is accredited as Burkholder and belongs to bacterium based on physio-biochemical characteristics and 16S rDNA analyses, it reaches 99% with Burkholderia gladioli homology.Quantification and qualification result shows that bacterial strain NJAU B8 have compared with strong solution Phos ability；Potted plant to show with field experiment result, bio-bacterial manure and biological organic fertilizer based on the bacterial strain development are able to effectively facilitate the potted plant growth of corn and increase the crop field yield of corn, Chinese cabbage and potato.

Description

One plant tool dissolving P capacity Burkholderia bacterium NJAU-B8 and its development it is micro- Bio-feritlizer
Technical field
The invention belongs to field of agricultural microorganism, and in particular to one plant of tool dissolving P capacity Burkholderia bacterium NJAU-B8 and its microbial manure of development.
Background technology
Phosphorus is one of main nutrient elements of limiting plant growth, the phosphate fertilizer this season crop utilization rate for applying be 5%~ 25%.Ca in most of phosphorus and soil2+、Fe3+、Fe2+、Al3+With reference to, formation insoluble phosphate (Abd-Alla M H, 1994), thus most of phosphate fertilizer accumulated in soil with the invalid state of indissoluble.Plant growth-promoting rhizobacteria (Plant Growth- Promoting Rhizobacteria, PGPR) refer to and survive in plant rhizosphere, Gen Biao and can directly or indirectly promote and adjust The microorganism (Ahmad et al, 2008) of plant growth, wherein phosphate solubilizing bacteria can be by insoluble phosphate, in ground phosphate rock Phosphorus is converted into water-soluble condition, so as to utilize (Zhu Ying etc., 2009) for plant absorption.Such bacterium can slightly solubility in activating soil Phosphorus element, can also increase root system of plant to around K, Ca, Mg, Fe, Zn by influenceing the type and quantity of secretions from plant roots Deng the absorption of nutrient, the resistance of plant is improved, plant is grown under the stress such as acid, scarce phosphorus habitat.
Therefore, it is necessary to screen efficient phosphate-solubilizing bacterial strain from the plant rhizosphere under natural conditions, and it is made phosphate solubilizing microorganism and has Machine fertilizer, with slightly solubility phosphorus element in activating soil while nutrition is provided, promotes absorption of the crops to phosphorus.
The content of the invention
It is an object of the invention to the practical problem and demand that are directed in production practices, there is provided one plant of tool phosphorus decomposing function, enter And effectively facilitate plant growth and increase the Burkholderia bacterium NJAU-B8 of crop yield.
The composite microbiological fertilizer prepared it is a further object of the present invention to provide the bacterium and its application.
The microbial bacterial agent prepared it is yet another object of the invention to provide the bacterium and its application.
The purpose of the present invention can be achieved through the following technical solutions：
One plant is used to promoting the growth-promoting bacterial strain NJAU-B8 of plant growth, and strain isolation is from Tonghua, Jilin Province maize field rhizosphere Soil, Classification And Nomenclature is Burkholderia bacterium (Burkholderia sp.), is preserved in Chinese microorganism strain preservation Administration committee's common micro-organisms center, preservation date is on April 11st, 2016, and deposit number is CGMCC No.12339.
Bacterium colonies of the bacterial strain NJAU-B8 on LB flat boards is smaller, in faint yellow, and can produce xanthein, edge-smoothing, tool It is a bit sticky, is difficult to provoke.Gram-negative, thalline uniform coloring.D-Fructose, maltose, arabinose, wood Sugar, lactose utilization are positive, and glucose fermentation produces acid not aerogenesis, 7%NaCl growths.With the 16S rDNA of NJAU-B8 bacterial strains The constructed development tree table of sequence (accession number is KX421207) is bright, bacterial strain NJAU-B8 and Burkholderia gladioli Bukhdderia gladioli (EU024168) homology reaches 99.1%.Morphological feature, physicochemical characteristicses with reference to bacterial strain are simultaneously tied 16SrDNA sequence analyses are closed, bacterial strain NJAU-B8 is initially identified as Burkholderia bacterium.
Burkholder of the present invention belongs to bacterium NJAU-B8 answering in solution Phos or promotion plant growth and volume increase With.
Burkholder of the present invention belongs to bacterium NJAU-B8 and is preparing the microbial bacterial agent of promotion plant yield-increasing or compound micro- Application in bio-feritlizer.
Belong to the microbial bacterial agent and/or composite microbiological fertilizer of bacterium NJAU-B8 preparations by Burkholder of the present invention.
Described microbial bacterial agent preferably mainly passes to lower section method and prepares：It is the Bai Ke of CGMCC No.12339 by preserving number Hall moral campylobacter bacteria NJAU-B8 is inoculated into LB fluid nutrient mediums, carries out liquid fermentation production, and the condition of fermenting and producing is： PH 7.0,30~35 DEG C of fermentation temperature scope, rotating speed is 170r/min, and fermentation time is 48h, makes bacteria containing amount >=1 in zymotic fluid ×1010CFU/mL.LB cultures liquid making method used is, as a example by preparing 1L culture mediums：Peptone 10g, dusty yeast 5g, NaCl 10g, agar 20g, are settled to 1000mL, pH natures, 121 DEG C of sterilizing 20min.
The zymotic fluid that described microbial bacterial agent will further preferably be obtained conventionally is prepared as other microorganisms Fertilizer common formulations, are preferably prepared as pulvis.
Application of the microbial bacterial agent of the present invention in solution Phos and promotion plant growth and volume increase.
Described promotion plant growth application, is preferably centrifuged 8~10min by described 6000~8000r/min of zymotic fluid Afterwards, bacteria suspension is prepared into using physiological saline (concentration 0.9%) resuspended thalline equal with fermentating liquid volume, is then fed directly to Plant rhizosphere.On the basis of reduces cost, the filling bacterium amount of bacteria suspension is 2.0~3.0 preferably by bacterium solution volume and soil quality ratio ×107CFU/g soil (butt) is applied, further preferably according to 2.5 × 107CFU/g soil (butt) is applied.
Described composite microbiological fertilizer preferably mainly passes to lower section method and prepares：By common organic fertilizer and amino acid hydrolyticsolution By 5:1(w:V, kg：L) mix, by 20:1(w：V, kg：L NJAU-B8 zymotic fluids) are added, by secondary fermentation, obtaining biology has Machine fertilizer, addition nutrient chemical to total nitrogen (N), full phosphorus (P2O5) and full potassium (K2O) nutrient is respectively 80gkg-1, 40gkg-1With 30g·kg-1Composite microbiological fertilizer is developed into, its living bacteria count >=0.5 hundred million/g fertilizer.Chemical fertilizer from urea (N, 46.6%), calcium superphosphate (P2O5, 12.0%), potassium chloride (K2O, 52.0%).
Described amino acid hydrolyticsolution is preferably prepared by the following method and forms：(1) will first die of illness poultry in closed container After fowl animal is transferred to closed hydrolytic decomposition pot automatically through automation crushing, all solid contents and liquid, in initial acid c (1/ 2H2SO4) it is 3-5mol L-1, hydrolyze 2-5 hours under 80-100 DEG C and 1-2 atmospheric pressure；(2) hydrolysis end treats molten in hydrolytic decomposition pot When liquid is cooled to less than 80 DEG C, stratification, the Freamine Ⅲ for collecting middle level is described amino acid hydrolyticsolution.Amino acid Hydrolyzate containing amino acid and various peptide matters about 10% (g/100ml) and more than.
Application of the composite microbiological fertilizer of the present invention in solution Phos and/or promotion plant growth and volume increase.
Described promotion crop yield application, field test continuously plant 3 season crop：Corn, Chinese cabbage and potato crop rotation.Often Season, biological organic fertilizer applied 3000kghm-2, points of 2 times administrations of fertilizer plant before seedling and apply the 1/2 of total nutrient as base manure (bag Include the chemical fertilizer of all of organic-inorganic compound mixed fertilizer or composite microbiological fertilizer and corresponding nutrient), remaining 1/2 nutrient is in just fruit Phase topdresses administration.
Beneficial effect：
It is of the invention main to belong to bacterium using the Burkholder that Phos and rhizosphere effectively colonize that solve for separating The administration of NJAU-B8, significantly improves the speed of growth of crops seedling stage, promotes crop to absorb nutrient in soil, so that Promote plant growth.The composite microbiological fertilizer being made from it, using field after, significantly improve the product of field Different Crop Amount.Promoting bacteria NJAU-B8 of the present invention 2 season pot experiment and 3 season field tests in show the growth-promoting and volume increase energy of stabilization Power.Growth-promoting with effect of increasing production substantially, is adapted to field crop producing region large area and uses after microbiology class fertilizer product application of the present invention.
Brief description of the drawings
Fig. 1 is based on the phylogenetic tree that the 16S rDNA gene orders of bacterial strain NJAU-B8 are set up using adjacent method
Fig. 2 the 1st season corn pot test effect figures
Fig. 3 the 2nd season corn pot test effect figures
Fig. 4 first season field experiment corn yields
Note：Different letters represent different significant difference (Duncan-test, P in same row in figure<0.05), similarly hereinafter.
Fig. 5 second season field experiment Chinese cabbage yield
Fig. 6 third quarter field experiment potato yields
Biomaterial preservation information
Bacterial strain NJAU-B8, Classification And Nomenclature is Burkholderia bacterium (Bukhdderia sp.), is preserved in China micro- Biological inoculum preservation administration committee common micro-organisms center, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, preservation date is on April 11st, 2016, and deposit number is CGMCC No.12339.
Specific embodiment
The separation and identification of embodiment 1, function stem
Corn root is gently rinsed into 2min under running water, removes rhizosphere surface mount soil.Then it is dry with aseptic water washing Only, root is cut, adds with bead and be equipped with the 250mL conical flasks of 100mL sterilized waters, 25 DEG C, 170r/min vibrations 20min forms soil supension, and 10 are drawn respectively-2、10-3、10-4、10-5The soil supension 0.1mL of extension rate, coats inorganic On phosphorus culture plate, 3 repetitions of each concentration, 28 DEG C culture 5 days after, selected on flat board around produce Soluble phosphorus circle bacterium colony it is pure Change more than 5 times, the bacterium colony that selection still has Soluble phosphorus circle goes to inclined-plane culture, and 4 DEG C of preservations are standby.
Strain after purification is inoculated on the PVK culture mediums of improvement, after being placed in 30 DEG C of incubator cultures 7 days, measurement Transparent circle and colony diameter, and calculate the ratio (D/d values) of transparent circle and colony diameter.
Selection transparent circle is relatively large in diameter or itself and the larger bacterial strain of colony diameter ratio, is inoculated into NBRIP culture mediums, 30 DEG C, 170r/min culture, 48h be measured by sampling nutrient solution in available phosphate concentration, after 7 days, determine nutrient solution in available phosphorus and full phosphorus Content, if not connecing bacterium control, treatment is all provided with 3 repetitions with compareing.
Isolation medium (the PVK culture mediums after improvement) compound method wherein used is, as a example by preparing 1L culture mediums：Portugal Grape sugar 10g, ammonium sulfate 0.5g, magnesium sulfate 0.3g, sodium chloride 0.3g, potassium chloride 0.2g, ferric sulfate 0.03g, manganese sulfate 0.03g, Tricalcium phosphate 5g, distilled water 1000mL, pH 6.5~7.0.115 DEG C of sterilizing 30min.
Screening and culturing medium used use Plant Res Internat B. V.'s phosphate growth medium (NBRIP) compound method for, with As a example by preparation 1L culture mediums：Glucose 10g, tricalcium phosphate 5g, magnesium chloride 5g, magnesium sulfate 0.25g, potassium chloride 0.2g, ammonium sulfate 0.1g, distilled water 1000mL, pH 7.0.115 DEG C of sterilizing 30min.
One plant of bacterium is finally obtained by qualitative and quantitative screening and is named as NJAU-B8, bacterial strain NJAU-B8 is on LB flat boards Bacterium colony it is smaller, in faint yellow, and xanthein, edge-smoothing, tool can be produced to be a bit sticky, be difficult to provoke.Gram It is negative staining, thalline uniform coloring.D-Fructose, maltose, arabinose, xylose, lactose utilization are positive, glucose fermentation Produce acid not aerogenesis, 7%NaCl growths.With the constructed hair of the 16S rDNA sequences (accession number is KX421207) of NJAU-B8 bacterial strains Educate tree table bright, it is same that bacterial strain NJAU-B8 belongs to bacterium Bukhdderia gladioli (EU024168) with Burkholderia gladioli Source property reaches 99.1%, not same with Vietnam bulkholderia cepasea on taxology, morphological feature, physics and chemistry with reference to bacterial strain Feature and 16S rDNA sequence analyses, Burkholderia bacterium is initially identified as by bacterial strain NJAU-B8.In being preserved in State's Microbiological Culture Collection administration committee common micro-organisms center, deposit number is CGMCC No.12339.
Embodiment 2, function bacterium fills the generation of bacterium bacteria suspension
Liquid fermentation is produced during bacterial strain NJAU-B8 is seeded into LB nutrient solutions, and the condition of fermenting and producing is：PH 7.0, hair 30~35 DEG C of ferment temperature range, rotating speed is 170r/min, and fermentation time is 48h, make bacteria containing amount >=1.0 in zymotic fluid × 1010CFU/mL；It is after zymotic fluid 6000r/min is centrifuged 8min, using the resuspended thalline of isometric 0.9% physiological saline to fill bacterium solution Obtain bacteria suspension.
LB cultures liquid making method used is, as a example by preparing 1L culture mediums：Peptone 10g, dusty yeast 5g, NaCl 10g, agar 20g, are settled to 1000mL, pH natures, 121 DEG C of sterilizing 20min.
The development of embodiment 3, composite microbiological fertilizer
The composite microbiological fertilizer of described promotion crop yield application, 5 are pressed by common organic fertilizer and amino acid hydrolyticsolution: 1(w:V, kg：L) mix, by 20Kg fertilizer:The ratio addition NJAU-B8 zymotic fluids of 1L NJAU-B8 zymotic fluids, by secondary hair Ferment, obtains biological organic fertilizer, addition nutrient chemical to total nitrogen (N), full phosphorus (P2O5) and full potassium (K2O) nutrient is respectively 80g kg-1, 40gkg-1And 30gkg-1Composite microbiological fertilizer is developed into, its living bacteria count >=0.5 hundred million/g fertilizer.Chemical fertilizer From urea (N, 46.6%), calcium superphosphate (P2O5, 12.0%), potassium chloride (K2O, 52.0%).Function bacterium be not inoculated with, is being adopted Nutrient is added in the mixture being fermented into same process and forms organic-inorganic compound mixed fertilizer to consistent with composite microbiological fertilizer.
The application effect of embodiment 4, growth-promoting bio-fertilizer on corn potted plant (the 1st season, the 2nd season)
Two season pot experiment on June 2nd, 2014 to October 24 in Jiangsu Province's solid organic castoff recycling Carried out in Hi-Tech Key Laboratory (Yixing) greenhouse, be Su Yu 29 for examination corn variety, purchased from academy of agricultural sciences of Jiangsu Province.Will Corn seed is immersed in 4h in normal-temperature water, is placed on the gauze of moistening vernalization 2-3 days in 30 DEG C of incubators, treats more than 80% After seed is sprouted, the seed that selection shows money or valuables one carries unintentionally moves to seedlings nursing plate nursery, and the consistent seedling replanting basin of growing way is selected after 2 true leaves are grown In.Experiment sets two treatment：1) CK, only applies common organic fertilizer；2) NJAU-B8 is processed, using common organic fertilizer and NJAU- The bacteria suspension of B8.Each 6 basin for the treatment of, the dress 2kg soil per basin, common organic fertilizer addition is 1.5% (mass percent, with 2kg soil On the basis of), the filling bacterium amount of the bacteria suspension of bacterial strain NJAU-B8 preferably presses 2.5 × 107CFU/g soil (butt) is applied.Conventional pipe Reason, determines that the plant height of plant, stem be thick, chlorophyll relative content measured value (Soil and plant respectively after 42d Analyzerdevelopment, SPAD), overground part fresh weight, overground part dry weight.
1st season corn growth-promoting effect such as Fig. 2, biomass situation as shown in table 1, after potted plant culture 42d, compared to not connecing bacterium Treatment, inoculating strain NJAU-B8 has growth-promoting effect substantially to maize seedling, and in addition to SPAD values, other growth indexes are significantly better than CK(P<0.05), its plant height, stem be thick, chlorophyll, overground part fresh weight, overground part dry weight increase by 40.3% compared with CK respectively, 18.7%, 1.3%th, 76.4%, 76.7%.
2nd season corn growth-promoting effect such as Fig. 3, biomass situation are as shown in table 2, after potted plant culture 42d, using common organic Caused after fertilizer and NJAU-B8 bacteria suspensions the overground part fresh weight and overground part dry weight of milpa be all remarkably higher than singly apply it is common organic Fertile (CK) treatment (p<0.05).Compared with CK, the treatment using Chicken dung and NJAU-B8 bacteria suspensions is thick, on the ground in plant height, stem Portion's fresh weight and overground part dry weight aspect, increased 15.3%, 1.9%, 6.2%, 9.9% respectively.
To sum up two seasons potted plant result shows, applies using common organic fertilizer and NJAU-B8 bacteria suspensions (NJAU-B8) treatment and singly The corn plant height of common organic fertilizer (CK) treatment, difference substantially, illustrates bacterial strain NJAU- between overground part fresh weight, overground part dry weight The growth-promoting effect of B8 is obvious.
Table 1, the 1st season corn growth-promoting pot experiment biomass situation
Note：SPAD：Chlorophyll relative content measured value；Data represent average value ± standard deviation in table.* represent different Significant difference (Duncan-test,P<0.05).Similarly hereinafter.
Table 2, the 2nd season the potted plant growth-promoting test organism amount situation of corn
The application effect of embodiment 5, growth-promoting bio-fertilizer on crop field corn
Go up pot experiment effect according to this, bacterial strain NJAU-B8 is chosen to be developed into composite microbiological fertilizer and further carries out field Between test.Field test is carried out in March, 2015 in April, 2016 in Haian County of Nantong City basic farmland, is for examination soil Yellow soil, pH：6.83, the content of organic matter：9.63g·kg-1, total nitrogen content：2.29g·kg-1, content of tatal phosphorus：4.08g·kg-1, available phosphorus contents：21.1mg·kg-1, quick-acting potassium content：719mg·kg-1.Continuous 3 season of the plantation crop of field test：Corn, Chinese cabbage and potato crop rotation.3 treatment are set altogether：1) chemical fertilizer control (CF)；2) organic-inorganic compound mixed fertilizer treatment (AOF):Do not connecing Function bacterium is planted, is supported using addition in the mixture being fermented into the composite microbiological fertilizers of NJAU-B8 containing bacterial strain treatment same process Divide to consistent with composite microbiological fertilizer and form organic-inorganic compound mixed fertilizer；3) composite microbiological fertilizers of NJAU-B8 containing bacterial strain treatment (NJAU-B8).Each 3 repetition for the treatment of, each plot area is 30m2。
As shown in figure 4, the 1st season corn field test carried out in March, 2015 in August, 2015.Organic-inorganic compound mixed fertilizer 3000kghm is applied with biological organic fertilizer-2, remaining nutrient urea, calcium superphosphate and potassium sulfate are pressed total nutrient polishing, are supported Distribution ratio is 8:4:3 (i.e. total nitrogen N, full phosphorus P2O5With full potassium K2O nutrients are respectively 80gkg-1, 40gkg-1And 30gkg-1)；Chemical fertilizer treatment directly applies urea, calcium superphosphate and the potassium sulfate of corresponding nutrient.2 administrations of fertilizer point, plant before seedling and apply always The 1/2 of nutrient as base manure (including chemical fertilizer of all of organic-inorganic compound mixed fertilizer or composite microbiological fertilizer and corresponding nutrient), Remaining 1/2 nutrient topdresses administration in first fruiting period.
1st season table of field test results it is bright：On the one hand, there was no significant difference for corn yield between respectively processing, it may be possible to soil Nutrient is higher, masks the effect of microbial manure；On the other hand, NJAU-B8 treatment is increased production respectively than AOF and CF treatment 0.82% and 3.37%, the production-increasing function of this functions bacterium starts to manifest.
The application effect of embodiment 6, growth-promoting bio-fertilizer on the Chinese cabbage of crop crop field
As shown in figure 5, the 2nd season Chinese cabbage field test carried out in September, 2015 in December, 2015.Organic and inorganic compound Fertilizer and biological organic fertilizer apply 3000kghm-2, remaining nutrient urea, calcium superphosphate and potassium sulfate press total nutrient polishing, Nutrient formula is 8:4:3；Chemical fertilizer treatment directly applies urea, calcium superphosphate and the potassium sulfate of corresponding nutrient.Fertilizer point is applied for 2 times With, plant before seedling apply the 1/2 of total nutrient as base manure (including all of organic-inorganic compound mixed fertilizer or composite microbiological fertilizer and The chemical fertilizer of corresponding nutrient), remaining 1/2 nutrient topdresses administration in the knot luxuriant phase.
2nd season table of field test results it is bright：The Chinese cabbage yield of NJAU-B8 treatment is significantly higher than CF and AOF treatment (Fig. 5), NJAU-B8 treatment increases production 3.92% and 5.74% respectively than AOF and CF treatment, and functions bacterium NJAU-B8 has significant volume increase Effect.
The application effect of embodiment 7, growth-promoting bio-fertilizer on the potato of crop crop field
As shown in fig. 6, the 3rd season potato field test carried out in December, 2015 in May, 2016.Organic and inorganic compound Fertilizer and biological organic fertilizer apply 3000kghm-2, remaining nutrient urea, calcium superphosphate and potassium sulfate press total nutrient polishing, Nutrient formula is 8:4:8；Chemical fertilizer treatment directly applies urea, calcium superphosphate and the potassium sulfate of corresponding nutrient.Fertilizer point is applied for 2 times With, plant before seedling apply the 1/2 of total nutrient as base manure (including all of organic-inorganic compound mixed fertilizer or composite microbiological fertilizer and The chemical fertilizer of corresponding nutrient), remaining 1/2 nutrient topdresses administration in initial bloom stage.
3rd season table of field test results it is bright：The potato yield of NJAU-B8 treatment is significantly higher than CF and AOF treatment (Fig. 6), NJAU-B8 treatment than AOF and CF treatment respectively increase production 12.73% and 35.89%, compared to the 2nd season rate of growth it is more significantly.

Claims (13)

1. one plant of burkholderia (Burkholderia) NJAU-B8, is preserved in Chinese microorganism strain preservation conservator Meeting common micro-organisms center, preservation date is on April 11st, 2016, and deposit number is CGMCC No.12339.
2. the burkholderia NJAU-B8 described in claim 1 is in solution Phos and/or promotion plant growth and volume increase Using；The preferred corn of plant, Chinese cabbage and potato.
3. the burkholderia NJAU-B8 described in claim 1 is preparing solution Phos, is promoting plant growth micro- with volume increase Application in bacteria agent or composite microbiological fertilizer；The preferred corn of plant, Chinese cabbage and potato.
4. the microbial bacterial agent that prepared by the burkholderia NJAU-B8 described in claim 1.
5. microbial bacterial agent according to claim 4, it is characterised in that be mainly prepared by the following method：It is by preserving number The burkholderia NJAU-B8 of CGMCC No.12339 is inoculated into LB fluid nutrient mediums, carries out liquid fermentation production, fermentation The condition of production is：PH 6.0~7.0,30~35 DEG C of fermentation temperature scope, mixing speed is 170r/min, and fermentation time is 48h, makes bacteria containing amount >=1 × 10 in zymotic fluid10CFU/mL。
6. microbial bacterial agent according to claim 5, it is characterised in that conventionally make it after obtaining zymotic fluid Standby is common formulations, is preferably prepared as pulvis, granule or suspending agent.
7. the composite microbiological fertilizer for being prepared as the burkholderia NJAU-B8 described in claim 1.
8. composite microbiological fertilizer according to claim 7, it is characterised in that be mainly prepared by the following method：By common Organic fertilizer presses 5 with amino acid hydrolyticsolution:1(kg：L composite fertilizer) is mixed to obtain, by composite fertilizer:NJAU-B8 zymotic fluid mass volume ratios It is 20:1(kg：L ratio), by secondary fermentation, obtains biological organic fertilizer to NJAU-B8 zymotic fluids are added in composite fertilizer, adds Plus nutrient chemical is to total nitrogen N, full phosphorus P2O5With full potassium K2O nutrients are respectively 80gkg-1, 40gkg-1And 30gkg-1It is made Composite microbiological fertilizer, wherein living bacteria count >=0.5 hundred million/g fertilizer.
9. application of the microbial bacterial agent described in claim 5 or 6 in solution Phos and/or promotion plant growth and volume increase； The preferred corn of plant, Chinese cabbage and potato.
10. application according to claim 9, it is characterised in that the zymotic fluid described in claim 5 is fed directly to plant Rhizosphere, or by described in claim 5 6000~8000r/min of zymotic fluid be centrifuged 8~10min after, using with fermentating liquid volume The resuspended thalline of equal physiological saline is prepared into bacteria suspension, is then fed directly to the filling bacterium of plant rhizosphere, zymotic fluid or bacteria suspension Amount is 2.0~3.0 × 10 preferably by bacterium solution volume and soil quality ratio7CFU/g soil (butt) is applied, and is further preferably pressed According to 2.5 × 107CFU/g soil (butt) is applied.
Composite microbiological fertilizer any one of 11. claim 7-8 is in solution Phos and/or promotion plant growth and increasing Application in product.
12. applications according to claim 11, its feature fertilization mode is composite microbiological fertilizer sowing amount 3000kg hm-2, points of 2 times administrations of fertilizer plant before seedling and apply the 1/2 of total nutrient as base manure, and remaining 1/2 nutrient topdresses in first fruiting period and applies With.
13. applications according to claim 11, it is characterised in that the plant is corn, Chinese cabbage and potato.
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CN113736705A (en) * 2021-09-22 2021-12-03 福建农林大学 Phosphate solubilizing bacteria RBC25 regulated and controlled by phosphate deficiency signal and application thereof
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