CN110317087A - Suitable for the phosphorus decomposing fixed nitrogen composite bacterial fertilizer of southern lean soil, preparation method and applications - Google Patents

Suitable for the phosphorus decomposing fixed nitrogen composite bacterial fertilizer of southern lean soil, preparation method and applications Download PDF

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CN110317087A
CN110317087A CN201910675673.XA CN201910675673A CN110317087A CN 110317087 A CN110317087 A CN 110317087A CN 201910675673 A CN201910675673 A CN 201910675673A CN 110317087 A CN110317087 A CN 110317087A
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soil
bacterial fertilizer
composite bacterial
fixed nitrogen
phosphorus decomposing
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王志康
洑香香
陈紫云
尚旭岚
徐子恒
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Nanjing Forestry University
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Nanjing Forestry University
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01B—SOIL WORKING IN AGRICULTURE OR FORESTRY; PARTS, DETAILS, OR ACCESSORIES OF AGRICULTURAL MACHINES OR IMPLEMENTS, IN GENERAL
    • A01B79/00—Methods for working soil
    • A01B79/02—Methods for working soil combined with other agricultural processing, e.g. fertilising, planting
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C21/00—Methods of fertilising, sowing or planting
    • A01C21/005—Following a specific plan, e.g. pattern
    • 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
    • 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

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  • Chemical & Material Sciences (AREA)
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  • General Health & Medical Sciences (AREA)
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Abstract

The present invention relates to the phosphorus decomposing fixed nitrogen composite bacterial fertilizer of southern lean soil, preparation method and applications, the composite bacterial fertilizer is made of at least two bacterium in bacillus megaterium, Pseudomonas fluorescens, blown-ball Azotobacter, Azospirillum brasilense.The present invention has screened the bacterial manure combination suitable for the barren yellowish soil in south, and discloses its cultural method, comprising: cultivates microbionation on improved LB culture medium；By the bacterium solution dilution after culture, then it is inoculated according to different bacterium combination and is cultivated wait improve in soil.The invention proposes a kind of green and economic and environment-friendly fertilization mode --- bio-bacterial manures suitable for the barren yellowish soil in south, by being optimized to many aspects such as bacteria combination, incubation times, a kind of preferable Fertilizer Combinations and fertilization time have been obtained according to soil nutrient characteristic dynamic change, system is improved for soil sustainability to lay a good foundation, while being also of great significance to the development and utilization of microbial bacteria.

Description

Suitable for the phosphorus decomposing fixed nitrogen composite bacterial fertilizer of southern lean soil, preparation method and its Using
Technical field
The present invention relates to the phosphorus decomposing fixed nitrogen composite bacterial fertilizer, the preparation method and applications that are suitable for southern lean soil, belong to Technical field of land improvement.
Background technique
Soil available nitrogen (N), phosphorus (P) content are to measure the important element index of soil fertility, plant vivo protein, phosphorus The metabolic processes such as the synthesis of acid and energy circulation all be unable to do without N, P element.Some barren mountainous regions in south China, the soil organism Content is low, and the nutrient utilized for plant absorption is less, has seriously affected plant growth and nutrient cycling and utilization.Soil is solved at present It is still that fertilize, but long-term unreasonable application chemical fertilizer can bring soil environment seriously that earth nitrogen stress, which lacks the widest method of phosphorus, Pollution and adverse effect, soil with organic matter content can be increased by applying organic fertilizer, but its fertilizer efficiency is slower.Therefore, seek one New ecological, environmental protective approach is planted to improve soil fertility, improve farm-forestry crop yield and quality and scientifically and rationally applies fertilizer and mentions High fertilizer efficiency is the emphasis of the future of agriculture research, while being also the important technical of development circulation agriculture and forestry.
Bio-bacterial manure can be divided into phosphate solubilizing bacteria, nitrogen-fixing bacteria and potassium solubilizing bacteria by application type as a kind of emerging green fertilizer, Generally soil can be effectively improved with mixed apply of organic fertilizer.In recent years, the utilization of bio-bacterial manure was rapidly developed, research Emphasis is mainly in the side such as single bacterium kind or the mechanism of action of finished product bacterial manure, the improvement of soil nutrient and the raising of site quality Face, wherein most widely used with phosphate solubilizing bacteria and nitrogen-fixing bacteria.The phosphate solubilizing bacteria separated from soil at present mainly has huge bud Spore bacillus (Bacillus megaterium) and Pseudomonas fluorescens (Pseudomonas fluorescens), nitrogen-fixing bacteria master There are blown-ball Azotobacter (Azotobacter chroococcum) and Azospirillum brasilense (Azospirillum Brasilence) etc..It is more single beneficial to thin by being inoculated with although existing much about the independent report of these bacteriums Bacterium evaluates its reclamation result, and the application about the mixing of a variety of bacterium is also more rare.Beneficial bacteria is inoculated in practical application In be also faced with some problems, for example the reclamation result Chang Yinwei Survival probability of bacteria of bacterial manure is lower and have a greatly reduced quality, single Single bacterium inoculation is also difficult to form a large amount of floras in the soil.
Soil improvement at present with to fertilize with organic fertilizer based on, but have that production energy consumption is big, applies at high cost, environmental pollution Seriously, the problems such as soil texture is hardened, soil available nitrogen and available phosphorus content are generally relatively low, which greatly limits agriculture and forestry can Ensured sustained development development, and these bio-bacterial manures have the characteristics that pollution-free, sustainability is high, can rapid, high volume produce, can be one Determine the use that chemical fertilizer is reduced in degree, it is considered to be realize the better method of soil green improvement.
Summary of the invention
The technical problem to be solved by the present invention is to change traditional fertilization mode, overcome the deficiencies of existing technologies simultaneously (bacterial manure unification) provides a kind of suitable for the phosphorus decomposing fixed nitrogen composite bacterial fertilizer of southern lean soil, preparation method and applications；Benefit Be suitable for bacterial manure combination, amount of application and the fertilization time of the barren mountain soil in south with the screening of four kinds of beneficial bacterias, can compared with The a large amount of bacterial manure of production in short time, which can effectively improve soil, and have the characteristics that pollution-free, sustainability is high, simultaneously Also it lays a good foundation for industrialized production and development circulation agriculture and forestry.
In order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention are as follows: the phosphorus decomposing suitable for southern lean soil Fixed nitrogen composite bacterial fertilizer, which is characterized in that the composite bacterial fertilizer is by bacillus megaterium Bacillus megaterium, fluorescence Pseudomonad Pseudomonas fluorescens, blown-ball Azotobacter Azotobacter chroococcum, azospirillum brasilense spiral shell At least two bacterium composition in bacterium Azospirillum brasilense.
It is furthermore preferred that the composite bacterial fertilizer is by Pseudomonas fluorescens Pseudomonas fluorescens, azospirillum brasilense Spirillum Azospirillum brasilense composition；Alternatively, the composite bacterial fertilizer is by bacillus megaterium Bacillus Megaterium, Pseudomonas fluorescens Pseudomonas fluorescens, blown-ball Azotobacter Azotobacter Chroococcum, Azospirillum brasilense Azospirillum brasilense composition；Again alternatively, the composite bacterial fertilizer by Bacillus megaterium Bacillus megaterium, Pseudomonas fluorescens Pseudomonas fluorescens composition.
Each bacteria concentration is 1 × 10 when phosphorus decomposing fixed nitrogen composite bacterial fertilizer described in single administration8~1 × 109CFU·mL-1(this is dense Degree refers to concentration of each independent bacterium in phosphorus decomposing fixed nitrogen composite bacterial fertilizer (that is, mixed bacteria liquid)；Consolidate when two kinds of bacterium are mixed to form phosphorus decomposing Nitrogen composite bacterial fertilizer, the volume ratio of two kinds of bacterium are 1:1；When four kinds of bacterium are mixed to form phosphorus decomposing fixed nitrogen composite bacterial fertilizer, the volume of four kinds of bacterium Than for 1:1:1:1.
In single administration phosphorus decomposing fixed nitrogen composite bacterial fertilizer, every yellowish-brown loam of 300g needs to apply at least 5mL mixed bacteria liquid, and Each bacteria concentration is 1 × 10 in mixed bacteria liquid8~1 × 109CFU·mL-1.For example, when the composite bacterial fertilizer is false single by fluorescence When born of the same parents bacterium Pseudomonas fluorescens, Azospirillum brasilense Azospirillum brasilense are formed, at least (wherein Pseudomonas bacteria concentration is 1 × 10 to 5mL mixed bacteria liquid8~1 × 109CFU·mL-1, Azospirillum brasilense concentration is 1 ×108~1 × 109 CFU·mL-1) be uniformly distributed in 300g yellowish soil and (be uniformly distributed and be achieved in the following ways: is mixed Combined bacteria liquid is applied to yellowish soil surface, mixes mixed bacteria liquid and certain thickness yellowish soil by turning over yellowish soil).
The lean soil is the barren yellowish soil in south.
The present invention also provides a kind of preparation method of phosphorus decomposing fixed nitrogen composite bacterial fertilizer suitable for southern lean soil, features It is, steps are as follows:
By at least two microbionations in phosphorus decomposing fixed nitrogen composite bacterial fertilizer above-mentioned on improved LB culture medium, culture 24-48h, wherein the formula of improved LB culture medium are as follows: pH:7.0, tryptone 10g/L, yeast extract 5g/L, NaCl 10g/ L, MnSO4·H2O 5mg/L,K2HPO4 0.8g/L,MgSO4·7H2O 0.1g/L,CaSO4·2H2O 0.1g/L,FeCl3 2mg/L；Improved LB culture medium is relative to common LB culture medium (tryptone 10g/L, yeast extract 5g/L, NaCl 10g/L) It additionally is added to microelement required for nitrogen-fixing bacteria and phosphate solubilizing bacteria and other nutrients, compound bacteria fast-growth can be promoted to expand It is numerous.Each bacterium solution can suitably dilute after culture, and the corresponding each bacterium solution of phosphorus decomposing fixed nitrogen composite bacterial fertilizer above-mentioned is mixed to form Mixed Microbes Liquid, the concentration of each bacterium is 1 × 10 in mixed bacteria liquid8~1 × 109CFU·mL-1.It is poor suitable for south that the invention further relates to one kind The screening technique of the phosphorus decomposing fixed nitrogen composite bacterial fertilizer of poor soil earth, which comprises the steps of:
1) two kinds of phosphate solubilizing bacterias-bacillus megaterium Bacillus megaterium, Pseudomonas fluorescens is chosen Pseudomonas fluorescens and two kinds of nitrogen-fixing bacteria-blown-ball Azotobacter Azotobacter chroococcum, Brazil are solid Nitrogen spirillum Azospirillum brasilense carries out soil improvement；
2) by four kinds of microbionations in step 1) on improved LB culture medium, 24-48h is cultivated, wherein improveing LB culture medium formula are as follows: pH:7.0, tryptone 10g/L, yeast extract 5g/L, NaCl 10g/L, MnSO4·H2O 5mg/ L, K2HPO4 0.8g/L,MgSO4·7H2O 0.1g/L,CaSO4·2H2O 0.1g/L,FeCl32mg/L；
3) four kinds of bacterium solutions after step 2) culture are diluted to according to bacterial concentration-OD value standard curve same dense Degree, the concentration range of each bacterium are 1 × 108~1 × 109CFU·mL-1, then it is inoculated according to different bacterium combination to conditioned soil In earth, bacteria combination mode are as follows: independent, combination of two, four kinds of bacteriums mixing；60d is cultivated under aseptic condition；Wherein, wait improve Soil is yellowish soil topsoil；
4) soil bacteria survival rate and change of soil nutrient are persistently measured, most suitable bacteria combination is comprehensively considered.
Screening technique above-mentioned, more specifically steps are as follows: 1) for the spy of the barren yellowish soil nitrogen stress in south, few available phosphorus Point has chosen two kinds of phosphate solubilizing bacterias (bacillus megaterium: Bacillus megaterium；Pseudomonas fluorescens: Pseudomonas ) and two kinds of nitrogen-fixing bacteria (blown-ball Azotobacters: Azotobacter chroococcum fluorescens；Azospirillum brasilense: Azospirillum brasilense) carry out soil improvement；2) by these four microbionations on improved LB culture medium, 24-48h is cultivated, wherein the formula of culture medium are as follows: pH:7.0, tryptone 10g/L, yeast extract 5g/L, NaCl 10g/L, MnSO4·H2O 5mg/L, K2HPO4 0.8g/L,MgSO4·7H2O 0.1g/L,CaSO4·2H2O 0.1g/L,FeCl3 2mg/ L；3) by the bacterium solution after culture be diluted to same concentration (concentration range be 1 × 108~1 × 109CFU·mL-1), then according to not Same bacteria combination (individually, combination of two, four kinds of bacterium mixing) is inoculated in wait improve in soil, cultivates 60d under aseptic condition；Its In, soil to be improved is derived from barren mountainous region (N31 ° 35 ' E119 ° 09 ') the 0-20cm upper soll layer soil in south, according to pedologic classification Standard is meager clay；4) soil bacteria survival rate and change of soil nutrient are persistently measured, most suitable bacteria combination is comprehensively considered.
The present invention also provides the method for administration of phosphorus decomposing fixed nitrogen composite bacterial fertilizer, and steps are as follows: phosphorus decomposing fixed nitrogen composite bacterial fertilizer applies In yellowish soil surface to be improved, yellowish soil is turned over after application, ploughed depth 5-20cm makes phosphorus decomposing fixed nitrogen composite bacterial fertilizer and 5- The yellowish soil of 20cm thickness mixes well；The single administration amount of phosphorus decomposing fixed nitrogen composite bacterial fertilizer are as follows: every yellowish-brown loam of 300g needs to apply At least 5mL phosphorus decomposing fixed nitrogen composite bacterial fertilizer (i.e. mixed bacteria liquid) and in mixed bacteria liquid each bacteria concentration be 1 × 108~1 × 109CFU·mL-1；Apply a phosphorus decomposing fixed nitrogen composite bacterial fertilizer again every 30-45d, facilitates its duration improvement soil.
A further object of the present invention is to provide phosphorus decomposing fixed nitrogen composite bacterial fertilizer or preparation method above-mentioned and is improveing barren Huang Purposes and method of administration above-mentioned or screening technique above-mentioned in brown earth are improveing the purposes in barren yellowish soil.
Compared with the existing technology, the invention has the benefit that the present invention is for existing Soil suppressiveness, (chemical fertilizer has Machine fertilizer) in the problems such as pollution is big, utilization rate is low, sustainability is poor, propose a kind of suitable for the green of the barren yellowish soil in south Color and economic and environment-friendly fertilization mode --- bio-bacterial manure, by being optimized to many aspects such as bacteria combination, incubation times, A kind of preferable Fertilizer Combinations and fertilization time have been obtained according to soil nutrient characteristic dynamic change, have been improved for soil sustainability System is laid a good foundation, while being also of great significance to the development and utilization of microbial bacteria.
Detailed description of the invention
A. available phosphorus contents of Fig. 1 different disposal when cultivating 30d and 60d, b. activity of acid phosphatase size.
Specific embodiment
Present invention is further described in detail in the following with reference to the drawings and specific embodiments.
The present invention has screened bacterial manure combination and fertilization time, mainly comprises the steps that
1, antagonistic effect: by different strain (bacillus megaterium Bacillus megaterium, Pseudomonas fluorescens Pseudomonas fluorescens, blown-ball Azotobacter Azotobacter chroococcum, Azospirillum brasilense Azospirillum brasilense) it is mixed two-by-two.Using radial diffusion method, it is placed in LB culture medium, is trained at 28 DEG C Supporting culture in case, for 24 hours, whether the growth population of different bacterium and inhibition zone are obvious in observation culture medium, to judge two kinds of bacteriums With the presence or absence of antagonistic effect.As a result following (table 1): by radial diffusion method by the collective media culture containing different bacterium for 24 hours After find, each processing is without obvious inhibition zone, and each bacterial growth quantity is unaffected, without obvious poor between two kinds of bacterial numbers It is different.Can thus estimate they grow in soil it is interference-free mutually, there is no antagonism between different bacterium.
1 bacterium of table is mixed antagonistic effect analysis
Note: M in table: bacillus megaterium, B: Azospirillum brasilense, C: blown-ball Azotobacter, F: Pseudomonas fluorescens, letter Combination represents corresponding bacterium solution mixed processing, similarly hereinafter.Lowercase indicates otherness between the interior bacterial number of processing
2, soil incubation: 12 processing are set according to different bacterium solutions, handles 4 including single culture, mixes two-by-two Strain handles 6, and four kinds of bacteriums mix 1, and control is the sterile water (table 2) for applying equivalent.Soil (i.e. barren yellowish soil) sample After high pressure sterilization (121 DEG C, 15min), air-dries sieving and be fitted into the culture box of bore 9.5cm, lower relative aperture 8cm, high 8.4cm； Every box claims native 300g, carries out high pressure sterilization again.It is (logical that bacterium solution (after cultivating in the LB culture medium after i.e. improved) will be activated Cross measurement OD value and standard concentration curve determine phosphobacteria liquid it is consistent with fixed nitrogen bacterial concentration be 1 × 108CFU·mL-1, wherein changing The formula of LB culture medium after good are as follows: pH:7.0, tryptone 10g/L, yeast extract 5g/L, NaCl 10g/L, MnSO4·H2O 5mg/L,K2HPO4 0.8g/L,MgSO4·7H2O 0.1g/L,CaSO4·2H2O 0.1g/L,FeCl32mg/L) according to difference Processing is mixed well with the soil in box, and distilled water, which is added, makes soil moisture be maintained at 60% of field capacity or so；Each Processing is repeated 4 times.28 DEG C of incubators are placed on after sealing and cultivate 60d under a dark condition, are periodically sprinkled during culture a certain amount of Sterile water is to guarantee that water content is basically unchanged.
The soil incubation of 2 bacterium of table combines and testing program
3, bacterial growth and its survival volume: the dynamic change of Soil Microorganism quantity: after incubation the 5th, 10,15,25, 30,45 and 60d weighs the fresh soil sample of 10g, adds and vibrates 30min in shaking table after 90ml sterile water, and 1mL mixed liquor is taken to be centrifuged to 10mL Pipe, adds 9mL sterile water, is successively diluted to 10 by this method4、105、106With 107, respectively take 0.2mL to be diluted painting plate, cultivate Count clump count afterwards for 24 hours.
Correlated results is following (table 3): on the whole, different disposal is changed greatly in the bacterial number of different incubation times. Wherein four kinds of bacterium mixed processing initial stage quantity are more (has reached 43.3 × 10 when culture 10d6CFU/g), the later period, which has, gradually drops (culture 60d is reduced to 3.7 × 10 to low trend6CFU/g)；And two kinds of bacterium mixed processings bacterial population when cultivating 45~60d Amount is more relative to other processing, and wherein MF processing is maximum, has reached 15.3 × 106CFU/g, and CB processing duration is best, It still is keeping increasing in 60d.It singly applies in processing, issue amount has certain growth after incubation by processing C and processing B, respectively Reach 17.3 × 10 in 60d and 45d6With 15.3 × 106CFU/g；And it handles F and handles what M bacterial number did not increased Trend shows as being gradually reduced instead, and the bacterial number for cultivating 60d reduces 78.31% He compared with cultivating 5d respectively 35.71%.It is found by variance analysis, processing MFCB is significant in culture early period and other processing differences, and bacterial number is opposite More, then issue amount is substantially less than other processing.
From such results, it can be seen that growth pattern is also different in the soil for different strain combination；It is 30-45 days or so big The bacterial number of part processing reaches the larger value；Mixed Shi Bidan, which applies effect, to get well, and mixed apply of 4 kinds of bacterium in a short time can shape At a large amount of floras；Phosphate solubilizing bacteria and nitrogen-fixing bacteria may have synergistic effect, can form cooperative relationship in the soil, and promotion is given birth to each other It is long.
3 bacterial number (× 10 of table6CFU·g–1Soil) with soil incubation time dynamic
Note: different capitalizations indicate that the bacterial number otherness of the different incubation times of same processing reaches p < 0.05 in table It is horizontal；Different lowercases indicate that the bacterial number otherness of same incubation time different disposal reaches the level of p < 0.05.
4, it the measurement of soil full carbon, full nitrogen, available nitrogen: in culture mid-term (30d) and latter stage (60d), weighs appropriate Soil, fresh soil is for measuring available nitrogen, and wind desiceted soil is for measuring the full carbon content of total soil nitrogen.
Main result is as follows: compared with control (applying the sterile water of equivalent), handling the soil of MF, FB and FC in 30d Total nitrogen content significantly improves, but the otherness in 60d between different disposal is not significant.The data of full carbon also with the variation class of full nitrogen Seemingly, in 60d handle between difference it is not significant, but when 30d compared to significant decrease.Such as processing M, C, MC are complete in 60d 12.1%, 7.4%, 16.1% is reduced respectively when carbon content ratio 30d.From the data of available nitrogen, all strain additions are all Soil available nitrogen content is improved, but individually applies nitrogen-fixing bacteria (such as processing C) and the promotion effect of available nitrogen is not obvious.
The content of full carbon, full nitrogen and available nitrogen when 4 soil incubation 30d and 60d of table
Note: different lowercases indicate to reach the water of p < 0.05 in the Nutrient Contents of same incubation time different disposal It is flat.* indicate that the content difference of the 30d and 60d of same processing are significant (p < 0.05)
5, it the measurement of soil quick-effective phosphor and related enzyme activity: in culture mid-term (30d) and latter stage (60d), weighs suitable Fresh soil is measured for measuring available phosphorus contents and activity of acid phosphatase.Main result is following (Fig. 1): what Fig. 1 a was indicated is different bacterium Variation of the soil available phosphorus in 30d and 60d after liquid processing；What Fig. 1 b was indicated is that soil acidity phosphatase exists after different bacterium solutions are handled The variation of 30d and 60d.It sees on the whole, available phosphorus contents (1~5mg/kg) within the scope of lower, although addition phosphorus decomposing Bacterium and nitrogen-fixing bacteria significantly improve content of soil available phosphor in 30d, but the level (figure before be reduced significantly in 60d again 1a).Similarly, activity of acid phosphatase also shows as first increasing reducing (Fig. 1 b) afterwards.Therefore, reasonable Fertilizer Combinations no doubt weigh Want, but fertilising interval time on also to pay attention to, i.e., timely supplement strain, guarantee its in the soil growth not by big It influences, keeps higher soil improvement efficiency.It was found that being the suitable time for supplementing microbial inoculum when 30-45d.
It summarizes: being improved for the barren yellowish soil in south that the present invention is studied, it has been found that (1) in the selection of strain combination On, the effect of mixed bacteria is got well than single bacterium effect；(2) from the point of view of bacteria living and growth result, it is within 30 days to 45 days or so Fertilising interval is more appropriate；(3) comprehensive every soil root system, proper processing have MF (two kinds of phosphate solubilizing bacteria mixing), FB (solution Phosphorus bacterium and nitrogen-fixing bacteria mixing) and MFCB (four kinds of bacterium mixing)；(4) culture medium prescription for being suitable for four kinds of bacteriums is pH: 7.0, tryptone 10g/L, yeast extract 5g/L, NaCl 10g/L, MnSO4·H2O 5mg/L,K2HPO4 0.8g/L, MgSO4· 7H2O 0.1g/L,CaSO4·2H2O 0.1g/L,FeCl3 2mg/L。
The above is only preferred Fertilizer Combinations and time of the invention, it is noted that for the common of the art For technical staff, without departing from the technical principles of the invention, several improvement and deformations can also be made, these improvement It also should be regarded as protection scope of the present invention with deformation.

Claims (9)

1. being suitable for the phosphorus decomposing fixed nitrogen composite bacterial fertilizer of southern lean soil, which is characterized in that the composite bacterial fertilizer is by huge bud Spore bacillus Bacillus megaterium, Pseudomonas fluorescens Pseudomonas fluorescens, blown-ball Azotobacter At least two bacterium groups in Azotobacter chroococcum, Azospirillum brasilense Azospirillum brasilense At.
2. the phosphorus decomposing fixed nitrogen composite bacterial fertilizer according to claim 1 for being suitable for southern lean soil, which is characterized in that described Composite bacterial fertilizer by Pseudomonas fluorescens Pseudomonas fluorescens, Azospirillum brasilense Azospirillum Brasilense composition；
Alternatively, the composite bacterial fertilizer is by bacillus megaterium Bacillus megaterium, Pseudomonas fluorescens Pseudomonas fluorescens, blown-ball Azotobacter Azotobacter chroococcum, Azospirillum brasilense Azospirillum brasilense composition；
Again alternatively, the composite bacterial fertilizer is by bacillus megaterium Bacillus megaterium, Pseudomonas fluorescens Pseudomonas fluorescens composition.
3. the phosphorus decomposing fixed nitrogen composite bacterial fertilizer according to claim 1 for being suitable for southern lean soil, it is characterised in that:
Described in single administration when phosphorus decomposing fixed nitrogen composite bacterial fertilizer, each single bacterium concentration is 1 × 108~1 × 109CFU·mL-1；When two kinds Bacterium is mixed to form phosphorus decomposing fixed nitrogen composite bacterial fertilizer, and the volume ratio of two kinds of bacterium is 1:1；When four kinds of bacterium are mixed to form phosphorus decomposing fixed nitrogen compound bacteria Fertilizer, the volume ratio of four kinds of bacterium are 1:1:1:1.
4. the phosphorus decomposing fixed nitrogen composite bacterial fertilizer according to claim 1 for being suitable for southern lean soil, it is characterised in that: described Lean soil be yellowish soil.
5. a kind of preparation method of the phosphorus decomposing fixed nitrogen composite bacterial fertilizer suitable for southern lean soil, which is characterized in that steps are as follows:
At least two microbionations in phosphorus decomposing fixed nitrogen composite bacterial fertilizer of any of claims 1 or 2 are cultivated in improved LB On base, 24-48h is cultivated, wherein the formula of improved LB culture medium are as follows: pH:7.0, tryptone 10g/L, yeast extract 5g/L, NaCl 10g/L,MnSO4·H2O 5mg/L,K2HPO4 0.8g/L,MgSO4·7H2O 0.1g/L,CaSO4·2H2O 0.1g/L, FeCl32mg/L；The corresponding each bacterium solution of phosphorus decomposing fixed nitrogen composite bacterial fertilizer of any of claims 1 or 2 is mixed to form mixed bacteria liquid, The concentration of each bacterium is 1 × 10 in mixed bacteria liquid8~1 × 109CFU·mL-1。
6. a kind of screening technique of the phosphorus decomposing fixed nitrogen composite bacterial fertilizer suitable for southern lean soil, which is characterized in that including as follows Step:
1) two kinds of phosphate solubilizing bacterias-bacillus megaterium Bacillus megaterium, Pseudomonas fluorescens Pseudomonas are chosen Fluorescens and two kinds of nitrogen-fixing bacteria-blown-ball Azotobacter Azotobacter chroococcum, Azospirillum brasilense Azospirillum brasilense carries out soil improvement；
2) by four kinds of microbionations in step 1) on improved LB culture medium, 24-48h is cultivated, wherein improved LB The formula of culture medium are as follows: pH:7.0, tryptone 10g/L, yeast extract 5g/L, NaCl 10g/L, MnSO4·H2O 5mg/L, K2HPO4 0.8g/L,MgSO4·7H2O 0.1g/L,CaSO4·2H2O 0.1g/L,FeCl32mg/L；
3) four kinds of bacterium solutions after step 2) culture are diluted to same concentration according to bacterial concentration-OD value standard curve, so It is inoculated in afterwards according to different bacterium combination wait improve in soil, bacteria combination mode are as follows: independent, combination of two, four kinds of bacteriums are mixed It closes；60d is cultivated under aseptic condition；Wherein, soil to be improved is yellowish soil topsoil；
4) soil bacteria survival rate and change of soil nutrient are persistently measured, most suitable bacteria combination is comprehensively considered.
7. the method for administration of phosphorus decomposing fixed nitrogen composite bacterial fertilizer described in claim 1-4 any one, which is characterized in that step is such as Under: phosphorus decomposing fixed nitrogen composite bacterial fertilizer is applied to yellowish soil surface to be improved, and turns over yellowish soil after application, ploughed depth 5-20cm, Mix well phosphorus decomposing fixed nitrogen composite bacterial fertilizer and the yellowish soil of 5-20cm thickness；The single administration amount of phosphorus decomposing fixed nitrogen composite bacterial fertilizer are as follows: Every yellowish-brown loam of 300g is applied to few 5mL mixed bacteria liquid, and each bacteria concentration is 1 × 10 in mixed bacteria liquid8~1 × 109CFU· mL-1；Apply a phosphorus decomposing fixed nitrogen composite bacterial fertilizer again every 30-45d.
8. preparation method described in phosphorus decomposing fixed nitrogen composite bacterial fertilizer described in claim 1-4 any one or claim 5 is changing Purposes in good barren yellowish soil.
9. method of administration as claimed in claim 7 or screening technique as claimed in claim 6 are improveing the use in barren yellowish soil On the way.
CN201910675673.XA 2019-07-25 2019-07-25 Suitable for the phosphorus decomposing fixed nitrogen composite bacterial fertilizer of southern lean soil, preparation method and applications Pending CN110317087A (en)

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CN114651548A (en) * 2022-03-09 2022-06-24 福建农林大学 Method for fixing nitrogen in soil in situ
CN115053915A (en) * 2022-06-24 2022-09-16 南京林业大学 Cultivation method for improving quality of cyclocarya paliurus leaf medicinal products by using bacillus

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CN1568296A (en) * 2001-08-13 2005-01-19 匈牙利农业－生物公司 Micro-organisms for the treatment of soil and process for obtaining them

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114651548A (en) * 2022-03-09 2022-06-24 福建农林大学 Method for fixing nitrogen in soil in situ
CN115053915A (en) * 2022-06-24 2022-09-16 南京林业大学 Cultivation method for improving quality of cyclocarya paliurus leaf medicinal products by using bacillus
CN115053915B (en) * 2022-06-24 2024-05-17 南京林业大学 Cultivation method for improving medicinal quality of cyclocarya paliurus leaves by using bacillus

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Application publication date: 20191011