CN108893421A - A kind of spindle lysine bacillus and its application in land reclamation in mining area ecological reconstruction - Google Patents
A kind of spindle lysine bacillus and its application in land reclamation in mining area ecological reconstruction Download PDFInfo
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Abstract
The invention discloses a kind of spindle lysine bacillus and its application in land reclamation in mining area ecological reconstruction, which is spindle lysine bacillus (Lysinibacillus fusiformis), and deposit number is CGMCC No.9799.The bacterial strain can be improved reclaimed minesoils nutrient content, to having remarkable result in the raising of reclaimed minesoils organic matter, alkali-hydrolyzable nitrogen, available potassium and available phosphorus content；And further experiment shows that the spindle lysine bacillus can also improve crop biomass, either for the culture of wheat or rape, can improve reclaimed minesoils crop biomass；Significantly the use of spindle lysine bacillus effectively and can significantly change the structure of community of edaphon, improve diversity of soil microorganism, effectively increase the quantity of microorganism, to accelerate the reparation of soil microbial community, the reparation of coal field Reclaimed Soil can be widely applied to.
Description
Technical field
The invention belongs to ecology reestablishing technology field, it is related to a kind of spindle lysine bacillus and its in land reclamation in mining area
Application in ecological reconstruction.
Background technique
Coal has very important status in China's production of energy and consumption, and specific gravity remains at 70% left side
It is right.For exploitation of coal resources while supporting socio-economic development, it is lasting and tight also to cause to land resource and ecological environment
The negative effect of weight.《Chinese Geological environment bulletin in 2007》It points out, national Mining Development occupies and the land area of damage is
165.8 ten thousand hectares, wherein tailing stacks 90.9 ten thousand hectares, and 52.2 ten thousand hectares of openpit, 20.3 ten thousand hectares of Collapse by Mining, and it is every
Year newly-increased about 4,000,000 mu of soil of damage, wherein 60% or more is arable land or other farming lands, contradiction is not or not causing coal field people
Disconnected aggravation, seriously threatens China's grain security and Sustainable Socioeconomic Development.
It actively develops land reclamation in mining area and ecological reconstruction is the system engineering with strategic importance.China is in 1988
Year promulgates《Land reclamation regulation》, and in clear stipulaties Resource Development Process land reclamation responsibilities and obligations, this rule
Fixed implementation can not only reduce soil loss, but also can preserve the ecological environment, and be the system engineering benefited the nation and the people, and have important
Economy and society meaning, but to the land reclamation in mining area rate in current China less than 10%.Land reclamation be not only be destroyed or
The simple recovery of disturbed landform utilizes, it is often more important that ecological recovery, reclaiming for ecological significance are only land reclamation in mining area
Final purpose.The key of ecological recovery is the recovery of ecosystem function and the building of Rational structure, carry out vegetation recovery and
The building of association of plants and animals and microbiologic population.
Currently, China, in the research of mining collapse area land reclamation, various regions are generally mostly based on traditional measures for building.
Its existing main problem is that do not have living organism substance, and Soil Microorganism lacks, and matter energy conversion is slow, to the object of soil
The soil ecosystems such as rationality matter, biological property improve and restore slow, and the period of reclaiming is long.Currently, many countries are all using first
Into microbe restoration technology carry out Reclaimed Land.Microbe restoration technology is the inoculation advantage using microorganism, to Subsided Reclamation Region soil
Earth carries out a biotechnology measure of comprehensive treatment and improvement.It is by the plant inoculating to new planting it is some it is beneficial,
The microorganism of growth-promoting excavates the potential fertility of reclaimation measures using the vital movement of microorganism and plant rhizosphere microbe itself,
Again restore and establish its syntaxial system, promote plant growth, accelerate the base material improvement of reclaimed land soil and holding for the ecosystem
Supervention exhibition improves ecology of mining areas governance quality, so as to shorten the period of reclaiming.However it is micro- with remarkable result in production practice
The shortage of biological inoculum becomes the bottleneck problem of microorganism remediation technology.
Summary of the invention
The existing land reclamation in mining area period is long, disadvantage of the structural difference of Reclaimed Soil it is an object of the invention to overcoming, and provides
A kind of spindle lysine bacillus that having both phosphorus decomposing, potassium decomposing and nitrogen fixation and its in land reclamation in mining area ecological reconstruction
Using can be effectively improved reclaimed minesoils nutrient levels, microbiologic population, promote plant growth, accelerate ecological reconstruction speed
Degree.
The present invention is to be achieved through the following technical solutions：
A kind of spindle lysine bacillus, the bacterium are spindle lysine bacillus
(Lysinibacillusfusiformis), deposit number CGMCCNo.9799.
The spindle lysine bacillus being capable of the area Ti Kuanggao Reclaimed Soil organic matter, alkali-hydrolyzable nitrogen, available phosphorus, quick-acting
The content of potassium.
The spindle lysine bacillus can be improved the biomass of the crop grown in reclaimed minesoils.
The spindle lysine bacillus can be improved Microbial Diversity in reclaimed minesoils.
A kind of spindle lysine micro-organisms bacillus preparation is to train spindle lysine bacillus through expanding
After supporting, with 2:The active constituent that 10~20 mass ratio culture obtains is sufficiently mixed with carrier, is stored at room temperature；
The carrier is solid carrier or liquid-carrier, and wherein solid carrier is mineral material, vegetable material, producing high-molecular
Close object or organic material；The mineral material is one or more of clay, kaolin, montmorillonite, zeolite and flyash；Institute
Platymiscium material is one or more of in corn flour, starch, bean powder and peanut powder；The high-molecular compound be polyvinyl alcohol,
One or both of polyglycols；The organic material is turf, humic acid, the one or more in organic fertilizer；
The active constituent is to expand the filter for cultivating resulting living cells, packet celliferous fermentation liquid, cell culture
One or more of liquid；
Following dosage form is made in the microorganism formulation：Liquid agent, emulsion, suspending agent, granule, wettable powder or water dispersion
Granula.
The spindle lysine bacillus is improving the application in reclaimed minesoils.
It is organic that the spindle lysine bacillus or its manufactured microorganism formulation can be improved reclaimed minesoils
Matter, alkali-hydrolyzable nitrogen, available phosphorus, quick-acting potassium content；And improve reclaimed minesoils Microbial Diversity.
Application of the spindle lysine bacillus in the microorganism formulation that preparation improves reclaimed minesoils.
The spindle lysine bacillus answers in the bio-fertilizer that preparation promotes reclaimed minesoils plant growth
With.
Compared with prior art, the invention has the following beneficial technical effects：
Spindle lysine bacillus provided by the invention is pure by separating from the soil that Reclaimed Soil of Mining Area acquires
Change, then passes sequentially through Phos Selective agar medium, the selective medium of solid silicate bacterium, solid nitrogen-free agar again
It is screened, is measured by bioassay, 16SrDNA serial analysis and phosphorus decomposing, ability of dissolving potassium, determine that screened microorganism is
New microorganism：Spindle lysine bacillus (Lysinibacillusfusiformis).The spindle lysine gemma
Bacillus shows extremely strong soil adaptability, is a kind of high-effective microorganism for having both phosphorus decomposing, potassium decomposing and nitrogen fixation；Experiment inspection
Survey display, can be improved reclaimed minesoils nutrient content, to reclaimed minesoils organic matter, alkali-hydrolyzable nitrogen, available potassium, effectively
There is remarkable result in the raising of phosphorus content；And further experiment shows that the spindle lysine bacillus can also
Crop biomass is improved, either for the culture of wheat or rape, can improve reclaimed minesoils crop biology
Amount；Significantly the use of spindle lysine bacillus can group's knot that is effective and significantly changing edaphon
Structure improves diversity of soil microorganism, the quantity of microorganism is effectively increased, to accelerate repairing for soil microbial community
It is multiple, it can be applied to improve reclaimed minesoils and promote reclaimed minesoils plant growth.
Detailed description of the invention
Fig. 1 is spindle lysine bacillus (Lysinibacillusfusiformis) of the invention in solid beef
48 hours forms are cultivated on cream peptone culture medium；
Fig. 2 is spindle lysine bacillus (Lysinibacillusfusiformis) in Solid nutritional agar culture
After being cultivated 24 hours on base after Gram's staining under optical microscopy oil mirror (10*100 times) form.
Preservation explanation
Spindle lysine bacillus (Lysinibacillusfusiformis) provided by the invention, the bacterial strain in
On October 21st, 2014 is stored in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), deposit number
For CGMCCNo.9799, depositary institution address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
Specific embodiment
Below with reference to specific embodiment, the present invention is described in further detail, it is described be explanation of the invention and
It is not to limit.
The present invention provides a kind of spindle lysine bacillus (Lysinibacillusfusiformis), wherein
The deposit number of the spindle lysine bacillus (Lysinibacillus fusiformis) is CGMCCNo.9799.
The present invention provides the spindle lysine bacillus or the microbial bacterial agent to improve land reclamation in mining area soil
Application in earth, promotion plant growth.
Further description is carried out to the present invention below by specific embodiment.
Embodiment 1
This example demonstrates that spindle lysine bacillus (Lysinibacillusfusiformis) of the invention
(CGMCCNo.9799) screening and identification process
1. sample acquires
Soil is acquired from Shanxi Province Typical Reclamation mining area, coal mining subsidence area is reclaimed from Shanxi Changzhi Xiangyuan County Western Hills bottom village respectively
The weeds clump (113 ° 00 ' 50.383 of 36 ° 28 ' 17.846 of N ", E ") in area acquires 0-20cm pedotheque, 4 DEG C of preservations；Shanxi filial piety
Weeds clump (111 ° 30 ' 65.453 of 37 ° 08 ' 25.485 of N ", E ") the acquisition 0-20cm pedotheque of the village Yi Kee refuse dump, 4
DEG C preservation.
2. the primary dcreening operation of bacterial strain
90mL is added equipped with bead and sterile water and sterilized 250mL tri- in the reclaimed minesoils for weighing 10g acquisition
In the bottle of angle, 20min is shaken under 30 DEG C, the constant-temperature table of 180r/min, draws 1mL bacterium solution to 9mL sterile water after standing 5min
In mix well that (dilution is calculated as 10 at this time-2), then filled in 9mL sterile water from absorption 1mL bacterium solution in this test tube to another
It mixes well, and so on, obtain 10-3、10-4、10-5、10-6The soil supension of different dilutions.Finally it is by diluted concentration
10-4、10-5、 10-6Bacteria suspension 0.1mL be coated on upper (10g/L containing glucose, Ca of solid inorganic phosphorus Selective agar medium3
(PO4)2The fine jade of 5g/L, magnesium chloride 5g/L, magnesium sulfate 0.25g/L, potassium chloride 0.25g/L, ammonium sulfate 0.15g/L and 18g/L
Rouge), three repetitions are arranged in the soil supension of each dilution gradient, cultivate 10d at 30 DEG C, screening obtain 8 plants can be formed it is larger
The bacterial strain of transparent circle, the transparent loop diameter (D) and colony diameter (d) ratio of this 8 plants of bacterial strains are equal>1.5.
The method that scribing line is respectively adopted in 8 plants of bacterial strains that screening obtains is inoculated into the selectivity culture of solid silicate bacterium
(Alexandria sieve husband's culture medium, 5g/L containing sucrose, magnesium sulfate 0.5g/L, Na on base2HPO42g/L, CaCO30.1g/L,
FeCL30.005g/L, soil mineral 1.0g/L, agar 18g/L), 28 DEG C, after cultivating 4d, choosing colony is big, transparent protrusion is high,
It adheres flexible bacterium colony, crosses repeatedly on sieve husband's culture medium of Alexandria, and do microscopy, be until obtaining pure culture
Only, 4 plants of pure culture bacterial strains are obtained altogether.
The method that scribing line is respectively adopted in 4 plants of bacterial strains that screening obtains is inoculated into (Ah Xu the shellfish culture of solid nitrogen-free agar
Base, 10g/L containing glucose, potassium dihydrogen phosphate 0.2g/L, magnesium sulfate 0.2g/L, sodium chloride 0.2g/L, calcium sulfate 0.1g/L, carbon
Sour calcium 5g/L, agar 18g/L, pH 7.0, agar 18g/L) on, 1 plant of well-grown bacterial strain is obtained altogether after cultivating 7d at 30 DEG C
G-1207, bacterial strain after purification are stored in beef extract peptone medium slant.
3. secondary screening
Phosphorus decomposing, ability of dissolving potassium measurement are carried out to the bacterial strain G-1207 filtered out.
The measurement of 3.1 dissolving P capacities：
Bacterial strain G-1207 is inoculated into Phos culture medium (10g/L containing glucose, Ca3(PO4)25g/L, magnesium chloride 5g/L,
Magnesium sulfate 0.25g/L, potassium chloride 0.25g/L and ammonium sulfate 0.15g/L) triangular flask in, make G-1207 content 109cfu/
ML is placed on shaking table, 150r/mim, 30 DEG C of shake culture 5d；It simultaneously will be from Chinese Academy of Agricultural Sciences's agricultural microbial preservation
The bacillus megaterium (ACCC11107) of heart purchase is inoculated in the same way as positive control；Bacterial strain G-1207 is inactivated
Inoculation is used as negative control afterwards, and each processing repeats three times；Obtained fermentation liquid 5000r/mim is centrifuged 30min, using molybdenum antimony
Available phosphorus content in anti-colorimetric method for determining supernatant, and phosphorus decomposing rate is calculated, with the pH value (testing result of acidometer measurement supernatant
As shown in table 1).
The phosphorus decomposing rate of table 1G-1207
Note：Same column difference lowercase is indicated in p<Significant difference on 0.05.
Testing result shows that the bacterial strain has significant phosphorus decomposing effect, and phosphorus decomposing rate reaches 23.18%.
The measurement of 3.2 ability of dissolving potassium：
1) prepared by seed liquor：
The inclined-plane lawn of the activated G-1207 of one ring of picking be linked into 30mL seed culture fluid (starch-containing 5.0g/L,
Yeast extract 1.0g/L, K2HPO42.0g/L, MgSO47H2O 0.5g/L, CaCO30.1 g/L, FeCl3·6H2O 5m g/L, pH
7.5-8.5), 180r/min shaking table culture, for 24 hours, thallus content is about 2 × 10 to 28 DEG C of shaken cultivations8cfu/mL.It will be from Chinese agriculture
The silicate bacteria (ACCC10013) of industry academy of sciences agricultural Organism Depositary purchase is inoculated in the same way as positive
Control.
2) the shaking flask culture of silicate bacteria：
Potassium decomposing culture medium (10.0g/L containing sucrose, MgSO that will be prepared4·7H2O 0.5g/L, (NH4)2·SO40.2g/L,
NaCl 0.1g/L, CaCO30.1g/L, soil mineral 0.1g/50ml culture solution, pH 7.2-7.5) 50mL is added to 250ml's
In conical flask, sterilizing cooling, every bottle connects the seed liquor that 3mL is prepared, and silicate bacteria (ACCC10013) is positive control, together
When equivalent inactivation G-1207 seed liquor is added as negative control, all processing repeat three times, in 180r/min after inoculation, 28
Under the conditions of DEG C, shaken cultivation 5 days.
3) research of silicate bacteria potassium decomposing rate
Whole fermentation liquids are poured into evaporating dish, heating is concentrated into 10ml or so in water-bath, and 2.0ml H is added2O2
Solution continues to evaporate, and is stirred continuously, and adds 20%H repeatedly2O2Solution digests completely to stickum.3500r/min centrifugation
Then 10min uses distilled water constant volume by supernatant collection into 50ml volumetric flask, measure in fermentation liquid on flame photometer
The content of water-soluble potassium, testing result are as shown in table 2.
The potassium decomposing rate of table 2G-1207
Note：Same column difference lowercase is indicated in p<Significant difference on 0.05.
The result shows that the bacterial strain potassium decomposing significant effect, the increment rate of water-soluble potassium reach 74.26%.
4. the identification of bacterial strain
According to《Common bacteria system identification handbook》With《Bacterial system principle, method and practice》In the experiment of middle record
Appearance and experimental method carry out strain morphology, the analysis mirror of Physiology and biochemistry and 16SrDNA to the bacterial strain for the G-1207 that screening obtains
It is fixed.
The morphological feature of the bacterial strain：Bacterium colony is circle, colorless and transparent, neat in edge, center protrusion, toughness, electricity
Under the microscope be it is rod-shaped (as shown in Figure 1), have gemma.
Further detection physiological and biochemical property is as shown in table 3.
The physiological and biochemical property of table 3G-1207
| Catalase | + | B- galactosidase | - | D-glucitol | + |
| Oxidizing ferment | + | Urase | - | PEARLITOL 25C | + |
| V.P test | - | Arginine dihydrolase | - | D-arabinose alcohol | + |
| M.R test | - | Lysine deshydroxy enzyme | - | Inositol | + |
| Nitrate reduction | - | Ornithine deshydroxy enzyme | + | L-arginine | - |
| Gelatin liquefaction | - | Glycerol | + | L-Alanine | - |
| Starch Hydrolysis | - | D-Glucose | + | L-Aspartic acid | + |
| H2S is generated | - | D- galactolipin | + | Pidolidone | + |
| Indoles generates | - | D-Fructose | + | 50 DEG C of growths | - |
| Citric acid growth | + | D-MANNOSE | + | ||
| 7%NaCl growth | + | D-Maltose | + |
The physiological and biochemical test result of the bacterial strain：Gram-positive (coloration result is as shown in Figure 2), catalase sun
Property, oxidase positive, acetonyl alcohol (V.P) is negative, and methyl red (M.R) is negative, nitrate reduction negative, gelatin liquefaction yin
Property, starch hydrolysis negative, production hydrogen sulfide is negative, and indole test is negative, and citric acid growth is positive, and 7%NaCl grows positive, b- half
Lactoside enzyme, urase, arginine dihydrolase, lysine deshydroxy enzyme are negative, ornithine deshydroxy enzyme positive, glycerol, D-Glucose,
D- galactolipin, D-Fructose, D-MANNOSE, D-Maltose fermentation and acid are positive, D-glucitol, PEARLITOL 25C, D- arabitol, flesh
Alcohol fermentation produces the acid positive, and L-arginine, l-Alanine fermentation and acid are negative, L-Aspartic acid, Pidolidone fermentation and acid yin
Property.
Further progress biology 16SrDNA serial analysis extracts bacterial genomes DNA and carries out PCR amplification later：
16SrDNA carries out PCR amplification using 27F/1492R primer, and primer sequence is as follows：
27F:5'-AGAGTTTGATCCTGGCTCAG-3',
1492R：5'-GGTTACCTTGTTACGACTT-3'
PCR reaction system (25 μ l)：Template<1μg,Primer1(10μM)1μl,Primer2(10μM) 1μl,2×
12.5 μ l of PfuPCRMasterMix, sterile purified water are mended to 25 μ l；
PCR reaction condition：93 DEG C of initial denaturations 5min, 94 DEG C of denaturation 18s, 56 DEG C of annealing 15s, 72 DEG C of extension 78s, circulation
30 times, 72 DEG C of extension 7min.After reaction, 5 μ l reaction products are taken, agarose gel electrophoresis detects amplified production.PCR sequencing
It is completed by Institute of Microorganism, Academia Sinica.
Sequencing obtains 16S rDNA sequence, and NCBI carries out BLAST tetraploid rice, and combining form observation and physiology are raw
Change test result, is ultimately determined to spindle lysine bacillus (Lysinibacillus fusiformis).
The bacterial strain is preserved in Chinese culture presevation administration committee common micro-organisms center (on November 5th, 2014
Location：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), deposit number is CGMCCNo. 9799.
Embodiment 2
This example demonstrates that the culture of spindle lysine bacillus of the invention and the preparation of microbial bacterial agent
A kind of spindle lysine micro-organisms bacillus preparation is to cultivate spindle lysine bacillus through expanding
Afterwards, with 2:The active constituent that 10~20 mass ratio culture obtains is sufficiently mixed with carrier, is stored at room temperature；
The carrier is solid carrier or liquid-carrier, and wherein solid carrier is mineral material, vegetable material, producing high-molecular
Close object or organic material；The mineral material is one or more of clay, kaolin, montmorillonite, zeolite and flyash；Institute
Platymiscium material is one or more of in corn flour, starch, bean powder and peanut powder；The high-molecular compound be polyvinyl alcohol,
One or both of polyglycols；The organic material is turf, humic acid, the one or two in organic fertilizer；
The active constituent is to expand the filter for cultivating resulting living cells, packet celliferous fermentation liquid, cell culture
One or more of liquid；
Following dosage form is made in the microorganism formulation：Liquid agent, emulsion, suspending agent, granule, wettable powder or water dispersion
Granula.
Specifically, being the spindle lysine bacillus for being CGMCC NO.9799 by deposit number
(Lysinibacillusfusiformis) it is inoculated into broth bouillon (the peptone and 5g/L of beef extract, 10g/L containing 5g/L
NaCl) in, culture is to strain density OD under 30 DEG C and 200r/min oscillations600Value is 0.6-0.8, obtains seed bacterium solution.
Above-mentioned seed bacterium solution is added in 100L fermentor by 5%-10% inoculum concentration (fermentation medium contains 10~
30mL/ yellow serofluid, the groundnut meal of 10~50g/L, the sucrose of 40~80g/L, the sodium chloride of 2~4g/L, 2~4g/L carbonic acid
The ferrous sulfate of calcium and 3~6g/L) 30 DEG C, it cultivates, is sampled during the cultivation process simultaneously under pH 7.2, ventilatory capacity 0.5vvm
It is observed by ascites method, until the viable count of the spindle lysine bacillus in every gram of culture solution is
109CFU。
After fermentation, by fermentation liquid (active constituent) according to 2:Turf (carrier) is added in 10~20 ratio, and room temperature is protected
It deposits to get the microbial bacterial agent of the present embodiment is arrived.
Embodiment 3
This example demonstrates that spindle lysine bacillus agent of the invention is improving reclaimed minesoils nutrient content
On effect
Rape, spring wheat are planted using pot-culture method, implantation time is respectively (60 days, 100 days).The substrate soil that potting uses
Earth, which comes from, is located at the Shanxi Province southeast, the sinking land in coalmining areas reclaimed land of Taihang Shan Xilu, Lu An group.
Cole soil content of organic matter 5.97g/kg, full nitrogen 0.50g/kg, full phosphorus 0.05g/kg, full potassium 20.00g/kg,
Alkali-hydrolyzable nitrogen 42.82mg/kg, available phosphorus 3.39mg/kg, available potassium 98.60mg/kg, pH8.29.
Spring wheat soil organic matter content 6.40g/kg, full nitrogen 0.40g/kg, full phosphorus 0.4g/kg, full potassium 19.80g/kg,
Alkali-hydrolyzable nitrogen 48.76mg/kg, available phosphorus 2.09mg/kg, available potassium 98.5mg/kg, pH8.37.
In this test, using the microbial inoculum in embodiment 2, while the restoration of the ecosystem microbial bacterial agent 1 for choosing market sale is (right
Ratio 1, the auspicious precious bioremediation agents in ground of lattice), restoration of the ecosystem microbial bacterial agent 2 (comparative example 2, pioneer's-Ao Ruigen microbial bacteria of exploring the way
Agent) and embodiment 2 in microbial inoculum inactivation microbial inoculum (comparative example 3) as control, be respectively added to cole soil, spring wheat soil
Middle progress effect comparison.The usage amount of microbial inoculum is 10g microbial inoculum/kg soil, and method of application is to fill basin after mixing well withing the soil, and is then planted
Crop.
Soil moisture keeps the 70% of water-retaining quantity among field of soil during plantation.To the shadow of soil nutrient content after plantation
Sound is shown in Table 4.
To soil nutrient content after 4 kinds of table plants
Note：Same column difference lowercase is indicated in p<Significant difference on 0.05.
As the result of table 4 as it can be seen that spindle lysine micro-organisms bacillus microbial inoculum provided by embodiment 2 is improving mine
There is remarkable result on area's Reclaimed Soil organic matter, alkali-hydrolyzable nitrogen, available potassium and available phosphorus content.
Embodiment 4
This example demonstrates that spindle lysine bacillus agent of the invention is improving the effect on crop biomass.
Rape, spring wheat are planted using pot-culture method, implantation time is respectively (60 days, 100 days).The substrate soil that potting uses
Earth, which comes from, is located at the Shanxi Province southeast, the sinking land in coalmining areas reclaimed land of Taihang Shan Xilu, Lu An group.
Cole soil content of organic matter 5.97g/kg, full nitrogen 0.50g/kg, full phosphorus 0.05g/kg, full potassium 20.00g/kg,
Alkali-hydrolyzable nitrogen 42.82mg/kg, available phosphorus 3.39mg/kg, available potassium 98.60mg/kg, pH8.29.
Spring wheat soil soil organic matter content 6.40g/kg, full nitrogen 0.40g/kg, full phosphorus 0.4g/kg, full potassium
19.80g/kg, alkali-hydrolyzable nitrogen 48.76mg/kg, available phosphorus 2.09mg/kg, available potassium 98.5mg/kg, pH8.37.
In this test, embodiment is the microbial inoculum in embodiment 2, while choosing the restoration of the ecosystem microbial bacterial agent of market sale
As a comparison；Comparative example 1, comparative example 2, comparative example 3 are same as Example 3.
The usage amount of microbial inoculum is 10g microbial inoculum/kg soil, and method of application is to fill basin after mixing well withing the soil, then long-term cropping.Kind
Soil moisture keeps 70% (the results are shown in Table 5 of crop biomass) of water-retaining quantity among field of soil during plant.
The raising of 5 crop biomass of table
Note：Same column difference lowercase is indicated in p<Significant difference on 0.05.
By the result of table 5 as it can be seen that embodiment has remarkable result on improving reclaimed minesoils crop biomass.
Embodiment 5
This example demonstrates that spindle lysine bacillus agent of the invention is improving reclaimed minesoils micropopulation
Fall the effect on structure diversity.
Rape, spring wheat are planted using pot-culture method, implantation time is respectively (60 days, 100 days).The substrate soil that potting uses
Earth, which comes from, is located at the Shanxi Province southeast, the sinking land in coalmining areas reclaimed land of Taihang Shan Xilu, Lu An group.
Cole soil content of organic matter 5.97g/kg, full nitrogen 0.50g/kg, full phosphorus 0.05g/kg, full potassium 20.00g/kg,
Alkali-hydrolyzable nitrogen 42.82mg/kg, available phosphorus 3.39mg/kg, available potassium 98.60mg/kg, pH8.29.The spring wheat soil soil organism
Content 6.40g/kg, full nitrogen 0.40g/kg, full phosphorus 0.4g/kg, full potassium 19.80g/kg, alkali-hydrolyzable nitrogen 48.76mg/kg, available phosphorus
2.09mg/kg, available potassium 98.5mg/kg, pH8.37.
In this test, embodiment is the spindle lysine micro-organisms bacillus microbial inoculum of embodiment 2, while choosing market
The restoration of the ecosystem microbial bacterial agent of sale is as a comparison；Comparative example 1, comparative example 2, comparative example 3 are same as Example 3.
The usage amount of microbial inoculum is 10g microbial inoculum/kg soil, and method of application is to fill basin after mixing well withing the soil, then long-term cropping.Kind
Soil moisture keeps the 70% of water-retaining quantity among field of soil during plant.
Soil microbial biomass is characterized using phospholipid fatty acid system (PLFA), analyzes edaphon after potted plant growth rape
Fall the variation (the results are shown in Table 6) of structure.
The variation of structure of soil microbial community after 6 potted plant growth rape of table
Note：Same column difference lowercase is indicated in p<Significant difference on 0.05.
Table 6 the result shows that, spindle lysine bacillus use can effectively and significantly change edaphon
Structure of community improves diversity of soil microorganism, significantly increases the quantity of microorganism, accelerate repairing for soil microbial community
It is multiple.
Effect based on the above spindle lysine bacillus and its microorganism formulation proposes following application：
Spindle lysine bacillus is improving the application in reclaimed minesoils.
Application of the spindle lysine bacillus in the microorganism formulation that preparation improves reclaimed minesoils.
Spindle lysine bacillus is applied in the bio-fertilizer that preparation promotes reclaimed minesoils plant growth.
The preferred embodiment of the present invention has been described above in detail, still, during present invention is not limited to the embodiments described above
Detail within the scope of the technical concept of the present invention can be with various simple variants of the technical solution of the present invention are made, this
A little simple variants all belong to the scope of protection of the present invention.
Claims (9)
1. a kind of spindle lysine bacillus, which is characterized in that the bacterium is spindle lysine bacillus
(Lysinibacillusfusiformis), deposit number CGMCCNo.9799.
2. spindle lysine bacillus as described in claim 1, which is characterized in that the spindle lysine gemma bar
Bacterium can the area Ti Kuanggao Reclaimed Soil organic matter, alkali-hydrolyzable nitrogen, available phosphorus, available potassium content.
3. spindle lysine bacillus as described in claim 1, which is characterized in that the spindle lysine gemma bar
Bacterium can be improved the biomass of the crop grown in reclaimed minesoils.
4. spindle lysine bacillus as described in claim 1, which is characterized in that the spindle lysine gemma bar
Bacterium can be improved Microbial Diversity in reclaimed minesoils.
5. a kind of spindle lysine micro-organisms bacillus preparation, which is characterized in that be by spindle lysine bacillus
After expanding culture, with 2:The active constituent that 10~20 mass ratio culture obtains is sufficiently mixed with carrier, is stored at room temperature；
The carrier is solid carrier or liquid-carrier, and wherein solid carrier is mineral material, vegetable material, high-molecular compound
Or organic material；The mineral material is one or more of clay, kaolin, montmorillonite, zeolite and flyash；Affiliated plant
Object material is one or more of in corn flour, starch, bean powder and peanut powder；The high-molecular compound is polyvinyl alcohol, poly- two
One or both of alcohol；The organic material is turf, humic acid, the one or more in organic fertilizer；
The active constituent is to expand in the filtrate for cultivating resulting living cells, the celliferous fermentation liquid of packet, cell culture
One or more；
Following dosage form is made in the microorganism formulation：Liquid agent, emulsion, suspending agent, granule, wettable powder or water-dispersible grain
Agent.
6. spindle lysine bacillus described in claim 1 is improving the application in reclaimed minesoils.
7. application as claimed in claim 6, which is characterized in that the spindle lysine bacillus or micro- life made of it
Object preparation can be improved reclaimed minesoils organic matter, hydrolyzable nitrogen, available potassium and available phosphorus content；And improve land reclamation in mining area
Community diversity in different habitats.
8. spindle lysine bacillus described in claim 1 is in the microorganism formulation that preparation improves reclaimed minesoils
Application.
9. the biology that spindle lysine bacillus described in claim 1 promotes reclaimed minesoils plant growth in preparation
It is applied in fertilizer.
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| CN107201320A (en) * | 2016-03-18 | 2017-09-26 | 兰州大学 | One plant can make tosca spindle lysine bacillus LZ1 and its application |
| CN107201320B (en) * | 2016-03-18 | 2021-09-17 | 兰州大学 | Lysinibacillus fusiformis LZ1 capable of depositing calcium carbonate and application thereof |
| CN109576171A (en) * | 2018-11-16 | 2019-04-05 | 广东植物龙生物技术股份有限公司 | Spindle lysine bacillus and its application |
| CN109576171B (en) * | 2018-11-16 | 2021-11-23 | 广东植物龙生物技术股份有限公司 | Lysinibacillus fusiformis and application thereof |
| CN111893074A (en) * | 2020-08-14 | 2020-11-06 | 贵州大学 | Bacillus fusiformis strain and application thereof |
| CN111893074B (en) * | 2020-08-14 | 2022-05-20 | 贵州大学 | Bacillus fusiformis strain and application thereof |
| CN112974512A (en) * | 2021-02-03 | 2021-06-18 | 宁夏大学 | Application of bacillus fusiformis with MICP function in tailing greening repair |
| CN116200308A (en) * | 2023-03-02 | 2023-06-02 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Bacillus fusiformis and application thereof |
| CN116573974A (en) * | 2023-05-09 | 2023-08-11 | 承德市农林科学院 | Soil conditioner based on industrial and agricultural wastes and preparation method thereof |
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