CN109762766A - One plant has absorption heavy metal, phosphorus decomposing and the prebiotic bacterium and its application of plant - Google Patents
One plant has absorption heavy metal, phosphorus decomposing and the prebiotic bacterium and its application of plant Download PDFInfo
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Abstract
The present invention relates to one plant of probiotic bacteria (Edaphovirga sp.) and its applications, and in particular to one plant of probiotic bacteria and its absorption heavy metal, phosphorus decomposing and prebiotic effect belong to the development and utilization field of microbial resources.The bacterium has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC No.15748.The bacterial strain has stronger heavy metal adsorption effect, phosphate solubilization and promotes plant growth, promotes plant seed germination effect.The present invention carries out pouring root to plant using the strain suspensions or bacterial strain fermentation liquor or bacterial strain fermentation liquor extracting solution of the bacterial strain or the solid fungicide containing the bacterial strain or spreads fertilizer over the fields processing.Can be used alone, can also be mixed with organic fertilizer bacterial manure use, this by for the high yield of planting industry, stable yields, it is high-quality take a firm foundation, to development green non-pollution agricultural production have very important significance.
Description
Technical field
The invention belongs to the development and utilization fields of microbial resources, and being related to one plant has absorption heavy metal, phosphorus decomposing and plant
The prebiotic bacterium of object (Edaphovirga sp.) and its application, and in particular to one plant of probiotic bacteria and its absorption heavy metal, phosphorus decomposing
And prebiotic effect.
Background technique
As the development and mankind's activity of social economy are gradually reinforced, exploitation of mineral resources, plating, steel and iron manufacturing, process hides,
The heavy metal release of high-content to natural environment is caused soil, water pollution problems more next by chemical process and the application of chemical fertilizer etc.
It is more serious.According to statistics, until 2016, China is had reached hectares up to ten thousand, Zhan Quan by the soil erosion of heavy metal pollution
The 1/6 of state's total area under cultivation.The exceeded heavy metal element of content mainly includes chromium in the soil of China's heavy metal pollution at present
(Cr), cadmium (Cd), lead (Pb) and mercury (Hg) etc. endanger human health by food chain accumulation.As the Cd of human body excess intake,
Bone density, which can be caused, to be reduced and fractures.And after Pb enters human body, immunity of organisms decline will be caused, when accumulation is to a certain amount of
Later, the symptoms such as headache, memory loss and abdomen pain will be caused, seriously threaten human health.Therefore, repairing heavy metal in soil
It is imperative to pollute.Mobility is very poor in the soil for heavy metal contaminants, and the residence time is long, with not destroying biologically
Property, therefore, finding the friendly method repairing heavy metal pollution of ecology is particularly important.Currently, heavy-metal contaminated soil reparation
Technical way include physics, chemistry and biological prosthetic.The repairing effect of first two method in a short time is preferable, but deposits
The problems such as project amount is big, costly, and it be easy to cause the secondary pollution of soil.In recent years, heavy metal biological repairs skill
Art is increasingly becoming research hotspot both domestic and external, and the plant and microbial association with the characteristics of green, environmentally friendly and is cheap are repaired
Recovering technology is concerned.And microbial remediation method utilizes absorption, conversion and metabolism of the microorganism to heavy metal, can not only reduce
The toxicity of heavy metal in soil, and the microenvironment of plant crop root system can be improved, to reduce rehabilitation cost, and it is not easy
Secondary pollution is caused, can also handle underground water and soil simultaneously, suffered time, limitation spatially are small, are remediating heavy metal dirts
Contaminate the important means of soil.The researchs such as Xiong Fen find the bases such as hydroxyl, carboxyl and the C-O-C in aspergillus fumigatus extracellular polymeric (EPS)
Group is and Pb2+In conjunction with essential groups.Manasi etc. is handled using one plant of Halomonas and is rich in Cd in electronics industry2+Waste water.
Phosphorus is one of three big nutrients necessary to higher plant grows, and is various metabolism in plant growth and development process
The indispensable element such as movable participant and formation Nuclear extract, lecithin.And in soil 95% phosphorus with organic or
Inorganic phosphide state exists, water-soluble low, it is difficult to be directly absorbed and utilized by plant, the quick-acting state phosphorus energy quilts of only seldom ratio
Plant directly utilizes, to cause the shortage of phosphorus element in world wide.Increased in agricultural production using a large amount of phosphate fertilizer are applied
The supply of soil phosphorus, however phosphate fertilizer easily with the Ca in soil2+、Al3+、Fe2+And Fe3+Chelating forms insoluble phosphate, phosphorus
Fertile utilization efficiency is only 5%-25%, and excessively can destroy soil texture using chemical fertilizer, pollutes environment.Therefore, it studies in soil
The decomposition and release of fixed unavailable phosphorus to the titanium pigment content improved in soil, adjust soil fertility with important
Meaning.And the phosphorus that plant can be difficult to absorb by phosphate solubilizing microorganism is converted into availability status, improve soil available phosphorus content,
Phosphate fertilizer utilization efficiency and promotion plant growth etc. have remarkable effect.History is sent out superfine and has screened 1 plant of dissolvable insoluble phosphorus
Bacterial strain P83 penicillium decumbens, can significantly improve soil available phosphorus level, have remarkable effect to corn growth and volume increase.Therefore,
Phosphate solubilizing microorganism resource will have more wide application prospect in the development process of the following sustainable agriculture and forestry.
Modern agriculture is overly dependent upon the use of chemical pesticide to the prevention and treatment of plant disease, not only has length to ecological environment
The destruction of phase, the problems such as also causing agricultural product quality decline, excessive pesticide residues, the drug resistance of pathogen.Plant is to disease
Harmful and other adverse circumstances resistances and phenylalanine lyase (PAL), superoxide dismutase (SOD), peroxidase (POD) etc.
A variety of Enzyme activities are related, and biocontrol microorganisms can induce these plant disease-resistant correlations defence enzyme activity and generate variation, enhance disease-resistant
Ability.Superoxide dismutase (SOD) is in critical role in active oxygen metabolism, and superoxide radical can be prevented to biological membrane system
The oxidation of system, activity height are the important symbols of plant cell itself anti-aging ability power.Peroxidase (POD) is not only
The polymerization reaction of lignin, and intracellular important endogenous activity oxygen scavenger are participated in, H can be catalyzed2O2Resolve into H2O and O2。
Woods Chen Qiang etc. discovery, Bacillus subtillis CS16 fermentation liquid and supernatant processing banana seedlings can induce SOD in Leaf of banana,
The variation of the Defense Enzyme Activities such as PAL and POD.Therefore, the activity of plant defense enzyme system is improved by biological method to improve plant
Resistance, the application for reducing chemical pesticide have very important significance to the production for developing green non-pollution agricultural.
Summary of the invention
It is an object of the invention to: the probiotic bacteria that one plant of offer is separated from Chinese soil, screened
(Edaphovirga sp.) bacterial strain, the bacterium have absorption heavy metal effect, phosphate solubilization and the benefit to vegetable seeds and plant
Raw effect.It can be used as bio-feritlizer use, achieve the purpose that improve the yield and quality, there is at low cost, no pollution to the environment, right
The characteristics of safety of human and livestock.
Probiotic bacteria of the present invention obtains by the following method:
Codonopsis clematidea Clarke's rhizosphere soil is taken, 5g is weighed after soil sample is removed stone, is put into the triangular flask for filling 45mL sterile water
In, 20min is vibrated, this i.e. 10-1Soil supension;It is diluted to 10 step by step-2、10-3、10-4、10-5Afterwards, each dilution is taken to apply respectively
It is distributed in separation plate, is inverted, 26 DEG C of 1~5d of culture.Culture observation, single colonie are transferred on LB culture medium after growing and cultivate, i.e.,
Obtain bacterial strain (Edaphovirga sp.).
The present invention also provides the probiotic bacteria absorption heavy metal, phosphorus decomposing and to vegetable seeds and plant it is prebiotic in
Effect.
The present invention can use the strain suspensions or bacterial strain fermentation liquor or strain fermentation of probiotic bacteria (Edaphovirga sp.)
Liquid extracting solution or solid fungicide containing the bacterial strain carry out pouring root to plant or spread fertilizer over the fields processing.
One, the strain suspensions preparation method: the bacterial strain culture presevation pipe is taken out, is carried out with LB solid medium
Recovery, 20-37 DEG C culture 24-48 hours, activated bacterial strain is prepared into bacteria suspension using distilled water, uses ultraviolet spectrometry light
Degree meter measures bacteria suspension concentration, the concentration of the bacteria suspension are as follows: 0.1-0.8 at wavelength 600nm.LB solid medium is matched
Side are as follows: peptone 1-5%, yeast extract 0.1-5%, NaCl 0.1-3%, agar 1.5-2%, water 1000ml.
Two, it the preparation method of the bacterial strain fermentation liquor or bacterial strain fermentation liquor extracting solution: takes out the bacterial strain strain and protects
Hiding pipe, recovered with LB solid medium, 20-37 DEG C culture 24-48 hours.Picking single bacterium colony is seeded on plate
In 50ml LB culture medium, 20-37 DEG C shake culture 24-72 hours in the incubator.Seed uses the inoculum concentration of 1-10%, connects
Kind expands culture 24-72 hours into LB culture medium.Bacterial concentration is greater than OD600=1 can terminate to ferment, and sterile filling obtains
Liquid end product, or powdery finished product is obtained by freeze-drying.Aforementioned resulting bacterial strain fermentation liquor is used into isometric ethyl acetate
Extraction collects concentrated extract, obtains bacterial strain fermentation liquor extracting solution, spare.
Three, the solid fungicide is prepared via a method which: above-mentioned resulting bacterial strain fermentation liquor is inoculated into solid bacterium
In agent culture medium, 20-40 DEG C of culture is aseptically stirred after cultivating 1-3d, is continued to cultivate 1-3d after stirring, be obtained
Solid fermentation microbial inoculum.The formula of solid fungicide culture medium, as follows in terms of g/ml, wheat bran 20-50%, corn flour 10-30%, bean powder
1-10%, potassium nitrate 0.1-1%, dipotassium hydrogen phosphate 0.1-1%, sodium chloride 0.4-0.7%, calcium carbonate 1-10%, distilled water
200ml。
Microbial inoculum made from step 1 and step 2 can be diluted 10-10000 times and used by the present invention, or step 3 is made
Microbial inoculum directly spread fertilizer over the fields, or be used in mixed way with other organic fertilizers.
Probiotic bacteria (Edaphovirga sp.) of the invention, it is general in China Committee for Culture Collection of Microorganisms
Logical microorganism center preservation, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, preservation
Number: CGMCC No.15748, the deposit date is on May 09th, 2018.
Detailed description of the invention
Fig. 1 is adsorption rate of the bacterial strain Edaphovirga sp.No.15748 to Pb, Cd;
Fig. 2 is the phosphorus content standard curve drawn under 700nm wavelength using molybdenum antimony resistance colorimetric method;
Fig. 3 is the phosphate solubilization on bacterial strain Edaphovirga sp.No.15748 liquid medium within.
Specific embodiment
It is described in detail With reference to embodiment, but it should be understood that protection scope of the present invention is not by specific
The limitation of embodiment.Experimental method used in following embodiments is conventional method unless otherwise specified.Following implementations
Material, reagent used in example etc., unless otherwise specified, is commercially available.
The effect of 1 bacterial strain of embodiment (Edaphovirga sp.) CGMCC No.15748 heavy metal adsorption
Heavy metal tolerance test: by bacterial strain (Edaphovirga sp.) CGMCC No.15748 heavy metal (lead,
Cadmium) concentration reaches final concentration of 200mg/L in solid LB, and the concentration gradient of 400mg/L, 800mg/L carry out plate tolerance test.
Heavy metal adsorption test: the fermentation liquid for being 60mg/L according to 2% ratio access heavy metal concentration by kind of liquid
In, 120-200r/min fermentation 48h, compares not connect the culture medium of bacterium under the conditions of 26-35 DEG C, and three groups are parallel, it takes for 24 hours, 48h's
Supernatant 3ml after fermentation liquid centrifugation, is added the concentrated nitric acid of 3ml, adds water and be diluted to 45ml, is i.e. sample to be tested.Utilize ICP-MS (electricity
Sense coupled plasma mass spectrometry) measure remaining total concentration of metal ions in fermentation liquid.
As a result as shown in Figure 1, bacterial strain (Edaphovirga sp.) CGMCC No.15748 reaches Pb adsorption rate for 24 hours
40 ± 1.63%, 37.5 ± 1.2%, 48h is reached to Cd adsorption rate, 70 ± 1.22% are reached to Pb adsorption rate, Cd adsorption rate is reached
To 67.5 ± 2.04%, the ability of very strong absorption heavy metal is shown.
2 bacterial strain of embodiment (Edaphovirga sp.) CGMCC No.15748 phosphate solubilization
The preparation of PKO culture medium: tricalcium phosphate 0.5%, sucrose 1-5%, ammonium sulfate 0.1-1%, yeast powder 0.01-
0.1%, sodium chloride 0.01-0.1%, anhydrous magnesium sulfate 0.01%g, 1% manganese sulfate of potassium chloride 0.02%g, 3ml, 3ml 1%
Ferrous sulfate, agar 18g, distilled water 1000ml, 7,121 DEG C of sterilizing 30min of pH value.
Soluble phosphorus circle method: bacterial strain (Edaphovirga sp.) CGMCC No.15748 point is inoculated on PKO culture medium, is set
3-4d is cultivated in 26-35 DEG C of incubator, observes transparent circle, primarily determines that the bacterium has dissolving P capacity.
Molybdenum antimony resistance colorimetric method: bacterial strain (Edaphovirga sp.) the CGMCC No.15748 liquid LB of 50ml is cultivated
Then base 120-200r/min fermentation under the conditions of 26-35 DEG C accesses PKO culture medium according to 1% inoculum concentration for 24 hours as kind of a liquid
It ferments 7 days under middle the same terms, control group (CK) accesses sterile water according to 1% inoculum concentration, and supernatant is collected by centrifugation after fermentation
Liquid, it is to be measured.It is accurate respectively to draw 5mg/L phosphorus (K2HPO 4) standard solution 0,2,4,6,8,10ml in 50ml volumetric flask, simultaneously
The blank solution isometric with sample solution used in chromogenic assay is added, dinitrophenol dinitrophenolate indicator 2-3 drop is added, is used in combination
100g/L sodium carbonate liquor be adjusted to solution just and be in it is yellowish, it is accurate that the anti-color developing agent of 5ml molybdenum antimony is added, shake up, add water constant volume, i.e.,
Obtain the standard solution that phosphorous (P) amount is respectively 0.0,0.2,0.4,0.6,0.8,1.0mg/L.It shakes up, is placed at room temperature for 30min.?
At wavelength 700nm, its absorbance is measured, using absorbance as ordinate, phosphorus concentration (mg/L) is abscissa, standard curve is drawn,
As a result as shown in Figure 2.Testing liquid is moved in right amount in 50ml volumetric flask, after being diluted with suitable quantity of water, dinitrophenol dinitrophenolate instruction is added
Agent 1~2 is dripped, and being adjusted to solution just with 100g/L sodium carbonate liquor or 50ml/L sulfuric acid solution is in yellowish, accurate plus 5ml molybdenum
The anti-color developing agent of antimony, shakes up, and adds water constant volume, is placed at room temperature for 30min.Ultraviolet specrophotometer, at wavelength 700nm, not connect thallus
Blank cultures be reference liquid zeroing after, measure its absorbance, establishing criteria curve calculates to obtain corresponding phosphorus content.
As a result as shown in figure 3, having in bacterial strain (Edaphovirga sp.) CGMCC No.15748 inoculation processing culture solution
Phosphorus content is imitated up to 525.33 ± 12.26mg/L, is significantly higher than CK, shows very strong phosphate solubilization.
The facilitation that 3 bacterial strain of embodiment (Edaphovirga sp.) CGMCC No.15748 sprouts Panax notoginseng seeds
Panax notoginseng seeds are cleaned, aseptically, rinsed with sterile water 3 times, sterile blotting paper blots spare behind surface.Make
The bacterial strain is prepared into 10 with sterile water7、105、103、101The bacteria suspension of cfu/ml, as a control group (CK) with distilled water,
It is spare.Every group of three repetitions, 30 seeds of each repetition, every group is impregnated 3-5h using bacteria suspension dilution or distilled water 10ml.
The screen to filtrate obtains seed, 20-25 DEG C moisturizing culture 16 days, statistics Panax notoginseng seeds germination rate surveys that root activity, bud be long, diameter
And Fresh Yuxincao, it the results are shown in Table 1, table 3.Root activity measuring method is triphenyltetrazolium chloride (TTC) method, referring to Zhang Zhiliang
" plant physiology experiment instructs the third edition ".
By bacterial strain (Edaphovirga sp.) No.15748 with the LB liquid medium of 50ml under the conditions of 26-35 DEG C
120-200r/min fermentation is for 24 hours as kind of a liquid, then according to fermenting 2 under the same terms in 1% inoculum concentration access LB culture medium
It, obtains OD600=0.3 fermentation liquid takes supernatant, dilution 10 after being centrifuged2、104With 106At times with above-mentioned test procedure
Panax notoginseng seeds are managed, the results are shown in Table 2, table 4.
By the above measurement index, significant difference analysis is carried out with Graphpad software, as a result such as table 1,2 institute of table
Show, be above control group using the processed Panax notoginseng seeds germination rate of Edaphovirga sp.No.15748 bacterial strain, and has significant
Sex differernce illustrates that the bacterial strain has facilitation to Panax notoginseng seeds sprouting.And such as table 3, shown in table 4, the bacteria suspension of the bacterial strain and
Fermentation liquid can significantly improve the biomass and root activity of Panax notoginseng seeds, play facilitation to the nutrient absorption of Radix Notoginseng, right
Panax notoginseng seeds have growth-promoting functions.
1. bacterial strain Edaphovirga sp.No.15748 bacteria suspension of table influences Panax notoginseng seeds germination rate (%)
Note: * *, P < 0.01;* *, P < 0.001.
2. bacterial strain Edaphovirga sp.No.15748 fermentation liquid of table influences Panax notoginseng seeds germination rate (%)
Note: *, P < 0.05;*, P < 0.01;
3. bacterial strain Edaphovirga sp.No.15748 bacteria suspension of table sprouts facilitation to Panax notoginseng seeds
Note: *, P < 0.05;*, P < 0.01;* *, P < 0.001.
4. bacterial strain Edaphovirga sp.No.15748 fermentation liquid of table sprouts facilitation to Panax notoginseng seeds
Note: *, P < 0.05;* *, P < 0.001.
4 bacterial strain of embodiment (Edaphovirga sp.) CGMCC No.15748 improves Radix Notoginseng plant root enzymatic activity
Bacterial strain (Edaphovirga sp.) CGMCC No.15748 solid fungicide handles Radix Notoginseng plant, not add bacterial strain
The solid fungicide culture medium that sterilizes is control group (CK), is sampled within the 3rd, 6,9,12,18 and 30 day after microbial inoculum processing respectively, at every group
Reason selects representative intact plant, each 3 groups of repetitions.Repeated flushing root carries out every root index in time after cleaning
Measurement.Root activity is measured using TTC method;Superoxide dismutase (SOD) activity is measured using nitroblue tetrazolium (NBT) method, with
Inhibition NBT photochemical reduction 50% is a unit of enzyme activity;Peroxidase (POD) activity is measured using guaiacol method, with
Variation 0.01 is 1 peroxidase activity unit in every min.
As a result as shown in table 5,6,7, after handling with the bacterial strain solid fungicide, compared with the control group, Radix Notoginseng can be significantly improved
Plant root vigor facilitates plant absorption nutrition and is metabolized;Defensive ferment SOD activity and POD activity can be significantly improved, it can be bright
The aobvious defence capability for improving plant itself.
Table 5.Edaphovirga sp.CGMCC No.15748 microbial inoculum enhances Radix Notoginseng plant root activity [μ g/ (gh)]
Effect
Note: *, P < 0.05;* *, P < 0.001.
Table 6.Edaphovirga sp.No.15748 microbial inoculum is to active (U/g) humidification of Radix Notoginseng plant SOD
Note: *, P < 0.05;* *, P < 0.001.
Table 7.Edaphovirga sp.No.15748 microbial inoculum is to active (U/g) humidification of Radix Notoginseng plant POD
Note: * * *, P < 0.001.
Prebiotic effect of 5 bacterial strain of embodiment (Edaphovirga sp.) the CGMCC No.15748 to vegetable seeds
Test method sprouts Panax notoginseng seeds with embodiment 3Edaphovirga sp.No.15748 bacterial strain bacteria suspension
Influence test, 20-25 DEG C moisturizing culture 1-5 days, measure root activity, the root system SOD and POD of cucumber, corn and romaine lettuce seed
Activity.
As a result as shown in table 8,9,10, the root of the processed seed of Edaphovirga sp.No.15748 bacteria suspension is used
It is vigor, SOD activity and POD activity, is above control group, illustrates that bacteria suspension has facilitation to plant seed germination.
Table 8.Edaphovirga sp.No.15748 bacteria suspension is to vegetable seeds root activity [μ g/ (gh)] humidification
Note: * * *, P < 0.001.
Table 9.Edaphovirga sp.No.15748 bacteria suspension is to active (U/g) humidification of vegetable seeds SOD
Note: *, P < 0.05;*, P < 0.01;* *, P < 0.001.
Table 10.Edaphovirga sp.No.15748 bacteria suspension is to active (U/g) humidification of vegetable seeds POD
Note: *, P < 0.05;*, P < 0.01;* *, P < 0.001.
Sequence table
<110>Shenyang Pharmaceutical University
<120>one plants have absorption heavy metal, phosphorus decomposing and the prebiotic bacterium and its application of plant
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1564
<212> DNA
<213> Edaphovirga sp.
<400> 1
gctgccttag agtttgatcc tggctcagat tgaacgctgg cggcaggcct aacacatgca 60
agtcgggcgg tagcacgggg gagcttgctc cctgggtgac gagcggcgga cgggtgagta 120
atgtctggga aactgcctga tggaggggga taactactgg aaacggtagc taataccgca 180
tgacctcgca agagcaaagt gggggacctt agggcctcac gccatcggat gtgcccagat 240
gggattagct agtaggtggg gtaatggctc acctaggcga cgatccctag ctggtctgag 300
aggatgacca gccacactgg aactgagaca cggtccagac tcctacggga ggcagcagtg 360
gggaatattg cacaatgggc gcaagcctga tgcagccatg ccgcgtgtgt gaagaaggcc 420
ttcgggttgt aaagcacttt cagcgaggag gaaggcactg ttcctaatag ggatggtgat 480
tgacgttact cgcagaagaa gcaccggcta actccgtgcc agcagccgcg gtaatacgga 540
gggtgcaagc gttaatcgga attactgggc gtaaagcgca cgcaggcggt caattaagtt 600
ggatgtgaaa tccccgggct taacctggga actgcattca aaactgattg gctagagtct 660
tgtagagggg ggtagaattc caggtgtagc ggtgaaatgc gtagagatct ggaggaatac 720
cggtggcgaa ggcggccccc tggacaaaga ctgacgctca ggtgcgaaag cgtggggagc 780
aaacaggatt agataccctg gtagtccacg ctgtaaacga tgtcgacttg gaggttgtgg 840
ccttgagccg tggcttccgg agctaacgcg ttaagtcgac cgcctgggga gtacggccgc 900
aaggttaaaa ctcaaatgaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa 960
ttcgatgcaa cgcgaagaac cttacctact cttgacatcc acgggatttg gcagagatgc 1020
cttagtgcct tcgggaaccg tgagacaggt gctgcatggc tgtcgtcagc tcgtgttgtg 1080
aaatgttggg ttaagtcccg caacgagcgc aacccttatc ctttgttgcc agcgcgtaat 1140
ggcgggaact caaaggagac tgccggtgat aaaccggagg aaggtgggga tgacgtcaag 1200
tcatcatggc ccttacgagt agggctacac acgtgctaca atggcatata caaagagaag 1260
caaactcgcg agagccagcg gacctcataa agtatgtcgt agtccggatt ggagtctgca 1320
actcgactcc atgaagtcgg aatcgctagt aatcgtggat cagaatgcca cggtgaatac 1380
gttcccgggc cttgtacaca ccgcccgtca caccatggga gtgggttgca aaagaagtag 1440
gtagcttaac cttcgggagg gcgcttacca ctttgtgatt catgactggg gtgaagtcgt 1500
aacaaggtaa ccgtagggga acctgcggct ggatcacctc ctaagggcag ctcaatcgcc 1560
ctat 1564
Claims (10)
1. one plant of bacterium (Edaphovirga sp.), which is characterized in that deposit number are as follows: CGMCC No.15748.
2. application of the bacterium (Edaphovirga sp.) described in claim 1 in absorption heavy metal, phosphorus decomposing.
3. bacterium (Edaphovirga sp.) described in claim 1 is promoting plant growth, is promoting in plant seed germination
Using.
4. application as claimed in claim 2 or claim 3, which is characterized in that use the strain suspensions of bacterium (Edaphovirga sp.)
Or bacterial strain fermentation liquor or bacterial strain fermentation liquor extracting solution or the solid fungicide containing the bacterial strain carry out pouring root to plant or spread fertilizer over the fields processing.
5. application according to claim 4, which is characterized in that the preparation of the strain suspensions: taking out the bacterium
(Edaphovirga sp.) culture presevation pipe, is recovered with LB solid medium, 20-37 DEG C culture 24-48 hours, use
Activated bacterial strain is prepared into bacteria suspension by distilled water, and it is dense to measure bacteria suspension at wavelength 600nm using ultraviolet specrophotometer
Degree, the concentration of the bacteria suspension are as follows: 0.1-0.8.
6. application according to claim 4, which is characterized in that the bacterial strain fermentation liquor or bacterial strain fermentation liquor extracting solution
Preparation: taking out described bacterium (Edaphovirga sp.) the culture presevation pipe, recovered with LB solid medium, and 20-37 DEG C
Culture 24-48 hours, picking single bacterium colony is seeded in 50ml LB culture medium on plate, in the incubator 20-37 DEG C of concussion
Culture 24-72 hours, seed use the inoculum concentration of 1-10%, are seeded in LB culture medium and expand culture 24-72 hours, bacterium solution is dense
Degree is greater than OD600=1 can terminate to ferment, and sterile filling obtains liquid end product, or obtains powdery finished product by freeze-drying;It will
Aforementioned resulting bacterial strain fermentation liquor is extracted using isometric ethyl acetate, is collected concentrated extract, is obtained bacterial strain fermentation liquor extracting solution.
7. application according to claim 4, which is characterized in that the preparation of the solid fungicide: will be described in claim 6
Bacterial strain fermentation liquor be inoculated into solid fungicide culture medium, 20-40 DEG C culture, cultivate 1-3d after aseptically stirred
It mixes, continues to cultivate 1-3d after stirring, obtain solid fermentation microbial inoculum.
8. according to application described in claim 5 or 6 or 7, which is characterized in that the formula of the LB culture medium are as follows: peptone
1-5%, yeast extract 0.1-5%, NaCl 0.1-3%, water 1000ml;The formula of LB solid medium are as follows: peptone 1-5%,
Yeast extract 0.1-5%, NaCl 0.1-3%, agar 1.5-2%, water 1000ml.
9. the use as claimed in claim 7, which is characterized in that the formula of the solid fungicide culture medium is, in terms of g/ml as
Under, wheat bran 20-50%, corn flour 10-30%, bean powder 1-10%, potassium nitrate 0.1-1%, dipotassium hydrogen phosphate 0.1-1%, chlorination
Sodium 0.4-0.7%, calcium carbonate 1-10%, distilled water 200ml.
10. the application as described in claim 2-9 any one, which is characterized in that the bacterium (Edaphovirga sp.)
Independent role is used in mixed way with other organic fertilizers.
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CN112280563A (en) * | 2020-10-28 | 2021-01-29 | 福建龙净环保股份有限公司 | Phosphate solubilizing bacteria mineralization conditioner, preparation method thereof and heavy metal contaminated soil remediation and improvement method |
CN116102369A (en) * | 2021-11-10 | 2023-05-12 | 沈阳药科大学 | Application of bacteria in heavy metal resistance |
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