CN106544306B - One plant of siderophore Producing Strain and its application in terms of farmland pollution heavy metal-polluted soil reparation - Google Patents
One plant of siderophore Producing Strain and its application in terms of farmland pollution heavy metal-polluted soil reparation Download PDFInfo
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Abstract
The invention belongs to the biological prosthetic fields of environmental pollution, specially one plant of siderophore Producing Strain and its application in terms of farmland pollution heavy metal-polluted soil reparation.One plant of siderophore Producing Strain, the bacterium be pseudomonas (PseudomonasSp. bacterial strain S17), for the bacterial strain on October 12nd, 2016 in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, number is CGMCC No.13104.Of the inventionPseudomonasSp.S17 bacterial strain has reached the superlative degree for producing siderophore bacterium, produce siderophore ability be it is most strong in domestic registration bacterial strain,PseudomonasSp.S17 is sensitive to common antibiotics, different degrees of tolerance or resistance is presented to 12 heavy metal species, and have both disease fungus knot resistance, phosphate solubilization and production 3-indolyl acetic acid ability.
Description
Technical field
The invention belongs to the biological prosthetic field of environmental pollution, specially one plant of siderophore Producing Strain and its in farmland pollution soil
Earth heavy metal repairs the application of aspect.
Background technique
The heavy metal pollution of soil has become global environmental problem, pays close attention to by society and the common people, it would be highly desirable to solve.I
Many regional (especially East China) soil of state are in different degrees of heavy metal pollution, in many area grains, veterinary antibiotics
The content of beary metal such as cadmium, arsenic, copper, lead, zinc are exceeded or close to critical value, seriously endanger people's health.
Heavy metal pollution of soil improvement is extremely urgent but difficult, main reason is that heavy metal is unlike organic dirt
Dye object can be biodegradable like that, and migration is poor in the soil for heavy metal.Traditional physics, chemical method such as soil moved in improve the original, leaching
Wash, ion exchange etc. is often only suitable for the soil remediation of small-scale serious pollution, and because its is at high cost, destroys soil texture, with
And easily causes the disadvantages of secondary pollution and endure to the fullest extent and denounce.In contrast, biological prosthetic, because having, at low cost, effect is good, simple easy
The advantages that capable and without secondary pollution and by favor.Wherein, microorganism-is plant combined repairs as biological prosthetic one kind, because
Its toxicity that heavy metal in soil can be effectively reduced adsorbs heavy metal and changes rhizosphere environment, and then improves plant counterweight
The absorption of metal or fixed efficiency, and implementation cost relative moderate, environmental-friendly, and can extensive in-situ immobilization the advantages that and
It enjoys great popularity, is the new technology for being used to administer large area heavy metal pollution of soil greatly developed, carried out in recent years.On however,
The efficiency for stating combined remediation technology depends on the biomass of rehabilitation plant and the bioavailability of heavy metal in soil.It changes
Yan Zhi, phytomass is bigger and heavy metal can be as often as possible absorbed from soil, then remediation efficiency is higher, effect
Better.How to improve phytoremediation efficiency becomes the hot spot of research and concern.Hyperaccumulative plant (such as Btassica, front yard mustard category
With penny cress category) usually biomass is lower and be difficult to large-scale plantation, thus its in actual repair using extremely limited.And
The big and plant with heavy metal accumulation ability of some fast growings, biomass is just being increasingly being applied to heavy metal pollution field
Ground reparation.As a kind of energy crop that can be used for producing bio-ethanol, sugar grass is strong with voltage endurance capability, growth is rapid, raw
The advantages that object amount is big becomes heavy metal rehabilitation plant most competitive at present.
Siderophore (Siderophore) is the organic compound that a kind of pair of iron that microorganism generates has superpower complexing power,
Its function is to provide iron attainment point to microbial cell, especially under low iron hoop border.The combination of siderophore and heavy metal is main
It is embodied on the biological function of siderophore, i.e., in the iron being concentrated in environment and while promote it to be transported into cell, iron is carried
Body can also influence the bioavailability of other metals by complexing, to reduce its toxicity.In addition, Microbial Iron carrier
It can also be by inhibiting the growth of pathogenic microorganism to promote the disease resistance of root system of plant.Siderophore producing strains are because it is with upper
Advantage, speciality are stated, is waited so becoming the popular of microbial portion in the plant combined repairing heavy metal pollution system of microorganism-
Choosing.
Currently, having many siderophore producing strains by technologies such as manually enrichment, screenings and being isolated from environment, and is real
Pure culture is showed.However, siderophore superior strain is limited in these reported bacterial strains；It is (such as more to have both a variety of advantages, speciality
Heavy metal resistance and disease fungus knot resistance) bacterial strain it is deficient；Have practical fortification of plants and absorbs heavy metal in soil ability
Bacterial strain it is very few.Thus, finding and obtaining the excellent siderophore superior strain of character becomes the hot spot of the research field
And focus.
Summary of the invention
A kind of in view of the deficiencies of the prior art, it is an object of the present invention to provide characters excellent, antibiotic sensitive siderophore
Producing Strain.
It is a further object to provide application of the above-mentioned strain in terms of farmland pollution heavy metal-polluted soil reparation, should
Representative heavy metal contaminated soil is repaired using sugar grass is mainly strengthened.
Above-mentioned technical problem of the invention is carried out by the following technical programs:
One plant of siderophore Producing Strain, the bacterium are the bacterial strain S17 of pseudomonas (Pseudomonas sp.), the bacterial strain in
On October 12nd, 2016 is in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, number CGMCC
No.13104, depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica.This hair
High yield siderophore bacterium Pseudomonas sp.S17 provided by bright is originated from experimental plot in Scientific and Technological Institutes Of Zhejiang, Hangzhou, Zhejiang province city
Soil screens through artificial enrichment culture, pressure and isolates and purifies to obtain.The bacterium is Gram-negative, contacts enzyme positive, oxygen
It is negative to change enzyme, V.P. negative, indole test is positive, and thalli morphology is rod-short, and bacterium colony is omited in faint yellow, round, edge
Aobvious burr, smooth wet, other physiological and biochemical properties are as shown in table 1.
The physiological and biochemical property of table 1, bacterial strain S17
Note: "+" indicates growth or positive reaction；"-" expression is not grown or negative reaction.
The optimum growth temperature of the bacterium is 29 DEG C.With 1/5th LB (Luria-Bertani) or industrial fermentation culture medium
(corn flour 10g/L, bean cake powder 8g/L, (NH4)2SO46g/L, corn pulp 6g/L, MgSO4·7H2O 1g/L, K2HPO4·3H2O
2g/L, MnSO40.05g/L, pH 7.0) after culture 24 hours, bacterium solution cell concentration is up to 6.5-9.2 × 109 CFU/mL
(CFU, Colony Forming Unit).
The bacterium to ampicillin, streptomysin, chloramphenicol, kanamycins and sensitive tetracycline or it is highly sensitive (be all to seek
Normal antibiotic).It is considered that: when thallus is released in natural environment, will not be generated because of anti-(resistance to) pharmacological property problem super
Bacterium.
In the test of pathogen antagonism, it is found that it is true the bacterial strain can inhibit mould, aspergillus niger and three kinds of cause of diseases of sclerotinia sclerotiorum
Bacterium growth.
The bacterium is to 12 kinds of test metal ion (Li+、Ag+、Cd2+、Cu2+、Hg2+、Ba2+、Pb2+、Zn2+、Mn2+、Ni2+、Fe3+
And Co4+) different degrees of tolerance (or resistance) is presented.In view of China's heavy-metal contaminated soil status --- various heavy is dirty
It contaminates and deposits, it is believed that --- for the more other bacterial strains for not having various heavy tolerance or resistance of the bacterial strain, implementing by force
Repairing effect is influenced small by heavy metal inhibition in change repair process.
The bacterial strain siderophore active unit (Siderophore unit, SU) is 99.2%, highest up to siderophore bacterium is produced,
It is to have reported to produce in siderophore bacterium, produces the strongest bacterial strain of siderophore ability.In addition, S17 is also equipped with certain Soluble phosphorus and produces 3- Yin
The ability of indolylbutyric acid.Using 1.5% glucose as carbon source, 0.2% ammonium chloride is nitrogen source, and 30 DEG C, pH7.2, incubation time 66h (just
Beginning inoculum concentration is that 2%), which is 120.35mg/L.To the general LB Liquid Culture for containing L-Trp (200mg/L)
The bacterium (inoculum concentration 1%) is accessed in base, 28 DEG C, 160r/min culture 3 days, the 3-indolyl acetic acid then measured in fermentation liquid contains
Amount, measured value 72.36mg/L.
Application of the siderophore Producing Strain in terms of farmland pollution heavy metal-polluted soil reparation a kind of described in.
Preferably, the application is to strengthen sugar grass restoration of soil polluted by heavy metal, method is the sugar grass in growth period
Root sprinkling contain the bacterium solution of the siderophore Producing Strain, the concentration of bacterium solution is 1-5 × 108 CFU/mL。
Preferably, the bacterium solution is the siderophore Producing Strain bacterium solution of logarithmic phase.
Preferably, the bacterium solution is by 1/5th LB (Luria-Bertani) or industrial fermentation culture medium culture
It is obtained after 22-24 hours, the formula of industrial fermentation culture medium are as follows: corn flour 10g/L, bean cake powder 8g/L, (NH4)2SO46g/L,
Corn pulp 6g/L, MgSO4·7H2O 1g/L, K2HPO4·3H2O 2g/L, MnSO4 0.05g/L。
Preferably, the Liquid Culture temperature is 25-30 DEG C, the pH of basic inorganic salt fluid nutrient medium is 6.8-
7.2。
Preferably, the Liquid Culture temperature is 29 DEG C, the pH of basic inorganic salt fluid nutrient medium is 7.0.
In conclusion the present invention compared with the existing technology has the advantages that Pseudomonas sp.S17 of the invention
Bacterial strain has reached the superlative degree for producing siderophore bacterium, and producing siderophore ability is most strong in domestic registration bacterial strain, Pseudomonas
Sp.S17 is sensitive to common antibiotics, different degrees of tolerance or resistance is presented to 12 heavy metal species, and have both disease fungus knot
Resistance, phosphate solubilization and production 3-indolyl acetic acid ability.And these advantages are to microorganism-plant combined repairing heavy metal pollution soil
It is significant for ground！Enable this reparation system it is more practical, more efficiently be applied to contaminated site reparation.
Specific embodiment
Below by specific embodiment, technical scheme of the present invention will be further explained in detail.It should be appreciated that this hair
Bright implementation is not limited by the following examples, and the accommodation in any form made to the present invention and/or changed will all be fallen
Enter the scope of the present invention.
In the present invention, if not refering in particular to, all parts, percentage are unit of weight, used equipment and raw material etc.
It is commercially available or commonly used in the art.Method in following embodiments is unless otherwise instructed the normal of this field
Rule method.
The separation of embodiment 1:Pseudomonas sp.S17 and siderophore generate aptitude tests
With experimental plot soil 100g in Scientific and Technological Institutes Of Zhejiang, general five point samplings method collection in worksite Hangzhou, Zhejiang province city, with nothing
Bacterium envelope contains, and takes back laboratory rapidly.
Universal CAS detects liquid and MSA fluid nutrient medium and reports that (Chen Shaoxing, Zhao Xiang, Shen Ping wait Gao Ling according to Chen Shaoxing etc.
The plate detection method microbiology of quick pseudomonad siderophore is notified to, and 2006,33:122-127) it prepares.Solid detects plate
It is then that the CAS detection liquid of the addition 5% in MSA fluid nutrient medium and 2% agar powder solidify.
10 times of diminishing methods of pedotheque are diluted, being coated on 1/5th LB solid plates, (i.e. all nutritional ingredients are
The 1/5 of general LB culture medium) on.25 DEG C of cultures, after there is bacterium colony, the bacterium colony of different shape is chosen one by one with sterile toothpick,
It is transferred on 1/5th new LB solid plates, until purifying is at single colonie.All pure bacterial strain number consecutivelies, and carry out routine
Culture presevation is deposited in the form of inclined-plane among 4 DEG C of refrigerators.
By above-mentioned purifying bacterial strain, it is forwarded on solid detection plate one by one, 28 DEG C of inversions are cultivated 2 days.Observe periphery of bacterial colonies
Whether generation discoloration is enclosed, and does detailed record, produces siderophore ability power using the diameter for the circle that changes colour as siderophore bacterium is produced
Preliminary judgement.In addition, it is negative control that Escherichia coli DH5 α bacterial strain must be arranged simultaneously.
Siderophore quantitative determination reports that (Chen Shaoxing, Zhao Xiang, Shen Ping wait the highly sensitive pseudomonad iron of to carry according to Chen Shaoxing etc.
The plate detection method microbiology of body is notified to, and 2006,33:122-127) it carries out, by respective strains tested culture solution with 1%
Amount be linked into 30mL MSA fluid nutrient medium, 28 DEG C, 200r/min is cultivated for 24 hours, and 10000r/min is centrifuged 15min, supernatant
It is uniformly mixed with 0.22 micron of filtering with microporous membrane, and with isometric CAS detection liquid.Under room temperature, it is surveyed after placing sufficiently
Determine the OD of each sample680(As) value compares zeroing with distilled water.Same procedure measurement is made with each fluid nutrient medium for not connecing bacterium
For the light absorption value of supernatant, as reference value (Ar).The concentration of siderophore indicates that every processing is repeated with siderophore active unit
4 times.Then, according to the report of Persmark et al. (Persmark M, Expert D, Neilands JB.Isolation,
characterization,and synthesis of chrysobactin,a compound with siderophore
activity from Erwinia chrysanthemi.The Journal of Biological Chemistry,1989,
264:3187-3193) siderophore ability is produced to each bacterial strain to be classified.
As a result, can be cultivated by amounting to obtain on 1/5th LB solid plates by 126 plants of bacterial strain, wherein 9 plants have siderophore
Generation ability (referring to table 2).In the bacterial strain for having siderophore generation ability, but it is most strong with the ability of S17 bacterial strain.
Table 2, siderophore producing strains produce siderophore capability for registration table
The antibiotic susceptibility test and disease fungus antagonistic ability of embodiment 2:Pseudomonas sp.S17 is tested
Antibiotic sensitivity test uses filter paper enzyme, selects ampicillin, tetracycline, chloramphenicol, kanamycins and chain
Five kinds of common antibiotics of mycin, with antibacterial circle diameter of each antibiotic filter paper on culture siderophore producing strains S17 plate
Size, the criterion as sensitive or resistance to (anti-) medicine.The test result that table 3 provides shows bacterial strain S17 to above-mentioned anti-for trying
Raw element is in different degrees of sensitivity response.
The antibiotic susceptibility test result of table 3, bacterial strain S17
Note: antibacterial circle diameter be greater than medium sensitivity range higher limit be it is highly sensitive, less than sensitive range lower limit value
For resistance.
This example demonstrates that when Pseudomonas sp.S17 thallus is released in natural environment, it will not be because resisting (resistance to)
Pharmacological property problem and become superbacteria, this just provides safety assurance for its subsequent practical application.
With 10 kinds of pathogens --- graw mold of tomato germ, early blight of tomato germ, wilt of eggplant germ, muskmelon are climing withered
Germ, sclerotinia sclerotiorum germ, mould, cotton wilt germ, aspergillus niger, wheat sharp eyespot germ and wheat leaf blight disease
Bacterium is indicator bacteria, utilizes the germ fungi antagonistic ability of tablet face-off method measurement S17.It is inoculated with and indicates in general PDA plate center
Bacterium, surrounding are inoculated with S17, and every group of test is repeated 5 times, and pass through the antagonistic ability of colony radius and the clear bacterial strain S17 of antibacterial bandwidth.
The results are shown in Table 4, and Pseudomonas sp.S17 is in for the mould of examination, aspergillus niger and sclerotinia sclerotiorum germ
Obvious Antagonism.
The antagonistic ability of table 4, Pseudomonas sp.S17
Note: feminine gender is denoted as "-"；Antibacterial circle diameter 0-5mm is denoted as "+"；Antibacterial circle diameter 5-10mm is denoted as " ++ "；Inhibition zone
Diameter 10-15mm is denoted as " +++ "；Antibacterial circle diameter is greater than being denoted as of 15mm " ++++".
The heavy metal minimum inhibitory concentration (MIC) of embodiment 3:Pseudomonas sp.S17 is tested
MIC experimental design carry out referring to the research of Filali et al. (Filali BK, Taoufik J, Zeroual Y,
Dzairi FZ, Talbi M, Blaghen M.Waste water bacterial isolates resistant to heavy
Metals and antibiotics.Current Microbiology, 2000,41:151-156), picking etc. is big, activated
The S17 single bacterium of processing is fallen in culture test tube, and 30 DEG C, 160r/min shaken cultivation 32 hours.MIC value is determined according to thallus growing way,
Every group sets 3 repetitions, tests the bacterial strain respectively to the tolerance situation of 12 metal ion species.The results are shown in Table 5, and bacterial strain tool is more
Heavy metal is resistant to (anti-) property, and character is excellent.
The MIC test of table 5, bacterial strain S17
Note: data are the average value of 3 measured values in table.
The Soluble phosphorus and production 3-indolyl acetic acid aptitude tests of embodiment 4:Pseudomonas sp.S17
A ring Pseudomonas sp.S17 is taken from test tube slant with oese, is inoculated in and fills 50mL liquid LB culture
In the triangular flask of base, 160r/min shaken cultivation for 24 hours, then takes 1mL (about 1 × 108CFU) culture is inoculated into Ca3
(PO4)2For 100mL Phos fermentation medium [the glucose 10g, (NH of unique phosphorus source4)2SO41g, NaCl 0.3g, KCl
0.3g, MgSO4·7H2O 0.3g, FeSO4·7H2O 0.002g, MnSO4·H2O 0.002g, Ca3(PO4)25g, distilled water are fixed
Hold to 1000mL, pH 7.5] in, 3 repetitions are arranged in 30 DEG C of shaken cultivation (160r/min) 48h.With the general anti-colorimetric of molybdenum antimony
Method measures phosphorus content --- and take 1mL culture solution in the centrifuge tube accordingly numbered, 5000r/min is centrifuged 10min.Accurately take 1mL
Supernatant adds the anti-developer of 2.5mL molybdenum antimony in the 25mL volumetric flask accordingly numbered, and then uses distilled water constant volume.In room temperature
After lower reaction 30min, using light absorption value at spectrophotometric determination 700nm wavelength, with mg P2O5/ L is indicated.Not connect bacterium
Processing is blank control, and experiment sets 3 repetitions.As a result, measuring Pseudomonas sp.S17 amount of phosphorus dissolved is 88.6 ± 2.26mg/
L。
The access Pseudomonas sp.S17 into the LB liquid medium containing L-Trp (200mg/L), 28 DEG C,
160r/min is cultivated 3 days.Then with the OD of spectrophotometry measurement bacteria suspension600Bacteria suspension 10000r/min, is then centrifuged by value
10min takes supernatant that isometric Salkowski color solution (50mL 35%HClO is added4+1mL 0.5mol/L FeCl3)
(Libbert E and Risch H.Interactions between plants and epiphytic bacteria
Regarding their auxin metabolism [J] .Physiologia Plantarum, 1969,22:51-58), it is protected from light
Standing 30min makes its colour developing, measures OD530Value.The content of 3-indolyl acetic acid in unit of account volume bacterium solution, is arranged 3 repetitions,
And the E.coli DH5 α not produce 3-indolyl acetic acid is negative control.As a result, measuring S17 bacterial strain 3-indolyl acetic acid yield and being
62.8 ± 0.86mg/L, and 3-indolyl acetic acid is not detected in negative control.
People have found during previous heavy metal pollution site remediation --- many reparation plant can be (special because of poor nutritional
Be not the absence of P elements), absorb limited, and the reasons such as growth is suppressed can not farthest play reparation absorption.
This example demonstrates that Pseudomonas sp.S17 has Soluble phosphorus and produces the ability of 3-indolyl acetic acid, and this is deposited to plant is strengthened
It is very practical and important for living, growth and its reparation effectiveness.
Embodiment 5:Pseudomonas sp.S17 strengthens the application that sugar grass absorbs heavy metal
It is as follows that soil used in testing picks up from the not vegetable garden by heavy metal pollution, main physical and chemical: organic matter 4.84%,
Total nitrogen 832.6mg/kg, total phosphorus 292.1mg/kg, total potassium 220.3mg/kg, available nitrogen 68.2mg/kg, rapid available phosphorus 18.1mg/kg,
Available potassium 58.3mg/kg, pH6.2.Content of beary metal is cadmium 0.21mg/kg, copper 58mg/kg, zinc 101mg/kg.Air-dry sieving
Afterwards, it is separately added into solution state caddy, zinc chloride and the copper sulphate of different content, makes Cd in soil2+Concentration be 25mg/kg,
Zn2+Concentration be 800mg/kg, Cu2+Concentration be 800mg/kg.Contaminated soil is mixed well, stablizes 2 under 60% damp condition
It is tested after a month.
The seed of rehabilitation plant sugar grass is purchased from Linan City river bridge retail sales.The rudiment of seed elder generation, it is long to the left side 10cm to seedling
When right, select the consistent seedling replanting of growing way enter the good heavy-metal contaminated soil plastic tub of 1.5kg previous steady (high 13cm, directly
Diameter 17cm) in, 1, every basin.It is normal to plant strain growth after transplanting 7 days, 1mL logarithmic phase Pseudomonas is accessed in respective root
Sp.S17 bacterium solution (1 × 108CFU/mL).To avoid the nutritional ingredient in culture medium from influencing test result, above-mentioned access bacterium solution is
After sterile water washing 3 times, the bacterium solution that is suspended again with sterile water.Test sets 4 processing: not planting plant and does not add the blank of bacterium
Control (C), only kind plant does not connect bacterium (P), does not plant plant but adds bacterium (M), and has both planted plant or added bacterium (PM), and each processing is set
5 Duplicate Samples.Periodically watering, harvests after keeping the illumination of daily 8h, sugar grass to grow 82 days under greenhouse, and root system is used after cleaning
The EDTA solution of 10mmol impregnates 30min, then is rinsed 2 times with deionized water.Plant dries to constant weight for 80 DEG C after 105 DEG C of water-removings,
Weigh the dry weight of root, overground part.Plant tissue clears up (HClO4∶HNO3=1: 4) after, measuring its content of beary metal, Whole Process is done
Blank control is to eliminate interference.
The results are shown in Table 6, and access Pseudomonas sp.S17 can significantly improve the biomass and a huge sum of money of sugar grass
Belong to uptake.Wherein, the dry weight of overground part and root has increased separately 30.8% and 32.0% (PM processing than not connecing the control of bacterium
Compared with P processing), and sugar grass overground part Cd2+、Zn2+And Cu2+Total content 65.4%, 71.1% and has been respectively increased
68.6%.It confirms that bacterial strain S17 has promote sugar grass growth, strengthen sugar grass absorption for the ability of test mass metal.
The Metal uptake amount of table 6, sugar grass overground part and root
Note: content of beary metal=biomass × heavy metal concentration；Extraction efficiency is the total metals (root that plant is extracted
The sum of with overground part) the ratio between with heavy metal total content in initial soil；* indicating the value, there are significant differences with corresponding control value
(P<0.05)。
Compared with the existing technology (Zhang Lu produces siderophore bacterium and strengthens sugar grass repairing heavy metal in soil pollution environmental science
With technology, 2014,37:74-79), the overground part dry weight of of the invention " plant+microorganism " processing, overground part Cd2+、Zn2+And Cu2+
Total content, and respectively extraction efficiency compared with it improve 8.5%, 11.0%, 11.6% and 18.0% and 11.7% respectively,
11.1% and 14.3%, better effect, advantage is significant.And (PM handles recovery rate/P processing and extracts in terms of recovery rate promotion
Rate), three heavy metal species recovery rates of the invention promote effect and improve 22.4%, 81.6% and 8.0% respectively compared with it, and advantage is aobvious
It writes.Furthermore, it is necessary to be intended that, Pseudomonas sp.S17 with the Pseudomonas sp.T07 in above-mentioned fine jade research not
It is same bacterium.Because obvious differences (referring to the table 1) in terms of physiological and biochemical property, is in particular in catalase, oxygen
On four change enzyme, glucose utilization ability and methyl red test test items.
Claims (5)
1. one plant of siderophore Producing Strain, the bacterium be pseudomonas (PseudomonasSp. bacterial strain S17), the bacterial strain in
On October 12nd, 2016 is in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, number CGMCC
No. 13104。
2. a kind of application of siderophore Producing Strain described in claim 1 in terms of farmland pollution heavy metal-polluted soil reparation.
3. application as claimed in claim 2, it is characterised in that: the application is to strengthen sugar grass restoration of soil polluted by heavy metal, side
Method is the bacterium solution for containing the siderophore Producing Strain in the sprinkling of the root of the sugar grass in growth period, the concentration of bacterium solution be 1-5 ×
108 CFU/mL。
4. application as claimed in claim 3, it is characterised in that: the bacterium solution is the siderophore Producing Strain bacterium solution of logarithmic phase.
5. application described in claim 3 or 4, it is characterised in that: the bacterium solution is by 1/5th LB(Luria-
Bertani it) or after industrial fermentation culture medium culture 22-24 hours obtains, the formula of industrial fermentation culture medium are as follows: corn flour 10
G/L, bean cake powder 8 g/L, (NH4)2SO46 g/L, corn pulp 6 g/L, MgSO4·7H2O 1 g/L, K2HPO4·3H2O 2 g/
L, MnSO4 0.05 g/L。
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| CN108856285B (en) * | 2018-07-16 | 2020-05-19 | 浙江工商大学 | Method for improving phytoremediation efficiency of zinc-contaminated soil |
| CN110452839B (en) * | 2019-07-25 | 2020-12-11 | 云南省农业科学院农业环境资源研究所 | Pseudomonas putida and application thereof |
| CN112725221B (en) * | 2020-12-15 | 2023-01-10 | 湘潭大学 | Pseudomonas fluorescens, method for preparing hydroxamic acid type siderophore by using pseudomonas fluorescens and application of pseudomonas fluorescens |
| CN114231459B (en) * | 2021-12-27 | 2023-05-26 | 中南大学 | Escherichia coli, microbial agent, composition and application |
| CN115820463B (en) * | 2022-08-24 | 2023-06-23 | 杭州秀川科技有限公司 | Preparation method of siderophores based on microbial fermentation |
| CN115557856B (en) * | 2022-10-14 | 2024-03-19 | 福建省微生物研究所 | Ferrate compound Frandiamine C, fermentation strain and fermentation method thereof |
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