CN105132315B - High mountain bacillus altitudinis YLX-5 and its application - Google Patents
High mountain bacillus altitudinis YLX-5 and its application Download PDFInfo
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Abstract
The invention discloses it is a kind of be isolated from Hubei China bestow favour selenium mining area diafiltration water system in bacterium seat deposit high mountain bacillus altitudinis YLX-5 and its application, in China typical culture collection center, deposit number is CCTCC No:M2013675 for the culture presevation.The strain has thermophilic selenium microorganism feature, can have potential application in the conversion of microorganism organic selenium, the synthesis of microorganism nanometer selenium, the activation of microorganism of ore containing selenium, selenium-enriched microbe fertilizer and selenium contamination environment remediation.
Description
Technical field
The present invention relates to a kind of thermophilic selenium microorganism and its application more particularly to a plant height mountain bacillus
Altitudinis YLX-5 and its microorganism conversion synthesize organic selenium, Microbe synthesis nanometer selenium, activation of microorganism selenium mine
Application in stone, selenium-enriched microbe fertilizer and selenium contamination environment remediation, belongs to field of biotechnology.
Background technique
Extreme microorganism refers to the microorganism kind for being suitble to live in extreme environment, including Thermophilic Bacteria, Halophiles, basophilic
Bacterium, acidophil, barophilic bacteria, psychrophile and anti-radiation, resistance to drying, anti-high-concentration metallic ions and extreme anaerobism microorganism
(Rothschild&Mancinelli,2001；Chen Jun etc., 2006), the research of extreme microorganism at present mainly mid-ocean rise,
The regions such as hot spring, salt lake, mine drainage.It lives in the hydrothermal solution environment of deep-sea mid-oceanic ridge a large amount of thermophilic, barophilic bacterias,
Fe, S, Zn, Cu of mid-oceanic ridge recycle and at play in mine important role (Taylor&Wirsen, 1997；Labrenz et
al.,2000；Kasama&Murakami,2001；Emerson&Mayer,2002；Kennedy et al.,2003；Peng Xiaotong
Deng 2007；Chen Shun etc., 2010).Geomicrobiology scholar is to U.S. Yellowstone (Susan et al., 1994,1996), ice
Island (Marteinsson et al., 2001；Thomas et al., 2007), Italy (Kvist et al., 2005), China
Yunnan (Song Zhao is neat etc., and 2008；Huang Qiuyuan etc., 2010) etc. numerous hot springs on ground are studied, and disclose that be up to 70 at these Celsius
Degree or more, some are even more than inside 100 degrees Celsius of hot spring, enliven a large amount of thermophilic microorganisms --- spring archaeal is expanded
To the imagination of the microbe survival limit before us.And Utah, U.S.A Great Salt lake (salinity 2.2%), the Dead Sea (salinity 2.5%), inner
Sea (many anti-hyperosmosis microorganisms that live in the hypersaline environments such as salinity (1.7%) (Antonio et al., 1998；
Vreeland et al.,1998；Oren et al.,2001).Inland maximum within Chinese territory positioned at northeast of Qinghai province is high
Former salt water lake, more due to its unique depositional environment, become geomicrobiology research natural laboratory (it is appropriate into just etc.,
2005).Largely there is acidophilus iron-oxidizing bacteria in acid mine drainage, such as Thiobacillus ferrooxidans (Acidithiobacillus
Ferrooxidans), largely participate in Fe, Mn, S etc. circulation (Baker&Banfield, 2003；Fortin&Langley,
2005；Lu Jianjun etc., 2005；Jiang Lei etc., 2006), extensive ecological problem is on the one hand caused, is on the other hand mine drainage
Improvement provides new thinking (Johnson&Kevin, 2003).It is millions of even for being sealed up for safekeeping under the thick ice sheet in the remote South Pole
Subglacial lake life system as long as year, our still in store keen interests (Bulat et al., 2009,2011).
But few people pay close attention to selenium-rich extreme environment in microorganism group at.It is in the earth's crust that this, which is primarily due to selenium,
Microelement, it is difficult at the environment of mine or the extreme selenium-rich of formation, strongly limit the research to thermophilic selenium microorganism.
Enshi fishing pool dam has unique Se fractionation in the world, is typical and unique extreme selenium-rich environment, in selenium
There is the Se content of high concentration, wherein the Se content in water body in the diafiltration water system in mining area and the diafiltration water system of selenium mine tailing
Up to 40.0-94.1 μ g/L, average value be 58.4 ± 16.8 μ g/L (Zhu&Zheng, 2001；Zhu et al., 2008), it is gram
330 times (Fordyce et al., 2000) of mountain lesion water body Se content are that the World Health Organization (WHO) recommends drinking water selenium
10 times (Presser, 1994) of upper content limit；Se content in deposit is up to 8.28-82.9mg/kg DW, and average value is
26.6±26.8mg/kg DW(Zhu&Zheng,2001；Zhu et al., 2008), it is US West's deposit Se content upper limit
It is worth 19 times (Presser, 1994) of (1.4mg/kg DW).Currently, only Zhu Jianming group is to enshi fishing pool height of dam selenium carbon
Microorganism diversity profiles in matter mud stone have carried out Primary Study (thunder of heap of stone etc., 2009), and have separated two plants and have selenous acid
The bacterium (Wang Mingyi etc., 2006) of salt reducing power.It is only " a kind of to be lived using microbial fermentation production in terms of patent of invention
Change the method for selenium mineral powder " (ZL201210166664.4) and " a method of prepare red elemental selenium using super resistance to selenium microorganism "
Selenium slag, soil in (application number 201210167650.6) using enshi selenium minerals carry out the related resistance to micro- life of selenium of separation
Object, and then be inoculated into selenium mineral powder or other media containing selenium, preparation activation selenium mineral powder or red elemental selenium.But in patent of invention simultaneously
Related resistance to selenium microorganism is not isolated and purified, does not identify microorganism kind and feature yet.
Summary of the invention
In order to solve the above technical problems, the present invention provides a plant height mountain bacillus altitudinis
YLX-5 and its application.
The technical scheme is that
The present invention provides a plant height mountain bacillus altitudinis YLX-5, the high mountain bacillus
Be from Chinese enshi selenium mining area diafiltration water system bacterium seat deposit in isolate and purify, with thermophilic selenium microorganism feature
Bacterium bacterial strain, entitled high mountain bacillus YLX-5 (the Bacillus altitudinis of the preservation of the high mountain bacillus
YLX-5), depositary institution is China typical culture collection center (CCTCC), preservation address are as follows: Wuhan, China Wuhan University is protected
The hiding date is on December 18th, 2013, and deposit number is CCTCC NO:M2013675.
The strain mycology feature of the high mountain bacillus is described as follows:
[morphological feature] the high mountain bacillus cultivates 24 hours bacterial strains observed in 28 degrees Celsius of TSA culture medium
Group is rose pink smooth circle, and has full edge convex surface (attached drawing 1)；Bacterial strain be it is rod-shaped, size be 0.5~0.8 × 8~
10 μm (attached drawing 2)；Gram's staining is carried out to the bacterial strain, is shown as gram-positive bacteria.
[thermophilic selenium feature] the high mountain bacillus is successively respectively connected to containing selenium (being configured with sodium selenite) 0,10,20,
50,100,200,500,1000,2000, in the TSB culture solution of 5000mg/kg, as the result is shown: in selenium concentration≤20mg/kg feelings
Under condition, bacterial growth is not inhibited, and apparent red is not presented for culture solution color.In selenium concentration >=200mg/kg, training
Nutrient solution color starts that apparent red is presented, and under the conditions of 2000mg/kg, the red in culture solution is still obvious, shows the bacterium
Kind Metabolism of Normal.It must be noted that even if strain can still be grown in the case where Se content is up to the extreme condition of 2000mg/kg,
Extreme Selenium endurance is embodied, but culture solution red obviously weakens.More than under 2000mg/kg concentration, the strain is not
It can growth.
[16S rRNA gene sequencing] extracts genomic DNA from the pure culture of bacterial strain of the present invention, utilizes
Universal primer 27f and 1492r carry out PCR amplification and sequencing, further by CLUSTAL X software and 4.0 software of Mega with
Neighbour-joining method constructs systematic evolution tree (attached drawing 3), carries out Phylogenetic Analysis.Bacterial strain belongs to as the result is shown
High mountain bacillus (Bacillus altitudinis) is named as high mountain bacillus YLX-5 (Bacillus
Altitudinis YLX-5), depositary institution is China typical culture collection center (CCTCC), preservation address are as follows: China is military
Chinese Wuhan University, the deposit date is on December 18th, 2013, deposit number was CCTCC NO:M2013675.
The motility feature detection of [biochemical reactions feature] bacterial strain shows that its motility is good, and it is suitable for model that pH, which is grown,
Enclose for 5-8,11% and concentrations above NaCl under the conditions of cannot grow；It can be raw as sole carbon source using inositol or D-glucitol
It is long, but (table 1) cannot be grown by sole carbon source of D- raffinose.
1 bacterial strain motility of table, growth tolerance condition and utilization of carbon source
| Number | Detection project | As a result |
| 1 | Bacterial motility | Movement |
| 2 | PH growth scope | 5-8 |
| 3 | 2%NaCl concentration growing state | Growth |
| 4 | 11%NaCl concentration growing state | It does not grow |
| 5 | 17%NaCl concentration growing state | It does not grow |
| 6 | Inositol is sole carbon source | Growth |
| 7 | D- gossypose is sole carbon source | It does not grow |
| 8 | D-glucitol is sole carbon source | Growth |
The Enzyme activity assay of the bacterial strain is as the result is shown: bacterial strain has alkaline phosphatase, esterase (C4), esterase lipase (C8), white ammonia
Sour arylamine enzyme, chymotrypsin, acid phosphatase, naphthols-AS-BI- phosphohydrolase, beta galactosidase, β-uronic acid
Glycosides enzyme, alpha-glucosidase, beta-glucosidase, alpha-Mannosidase reaction are positive (table 2).
2 bacterial strain enzymatic activity characteristic of table
| Number | The enzyme and physiological property of measurement detect | As a result |
| 1 | Blank | - |
| 2 | Alkaline phosphate enzyme | + |
| 3 | Esterase (C4) | + |
| 4 | Esterase lipase (C8) | + |
| 5 | Lipase (C14) | - |
| 6 | Leucine arylamine enzyme | + |
| 7 | Valine arylamine enzyme | - |
| 8 | Cystine arylamine enzyme | - |
| 9 | Trypsase | - |
| 10 | Chymotrypsin | + |
| 11 | Acid phosphatase | + |
| 12 | Naphthols-AS-BI- phosphohydrolase | + |
| 13 | Alpha-galactosidase | - |
| 14 | Beta galactoside enzyme | + |
| 15 | β-uronic acid glucoside enzyme | + |
| 16 | Alpha-gluconase activity enzyme | + |
| 17 | β-glucosaccharase | + |
| 18 | N- acetyl-glucosaminidase | - |
| 19 | α-mannosidase | + |
| 20 | β-fucose glucoside enzyme | - |
Bacterial strain antibiotic resistance testing result is shown: bacterial strain to amikacin, Amoxicillin, ampicillin, cefoperazone,
Chloramphenicol, Ciprofloxacin, Compound New Nomin, Norfloxacin, ovobiocin, penicillin are sensitivity, only to lincomycin
Resistant (table 3).
3 bacterial strain antibiotic resistance of table
| Number | Antibiotic | As a result |
| 1 | Amikacin | S |
| 2 | Amoxicillin | S |
| 3 | Ampicillin | S |
| 4 | Cefoperazone | S |
| 5 | Chloramphenicol | S |
| 6 | Ciprofloxacin | S |
| 7 | Compound New Nomin | S |
| 8 | Lincomycin | R |
| 9 | Norfloxacin | S |
| 10 | Ovobiocin | S |
| 11 | Penicillin | S |
Indicate: wherein " R " indicates resistance, and " S " indicates sensitive.
The invention also discloses high mountain bacillus altitudinis YLX-5 in preparation microorganism conversion
The application of organic selenium.High mountain bacillus altitudinis YLX-5 of the present invention has thermophilic selenium microorganism special
Sign, the soluble state inorganic selenium (Se in culture solution4+、Se6+) content be no more than 20mg/L under conditions of, can be by inorganic selenium height
Effect is converted into containing selenoaminoacid (such as methylselenocysteinefrom), further can be using these organic seleniums as selenium deficiency crowd
Selenium-supply source.
The invention also discloses high mountain bacillus altitudinis YLX-5 in preparation microorganism conversion
Application in nanometer selenium.High mountain bacillus altitudinis YLX-5 of the present invention has thermophilic selenium microorganism
Feature, the soluble state inorganic selenium (Se in culture solution4+、Se6+) content under conditions of 200mg/L~2000mg/L, can will
Inorganic selenium Efficient Conversion is nanometer selenium, and the size of the nanometer selenium is 100~200nm, may further have biology using these
Selenium-supply source of the active nanometer selenium as selenium deficiency crowd.
The invention also discloses high mountain bacillus altitudinis YLX-5 in activation ore containing selenium
Application.High mountain bacillus altitudinis YLX-5 of the present invention has thermophilic selenium microorganism feature, can
To carry out activation of microorganism to ore containing selenium, and then activating the not bioavailable selenium in ore containing selenium is bioavailable
Selenium, can will naturally bioavailable Se content improves 4~5 times in ore containing selenium, may further be by the mine containing selenium after activation
Stone is processed as Selenium in Soil modifying agent, is applied in selenium deficiency soil and plants selenium-enriched crop.
The invention also discloses high mountain bacillus altitudinis YLX-5 to prepare selenium-enriched microbe
Application in fertilizer.High mountain bacillus altitudinis YLX-5 of the present invention has thermophilic selenium microorganism special
Sign, can directly be made as microorganism freeze-dried powder, and rehydration is brought back to life after being administered in soil, and the selenium in activating soil improves soil
The bioavailability of selenium, to improve crop to the cumulative efficiency of selenium；Microorganism can also be made with organic fertilizer composite fermentation
Organic fertilizer is applied in soil and plants selenium-enriched crop.
The invention also discloses high mountain bacillus altitudinis YLX-5 in selenium contamination environment remediation
In application.High mountain bacillus altitudinis YLX-5 of the present invention has thermophilic selenium microorganism feature,
When Se content is no more than 500 μ g/L in polluted-water, microorganism conversion, accumulation can be carried out to the selenium in selenium contamination environment,
These microorganisms may further be collected, the Se content in environment is on the one hand substantially reduced, achieve the purpose that repairing environment, it is another
Aspect can use the microorganism selenium of conversion, accumulation, as the source of microorganism organic selenium or microorganism nanometer selenium, be used for selenium deficiency
The selenium of crowd supplements source, to the elimination efficiency of selenium in selenium contamination water body up to 50~60%.
According to the above aspect of the present invention, the present invention has at least the following advantages: high mountain bacillus of the present invention
Altitudinis YLX-5 has thermophilic selenium microbiologic properties, can convert, the synthesis of microorganism nanometer selenium, contain in microorganism organic selenium
It is had potential application in selenium ore activation of microorganism, selenium-enriched microbe fertilizer and selenium contamination environment remediation.
The above description is only an overview of the technical scheme of the present invention, in order to better understand the technical means of the present invention,
And can be implemented in accordance with the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention and the accompanying drawings.
Detailed description of the invention
Fig. 1 is the morphological feature of bacterial strain of the present invention；
Fig. 2 is the morphological feature of bacterial strain of the present invention under the microscope；
Fig. 3 is the phylogenetic tree schematic diagram of the 16S rRNA gene order of bacterial strain of the present invention.
Specific embodiment
With reference to the accompanying drawings and examples, specific embodiments of the present invention will be described in further detail, implements below
Example is not intended to limit the scope of the invention for illustrating the present invention.
Bacteria selection separation is carried out as steps described below:
(1) primary dcreening operation: aseptic collection Hubei China bestow favour fishing pool dam selenium mining area diafiltration water system in bacterium seat sediment sample
About 20g is in the sealed bottle of the 25mL of sterilization treatment, 3 parts of parallel acquisition, takes back 4 DEG C of laboratory cryo-conservation.Then it is testing
3 parts of parallel samples are carried out in the aseptic operating platform of room uniformly mixed, takes 5g to be placed in the conical flask of 50mL sterile purified water, room temperature
Under the conditions of 150r/min shake 15min, stand 30 seconds after take 1mL bacterium solution echelon to be diluted to 10-3、10-4、10-5、10-6, then connect
Kind of 0.05mL is spread evenly across on isolation medium that (isolation medium composition: the TSA of the 200 μ g/kg containing selenium, selenium source is selenous acid
Sodium), 4 groups of repetitions are arranged in each processing.It is cultivated 2-3 days under room temperature, typically single bacterium colony carries out a little meeting purifying 2-3 picking form
It is secondary to obtain pure culture.
(2) secondary screening: by pure culture obtained in (1) be sequentially ingressed into containing selenium (being configured with sodium selenite) 0,10,20,50,
100,200,500,1000,2000, in the TSB culture solution of 5000mg/kg, under the conditions of selenium concentration≤2000mg/kg, the bacterial strain
Energy normal growth shows that the pure culture has thermophilic selenium ability, is needed thermophilic selenium microorganism, which is named as
YLX-5 is preserved in storage medium.
It is raw that morphological feature observation, molecular biology identification and physiology are carried out to the above-mentioned high mountain bacillus obtained through secondary screening
Change response feature test.
Morphological feature observation: the high mountain bacillus is cultivated in 28 degrees Celsius of TSA culture medium to be observed for 24 hours
Bacterial strain group is rose pink smooth circle, and has full edge convex surface (attached drawing 1)；The bacterium observed under transmission electron microscope
Strain be it is rod-shaped, size be 0.5~0.8 × 8~10 μm (attached drawing 2)；Gram's staining is carried out to the bacterial strain, is shown as gram sun
Property bacterium.
Molecular biology identification: genomic DNA is extracted from the pure culture of bacterial strain of the present invention, utilizes universal primer
27f and 1492r carries out PCR amplification and sequencing, further by CLUSTAL X software and 4.0 software of Mega with Neighbour-
Joining method constructs systematic evolution tree (attached drawing 3), carries out Phylogenetic Analysis.Bacterial strain belongs to high mountain gemma bar as the result is shown
Bacterium (Bacillus altitudinis) is named as high mountain bacillus YLX-5 (Bacillus altitudinis YLX-5),
Depositary institution is China typical culture collection center (CCTCC), preservation address are as follows: Wuhan, China Wuhan University, preservation date
For on December 18th, 2013, deposit number was CCTCC NO:M2013675.
Biochemical reactions characteristic test: the motility feature detection of YLX-5 bacterial strain shows that its motility is good, pH growth
Optimum range is 5-8,11% and concentrations above NaCl under the conditions of cannot grow；It can be unique with inositol or D-glucitol
Carbon source for growth, but (table 1) cannot be grown by sole carbon source of D- raffinose.The Enzyme activity assay of the bacterial strain is as the result is shown: bacterial strain
With alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamine enzyme, chymotrypsin, acid phosphatase, naphthalene
Phenol-AS-BI- phosphohydrolase, beta galactosidase, β-glycuronide enzyme, alpha-glucosidase, beta-glucosidase, α-are sweet
Dew glucosides enzyme reaction is positive (table 2).Bacterial strain antibiotic resistance testing result is shown: bacterial strain is to amikacin, Amoxicillin, ammonia
Benzyl XiLin, cefoperazone, chloramphenicol, Ciprofloxacin, Compound New Nomin, Norfloxacin, ovobiocin, penicillin are quick
Sense, only (table 3) resistant to lincomycin.
Embodiment 1
Application of the microorganism of the present invention in synthesis organic selenium.
By microorganism YLX-5 strain inoculated of the present invention to the TSB culture solution for containing selenium (sodium selenite configuration) 10mg/L
In, 200r/min is cultivated 48 hours under room temperature, and thalline were collected by centrifugation and supernatant.Then liquid chromatogram-atomic fluorescence light is utilized
It composes combined instrument (LC-GA-AFS) and detects supernatant and Se content and form in thallus respectively, as the result is shown in supernatant almost
It is Se4+, and the overwhelming majority is methylselenocysteinefrom (SeMeCys) (table 4) in thallus.This shows YLX-5 bacterial strain energy
Under the conditions of 10mg Se/L, by the inorganic selenium (Se in culture solution4+) be converted into organic selenium (SeMeCys), and accumulate and arrive thallus
It is interior.Further collecting thallus can be used as the organic selenium addition of animal feed etc., can also do further seleno-amino acids purifying
Raw material, the organic selenium selenium-supply source as selenium deficiency crowd.
The Se form feature of supernatant and thallus in 4 reaction system of table
Note: "-" expression does not detect；" * " indicates no sample.
Embodiment 2
Application of the microorganism of the present invention in synthesis organic selenium.
By microorganism YLX-5 strain inoculated of the present invention to the TSB culture solution for containing selenium (sodium selenite configuration) 20mg/L
In, 200r/min is cultivated 72 hours under room temperature, and thalline were collected by centrifugation and supernatant.Then liquid chromatogram-atomic fluorescence light is utilized
It composes combined instrument (LC-GA-AFS) and detects supernatant and Se content and form in thallus respectively, as the result is shown in supernatant almost
It is Se4+, and the overwhelming majority is methylselenocysteine (SeMeCys) in thallus, has few portion in the growth stage of stable development of bacterial strain
Divide Se4+(table 5).This shows that YLX-5 bacterial strain can be under the conditions of 20mg Se/L, by the inorganic selenium (Se in culture solution4+) be converted into
Organic selenium (SeMeCys), and accumulate into thallus.Further collecting thallus can be used as the organic selenium addition of animal feed etc.,
Further seleno-amino acids purified feed stock can be done, the organic selenium selenium-supply source as selenium deficiency crowd.
The Se form feature of supernatant and thallus in 5 reaction system of table
Note: "-" expression does not detect；" * " indicates no sample.
Embodiment 3
Application of the microorganism of the present invention in synthesis nanometer selenium.
By microorganism YLX-5 strain inoculated of the present invention to the TSB culture solution for containing selenium (sodium selenite configuration) 200mg/L
In, 200r/min is cultivated 24 hours under room temperature, and solution occurs apparent red, muddy.Culture 48 hours, thalline were collected by centrifugation and
Supernatant, what discovery collection obtained is precipitated as red suspended matter.Then liquid chromatogram-atomic fluorescence spectrophotometry combined instrument (LC- is utilized
GA-AFS) and Atomic Fluorescence Spectrometer, identify that the ingredient of red suspended matter is almost elemental selenium using minusing.This microorganism is closed
At nanometer selenium can be used as the selenium-supply source of selenium deficiency crowd, can also be used as selenium fertilizer or animal feed additive.
Embodiment 4
Application of the microorganism of the present invention in synthesis nanometer selenium.
Microorganism YLX-5 strain inoculated of the present invention is cultivated to the TSB containing selenium (sodium selenite configuration) 2000mg/L
In liquid, 200r/min is cultivated 12 hours under room temperature, and solution occurs apparent red, muddy.Culture 48 hours, thalline were collected by centrifugation
And supernatant, what discovery collection obtained is precipitated as red suspended matter.Then liquid chromatogram-atomic fluorescence spectrophotometry combined instrument is utilized
(LC-GA-AFS) and Atomic Fluorescence Spectrometer, identify that the ingredient of red suspended matter is almost elemental selenium using minusing.This micro- life
The nanometer selenium of object synthesis can be used as the selenium-supply source of selenium deficiency crowd, can also be used as selenium fertilizer or animal feed additive.
Embodiment 5
Application of the microorganism of the present invention in activation selenium ore.
The selenium ore for picking up from enshi selenium mining area is crushed to 100 mesh, then measuring its total Se content is about 3000mg/
Kg measures the water soluble speciation about accounting 1% in selenium ore, exchangeable species selenium about accounting further by continuous extraction method
3%, Fe-Mn oxide selenium about accounting 5%, organically combine state selenium about accounting 10%, residual selenium accounting about 81%.With this selenium mine
Powder is selenium source, adds the 100g selenium mineral powder into the TSB culture solution of 300mL, one group is not inoculated with YLX-5 (8 Duplicate Samples), another
Group inoculation YLX-5 (8 Duplicate Samples).200r/min is cultivated 90 days at room temperature, and is sampled according to certain time interval
Total Se content (table 6) in supernatant is detected, be inoculated in YLX-5 bacterial strain group obviously there are more soluble states compared with control group as the result is shown
Selenium enters in solution, and Se content is 4~5 times of control group or so.This shows that YLX-5 bacterial strain can will can not give birth in selenium ore
Object is converted into bioavailable selenium using selenium (such as exchange state selenium, Fe-Mn oxide selenium), and efficiency is higher.It is further available
This characteristic of YLX-5 activates natural selenium ore, and product can be used as Selenium in Soil modifying agent, for changing for selenium deficiency soil
It is good.
Se content feature in 6 reaction system supernatant of table
Embodiment 6
Application of the microorganism of the present invention in the preparation of selenium-enriched microbe fertilizer.
Microorganism YLX-5 bacterial strain of the present invention is carried out 50L scale fermented and cultured 3-5 days in TSB culture solution, into
One step prepares microorganism freeze-dried powder using freeze drying technology.The freeze-dried powder can be applied directly in romaine lettuce crop root soil, with
Watering can be such that freeze-dried microorganisms rehydration brings back to life afterwards.Normal cultivation harvests after 30 days, and detection shows that the Se content in its Chinese leaf is
8-11 times for not applying the Chinese leaf sample of the microorganism freeze-dried powder.
Embodiment 7
Application of the microorganism of the present invention in the preparation of selenium-enriched microbe fertilizer.
By microorganism YLX-5 strain inoculated of the present invention into organic fertilizer raw material, normal organic fertilizer heap is then carried out
Fertilizer fermentation, obtains microbial organic fertilizer.The microbial organic fertilizer is administered to romaine lettuce root, and normal culture harvests after 30 days, detects
Show that the Se content in its Chinese leaf is 5-6 times of Chinese leaf sample for not applying the microorganism freeze-dried powder.
Embodiment 8
Application of the microorganism of the present invention in selenium contamination environment remediation.
Measuring total Se content in the leakage water of enshi selenium mining area is about 85 μ g/L, and as selenium contamination environment water
Representative, YLX-5 strain is accessed in the 300mL water body (8 Duplicate Samples), and glucose is added as carbon source, is arranged simultaneously
Control group, 200r/min is cultivated 10 days at room temperature, and according to total in certain time interval sample detection supernatant
Se content (table 7) is not inoculated with the Se content in the control group of YLX-5 bacterial strain in solution as the result is shown and is basically unchanged, and is inoculated with
Selenium in solution is obviously absorbed during strain growth, is utilized by YLX-5 bacterial strain processing group, is greatly reduced in solution
Se content, clearance rate reach 50~60% or so.Therefore, further using this characteristic of YLX-5 to natural selenium contamination
Water body in area's (such as bestowing favour) or artificial selenium contamination environment (such as manganese smelting region) carries out repair process.
Se content feature in 7 reaction system supernatant of table
The above is only a preferred embodiment of the present invention, it is not intended to restrict the invention, it is noted that for this skill
For the those of ordinary skill in art field, without departing from the technical principles of the invention, can also make it is several improvement and
Modification, these improvements and modifications also should be regarded as protection scope of the present invention.
Claims (7)
1. a kind of high mountain bacillus (Bacillus altitudinis) YLX-5, it is characterised in that: the high mountain bacillus
The entitled high mountain bacillus altitudinis YLX-5 of preservation, depositary institution is China typical culture collection
Center, the deposit date is on December 18th, 2013, deposit number was CCTCC NO:M2013675.
2. a kind of application of strain CCTCC NO:M2013675 as described in claim 1, it is characterised in that: the high mountain gemma
Application of the bacillus Bacillus altitudinis YLX-5 in preparation microorganism conversion organic selenium, the high mountain bacillus
Bacillus altitudinis YLX-5 can inhale the soluble state inorganic selenium when soluble state Se content is lower than 20mg/L
Receipts are converted into organic selenium, wherein the soluble state selenium is soluble state inorganic selenium Se4+With soluble state inorganic selenium Se6+, wherein described have
Machine selenium is containing selenoaminoacid.
3. a kind of application of strain CCTCC NO:M2013675 as described in claim 1, it is characterised in that: the high mountain gemma
Application of the bacillus Bacillus altitudinis YLX-5 in preparation microorganism conversion nanometer selenium.
4. application according to claim 3, it is characterised in that: the high mountain bacillus altitudinis
YLX-5 can convert the soluble state inorganic selenium to biologically active when soluble state Se content is 200~2000mg/L
Nanometer selenium.
5. a kind of application of strain CCTCC NO:M2013675 as described in claim 1, it is characterised in that: the high mountain gemma
Application of the bacillus Bacillus altitudinis YLX-5 in activation ore containing selenium.
6. a kind of application of strain CCTCC NO:M2013675 as described in claim 1, it is characterised in that: the high mountain gemma
Bacillus Bacillus altitudinis YLX-5 is preparing the application in selenium-enriched microbe fertilizer.
7. a kind of application of strain CCTCC NO:M2013675 as described in claim 1, it is characterised in that: the high mountain gemma
Application of the bacillus Bacillus altitudinis YLX-5 in selenium contamination environment remediation.
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| CN201510547672.9A CN105132315B (en) | 2015-08-31 | 2015-08-31 | High mountain bacillus altitudinis YLX-5 and its application |
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| CN109402007B (en) * | 2018-11-12 | 2021-12-24 | 中国科学院合肥物质科学研究院 | Biological nano-selenium producing strain and method for preparing biological nano-selenium by using strain |
| CN109777749B (en) * | 2018-12-18 | 2021-03-30 | 广西大学 | Selenium-enriched bacillus pumilus strain D1-019 and separation and screening method and application thereof |
| CN111100818B (en) * | 2020-01-10 | 2023-05-12 | 新疆根力多生物科技有限公司 | Geobacillus altitudinalis SWY137 and application thereof |
| CN112759453A (en) * | 2020-12-31 | 2021-05-07 | 苏州硒谷科技有限公司 | Special selenium-rich organic fertilizer for fruit trees and preparation method thereof |
| CN115232760B (en) * | 2022-05-24 | 2023-10-20 | 武汉轻工大学 | Bacillus alpine and application thereof |
| CN114990033B (en) * | 2022-07-19 | 2022-12-06 | 广东省科学院动物研究所 | Heavy metal resistant bacillus altitudinis and application thereof |
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