CN109182222A - The method of heavy metal cadmium in selenium-rich microbial agent preparation and its degradation soil and crops - Google Patents
The method of heavy metal cadmium in selenium-rich microbial agent preparation and its degradation soil and crops Download PDFInfo
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- CN109182222A CN109182222A CN201811251083.6A CN201811251083A CN109182222A CN 109182222 A CN109182222 A CN 109182222A CN 201811251083 A CN201811251083 A CN 201811251083A CN 109182222 A CN109182222 A CN 109182222A
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- 239000011669 selenium Substances 0.000 title claims abstract description 103
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 title claims abstract description 101
- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 101
- 238000002360 preparation method Methods 0.000 title claims abstract description 61
- 239000002689 soil Substances 0.000 title claims abstract description 53
- 229910052793 cadmium Inorganic materials 0.000 title claims abstract description 42
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 28
- 229910001385 heavy metal Inorganic materials 0.000 title claims abstract description 17
- 239000003795 chemical substances by application Substances 0.000 title abstract description 6
- 230000015556 catabolic process Effects 0.000 title abstract description 5
- 238000006731 degradation reaction Methods 0.000 title abstract description 5
- 230000000813 microbial effect Effects 0.000 title abstract description 3
- 241000894006 Bacteria Species 0.000 claims abstract description 114
- 238000000855 fermentation Methods 0.000 claims abstract description 29
- 230000004151 fermentation Effects 0.000 claims abstract description 29
- 230000000243 photosynthetic effect Effects 0.000 claims abstract description 24
- 238000007598 dipping method Methods 0.000 claims abstract description 9
- 239000001963 growth medium Substances 0.000 claims abstract description 5
- 239000000243 solution Substances 0.000 claims description 33
- 238000010790 dilution Methods 0.000 claims description 17
- 239000012895 dilution Substances 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 238000002955 isolation Methods 0.000 claims description 16
- 239000002609 medium Substances 0.000 claims description 16
- 238000005286 illumination Methods 0.000 claims description 13
- 239000001888 Peptone Substances 0.000 claims description 11
- 108010080698 Peptones Proteins 0.000 claims description 11
- BJEPYKJPYRNKOW-UHFFFAOYSA-N malic acid Chemical compound OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 claims description 11
- 235000019319 peptone Nutrition 0.000 claims description 11
- 239000007921 spray Substances 0.000 claims description 10
- 239000003337 fertilizer Substances 0.000 claims description 9
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 8
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 8
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 8
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 7
- 229940041514 candida albicans extract Drugs 0.000 claims description 7
- 238000012216 screening Methods 0.000 claims description 7
- 239000001632 sodium acetate Substances 0.000 claims description 7
- 235000017281 sodium acetate Nutrition 0.000 claims description 7
- 238000005507 spraying Methods 0.000 claims description 7
- 239000012138 yeast extract Substances 0.000 claims description 7
- 229910018143 SeO3 Inorganic materials 0.000 claims description 6
- 238000003973 irrigation Methods 0.000 claims description 6
- 230000002262 irrigation Effects 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 6
- 239000011734 sodium Substances 0.000 claims description 6
- 229910003424 Na2SeO3 Inorganic materials 0.000 claims description 5
- 239000011781 sodium selenite Substances 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- 239000001110 calcium chloride Substances 0.000 claims description 4
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 238000012545 processing Methods 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 3
- 241000209504 Poaceae Species 0.000 claims description 2
- 241000589516 Pseudomonas Species 0.000 claims description 2
- 241000191023 Rhodobacter capsulatus Species 0.000 claims description 2
- 230000000844 anti-bacterial effect Effects 0.000 claims description 2
- 230000000593 degrading effect Effects 0.000 claims description 2
- 238000009313 farming Methods 0.000 claims description 2
- 229910052757 nitrogen Inorganic materials 0.000 claims description 2
- 241000605008 Spirillum Species 0.000 claims 1
- 238000011081 inoculation Methods 0.000 claims 1
- 235000015097 nutrients Nutrition 0.000 claims 1
- 238000010521 absorption reaction Methods 0.000 abstract description 7
- 230000002378 acidificating effect Effects 0.000 abstract description 2
- 238000012258 culturing Methods 0.000 abstract description 2
- 239000000725 suspension Substances 0.000 abstract description 2
- 241000196324 Embryophyta Species 0.000 description 17
- 241000209094 Oryza Species 0.000 description 10
- 235000007164 Oryza sativa Nutrition 0.000 description 10
- 235000009566 rice Nutrition 0.000 description 10
- 230000000694 effects Effects 0.000 description 5
- 239000002253 acid Substances 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 2
- 241000190984 Rhodospirillum rubrum Species 0.000 description 2
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- WLZRMCYVCSSEQC-UHFFFAOYSA-N cadmium(2+) Chemical compound [Cd+2] WLZRMCYVCSSEQC-UHFFFAOYSA-N 0.000 description 2
- 238000003113 dilution method Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 235000008733 Citrus aurantifolia Nutrition 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000006587 Glutathione peroxidase Human genes 0.000 description 1
- 108700016172 Glutathione peroxidases Proteins 0.000 description 1
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 1
- 241000190932 Rhodopseudomonas Species 0.000 description 1
- 241001062472 Stokellia anisodon Species 0.000 description 1
- 102000013090 Thioredoxin-Disulfide Reductase Human genes 0.000 description 1
- 108010079911 Thioredoxin-disulfide reductase Proteins 0.000 description 1
- 235000011941 Tilia x europaea Nutrition 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 229910052785 arsenic Inorganic materials 0.000 description 1
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 description 1
- 230000037358 bacterial metabolism Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 208000005882 cadmium poisoning Diseases 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 150000001722 carbon compounds Chemical class 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 239000004571 lime Substances 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 238000005065 mining Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229940072033 potash Drugs 0.000 description 1
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Substances [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 1
- 235000015320 potassium carbonate Nutrition 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000005067 remediation Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 238000003971 tillage Methods 0.000 description 1
- 229910021654 trace metal Inorganic materials 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000009333 weeding Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protection of plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Forests & Forestry (AREA)
- Microbiology (AREA)
- Ecology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Biomedical Technology (AREA)
- Environmental Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Toxicology (AREA)
- Medicinal Chemistry (AREA)
- Environmental & Geological Engineering (AREA)
- Soil Sciences (AREA)
- Mycology (AREA)
- Botany (AREA)
- Soil Conditioners And Soil-Stabilizing Materials (AREA)
- Processing Of Solid Wastes (AREA)
Abstract
The invention discloses the methods of heavy metal cadmium in selenium-rich microbial agent preparation and its degradation soil and crops, are related to soil and educate technical field again；Using Selenium-enriched fermentation culture medium to gained selenium-rich bacteria preparation after photosynthetic bacteria culture, the selenium-rich bacteria preparation is applied in degradation soil in cadmium element, selenium-rich bacteria preparation is repeatedly sprayed by root dipping, pouring root and in plant growing process, can effectively prevent crops to the content of cadmium in the absorption of cadmium and soil；The present invention filters out resistance to selenium combined bacteria strain, a kind of photosynthetic bacteria suspension of selenium-rich is produced by selenium-rich culturing, it is applied in acidic cadmium polluted soil, the available Cd in soil can be reduced, contaminated soil is repaired, absorption of the plant to cadmium can be prevented again, while improving the Se content in plant, realize that the selenium-rich of crops or other industrial crops is planted safely in cadmium pollution soil.
Description
Technical field
The present invention relates to soil to educate technical field again, is related to a kind of removing cadmium in acid soil using biotechnology
The method of content more particularly relates to a kind of preparation method of selenium-rich bacteria preparation and its in soil except the application in cadmium, Yi Jisuo
The selenium-rich bacteria preparation stated is applied to prevent crops to the method for Cd uptake.
Background technique
Cadmium is to plant and animal toxicity most by force to one of trace metal element, mankind's industrial or agricultural activity such as mining, smelt,
Fertilising, pesticide, agricultural sludge and sewage irrigation etc. cause many Regional field soil in the world different degrees of cadmium pollution occur, and
The cadmium of available state is easily absorbed by plants in agricultural land soil, is entered its edible portion from root system of plant, is caused human body cadmium poisoning,
And cadmium content is positively correlated with Cadmium Pollution in Soils degree in crop, therefore how to control Cadmium Pollution in Soils and realize crop safety kind
Plant has become very important project instantly.
The method of currently used processing heavy metal-polluted soil has: overturning dilution method, chemical removal method and vegetation recovery method.
It overturns dilution method predominantly to mix contaminated soil turning, but in fact there are still have heavy metal not really from soil for soil
Earth removal.Chemical removal method predominantly utilizes dilute acid soln or chelating agent and contaminated soil to act on；After chemical removal method,
Although heavy metal can be removed from contaminated soil, the soil texture and fertility are deteriorated after effect, to influence that soil is subsequent to answer
With.Vegetation recovery method is mainly planting plants in contaminated soil, and penetrates plant absorption heavy metal；However, heavy metal with
Soil particle is respectively provided with positive charge and negative electrical charge and attracts each other, therefore heavy metal is not easily shifted so that the regulation that must expend
Time is long.In terms of another, after plant absorption heavy metal, heavy metal focuses mostly in plant root, thus must remove whole plant beginning energy
Removing heavy metals are removed from soil.If the contaminated degree of soil seriously must repeatedly carry out vegetation recovery method excessively, every time must be again
Planting plants simultaneously come into leaves to plant, bloom or result.In this way, more extend the regulation time repeatedly integrally expended.
Photosynthetic bacterium growth is using light as the energy, with CO2Or organic carbon compound is that carbon source carries out photosynthesis, it can be in anaerobism light
According to being proliferated in, aerobic illumination or even aerobic dark surrounds, studies have shown that photosynthetic bacteria can pass through absorption and reduction is inhaled
Conversion heavy metal cadmium ion is received, the cadmium ion of soil available is removed, prevents its pollution to plant, but under natural environment
Photosynthetic bacteria its remove cadmium low efficiency, not can effectively prevent enrichment of the cadmium in plant；It is a kind of new it is therefore desirable to propose
Soil educates method again to solve currently used method problem encountered.
Summary of the invention
The present invention overcomes the shortcomings of the prior art, provides a kind of preparation method of selenium-rich bacteria preparation, passes through this method
The selenium-rich bacteria preparation of preparation can effectively degrade the heavy metal cadmium in soil.The present invention is achieved through the following technical solutions.
A kind of preparation method of selenium-rich bacteria preparation, comprising the following steps:
A) resistance to selenium combined bacteria primary dcreening operation: photosynthetic bacteria being inoculated in isolation medium and carries out primary dcreening operation, filters out resistance to selenium combined bacteria；Institute
The isolation medium stated are as follows: DL-malic acid 2~5g/L, (NH4)2SO4 0.5~1.5g/L, 2~5g/L of yeast extract, peptone
2~5g/L, agar 10~20g, Na2SeO340~120mg/ L adds water to 1L, adjusts pH6.5~7.0.
B) the 3-5 resistance to selenium combined bacteria bacterium colonies filtered out photosynthetic bacteria Selenium-enriched fermentation: are taken from the isolation medium
It is seeded in resistance to selenium combined bacteria culture solution, Na is added in the culture solution when cultivating 2-3d2SeO3Aqueous solution makes the culture
The selenium concentration of liquid is 1-100mg/L, and 1200~1500lux intensity of illumination, 400-800nm illumination are then placed at 28-32 DEG C
Illumination cultivation 3-10d is carried out under wave band obtains selenium-rich bacteria preparation when the culture solution OD value reaches 1-2.
The resistance to selenium combined bacteria culture solution is 2~5g/L of DL-malic acid, (NH4)2SO4 0.5~1.5g/L, MgSO4
100~150mg/L, CaCl2 60~85mg/L, 15~30mg/L of sodium acetate, 2~5g/L of yeast powder, 2~5g/L of peptone add
Water adjusts pH6.5~7.0 to 1L.
Preferably, the condition of resistance to selenium combined bacteria primary dcreening operation is to be inverted isolation medium plate in the step a, is trained in constant temperature
It supports in case and cultivates 2-3d at 28-32 DEG C.
Preferably, secondary fermentation and secondary screening separation are successively carried out after the resistance to selenium combined bacteria primary dcreening operation step；By secondary screening
Resistance to selenium combined bacteria bacterium colony obtained after separation is used for the photosynthetic bacteria Selenium-enriched fermentation；The secondary fermentation is by resistance to selenium
The bacterium colony that combined bacteria primary dcreening operation obtains when cultivating 0-1d, is added in the second order fermentation culture solution and adds in second order fermentation culture solution
Enter Na2SeO3Aqueous solution makes the selenium concentration 1-100mg/L of the second order fermentation culture solution, cultivates 2-3d at 28-32 DEG C.
The second order fermentation culture solution be 2~5g/L of DL-malic acid, 15~30mg/L of sodium acetate, yeast extract 2~
5g/L, 2~5g/L of peptone add water to 1L, adjust pH6.5~7.0.
The photosynthetic bacteria is Rhodospirillum rubrum (Rhodospirillum rubrum), the red bacterium of fixed nitrogen
(Rhodotacter azotoformans), red pseudomonas (Rhodopseudomonas), Spherical red antibacterial
The one of which of (Rhodotacter sphaeroides), Rhodobacter capsulatus (Rhodotacter capsulatus) or its
Meaning combination.
A kind of method of cadmium element in degradation soil, comprising the following steps: add the water to be according to volume ratio selenium-rich bacteria preparation
1-5:100 dilution, is prepared into bacteria preparation dilution, pours according to 18-28t/ mus to soil；The selenium-rich bacteria preparation is
Using Selenium-enriched fermentation culture medium to gained mixture after photosynthetic bacteria culture.
A method of it prevents crops to Cd uptake, including nursery, basal dressing in planting process, further includes following step
It is rapid:
A) root dipping: by selenium-rich bacteria preparation add water according to volume ratio be 1-5:100 dilute, bacteria preparation dilution is prepared into, to farming
Object seedling carries out root dipping processing, is transplanted later；
B) bacteria preparation dilution pouring root: is subjected to root irrigation to the seedling after transplanting；
C) it spraying fertilizer and sprays for the first time: spraying fertilizer after turning green, and spray bacteria preparation dilution；
D) in the entire growth course of crops, then the bacteria preparation dilution is sprayed several times；
The selenium-rich bacteria preparation is using Selenium-enriched fermentation culture medium to gained mixture after photosynthetic bacteria culture.
The crops that reduce in planting process are suitable for the grass family class such as rice, wheat work to the method for Cd uptake
The plantation of object.
This technology is more suitable for cadmium serious pollution, i.e., the full crop field below cadmium concentration≤0.9mg/kg in soil.
For irrigation, fertilising, tillage and weeding and prevention and treatment disease pest etc. cultivate management measure in planting process and conventional method is complete
It is exactly the same.
The present invention is generated compared with the existing technology to be had the beneficial effect that.
Selenium is the essential trace element of human and animal, can mitigate the toxicity of the various heavies such as cadmium, arsenic, mercury, and
It and is the activated centre of anti-oxidant selenium enzyme (glutathione peroxidase and thioredoxin reductase), richness of the present invention
Selenium photosynthetic bacteria enhances light after mainly passing through selenium-rich for being better than common photosynthetic bacteria to the significant effect for dropping cadmium in soil
The physiological activity for closing the desulfurase that bacterial metabolism goes out, makes the heavy metal cadmium in soil generate sulfide-cadmium precipitation with this condition, from
And the heavy metal cadmium in soil of degrading, prevent the absorption of plant pair cadmium.
The present invention filters out resistance to selenium combined bacteria strain, produces a kind of photosynthetic bacteria suspension of selenium-rich by selenium-rich culturing, is applied
In acidic cadmium polluted soil, the available Cd in soil can be reduced, soil pH is promoted, contaminated soil is repaired, again
Can prevent absorption and enrichment of the plant to cadmium, while improve the Se content in plant, realize cadmium pollution soil in crops or its
The selenium-rich of its industrial crops is planted safely.
Specific embodiment
In order to which technical problems, technical solutions and advantages to be solved are more clearly understood, in conjunction with reality
Example is applied, the present invention will be described in further detail.It should be appreciated that specific embodiment described herein is only used to explain this
Invention, is not intended to limit the present invention.Below with reference to the embodiment technical solution that the present invention will be described in detail, but protection scope not by
This limitation.
Embodiment 1
A kind of preparation method of selenium-rich bacteria preparation, comprising the following steps:
A) resistance to selenium combined bacteria primary dcreening operation: photosynthetic bacteria being inoculated in isolation medium and carries out primary dcreening operation, filters out resistance to selenium combined bacteria；Institute
The isolation medium stated are as follows: DL-malic acid 2g/L, (NH4)2SO4 1.5g/L, yeast extract 2g/L, peptone 5g/L, agar
10g, Na2SeO3120mg/ L adds water to 1L, adjusts pH6.5.The condition of resistance to selenium combined bacteria primary dcreening operation is to be inverted isolation medium to put down
Plate cultivates 3d at 32 DEG C in constant incubator.
B) photosynthetic bacteria Selenium-enriched fermentation: 3 resistance to selenium combined bacteria bacterium colonies filtered out is taken to connect from the isolation medium
Na is added in the culture solution when cultivating 2d in resistance to selenium combined bacteria culture solution in kind2SeO3Aqueous solution makes the culture solution
Selenium concentration is 100mg/L, and 1500lux intensity of illumination is then placed at 28 DEG C, carries out illumination cultivation under 800nm illumination wave band
3d obtains selenium-rich bacteria preparation when the culture solution OD value reaches 1.
The resistance to selenium combined bacteria culture solution is DL-malic acid 2g/L, (NH4)2SO4 1.5g/L, MgSO4 150mg/L,
CaCl2 85mg/L, sodium acetate 15mg/L, yeast powder 2g/L, peptone 2g/L add water to 1L, adjust pH7.0.
Embodiment 2
A) resistance to selenium combined bacteria primary dcreening operation: photosynthetic bacteria being inoculated in isolation medium and carries out primary dcreening operation, filters out resistance to selenium combined bacteria；Institute
The isolation medium stated are as follows: DL-malic acid 5g/L, (NH4)2SO4 0.5g/L, yeast extract 5g/L, peptone 2g/L, agar
20g, Na2SeO3100mg/ L adds water to 1L, adjusts pH7.The condition of resistance to selenium combined bacteria primary dcreening operation is to be inverted isolation medium plate,
2d is cultivated at 30 DEG C in constant incubator.
B) secondary fermentation and secondary screening separation；The bacterium colony that resistance to selenium combined bacteria primary dcreening operation is obtained is in second order fermentation culture solution in training
When supporting 1d, Na is added in the second order fermentation culture solution2SeO3Aqueous solution keeps the selenium of the second order fermentation culture solution dense
Degree is 100mg/L, cultivates 3d at 32 DEG C.
The second order fermentation culture solution is DL-malic acid 5g/L, sodium acetate 30mg/L, yeast extract 2g/L, peptone
5g/L adds water to 1L, adjusts pH7.0.
The bacterium colony turned out in secondary fermentation culture solution is taken to carry out secondary screening separation according to the method for step a.
C) 5 resistance to selenium combined bacteria bacterium filtered out photosynthetic bacteria Selenium-enriched fermentation: are taken from the secondary screening isolation medium
It falls and is seeded in resistance to selenium combined bacteria culture solution, Na is added in the culture solution when cultivating 3d2SeO3Aqueous solution makes the culture
The selenium concentration of liquid is 50mg/L, and 1200lux intensity of illumination is then placed at 32 DEG C, carries out illumination training under 400nm illumination wave band
It supports 10d and obtains selenium-rich bacteria preparation when the culture solution OD value reaches 2.
The resistance to selenium combined bacteria culture solution is DL-malic acid 5g/L, (NH4)2SO4 1g/L, MgSO4 100mg/L,
CaCl2 60mg/L, sodium acetate 30mg/L, yeast powder 3g/L, peptone 4g/L add water to 1L, adjust pH6.5.
Embodiment 3
It is applied in rice field using selenium-rich bacteria preparation prepared by embodiment 1: the nursery sprinkling irrigation method of use are as follows: educate
--- --- --- transplanting --- pouring root --- tillering stage top dressing simultaneously sprays the selenium-rich bacterium system to root dipping to basal dressing to seedling for the first time
Agent --- jointing stage sprays selenium-rich bacteria preparation for the second time --- pustulation period third time sprays selenium-rich bacteria preparation.
Nursery: rice transplanting is educated using disk.
Basal dressing: transplanting a few days ago basal dressing, 20% compound fertilizer 50kg/ mu, lime 90kg/ mus of addition.
Root dipping: being watered dilution for selenium-rich bacteria preparation and be made bacteria preparation dilution, and the weight ratio of selenium-rich bacteria preparation and water is 1:
100, root dipping processing is carried out to seedling.
Transplanting: root dipping moves back sanction, and planting density is 13340 root and stem of certain plants/mu or so.
Pouring root: carrying out root irrigation for above-mentioned bacteria preparation dilution, and the usage amount of pouring root is 28t/ mus, is guaranteed biological prosthetic
The soil horizon in 25cm that agent covering heavy metal cadmium easily accumulates.
Spraying fertilizer simultaneously sprays for the first time: spraying fertilizer after turning green, every 667m2Spraying fertilizer essence 12.5kg, potash fertilizer 5kg are sprayed dilute
Bacteria preparation dilution after releasing, dosage: 60L/ mus.
Jointing stage sprays selenium-rich bacteria preparation for the second time, dosage: 30~60L/ mus.
Pustulation period third time sprays selenium-rich bacteria preparation, dosage: 30~60L/ mus.
Harvest: after the rice harves of above-mentioned plantation, the cadmium in rice is measured, final result is obtained.
Cadmium effect drops in rice: in treated rice, cadmium decreases by 33%, lower than in GB2762-2012 about rice cadmium
The limit value (0.20mg/kg) of content.
Embodiment 4
The selenium-rich bacteria preparation prepared using embodiment 1 is poured on the experimental plot of plantation wheat, specially by selenium-rich bacterium system
Agent add water according to volume ratio be 5:100 dilute, be prepared into bacteria preparation dilution, soil poured according to 20t/ mus.
Soil remediation effect: having sprayed the soil of selenium-rich bacteria preparation, and pH and cadmium content change significantly, wherein soil
PH improve 0.41, cadmium fall is respectively 35.29%.
In conclusion, using selenium-rich bacteria preparation, being imitated to rice with good volume increase in cadmium pollution Acid Paddy soil
Fruit and economic benefit, while can reduce accumulation of the cadmium in rice, soil pH is promoted, containing for soil available Cd is reduced
Amount, it is proposed that be widely applied.
The above content is combine specific preferred embodiment to the further description done of the present invention, and it cannot be said that
A specific embodiment of the invention is only limitted to this, for those of ordinary skill in the art to which the present invention belongs, is not taking off
Under the premise of from the present invention, several simple deduction or replace can also be made, all shall be regarded as belonging to the present invention by being submitted
Claims determine scope of patent protection.
Claims (6)
1. a kind of preparation method of selenium-rich bacteria preparation, which comprises the following steps:
A) resistance to selenium combined bacteria primary dcreening operation: photosynthetic bacteria being inoculated in isolation medium and carries out primary dcreening operation, filters out resistance to selenium combined bacteria；Institute
The isolation medium stated are as follows: DL-malic acid 2~5g/L, (NH4)2SO4 0.5~1.5g/L, 2~5g/L of yeast extract, peptone
2~5g/L, agar 10~20g, Na2SeO340~120mg/ L adds water to 1L, adjusts pH6.5~7.0；
B) the 3-5 resistance to selenium combined bacteria colony inoculations filtered out photosynthetic bacteria Selenium-enriched fermentation: are taken from the isolation medium
In resistance to selenium combined bacteria culture solution, Na is added in the culture solution when cultivating 2-3d2SeO3Aqueous solution makes the culture solution
Selenium concentration is 1-100mg/L, and 1200~1500lux intensity of illumination, 400-800nm illumination wave band are then placed at 28-32 DEG C
Lower progress illumination cultivation 3-10d obtains selenium-rich bacteria preparation when the culture solution OD value reaches 1-2；
The resistance to selenium combined bacteria culture solution is 2~5g/L of DL-malic acid, (NH4)2SO4 0.5~1.5g/L, MgSO4 100~
150mg/L, CaCl2 60~85mg/L, 15~30mg/L of sodium acetate, 2~5g/L of yeast powder, 2~5g/L of peptone are added water to
1L adjusts pH6.5~7.0.
2. a kind of preparation method of selenium-rich bacteria preparation according to claim 1, it is characterised in that: resistance in the step a
The condition of selenium combined bacteria primary dcreening operation is to be inverted isolation medium plate, cultivates 2-3d at 28-32 DEG C in constant incubator.
3. a kind of preparation method of selenium-rich bacteria preparation according to claim 1, it is characterised in that: closed in the resistance to selenium
Secondary fermentation and secondary screening separation are successively carried out after bacterium primary dcreening operation step；Secondary screening resistance to selenium combined bacteria bacterium colony obtained after separation is used for institute
The photosynthetic bacteria Selenium-enriched fermentation stated；The secondary fermentation is the bacterium colony that obtains resistance to selenium combined bacteria primary dcreening operation in second order fermentation culture
In liquid when cultivating 0-1d, Na is added in the second order fermentation culture solution2SeO3Aqueous solution trains the second order fermentation
The selenium concentration of nutrient solution is 1-100mg/L, cultivates 2-3d at 28-32 DEG C；
The second order fermentation culture solution be 2~5g/L of DL-malic acid, 15~30mg/L of sodium acetate, 2~5g/L of yeast extract,
2~5g/L of peptone adds water to 1L, adjusts pH6.5~7.0.
4. a kind of preparation method of selenium-rich bacteria preparation according to claim 1, it is characterised in that: the photosynthetic bacteria is deep
The red bacterium of red spirillum, fixed nitrogen, red pseudomonas, Spherical red antibacterial and one of which of Rhodobacter capsulatus or any combination thereof.
5. a kind of method for heavy metal in soil cadmium of degrading, which comprises the following steps: add water to press selenium-rich bacteria preparation
It is 1-5:100 dilution according to volume ratio, is prepared into bacteria preparation dilution, soil is poured according to 18-28t/ mus；The richness
Selenium bacteria preparation is using Selenium-enriched fermentation culture medium to gained mixture after photosynthetic bacteria culture.
6. a kind of prevent gramineae farm crop from existing the method for Cd uptake, including nursery, basal dressing, feature in planting process
In further comprising the steps of:
A) root dipping: by selenium-rich bacteria preparation add water according to volume ratio be 1-5:100 dilute, bacteria preparation dilution is prepared into, to farming
Object seedling carries out root dipping processing, is transplanted later；
B) bacteria preparation dilution pouring root: is subjected to root irrigation to the seedling after transplanting；
C) it spraying fertilizer and sprays for the first time: spraying fertilizer after turning green, and spray bacteria preparation dilution；
D) in the entire growth course of crops, then the bacteria preparation dilution is sprayed several times；
The selenium-rich bacteria preparation is using Selenium-enriched fermentation culture medium to gained mixture after photosynthetic bacteria culture.
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| CN114317373A (en) * | 2022-01-14 | 2022-04-12 | 湘潭大学 | A kind of Sphingomonas PAH02, microbial preparation and its application as functional conditioner for reducing cadmium and enriching selenium in crops |
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| CN111518564A (en) * | 2020-06-24 | 2020-08-11 | 代彦霞 | Material for reducing migration-state lead ions in soil and preparation method thereof |
| CN112872002A (en) * | 2021-02-02 | 2021-06-01 | 华中农业大学 | Application of selenium combined pseudomonas aeruginosa in strengthening phytoremediation of cadmium-nonylphenol composite contaminated soil |
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