CN101735997B - Microorganism bactericide capable of reinforcing plants to restore fields polluted by arsenic and preparation and application methods thereof - Google Patents
Microorganism bactericide capable of reinforcing plants to restore fields polluted by arsenic and preparation and application methods thereof Download PDFInfo
- Publication number
- CN101735997B CN101735997B CN200910242389XA CN200910242389A CN101735997B CN 101735997 B CN101735997 B CN 101735997B CN 200910242389X A CN200910242389X A CN 200910242389XA CN 200910242389 A CN200910242389 A CN 200910242389A CN 101735997 B CN101735997 B CN 101735997B
- Authority
- CN
- China
- Prior art keywords
- arsenic
- fermentation
- plants
- restore
- reinforcing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Processing Of Solid Wastes (AREA)
Abstract
The invention provides a microorganism bactericide capable of reinforcing plants to restore fields polluted by arsenic and preparation and application methods thereof, belonging to the technical field of environment restoration engineering. The strain utilized by the bactericide is Agrobacteriumsp. C13. The bactericide adopts a composite carrier which comprises turfy soil, saw dust, rice hulls and active carbon with the mass percentage being 30:25-30:25-30:15; the microorganism fermentation liquor is mixed with the composite carrier according to the mass ratio of 0.5-1:1; and the adsorption amount of the microorganisms of the composite carrier reaches 109-1012 per gram. The prepared bactericide is applied to the arsenic-polluted fields restored by ciliate desert-grass according to the useamount of 5g/kg, the biomass liveweight of the overground part of the restored plants is improved by 16-17%, and the removal rate of arsenic in the fields is improved by 40-41% in comparison. The carrier of the bactericide in the invention has rich raw materials, simple preparation method and easy realization of large-scale production.
Description
Technical field
The present invention relates to a kind of high-effective microorganism microbial inoculum and methods for making and using same, concrete is a kind of microbiobacterial agent that can promote arsenic super enriching plant-Herba pteridis vittatae absorption arsenic in soil, belongs to the environment remediation field of engineering technology.
Background technology
Character such as that the compound of arsenic has is carcinogenic, teratogenesis, mutagenesis.Arseniasis is the systemic disease based on skin, the skin that it can harmful to human, breathing, digestion, uropoiesis, cardiovascular, neural, hemopoietic system etc.Because the phreatic exploitation of arsenic contamination, the use that contains the arsenic agriculture chemical, Metal smelting and mining cause a large amount of soils of China serious arsenic contamination to occur.Arsenic in the edatope enters human body by food, tap water and breathing, and surpassing limits the quantity of can cause human body arseniasis.Arsenic contamination and arseniasis incident all appearred in ground such as China Hunan, Guangxi, Yunnan in recent years, and the arsenic in soil pollution problem has become one of important environmental problem of China.
At present, the research hot issue that has become various countries is repaired in the arsenic contamination place.The phytoremediation in arsenic contamination place is to administer one of arsenic contamination effective technology means.Arsenic super enriching plant-Herba pteridis vittatae that the research report is found is subjected to paying close attention to widely with great biomass with extremely strong rich arsenic characteristic.Chen Tongbins in 2003 etc. disclose a kind of method (CN1397390A) that arsenic in soil pollutes of administering, and this method absorbs a large amount of arsenic and upwards carries and be transported to overground part by the Herba pteridis vittatae root.Old Bao Dong in 2005 etc. disclose a kind of method (CN1633834A) that improves As polluted soil phytoremediation efficient, and this method is to pteridophyte Herba pteridis vittatae inoculation bush mycorrhizal fungi, makes its Mycorrhizal.The brave grade of Liao Xiao in 2007 discloses the method (CN1981947A) of utilizing the phytoremediation As polluted soil, and this method includes and grows seedlings, planting density and cradle the complete set technology of combination, can improve the remediation efficiency of the reparation As polluted soil of Herba pteridis vittatae.Through effort for many years, the soil that utilizes Herba pteridis vittatae to repair arsenic contamination has been obtained success (Liao Xiaoyong etc., ACTA Scientiae Circumstantiae, 2004) in practice.At present, how further to improve the Herba pteridis vittatae remediation efficiency, expanding the phytoremediation technology application power is current domestic and international research focus.
Summary of the invention
Technology of the present invention is dealt with problems: the deficiency that overcomes existing phytoremediation technology, a kind of microbiobacterial agent and methods for making and using same thereof of reinforcing plants to restore fields polluted by arsenic are provided, can improve the Herba pteridis vittatae remediation efficiency greatly, expand the phytoremediation technology application power.
Technical solution of the present invention: a kind of microbiobacterial agent of reinforcing plants to restore fields polluted by arsenic, described microbiobacterial agent is mixed with forms by mass percentage for the bacterium liquid of Agrobacterium Agrobacteriumsp.C13 fermentation and complex carrier at 0.5～1: 1; Described complex carrier is on the peat composed of rotten mosses, rice husk, saw art and gac, and their fineness are 60 orders, mix in 30: 25 by mass percentage～30: 25～30: 15.The microorganism adsorptive capacity of complex carrier reaches 10
9～10
12Individual/gram.
Described Agrobacterium Agrobacteriumsp.C13 zymocyte liquid is the 18～24h that ferments in the Gause I substratum, fermentation culture is through primary inclined plane culture medium culturing, secondary triangular flask liquid culture, the fermentation of three grade fermemtation jar, and the bacterium number is 10 in the bacterium liquid of fermentation back
9～10
12Individual/mL.
A kind of preparation method of microbiobacterial agent of reinforcing plants to restore fields polluted by arsenic, step is as follows:
(1) strain fermentation
A. primary inclined plane substratum
Peptone 0.5%, extractum carnis 0.3%, sodium-chlor 0.5%, agar 1.5%, pH7.0～7.2 are with the substratum test tube of packing into; Aseptic; Agrobacterium Agrobacteriumsp.C13 is inoculated; Cultivating 18-24h down at 28 ℃ preserves standby;
B. secondary triangular flask liquid culture
Remove agar in the primary inclined plane substratum major ingredient in step a, cultivate major ingredient as secondary, pH7.0～7.2 with the substratum triangular flask of packing into, after the sterilization cooling, are seeded to liquid nutrient medium to inclined-plane bacterium liquid; At 28 ℃ of following shaking culture 18-24h, stop cultivating, standby;
C. three grade fermemtation jar fermentation
It is identical with secondary cultivation major ingredient to cultivate major ingredient in the fermentor tank, the secondary nutrient solution is inserted in the fermentor tank by 1～5% inoculum size, carry out fermentation culture, condition is: jar 30～35 ℃ of temperature, tank pressure 0.02～0.05MPa air flow 1: 1v/v/min, stirring velocity 300～350 commentaries on classics/min, fermentation time 24～36h, and the fermented liquid of producing is standby;
(2) with turfy soil, rice husk, sawdust, gac, pulverized 60 orders with high speed disintegrator, mixed according to 30: 25 by mass percentage～30: 25～30: 15 then;
(3) with the fermented liquid that obtains in the step (1) and the complex carrier in the step (2) 0.5～1: 1 ratio uniform mixing by mass percentage.
A kind of application method of microbiobacterial agent of reinforcing plants to restore fields polluted by arsenic, with prepare microbiobacterial agent be applied to by 5g/kg soil consumption and use in the arsenic contamination place that Herba pteridis vittatae repairs.
The present invention's advantage compared with prior art:
(1) the present invention is a carrier with turfy soil, sawdust, rice husk, 4 kinds of materials of gac, they all have advantages such as wide material sources, with low cost, easy to use, preservation term are long, the preparation method is simple, is easy to realize scale operation, thereby can be by wide popularization and application.
(2) the present invention has improved the quality of microbiobacterial agent, the complex carrier that screens is compared the living bacteria count showed increased with existing single microbial inoculum carrier.
(3) growth, fecundity and the adaptability selected for use of the present invention is stronger, and can guarantee that in the arsenic contamination environment Herba pteridis vittatae being inhaled the arsenic ability has bigger promoter action.
(4) the present invention is applied to the microbial inoculum for preparing in the arsenic contamination place of using the Herba pteridis vittatae reparation by 5g/kg soil consumption, and rehabilitation plant the upperground part biomass has improved 16-17%, and the comparison of the clearance of arsenic is according to having improved 40-41% in the place.
(5) the invention solves all inconvenient problem of liquid microbial inoculum storage and transport, microbial preparation is made solid fungicide, so that engineering is used.
Description of drawings
Fig. 1 be the different carriers bacterial content over time;
Fig. 2 is Herba pteridis vittatae the upperground part biomass;
Fig. 3 is the removal amount of arsenic in the soil.
Embodiment
Below by embodiment the present invention is described in further details
(1) strain fermentation:
A. primary inclined plane is cultivated
Peptone 0.5%, extractum carnis 0.3%, sodium-chlor 0.5%, agar 1.5%, pH are 7.2, with the substratum test tube of packing into; Aseptic; Agrobacterium Agrobacteriumsp.C13 is inoculated; Cultivating 24h down at 28 ℃ preserves standby;
B. secondary triangular flask liquid culture
Remove agar in the primary inclined plane substratum major ingredient in step a, cultivate major ingredient as secondary, pH is 7.2, with the substratum triangular flask of packing into, after the sterilization cooling, inclined-plane bacterium liquid is seeded to liquid nutrient medium; At 28 ℃ of following shaking culture 24h, stop cultivating, standby;
C. three grade fermemtation jar fermentation
It is identical with secondary cultivation major ingredient to cultivate major ingredient in the fermentor tank, the secondary nutrient solution is inserted in the fermentor tank by 1～5% inoculum size, carry out fermentation culture, condition is: jar temperature 30 ℃, tank pressure 0.03MPa air flow 1: 1v/v/mi n, stirring velocity 300 commentaries on classics/min, fermentation time 36h, and the fermented liquid of producing is standby;
(2) support material is obtained: turfy soil, rice husk, gac are available from flowers market, Beijing, and sawdust is taken from Fangshan District, Beijing.
(3) the different composite carrier is relatively: turfy soil, rice husk, sawdust, gac, pulverized 60 orders with high speed disintegrator.According to the described mixed of table 1, it is standby to sterilize.With fermented liquid and each ratio complex carrier 0.5 ~ 1: 1 ratio uniform mixing by mass percentage.
In different time sections, detect different proportioning complex carrier living bacteria count amounts.The result as shown in Figure 1,5 kinds of overall Changing Patterns of complex carrier bacterial content are consistent, promptly all show in time to reduce afterwards earlier to increase.The reason that produces this phenomenon be because since microorganism from liquid environment to the solid environment, microorganism is in the cessation of growth cessation phase, causes that the bacterium number descends rapidly in two days that connect behind the bacterium.Breeding rapidly after microorganism adapts to the carrier environment gradually, bacterial content sharply increases, and at this moment microorganism is in fast growing period, and growth regulation tended towards stability in six days.Complex carrier 4 and 5 living bacteria count are maximum, and effect is best.The mass percent that is turfy soil in the carrier, sawdust, rice husk, gac is 30: 25 ~ 30: 25 ~ 30: 15th, optimum proportion.
The proportioning of table 1 complex carrier
(4) fungicide preparation: the optimum proportion carrier is proportionally mixed with fermented liquid, and aseptic condition leeward solid carbon dioxide branch remains on about 30%, is not as the criterion with loose the caking, and packing is standby.
Microbial inoculum with method for preparing carries out the indoor pot experiment.Effective viable bacteria amount 10 in the microbial inoculum
9～10
12Individual/gram.
Specific implementation method is: apply the 5g microbial inoculum in the high arsenic soil of 1kg, after mixing, in the basin of packing into, transplant the Herba pteridis vittatae seedling.Plantation is gathered in the crops after two months.
As can be seen from Figure 2, apply microbial inoculum after Herba pteridis vittatae the upperground part biomass dry weight reach 2.1g, comparison is according to improving 16%.
From as Fig. 3 as can be seen Herba pteridis vittatae overground part arsenic enriching quantity reach 3.1mg, comparison is according to having improved 41%.
From above experimental result as can be seen, apply the microbial inoculum of anti-arsenic, obviously promoted the growth of Herba pteridis vittatae, and the also obviously increase of the amount of Herba pteridis vittatae overground part enrichment arsenic, the phytoremediation efficient of As polluted soil obviously improves.
Claims (4)
1. the microbiobacterial agent of a reinforcing plants to restore fields polluted by arsenic is characterized in that: described microbiobacterial agent is that the bacterium liquid of Agrobacterium (Agrobacterium sp.) C13 fermentation and complex carrier are mixed with form by mass percentage at 0.5～1: 1; Described complex carrier is turfy soil, rice husk, sawdust and gac, and their fineness are 60 orders, mixes in 30: 25 by mass percentage～30: 25～30: 15.
2. the microbiobacterial agent of reinforcing plants to restore fields polluted by arsenic according to claim 1, it is characterized in that: described Agrobacterium (Agrobacterium sp.) C13 beef-protein medium fermentation culture, fermentation culture is through primary inclined plane culture medium culturing, secondary triangular flask liquid culture, the fermentation of three grade fermemtation jar, and the bacterium number is 10 in the bacterium liquid of fermentation back
9～10
12Individual/mL.
3. the preparation method of the microbiobacterial agent of a reinforcing plants to restore fields polluted by arsenic is characterized in that step is as follows:
(1) strain fermentation
A. primary inclined plane substratum major ingredient prescription is
Peptone 0.5%, extractum carnis 0.3%, sodium-chlor 0.5%, agar 1.5%, pH7.0～7.2 are with the substratum test tube of packing into; Aseptic; Agrobacterium (Agrobacterium sp.) C13 is inoculated; Cultivating 18-24h down at 28 ℃ preserves standby;
B. secondary triangular flask liquid culture
Remove agar in the primary inclined plane substratum major ingredient in step a, cultivate major ingredient as secondary, pH7.0～7.2 with the substratum triangular flask of packing into, after the sterilization cooling, are seeded to liquid nutrient medium to inclined-plane bacterium liquid; At 28 ℃ of following shaking culture 18-24h, stop cultivating, standby;
C. three grade fermemtation jar fermentation
It is identical with secondary cultivation major ingredient to cultivate major ingredient in the fermentor tank, the secondary nutrient solution is inserted in the fermentor tank by 1～5% inoculum size, carry out fermentation culture, condition is: jar 30～35 ℃ of temperature, tank pressure 0.02～0.05MPa air flow 1: 1v/v/min, stirring velocity 300～350 commentaries on classics/min, fermentation time 24～36h, and the fermented liquid of producing is standby;
(2) with turfy soil, rice husk, sawdust, gac, pulverized 60 orders with high speed disintegrator, mixed according to 30: 25 by mass percentage～30: 25～30: 15 then;
(3) with the fermented liquid that obtains in the step (1) and the complex carrier in the step (2) 0.5～1: 1 ratio uniform mixing by mass percentage.
4. the application method of the microbiobacterial agent of a reinforcing plants to restore fields polluted by arsenic is characterized in that: the described microbiobacterial agent of the claim 1 for preparing is applied in the arsenic contamination place of using the Herba pteridis vittatae reparation by 5g/kg soil consumption.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910242389XA CN101735997B (en) | 2009-12-10 | 2009-12-10 | Microorganism bactericide capable of reinforcing plants to restore fields polluted by arsenic and preparation and application methods thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910242389XA CN101735997B (en) | 2009-12-10 | 2009-12-10 | Microorganism bactericide capable of reinforcing plants to restore fields polluted by arsenic and preparation and application methods thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101735997A CN101735997A (en) | 2010-06-16 |
| CN101735997B true CN101735997B (en) | 2011-12-14 |
Family
ID=42460027
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910242389XA Expired - Fee Related CN101735997B (en) | 2009-12-10 | 2009-12-10 | Microorganism bactericide capable of reinforcing plants to restore fields polluted by arsenic and preparation and application methods thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101735997B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104004691B (en) * | 2014-06-16 | 2016-08-24 | 中南大学 | A kind of bacterial strain repaired for As polluted soil and application process thereof |
| CN104096710B (en) * | 2014-07-02 | 2016-03-30 | 西南科技大学 | A kind of method of cultivating purple striae plumage volume load bacterium mycoderma original position removal heavy metal in soil |
| CN105777231A (en) * | 2016-03-17 | 2016-07-20 | 李军 | Method of utilizing mushroom bran to make organic mushroom fertilizer |
| CN106807738A (en) * | 2016-12-23 | 2017-06-09 | 镇江华域环保设备制造有限公司 | A kind of in-situ remediation method of organic material contaminated soil |
| CN106995704A (en) * | 2017-05-05 | 2017-08-01 | 千人计划土壤治理研究院（唐山）有限公司 | A kind of renovation agent, its preparation method and the method with its restoration of soil polluted by heavy metal |
| CN107217147B (en) * | 2017-05-23 | 2018-11-02 | 覃淑兰 | A method of removing and absorption ciliate desert-grass heavy metal |
| CN108672492B (en) * | 2018-06-22 | 2020-09-11 | 中国石油大学(华东) | Construction method of organic fertilizer nutrition system for biostimulation remediation of petroleum-contaminated soil |
| CN110449451B (en) * | 2019-01-18 | 2021-12-28 | 湖南高岭环保科技有限公司 | Device and method for repairing mine soil polluted by arsenic |
| CN112063385B (en) * | 2020-09-16 | 2021-04-30 | 南京大学 | Passivating agent for efficiently fixing trivalent arsenic under anaerobic condition, and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1397390A (en) * | 2001-07-18 | 2003-02-19 | 中国科学院地理科学与资源研究所 | Method for treating As polluted soil |
| CN1633834A (en) * | 2003-12-26 | 2005-07-06 | 中国科学院生态环境研究中心 | Method for promoting plant restoration efficiency of arsenic polluted soil |
| CN1981947A (en) * | 2005-12-15 | 2007-06-20 | 中国科学院地理科学与资源研究所 | Method for repairing soil polluted by arsenic by plant |
-
2009
- 2009-12-10 CN CN200910242389XA patent/CN101735997B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1397390A (en) * | 2001-07-18 | 2003-02-19 | 中国科学院地理科学与资源研究所 | Method for treating As polluted soil |
| CN1633834A (en) * | 2003-12-26 | 2005-07-06 | 中国科学院生态环境研究中心 | Method for promoting plant restoration efficiency of arsenic polluted soil |
| CN1981947A (en) * | 2005-12-15 | 2007-06-20 | 中国科学院地理科学与资源研究所 | Method for repairing soil polluted by arsenic by plant |
Non-Patent Citations (1)
| Title |
|---|
| 涂书新.我国生物修复技术的现状与展望.《地理科学进展》.2004,第23卷(第6期),全文. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101735997A (en) | 2010-06-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101735997B (en) | Microorganism bactericide capable of reinforcing plants to restore fields polluted by arsenic and preparation and application methods thereof | |
| CN104450597B (en) | Preparation method of petroleum degrading bacteria solid microbial inoculum and method for repairing petroleum-polluted soil by using solid microbial inoculum prepared by preparation method | |
| CN105695354B (en) | The technique and application of superhigh temperature aerobic composting fermentation processing municipal sludge | |
| CN103952349B (en) | A Shi genus bacillus and microbiobacterial agent and their application | |
| CN102747002B (en) | Agrobacteriumsp. having free-living nitrogen fixing ability, and applications thereof | |
| CN102409014B (en) | Ziziphus jujube growth-promoting rhizobacteria Bacillus subtilis and its uses | |
| CN102513340A (en) | Method for repairing soils polluted by heavy metal Cd | |
| CN106180175B (en) | A method of plantation rape restoration of soil polluted by heavy metal | |
| CN101153273B (en) | Copper activation bacterium and method for plants renovation of soil pollution by heavy metal | |
| CN105290103A (en) | Method for utilizing cadmium-resisting fungus aspergillus aculeatus for promoting festuca arundinacea to remedy cadmium contaminated soil | |
| CN112573974A (en) | Biological salt-decomposing bacterial fertilizer and preparation method thereof | |
| CN104707864A (en) | Compound photosynthetic bacteria preparation for enhancing phytoremediation for heavy metal pollution of soil and preparation method thereof | |
| CN109593686A (en) | A kind of Shewanella and its application in control vegetable field soil nitrogen pollution | |
| CN101695711A (en) | Chemical fixing material suitable for arsenic in soil and application thereof | |
| CN102408259B (en) | Solid waste co-composting stepwise regulation and control system and method | |
| CN103551379A (en) | Method for carrying out ecological remediation in cadmium-contaminated soil by using cadmium resistant fungi | |
| CN109536410B (en) | Salt-tolerant growth-promoting composite microbial inoculum and preparation method and application thereof | |
| CN105505843B (en) | One plant of Photosynthetic bacterium strain, the Liquid Fertilizer containing the bacterial strain and preparation method, application | |
| CN112854250B (en) | Method for solidifying road slope soil by combining microorganisms and plants | |
| CN103650977B (en) | Method for improving survival rate of afforestation of non-woven fabric container seedlings in arid mountains | |
| CN109468343A (en) | A kind of stalk anaerobic fermentation produces methane accelerator and its preparation method and application | |
| CN114591111A (en) | Composite functional organic bacterial fertilizer and preparation method and application thereof | |
| CN111172073B (en) | Bacillus subtilis strain and application thereof in plant growth | |
| CN110683870A (en) | Preparation method and application of biofertilizer prepared from kelp residues and bacillus | |
| CN104496724B (en) | Organic fungi-manure for manganese ore contaminated soil remediation and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20100616 Assignee: Beijing CAS Jiuzhou Environmental Engineering Ltd. Assignor: Institute of Geographic Sciences and Natural Resources Research, CAS Contract record no.: 2013110000015 Denomination of invention: Microorganism bactericide capable of reinforcing plants to restore fields polluted by arsenic and preparation and application methods thereof Granted publication date: 20111214 License type: Exclusive License Record date: 20130416 |
|
| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20111214 Termination date: 20211210 |