CN102321539B - Fermentation and preparation of composite microbial agent for degrading sulfonylurea herbicide with high efficiency - Google Patents
Fermentation and preparation of composite microbial agent for degrading sulfonylurea herbicide with high efficiency Download PDFInfo
- Publication number
- CN102321539B CN102321539B CN 201110247715 CN201110247715A CN102321539B CN 102321539 B CN102321539 B CN 102321539B CN 201110247715 CN201110247715 CN 201110247715 CN 201110247715 A CN201110247715 A CN 201110247715A CN 102321539 B CN102321539 B CN 102321539B
- Authority
- CN
- China
- Prior art keywords
- bacterium
- bacillus
- gram
- ratio
- micro organism
- 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
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a fermentation and preparation method of a composite microbial agent for degrading sulfonylurea herbicides in soil and water with high efficiency, and the composite microbial agent is fermented compositely by using bacillus benzoevorans ACCC 10244 and bacillus chitinivorous ACCC 10227 as bacterial sources. After fermentation, a carrier substance of alginic acid (in a form of micro-colloid in aqueous solutions), a electrolyte (potassium nitrate) and an antifreeze agent (glycerin) are added into the fermented bacterial liquid; the bacterial number ratio of bacillus benzoevorans ACCC 10244 and bacillus chitinivorous ACCC 10227 in the prepared bacterial agent is 1:1, and the bacterial number contents of both bacteria in the bacterial agent are up to above 1 billion/ml. The bacterial agent can degrade the followings with high efficiency: nicosulfuron, suifometuron-methyl, monosulfuron, bensulfuron-methyl, thifensulfuron-methyl, tribenuron-methyl, metsulfuron-methyl, chlorsulfuron, methyl sulfuron, etc.
Description
Technical field
The present invention relates to a kind of fermentation and preparation of complex micro organism fungicide, this microbial inoculum can efficient degradation soil and water body in sulfonylurea herbicide,, mesosulfuron grand such as: nicosulfuron, sulfometuronmethyl, monosulfmeturon, benbbensulfuronmethyl, thifensulfuronmethyl, tribenuron-methyl, metsulfuronmethyl, green sulphur etc.
Technical background
Sulfonylurea herbicide is a present in the world class weedicide of usage quantity maximum, it is to be studied successfully in 20 century 70s by du pont company, and it is grand to develop first cornfield herbicidal chlorine sulphur in nineteen eighty-two, develops again afterwards many efficient, wide spectrums, low toxicity, highly selective sulfonylurea herbicide.The sulfonylurea herbicide kind of the present widespread use of China mainly contains pyrazosulfuronmethyl, nicosulfuron, ethametsulfuron, tribenuron-methyl, chlorimuronethyl, benbbensulfuronmethyl etc., can prevent and kill off most of Gramineae and broadleaf weeds.
Sulfonylurea herbicide is since coming out, for huge contribution has been made in the agriculture production in the world, because it has high residual activity and selectivity, some difficult problems in its use procedure, have been run into, the long residual drug injury problem that the most outstanding is exactly to succession crop, since the migration leaching of such weedicide a little less than, will cause poisoning when it is residual to succession crop after reaching doses.The situation of herbicide residue amount accumulation has also appearred in the field of the long residual effect weedicide of some life-time service, has a strong impact on the growth of succession crop, has formed the phenomenon in " cancer field ".
Studies show that in a large number the multiple-microorganism that exists in the physical environment plays an important role aspect degradation of pesticide, researcher has been found many microorganisms that can degrading pesticide by technology such as enrichment culture, separation screenings.Studies show that now the microorganism of degrading pesticide mainly is bacterium, fungi, actinomycetes and algae, wherein bacterium is owing to easily bringing out sudden change and adaptable occupying the majority.Microorganism is owing to its special adaptive faculty, and in general the same bacterium may have Degradation to Multiple Pesticides, and a certain agricultural chemicals often has multiple degradation bacteria simultaneously simultaneously.
Summary of the invention
The present invention conciliates chitin genus bacillus ACCC10227 with bacillus benzoevorans ACCC10244, and (above bacterial classification all derives from Chinese agriculture microbial strains preservation administrative center, see " Chinese agriculture bacterial classification catalogue " China, Chinese agriculture microbial strains preservation administrative center compiles, version in 2005, Scientia Agricultura Sinica technology press) produce respectively fermentation for the bacterium source, it is 2.0 * 10 that fermentation obtains living bacteria count
9The bacterium liquid of individual/ml is that 1: 1 ratio is mixed with composite fungus agent with the latter two according to volume ratio.It is better to the degradation effect of sulfonylurea herbicide that this composite fungus agent is compared single strain fermentation product, and the two has synergy.
Preferably, above-mentioned composite fungus agent is carried out complex ferment again, in the fermented liquid after the fermentation bacterium of the two to count when content all reaches hundred million/ml of 15-20 be qualified bacterium liquid, add subsequently carrier substance Lalgine, ionogen and antifreezing agent, described Lalgine form with idiozome in the aqueous solution exists, described ionogen is saltpetre, and described antifreezing agent is glycerine.To conciliate the bacterium ratio of counting of chitin genus bacillus ACCC 10227 be about 1: 1 for bacillus benzoevorans ACCC 10244 in the microbial inoculum for preparing, the two bacterium in microbial inoculum count content all reach 1,000,000,000/more than the ml.
But the present invention relates to kind of the complex micro organism fungicide of efficient degradation sulfonylurea herbicide, (living bacteria count is 2.0 * 10 to two genus bacillus of wherein adopting
9Individual/ml) be 1: 1 o'clock in volume ratio, can give full play to each other Synergistic degradation and the effect that mutually remedies, secrete multiple degraded enzyme, the sulfonylurea herbicide of can effectively degrading, this composite fungus agent can be diluted to 1000-2000 times of liquid when concrete the use, the effect of degraded is still very obvious.
One, the cultural method of complex micro organism fungicide involved in the present invention
1, the preparation of nutrition gravy protein culture medium
Culture medium prescription: peptone-5 gram; The NaCl-5 gram; Extractum carnis-3 gram; Yeast extract paste-3 gram; Oxyenadenine 0.005 gram; Alkene acyl VITAMIN B4 0.005 gram; MnSO
4H
2O 0.005 gram; Nutrient agar medium-15 gram; Add distilled water to 1000 milliliter; PH value 6.57.2.121 ℃ of autoclavings 30 minutes.Taking-up is cooled to about 60 ℃, on super clean bench, under aseptic condition, pour in the culture dish of the bacterium of going out, respectively bacillus benzoevorans ACCC10244 reconciliation chitin genus bacillus ACCC10227 is forwarded in the nutrition gravy protein culture medium and cultivate, in 30 ℃ of incubators, cultivated 72 hours, then carry out biological degradation and measure.
2, biological degradation is measured
Cultured bacillus benzoevorans ACCC10244 is conciliate chitin genus bacillus ACCC10227, and being mixed with respectively living bacteria count is 2.0 * 10
9100 milliliters of the bacterium liquid of individual/ml, and then be that 1: 1 ratio is mixed with the above-mentioned microbial inoculum that composite fungus agent will prepare by volume, be used for respectively measuring the degradation effect of the sulfonylurea herbicides such as it is grand to nicosulfuron, sulfometuronmethyl, monosulfmeturon, benbbensulfuronmethyl, thifensulfuronmethyl, tribenuron-methyl, metsulfuronmethyl, green sulphur, mesosulfuron.(degradation effect sees Table 1).
Table 1. bacillus benzoevorans ACCC10244 and the degradation effect of separating chitin genus bacillus ACCC10227 (adopt transparent bacterium circle method to measure degradation rate, represent with %)
Bacillus benzoevorans ACCC10244 conciliates the bacterium liquid that chitin genus bacillus ACCC10227 is mixed with as can be seen from Table 1, degradation rate is not very high when routine tests, but through after compound, the effect that the two performance is collaborative with complementary, the test effect of common degraded has obtained significant raising.
3, antagonistic effect
Bacillus benzoevorans ACCC10244 is conciliate chitin genus bacillus ACCC10227 on nutrition gravy protein culture medium, 2 placements are at a distance of 4cm, and each makes up triplicate, intersect antagonistic effect, there is not antagonism each other in the result person of showing, can jointly grow.Be that (living bacteria count was 2.0 * 10 in 1: 1 by volume with the two
9Individual/ml) ratio is hybridly prepared into compound microbial inoculum, co-cultivation on nutrition gravy protein culture medium then, and test the person of showing can grow jointly equably.
, complex micro organism fungicide fermentation and preparation
1, the preparation of complex microorganism bacterial classification
Nutrition gravy peptone nutrient solution prescription: peptone-5 gram; The NaCl-5 gram; Extractum carnis-3 gram; Yeast extract paste-3 gram; Oxyenadenine-0.005 gram; Alkene acyl VITAMIN B4-0.005 gram; MnSO
4H
2The O-0.005 gram; Nutrient agar medium-15 gram; Add distilled water to 1000 milliliter; PH value 6.5-7.2.
After by above-mentioned prescription nutrition gravy protein culture medium being prepared, pour in the triangle shaking flask, then 121 ℃ of autoclavings are 30 minutes, after cooling on Bechtop, under the condition of aseptic technique, with preserving the bacterial classification of two bacterial strains of cultured bacillus benzoevorans ACCC10244 reconciliation chitin genus bacillus ACCC10277, be inoculated into respectively in the triangle shaking flask, (living bacteria count all reaches 2.0 * 10 after 72 hours in shaking culture on the shaking table under 30 ℃ of conditions
9Individual/as ml), the bacterium liquid of two bacterial strains to be mixed shaking up with 1: 1 ratio of volume ratio, then the ratio with volume ratio 10 % by weight is inoculated into fermentor tank.
2, the complex ferment of microbiobacterial agent
Fermentor tank nutrient solution prescription: peptone-0.5%; NaCl-0.5%; Extractum carnis-0.3%; Yeast extract paste-0.3%; Oxyenadenine-0.5 ‰; Alkene acyl VITAMIN B4 0.5 ‰; MgS O
4H
2O-0.5 ‰; PH value 6.5-7.2.
At 121 ℃, sterilization is 30 minutes under the 0.12MPa behind the tank body material loading.Ratio in 3 % by weight after being cooled to about 30 ℃ is inoculated: 30 ℃ of culture temperature, air flow begin at 1: 1 (v.v.m), cultivate and increase to 1: 1.5 (v.v.m) after 24 hours, are down to 1: 1.2 (v.v.m) after 48 hours.Fermentation to 72 hour fermentation is complete, conciliate the bacterium of chitin genus bacillus ACC10227 through detecting bacillus benzoevorans ACCC10244 and count ratio about 1: 1, in the fermented liquid bacterium of the two to count when content all reaches hundred million/ml of 15-20 be qualified bacterium liquid.
3, the preparation of complex micro organism fungicide
In order to prolong the quality guaranteed period of microbial inoculum; guarantee activity and the effect of microbial inoculum; in zymocyte liquid, add the colloidalmaterial (Lalgine of 30 % by weight; form with idiozome in the aqueous solution exists; microbial bacteria preferably suspending carrier and Protective substances in liquid preparation; can also prevent effectively that uviolizing is on the impact of microbial inoculum activity); 3 % by weight ionogen (saltpetre); 3 % by weight antifreezing agents (glycerine); to conciliate the bacterium ratio of counting of chitin genus bacillus ACCC10227 be about 1: 1 to bacillus benzoevorans ACCC10244 in the microbial inoculum for preparing, the two bacterium in microbial inoculum count content all reach 1,000,000,000/more than the ml.
In the preparation of the application's complex micro organism fungicide and relevant substratum process for preparation, " % " that occur is " weight percent ".
Embodiment
Complex micro organism fungicide involved in the present invention, can secrete multiple degraded enzyme, can efficient degradation soil and water body in sulfonylurea herbicide, spray or execute with the irrigation water punching after the finished product behind the metering packings such as, mesosulfuron grand such as: nicosulfuron, sulfometuronmethyl, monosulfmeturon, benbbensulfuronmethyl, thifensulfuronmethyl, tribenuron-methyl, metsulfuronmethyl, green sulphur can dilute 1000 times.
Example 1, the microbial inoculum for preparing is diluted to 1000 times, getting 1m l, join 100ml for trying the water in the sample, is 2.3mg/L for the starting point concentration of sulfometuronmethyl in the sample that tries the water, after 72 hours in the water sample residual concentration of sulfometuronmethyl be lower than detection limit, i.e. inspection does not measure residual nicosulfuron.
Example 2, the microbial inoculum for preparing is diluted to 1000 times, getting 10ml, evenly spray and puddle and join 1kg in the upper earth sample of examination, is 2.0mg/kg for the residual quantity of tribenuron-methyl in the examination soil, after 72 hours in the soil sample residual quantity of tribenuron-methyl be lower than detection limit, i.e. inspection does not measure residual tribenuron-methyl.
Example 3, get the microbial inoculum 10kg for preparing, execute the confession examination field of 0.07 hectare with the irrigation water punching, initial residual amount for nicosulfuron in the upper earth of examination field is 1.6mg/kg, residual quantity for nicosulfuron in the soil of examination field after 72 hours is lower than detection limit, i.e. inspection does not measure residual nicosulfuron.
Claims (5)
1. complex micro organism fungicide is characterized in that: described complex micro organism fungicide is conciliate chitin genus bacillus ACCC10227 take bacillus benzoevorans ACCC10244 respectively and is produced as the bacterium source that fermentation obtains, and it is 2.0 * 10 that fermentation obtains living bacteria count
9The bacterium liquid of individual/mL is that 1: 1 ratio is mixed with composite fungus agent with the latter two according to volume ratio.
2. according to claim 1 complex micro organism fungicide, it is characterized in that: described complex micro organism fungicide is carried out complex ferment again, it is qualified bacterium liquid that the bacterium that bacillus benzoevorans ACCC10244 conciliates chitin genus bacillus ACCC10227 in the fermented liquid after the fermentation is counted when content all reaches hundred million/mL of 15-20, add subsequently carrier substance Lalgine, ionogen and antifreezing agent, described Lalgine form with idiozome in the aqueous solution exists, described ionogen is saltpetre, and described antifreezing agent is glycerine; To conciliate the bacterium ratio of counting of chitin genus bacillus ACCC10227 be 1: 1 to bacillus benzoevorans ACCC10244 in the microbial inoculum for preparing, the two bacterium in microbial inoculum count content all reach 1,000,000,000/more than the mL.
3. the application in the sulfonylurea herbicide of the complex micro organism fungicide described in the claim 1 or 2 in degraded soil and water body, wherein said sulfonylurea herbicide are nicosulfuron, sulfometuronmethyl, monosulfmeturon, benbbensulfuronmethyl, thifensulfuronmethyl, tribenuron-methyl, metsulfuronmethyl, green sulphur is grand or mesosulfuron.
4. the preparation method of the complex micro organism fungicide described in the claim 1 is characterized in that: this complex micro organism fungicide adopts following method to make:
(1) preparation nutrition gravy protein culture medium: with peptone 5 grams, NaCl5 gram, extractum carnis 3 grams, yeast extract paste 3 grams, oxyenadenine 0.005 gram, alkene acyl VITAMIN B4 0.005 gram, MnSO
4.H
2O0.005 gram, nutrient agar medium 15 grams mix mutually, add distilled water to 1000 milliliter, and pH value transfers to 6.5-7.2, and 121 ℃ of autoclavings 30 minutes take out and are cooled to about 60 ℃;
(2) on super clean bench, under aseptic condition, above-mentioned nutrition gravy protein culture medium is poured in the culture dish of the bacterium of going out, bacillus benzoevorans ACCC10244 is conciliate chitin genus bacillus ACCC10227 forward respectively cultivation in the above-mentioned culture dish to, in 30 ℃ of incubators, cultivated 72 hours;
(3) cultured bacillus benzoevorans ACCC10244 is conciliate chitin genus bacillus ACCC10227, being mixed with respectively living bacteria count is 2.0 * 10
9100 milliliters of the bacterium liquid of individual/mL, and then be that 1: 1 ratio is mixed with composite fungus agent by volume.
5. the preparation method of complex micro organism fungicide claimed in claim 2 is characterized in that, this complex micro organism fungicide adopts following method to make:
(1) preparation nutrition gravy protein culture medium: with peptone 5 grams, NaCl5 gram, extractum carnis 3 grams, yeast extract paste 3 grams, oxyenadenine 0.005 gram, alkene acyl VITAMIN B4 0.005 gram, MnSO
4.H
2O0.005 gram, nutrient agar medium 15 grams mix mutually, add distilled water to 1000 milliliter, and pH value transfers to 6.5-7.2,121 ℃ of autoclavings 30 minutes;
(2) after cooling on Bechtop, under the condition of aseptic technique, the bacterial classification of two bacterial strains of cultured bacillus benzoevorans ACCC10244 reconciliation chitin genus bacillus ACCC10277 will be preserved, be inoculated into respectively in the above-mentioned nutrition gravy protein culture medium, under 30 ℃ of conditions in shaking culture on the shaking table after 72 hours, the bacterium liquid of two bacterial strains is shaken up with the mixing of 1: 1 ratio of volume ratio, and then the ratio with volume ratio 10% is inoculated into fermentor tank;
(3) complex ferment of microbiobacterial agent: fermentor tank nutrient solution prescription: peptone 0.5%; NaCl0.5%; Extractum carnis 0.3%; Yeast extract paste 0.3%; Oxyenadenine 0.5 ‰; Alkene acyl VITAMIN B4 0.5 ‰; MgSO
4.H
2O0.5 ‰; PH value transfers to 6.5-7.2; At 121 ℃, sterilization is 30 minutes under the 0.12MPa behind the tank body material loading, after being cooled to about 30 ℃ in 3% ratio inoculation; 30 ℃ of culture temperature, air flow begins 1: 1v.v.m, cultivate and increase to 1 after 24 hours: 1.5v.v.m, be down to 1 after 48 hours: 1.2v.v.m, fermentation to 72 hour fermentation is complete, conciliate the bacterium of chitin genus bacillus ACCC10227 through detecting bacillus benzoevorans ACCC10244 and count ratio about 1: 1, in the fermented liquid bacterium of the two to count when content all reaches hundred million/mL of 15-20 be qualified bacterium liquid;
(4) preparation of complex micro organism fungicide: add subsequently 30% Lalgine, 3% saltpetre, 3% glycerine, above percentage ratio is weight percentage.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110247715 CN102321539B (en) | 2011-08-26 | 2011-08-26 | Fermentation and preparation of composite microbial agent for degrading sulfonylurea herbicide with high efficiency |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110247715 CN102321539B (en) | 2011-08-26 | 2011-08-26 | Fermentation and preparation of composite microbial agent for degrading sulfonylurea herbicide with high efficiency |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102321539A CN102321539A (en) | 2012-01-18 |
| CN102321539B true CN102321539B (en) | 2013-05-29 |
Family
ID=45449379
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110247715 Expired - Fee Related CN102321539B (en) | 2011-08-26 | 2011-08-26 | Fermentation and preparation of composite microbial agent for degrading sulfonylurea herbicide with high efficiency |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102321539B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103756927A (en) * | 2013-11-12 | 2014-04-30 | 张越 | Bacillus subtilis for efficient degradation of nicosulfuron and application thereof |
| CN103749511B (en) * | 2013-12-26 | 2016-10-05 | 广东中迅农科股份有限公司 | A kind of containing flucarbazone with the Herbicidal combinations of monosulfmeturon |
| CN105132344A (en) * | 2015-10-13 | 2015-12-09 | 黑龙江中农欣欣农业科技发展有限公司 | Herbicide phytotoxicity biological antidote and preparing method and application thereof |
| CN108853864A (en) * | 2018-07-09 | 2018-11-23 | 沈阳工业大学 | Pass through the method for composite microbial system degradation sulfonylurea herbicide |
| CN109758713B (en) * | 2019-01-26 | 2021-02-05 | 吉林省农业科学院 | Biodegradation method of sulfonylurea herbicide |
| CN111088180B (en) * | 2019-12-16 | 2023-09-12 | 南阳师范学院 | Pyrazosulfuron-ethyl degradation strain BI-1 and application thereof |
| CN113583891B (en) * | 2021-06-04 | 2023-04-07 | 广东省农业科学院农业资源与环境研究所 | Bacillus xiamenensis BMS19 and application thereof |
| CN114806955A (en) * | 2022-04-25 | 2022-07-29 | 西北工业大学深圳研究院 | Compound microbial agent for degrading herbicide as well as preparation method and application of compound microbial agent |
| CN115353997B (en) * | 2022-08-29 | 2023-08-22 | 山西农业大学高粱研究所 | Composite microbial agent and application thereof in degrading nicosulfuron in soil |
-
2011
- 2011-08-26 CN CN 201110247715 patent/CN102321539B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN102321539A (en) | 2012-01-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102321539B (en) | Fermentation and preparation of composite microbial agent for degrading sulfonylurea herbicide with high efficiency | |
| CN104498399B (en) | Rhodopseudomonas palustris bacterial strain, biocontrol agent and its preparation method and application | |
| Moon et al. | Bacillus velezensis CE 100 inhibits root rot diseases (Phytophthora spp.) and promotes growth of Japanese cypress (Chamaecyparis obtusa Endlicher) seedlings | |
| CN106135285B (en) | Application of the Rhodopseudomonas palustris biocontrol agent in prevention and control rice blast | |
| CN102212485A (en) | Trichoderma longibrachiatum and application thereof to preventing and treating vegetable diseases | |
| Hassan et al. | Natural plant extracts and microbial antagonists to control fungal pathogens and improve the productivity of zucchini (Cucurbita pepo L.) in vitro and in greenhouse | |
| CN102757915A (en) | Chloro acetamide herbicide degrading bacteria as well as bactericide prepared thereby and application thereof | |
| CN102217656A (en) | Application of alternaria alternata metabolite to controlling Botrytis cirerea | |
| CN101654665B (en) | Method for preparing Bacillus subtilis | |
| CN108048332A (en) | A kind of green alga viable bacteria compound and its preparation method and application | |
| CN106635884B (en) | The separation and its application of a kind of Kiwi berry root endogeny rayungus | |
| CN105794853A (en) | Application of rhodopseudomonas palustris fermentation liquor to rice blast prevention and control | |
| Zhang et al. | Study on the preparation and effect of tomato seedling disease biocontrol compound seed-coating agent | |
| CN104673704B (en) | The pseudomonas aeruginosa strains of one plant of degraded agricultural chemicals containing ester bond | |
| CN101019556A (en) | Prepn process of pesticidal composite Bt microbe prepn | |
| CN103805519A (en) | Aspergillus niger capable of efficiently degrading nicosulfuron and application of aspergillus niger | |
| CN104004689B (en) | Can degrade 2,4 – d butyl esters and DDVP bacterial strain and prepare the method for remediation microbial inoculum | |
| CN102210329B (en) | Application of Aspergillus restrictus 28-generation product on preventing and curing cucumber Rhizoctonia solani | |
| CN103222479B (en) | Use of Streptomyces sioyaensis metabolite in prevention and control of Colletotrichum lindemuthianum | |
| CN112708565A (en) | Locust microsporidia strain and preparation and use method thereof | |
| CN102191185B (en) | Biological control endophytic fungi - Aspergillus restrictus | |
| CN102352326B (en) | Method of removing bloom-forming cyanobacteria by using Aeromonas sp. | |
| CN104726362A (en) | Streptomyces murinus and application thereof | |
| CN104542135A (en) | Method for prevention and control of vegetable root-knot nematodiasis with mixed Sr18 biological nematocide | |
| CN103222481B (en) | Use of Streptomyces sioyaensis metabolite in prevention and control of Thanatephorus cucumeris |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130529 Termination date: 20180826 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |