CN105602884A - Method for alleviating toxicity of heavy metal cadmium to root system growth-promoting bacteria - Google Patents
Method for alleviating toxicity of heavy metal cadmium to root system growth-promoting bacteria Download PDFInfo
- Publication number
- CN105602884A CN105602884A CN201610088623.8A CN201610088623A CN105602884A CN 105602884 A CN105602884 A CN 105602884A CN 201610088623 A CN201610088623 A CN 201610088623A CN 105602884 A CN105602884 A CN 105602884A
- Authority
- CN
- China
- Prior art keywords
- heavy metal
- cadmium
- fermentation
- metal cadmium
- growth
- 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.)
- Pending
Links
Classifications
-
- 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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for alleviating the toxicity of heavy metal cadmium to root system growth-promoting bacteria. According to the method, rare earth element ceric ammonium nitrate with the suitable concentration is added in the fermentation process in a specific time period to ease the influence of the heavy metal cadmium on B. amyloliquefaciens growth. Experimental results show that the heavy metal cadmium has the inhibiting effect on the growth of bacillus amyloliquefaciens, and strains need longer time to adapt to the high concentration of a cadmium chloride solution with the increase of concentration. 1 mg/L of a ceric ammonium nitrate solution can promote the growth of the bacillus amyloliquefaciens. The ceric ammonium nitrate solution added during initial fermentation can alleviate the toxic action of cadmium chloride added when fermentation is performed for 8 hours to the bacillus amyloliquefaciens.
Description
Obtain-Tianjin of the present invention Natural Science Fund In The Light 13JCYBJC25500 subsidizes.
Technical field
The invention belongs to fermentation engineering field, relate to a kind of side of heavy metal cadmium to root system growth promotion mushroom toxin of alleviatingMethod.
Background technology
Bacillus amyloliquefaciens can promote the growth of plant. Root system of plant growth-promoting bacterium can help plant heavy metal-pollutedIn the environment dying, reduce the toxic action of heavy metal on plants. Bacillus amyloliquefaciens is exactly one in root system of plant growth-promoting bacteriumKind. It can affect growing of root system of plant. For example: bacillus amyloliquefaciens SQR9, in the root system of cucumber, Neng GoubaoProtect the invasion and attack that plant avoids pathogen. The plant restoration method relevant to microorganism is widely used in reducing the dirt of heavy metal environmentDye the impact on plant products, the heavy metal cadmium in root system of plant growth-promoting fungi degradation bacillus amyloliquefaciens energy adsorption fouling environment, withTime cadmium the growth of bacterial strain is had to certain inhibitory action, in order to allow bacterial strain better adsorb cadmium, and promote plant growth andSuppress the breeding of pathogen, so in this experiment, heavy metal cadmium is malicious along with concentration increases to the growth of bacillus amyloliquefaciensEvil effect strengthens gradually. And the rare-earth element cerium of suitable concentration can promote the growth of bacterial strain, so utilize both characteristicsThe cerium that adds suitable concentration is alleviated the impact of cadmium on root system of plant growth-promoting fungi degradation bacillus amyloliquefaciens, and current technology is mainlyDo not use the rare earth element that China abounds with, and rare earth element own concerning ambient influnence compared with chemical pesticide, to environmentPollution level is lower again, can prevent from reducing the pollution to environment in promoting crop growth.
Summary of the invention
Due to the soil of current heavy metal, a large amount of existence in water, wherein the environment of heavy metal cadmium is more outstanding, instituteAlleviate the impact of cadmium on plant growth-promoting bacteria to propose this idea.
For achieving the above object, the invention discloses a kind of method of heavy metal cadmium to root system growth promotion mushroom toxin of alleviating, compriseFollowing steps:
(1) seed culture: bacillus amyloliquefaciens (Bacillusamyloliquefaciens) HM618 deposit number is(CGMCC7097) in seed culture medium, cultivate, cultivate 24h. Described seed culture based component is in g/L: glucose60, dusty yeast 10, peptone 10, sodium chloride 5,10,121 DEG C of steam sterilizing 20min of beef extract;
(2) fermented and cultured: by the initial 0D value after switching fermentation be 0.1 be basic vaccination in the fermentation medium of 100ml,Wherein heavy metal cadmium and rare-earth element cerium are pressed final concentration preparation mother liquor, and temperature is 37 DEG C, and rotating speed is 200rpm condition bottom fermentationCultivate, different experiments group is added the asynchronism(-nization) of ammonium ceric nitrate and cadmium chloride solution, is respectively fermentation 0h and adds suitable concentrationCeric ammonium nitrate solution, 8h adds the heavy metal cadmium solution of high concentration 40,80,160mg/L, and fermentation 8h adds suitable concentrationCeric ammonium nitrate solution, fermentation 0h adds the ceric ammonium nitrate solution of suitable concentration and 8h and adds high concentration 40,80,160mg/LHeavy metal cadmium solution, and heavy metal cadmium solution and fermentation 8h that 8h adds high concentration 40,80,160mg/L add suitable denseThe ceric ammonium nitrate solution of degree. In the time that in fermentation medium, concentration of glucose is reduced to 0g/L, fermentation ends while fermenting 96h.Described fermentation medium is made up of in g/L following compositions: glucose 60, dusty yeast 5, peptone 5, sodium chloride 5, phosphoric acid hydrogenDipotassium 3, potassium dihydrogen phosphate 3, potassium chloride 0.5, magnesium sulfate 0.5, pH nature.
The bacillus amyloliquefaciens (Paenibacilluspolymyxa) the present invention relates to, called after BacillusAmyloliquefaciensHM-618. And be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center,(ChinaGeneralMicrobiologicalCultuerCollectionCentre) carry out preservation, preservation address: northNo. 3, No. 1, North Star West Road, Jing Shi Chaoyang District institute, Institute of Microorganism, Academia Sinica, postcode: 100101, preserving number:CGMCC7097, preservation time 2013-01-08.
The present invention further discloses alleviation heavy metal cadmium root system growth promotion mushroom toxin method is being added to rare-earth element cerium alleviationThe application of the murder by poisoning aspect of high concentration heavy metal cadmium to bacillus amyloliquefaciens. Wherein add rare earth element and refer to 1mg/L'sThe application of ceric ammonium nitrate solution aspect the growth of promotion bacillus amyloliquefaciens. Experimental result shows that heavy metal cadmium is to separating starch budThe growth of spore bacillus is inhibited, and with the rising of concentration, bacterial strain need to adapt to the caddy of high concentration for more timeSolution. The ceric ammonium nitrate solution of 1mg/L can promote the growth of bacillus amyloliquefaciens. The initial ammonium ceric nitrate adding of fermentationSolution can be alleviated the toxic action of caddy to bacillus amyloliquefaciens that fermentation is added for 8 hours.
The present invention has utilized 0.5 ~ 50mg/LCe4+Can promote the growth of bacillus amyloliquefaciens, and 40,80,160mg/LCdCl2Can suppress the growth of bacterial strain, strain growth impaired development, will affect growing of cereal crops, institute simultaneouslyAdd the murder by poisoning of rare-earth element cerium alleviation high concentration heavy metal cadmium to bacillus amyloliquefaciens to attempt external source, this research has been foundThe optimal time (cadmium adds after 8 hours in fermentation, and ammonium ceric nitrate is in the initial interpolation of fermentation) of cadmium and cerium, improves and alleviates effectRate.
It is long-pending that alleviation heavy metal cadmium disclosed by the invention compared with prior art had the method for root system growth promotion mushroom toxinUtmost point effect is:
The present invention has utilized the rare earth element ammonium ceric nitrate of extensive existence to alleviate heavy metal cadmium root system of plant growth-promoting fungi degradation is formed sedimentThe toxic action of powder bacillus, the ammonium ceric nitrate of 1mg/L can effectively promote the growth of bacterial strain, heavy metal cadmiumIn soil environment, add rare earth element ammonium ceric nitrate and can alleviate cadmium to cereal crops root system as a kind of novel methodThe toxic action of root system growth-promoting fungi degradation bacillus amyloliquefaciens, thus the output of cereal crops improved, substitute people at ordinary times and usedChemicals agricultural chemicals, can alleviate agricultural chemicals use after polluting again soil environment.
Brief description of the drawings
The impact of the heavy metal cadmium solution of Fig. 1 variable concentrations on bacillus amyloliquefaciens growth;
The impact of the tetravalence ceric ammonium nitrate solution of Fig. 2 variable concentrations on bacillus amyloliquefaciens growth;
The ferment impact of bacillus amyloliquefaciens growth under the heavy metal cadmium environment adding for 8 hours of Fig. 3;
Fig. 4 ferments cadmium that the initial ammonium ceric nitrate adding adds fermenting for 8 hours to the bacillus amyloliquefaciens relieving effect of growing;
Fig. 5 ferment cadmium that the ammonium ceric nitrate that adds for 8 hours adds fermenting for 8 hours to bacillus amyloliquefaciens growth alleviate effectShould.
Detailed description of the invention
Below by specific embodiment narration the present invention. Unless stated otherwise, in the present invention, technological means used isMethod known in those skilled in the art. In addition, embodiment is interpreted as illustrative, and unrestricted model of the present inventionEnclose, the spirit and scope of the invention are only limited by claims. To those skilled in the art, do not deviating from thisUnder the prerequisite of bright essence and scope, various changes that the material component in these embodiments and consumption are carried out or change also belong toIn protection scope of the present invention.
Embodiment 1
Alleviate the method for heavy metal cadmium to root system growth promotion mushroom toxin, comprise the steps:
(1) preparation of culture medium
Seed culture based component is in g/L: glucose 60, and dusty yeast 10, peptone 10, sodium chloride 5, beef extract 10,121 DEG C of steam sterilizing 20min; Fermented and cultured based component is in g/L: glucose 60, dusty yeast 5, peptone 5, sodium chloride5, dipotassium hydrogen phosphate 3, potassium dihydrogen phosphate 3, potassium chloride 0.5, magnesium sulfate 0.5, pH nature, 121 DEG C of steam sterilizing 20min;
(2) shaking flask is cultivated seed culture
By in the seed culture medium of BacillusamyloliquefaciensHM618 glycerol stock access 100mL, in temperature 37DEG C, under rotating speed 200rpm condition, cultivate 24h, make seed culture fluid;
(3) fermented and cultured
The inoculum density that is 0.1 by initial OD accesses seed in the shaking flask of 100ml, and rotating speed is 200rpm, and temperature is 37DEG C, fermentation access when 0h variable concentrations cadmium chloride solution (Cd0,5,10,20,40,80,160mg/L add respectively 0,0.25,0.5, the caddy mother liquor of the 2g/L of 1ml, 0.125,0.25, the caddy mother liquor of the 32g/L of 0.5ml), by oneThe fixed time interval samples, and uses the OD value of spectrophotometric determination bacterial strain, make chart, can obtain variable concentrationsThe affect (see figure 1) of cadmium solution on bacillus amyloliquefaciens growth.
Embodiment 2
Alleviate the method for heavy metal cadmium to root system growth promotion mushroom toxin, comprise the steps:
(1)～(2) are with embodiment 1 step (1)～(2);
(3) inoculum density that is 0.1 by initial OD accesses seed in the shaking flask of 100ml, and rotating speed is 200rpm, and temperature is 37DEG C, access the tetravalence ceric ammonium nitrate solution (Ce of variable concentrations when 0h in fermentation4+0,0.5,1,50mg/L add respectively 0,0.1, the ammonium ceric nitrate mother liquor of the 0.5g/L of 0.2ml, the ammonium ceric nitrate mother liquor of the 10g/L of 0.5ml), by the regular hourInterval samples, and uses the OD value of spectrophotometric determination bacterial strain, makes chart, can obtain the tetravalence nitric acid of variable concentrationsThe affect (see figure 2) of cerium ammonium on bacillus amyloliquefaciens growth.
Embodiment 3
Alleviate the method for heavy metal cadmium to root system growth promotion mushroom toxin, comprise the steps:
(1)～(2) are with embodiment 1 step (1)～(2);
The inoculum density that is 0.1 by initial OD accesses seed in the shaking flask of 100ml, and rotating speed is 200rpm, and temperature is 37DEG C, experiment need arrange Cd40, tetra-caddy concentration of 80,160,320mg/L, and HM bacterial strain is only added in contrast, and CK arrangesOnly add Ce4+0h-1mg/L, adds at 0h, then has at fermentation 8h interpolation 8h-Cd40,80,160,320mg/L,After fermentation 8h, add Ce4+8h-1mg/L，Ce4+0h-1mg/L&8hCd40、80、160、320mg/L，Ce4+8H-1mg/L8h&Cd40,80,160,320mg/L, sample by certain time interval, uses spectrophotometerMeasure the OD value of bacterial strain, make chart, can obtain suitable tetravalence ammonium ceric nitrate and alleviate heavy metal cadmium to separating starch gemma barBacterium evil (seeing Fig. 3,4,5).
Conclusion:
(1) heavy metal cadmium is inhibited to the growth of bacillus amyloliquefaciens, wherein 5,10, the caddy of 20mg/L is moltenLiquid inhibitory action lower than 40,80, the cadmium chloride solution of 160mg/L. And with the rising of concentration, bacterial strain needs the longer time to fitAnswer the cadmium chloride solution of high concentration.
(2) ceric ammonium nitrate solution of 1mg/L can promote the growth of bacillus amyloliquefaciens.
(3) caddy that the initial ceric ammonium nitrate solution adding of fermentation can be alleviated fermentation interpolation in 8 hours is to separating starch gemma barThe toxic action of bacterium.
Claims (5)
1. alleviate the method for heavy metal cadmium to root system growth promotion mushroom toxin, it is characterized in that comprising the steps:
(1) seed culture: be (CGMCC by bacillus amyloliquefaciens (B.amyloliquefaciens) HM618 deposit number7097) in seed culture medium, cultivate, cultivate 24h; Described seed culture based component is in g/L: glucose 60, yeastPowder 10, peptone 10, sodium chloride 5,10,121 DEG C of steam sterilizing 20min of beef extract;
(2) fermented and cultured: be 0.1 for basic vaccination is in the fermentation medium of 100ml by the initial OD values after switching fermentation,Wherein heavy metal cadmium and rare earth element ammonium ceric nitrate are pressed final concentration preparation mother liquor, and temperature is 37 DEG C, and rotating speed is 200rpm conditionBottom fermentation is cultivated, and different experiments group is added the asynchronism(-nization) of ammonium ceric nitrate and cadmium chloride solution, when glucose in fermentation mediumWhen concentration is reduced to 0g/L, fermentation ends while fermenting 96h.
2. method claimed in claim 1, is characterized in that described heavy metal cadmium and rare earth element ammonium ceric nitrate are by final concentrationPreparation mother liquor refers to both actual interpolation concentration C e4+0.5、1、50mg/L、Cd5、10、20、40、80、160mg/L，Ce4+It is best that the interpolation scheme of 8h1mg/L&Cd8h40mg/L is alleviated cadmium poisoning effect.
3. method claimed in claim 1, is characterized in that described fermentation medium is made up of in g/L following compositions: glucose60, dusty yeast 5, peptone 5, sodium chloride 5, dipotassium hydrogen phosphate 3, potassium dihydrogen phosphate 3, potassium chloride 0.5, magnesium sulfate 0.5,PH nature.
4. described in claim 1, alleviating heavy metal cadmium alleviates high dense to root system growth promotion mushroom toxin method at interpolation rare-earth element ceriumThe application of the murder by poisoning aspect of degree heavy metal cadmium to bacillus amyloliquefaciens.
5. application claimed in claim 3, the ceric ammonium nitrate solution that wherein said interpolation rare earth element refers to 1mg/L existsPromote the application of bacillus amyloliquefaciens growth aspect.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610088623.8A CN105602884A (en) | 2016-02-17 | 2016-02-17 | Method for alleviating toxicity of heavy metal cadmium to root system growth-promoting bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610088623.8A CN105602884A (en) | 2016-02-17 | 2016-02-17 | Method for alleviating toxicity of heavy metal cadmium to root system growth-promoting bacteria |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105602884A true CN105602884A (en) | 2016-05-25 |
Family
ID=55983233
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610088623.8A Pending CN105602884A (en) | 2016-02-17 | 2016-02-17 | Method for alleviating toxicity of heavy metal cadmium to root system growth-promoting bacteria |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105602884A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110669811A (en) * | 2019-10-21 | 2020-01-10 | 天津大学 | Method for improving surfactant yield |
| CN110699408A (en) * | 2019-10-21 | 2020-01-17 | 天津大学 | Method for improving surfactant yield by mixed fermentation |
| CN110904002A (en) * | 2019-11-25 | 2020-03-24 | 天津大学 | Method for biologically removing tetracycline antibiotics |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103539535A (en) * | 2013-10-09 | 2014-01-29 | 南京农业大学 | Active biological matrix product specially used for culture of cucumber seedlings |
-
2016
- 2016-02-17 CN CN201610088623.8A patent/CN105602884A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103539535A (en) * | 2013-10-09 | 2014-01-29 | 南京农业大学 | Active biological matrix product specially used for culture of cucumber seedlings |
Non-Patent Citations (9)
| Title |
|---|
| 刘秀花: "《芽孢杆菌生物学及其应用》", 30 September 2007, 河南大学出版社 * |
| 宋农: "稀土促细菌生长作用研究进展", 《中国公共卫生》 * |
| 徐书显 等: "《临床医学与预防医学》", 31 May 1995, 四川科学技术出版社 * |
| 杜春梅: "《芽孢杆菌在农业中的研究与应用》", 31 May 2013, 黑龙江大学出版社 * |
| 杜瑞英: "镉胁迫对节杆菌(Arthrobacter sp.)和芽孢杆菌(Bacillus sp.) 生理生化特性的影响", 《安全与环境学报》 * |
| 柴瑞娟: "镧和铈对蜡状芽孢杆菌抗性、生长及胞内核酸的影响", 《微生物学通报》 * |
| 罗玉明 等: "稀土元素铈缓解镉对菱叶的毒害效应研究", 《土壤学报》 * |
| 胡开辉: "《微生物学实验》", 31 August 2004, 中国林业出版社 * |
| 陶永清 等: "《发酵工程原理与技术》", 31 May 2015, 中国水利水电出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110669811A (en) * | 2019-10-21 | 2020-01-10 | 天津大学 | Method for improving surfactant yield |
| CN110699408A (en) * | 2019-10-21 | 2020-01-17 | 天津大学 | Method for improving surfactant yield by mixed fermentation |
| CN110904002A (en) * | 2019-11-25 | 2020-03-24 | 天津大学 | Method for biologically removing tetracycline antibiotics |
| CN110904002B (en) * | 2019-11-25 | 2022-04-29 | 天津大学 | Method for biologically removing tetracycline antibiotics |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103911311B (en) | The microbial inoculum of a kind of efficient phosphate-solubilizing bacterium and production thereof | |
| CN103320369A (en) | Composite inoculant for promoting decomposing of cellulose organic wastes, and preparation method thereof | |
| CN103740634A (en) | Trichoderma asperellum chlamydospore powder as well as preparation method thereof and application | |
| CN105602884A (en) | Method for alleviating toxicity of heavy metal cadmium to root system growth-promoting bacteria | |
| CN104988082A (en) | Debaryomyces hansenii strain and applications thereof | |
| CN112831441A (en) | Compound microbial agent for preventing and treating basal rot of passion fruit stems | |
| CN105602883A (en) | Method for utilizing cerium chloride to relieve toxicity on root system growth-promoting bacteria caused by heavy metal cadmium | |
| CN104250624B (en) | A kind of preparation method of the active flora of HyM soil remediations | |
| CN104152373B (en) | Bacterial strain capable of efficiently degrading pendimethalin and application thereof | |
| CN106348887A (en) | Method for preparing amino acid liquid water-soluble fertilizer through compound stepped fermentation of seaweed | |
| CN109516869A (en) | Complex function microorganism formulation and its preparation method and application | |
| CN103045500B (en) | Mesorhizobium KDRM295 and application thereof | |
| CN105950480B (en) | A kind of method of herbage degradation volume reduction | |
| CN105802874A (en) | Mixed bacterium for efficiently degrading atrazine and fermental culture method | |
| CN104928198A (en) | Candida tropicalis strain and application thereof | |
| CN102433276B (en) | Sinorhizobium sp. and application thereof | |
| CN113881601A (en) | In-situ efficient compound rhizobium group screening method | |
| CN114644994A (en) | Petroleum hydrocarbon degradation compound microbial agent and preparation method and application thereof | |
| CN108865926B (en) | Tannin and saponin degradation microbial agent | |
| CN106348884A (en) | Production method of bacillus megaterium and humic acid containing liquid water-soluble fertilizer | |
| CN110669704A (en) | Efficient composite phosphorus-dissolving microbial inoculum and preparation method and application thereof | |
| CN105647812A (en) | Schizophyllum commune strain and application thereof in production of Pu'er tea | |
| CN103013856B (en) | Mesorhizobium KDRM611 and application thereof | |
| CN102978138B (en) | Mesorhizobium KDRM185 and application thereof | |
| CN103013857B (en) | Mesorhizobium KDRM 565 and application 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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160525 |