CN102191198A - Biocontrol strain JH21 for controlling rice sheath blight and rice blast and biocontrol bacterial agent thereof - Google Patents
Biocontrol strain JH21 for controlling rice sheath blight and rice blast and biocontrol bacterial agent thereof Download PDFInfo
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Abstract
The invention relates to a biocontrol strain JH21 for controlling rice sheath blight and rice blast, belonging to the technical field of plant protection. The bacteria are Pantoea agglomerans. The preparation method of the bacterial agent thereof comprises the following steps: using the bacteria to perform shaking cultivation in potato dextrose agar(PDA) culture solution at 30 DEG C in 180rpm for 20-24 hours, then centrifuging in 6000rpm for 10 minutes, and using wet bacteria and the preservative fluid of the laboratory to prepare the bacterial agent according to a ratio of 1:40 (g/ml), wherein the total concentration of the live bacteria in the finished product is 3*10<9>-3*10<12>CFU/ml. The control rate of the JH21 bacterial agent to rice blast is about 65-75%, the yield can be increased by 40-50%; and the control rate to rice sheath blight is more than 60-70% and the yield can be increased by 40-60%. The JH21 bacterial agent can control rice sheath blight and rice blast simultaneously in one field; and the bacterial agent can obviously promote the growth of rice and increase the yield of rice. The bacterial agent can be used through soaking during seeding and after rice is transplanted, the bacterial agent is sprayed to control diseases.
Description
One, technical field
The present invention relates to a kind of biocontrol strain JH21 and biocontrol fungicide thereof of preventing and treating rice sheath blight disease and rice blast, belong to the farm crop technical field of preventing and curing diseases, be exclusively used in of the control of paddy rice main producing region rice blast and rice sheath blight disease.
Two, background technology
Paddy rice is one of China's staple food crop, and cultivated area accounts for 1/4 of national cultivated area at present, and annual production is about 1/2 of national total output of grain.Yet the lasting generation of rice disease is having a strong impact on the production of paddy rice always.Prevent and treat under the condition existing, the whole nation reaches 20,000,000,000 kg because of various rice diseases cause the paddy underproduction every year on average, and as not preventing and treating, the underproduction can reach 30,000,000,000 kg.Therefore, it is very urgent to study and prevent and treat rice disease.In rice disease, rice blast and banded sclerotial blight generation area are big, and popular strong, harm is serious, and difficulty is bigger in the control, is topmost two big diseases in the Rice Production.
Rice blast claims rice blast again, the pest etc. of kowtowing, it is a kind of worldwide fungal disease, global Rice Production is caused very big harm, be listed in first of the paddy rice three big diseases, be rice seedling and heading stage most important disease, in all rice districts of China generation is arranged all, generally cause the underproduction 10%～20% in the popular time of disease, the time underproduction 40%～50% that disease is heavy, even No kernels or seeds are gathered, as in a year of scarcity.Since the 90's of 20th century, area takes place all at 3,800,000 hm in the year of China's rice blast
2More than, year loss paddy reaches several hundred million kilograms.China mainly is that the applied chemistry agricultural chemicals is prevented and treated rice blast at present on producing, and generally adopts chemical bactericide seed soaking control seedling pest when seedling stage, uses foliar spray control leaf pest and panicle blast behind the transplanting Honda.Present stage produce to go up main use kasugamycin, tricyclazole or spray at seedling stage and heading stage with the composite composite bactericide of tricyclazole and sulphur, tricyclazole and iprobenfos prevent and treat rice blast.But rice blast be subjected to kind, weather effect bigger, occurrence degree, the prediction precision of emergence period to rice blast are not high, applied chemistry pesticide control rice blast, because of medicament itself does not have interior absorption, be difficult in and prevent and treat the optimum period medication, and, make preventive effect constantly descend because germ strengthens gradually to medicament commonly used is drug-fast.The use at double of chemical agent in addition, farmland ecosystem is seriously damaged, and the rice pesticide residue that the results post-treatment becomes are serious, bring the very important serious danger side of body to health of people.
By rhizoctonia solani
(Rhizoctonia solani)The rice sheath blight disease that causes is one of rice fungus diseases evil of the refractory reason of paddy rice.Along with the popularizing planting of short-stalked variety and hybridisation rice and the raising of cultivation management level, banded sclerotial blight takes place serious day by day.Wherein, be injured the heaviest with the high-yield rice district especially.Banded sclerotial blight causes that mainly sheath is withered and leaf is withered, and the paddy rice setting percentage is reduced, and rate of immatured grain increases, and grain heavily descends, and serious underproduction 40-50% takes place general underproduction 10-20%.And, do not find high anti-kind both at home and abroad so far as yet to this disease because the saprophytic ability of sheath blight fungus is strong and host range is wide; Main control medicament---jingganmycin has produced tangible resistance to this disease to domestic, and per season medication number of times is increased to present 4～7 times for 2 times before 20 years, dosage also from before 1.5kg/ hm
2Be increased to present 4.5～6.0kg/ hm
2Cause the problem that natural ecological environment pollutes and the rice pesticide residue increases also more and more outstanding because of preventing and treating the long-term a large amount of agricultural chemicals that use of banded sclerotial blight.
Chemical pesticide is to the havoc of farmland ecosystem, and pathogenic bacteria is more and more obvious to the resistance of medicament.Simultaneously, chemical pesticide is difficult for degraded, and high residue is absolutely unsafe to people and animals and has all seriously limited use in agriculture production in the future.Therefore, property prophylactico-therapeutic measures has non-chemically become the core mode of preventing and treating rice disease at present, and biological control then is the most promising method wherein.In the prior art according to this breadboard preservative fluid for thallus patent (publication CN1358838, open day: on July 17th, 2002) method, but thalline room temperature preservation 2～3 years.
Three, summary of the invention
Technical problem:
The objective of the invention is at rice blast in the present agriculture production and rice sheath blight disease controlling difficulty big, the cost height, the chemical pesticide resistance becomes increasingly conspicuous, problems such as pesticide residue height, screen efficient biocontrol strain and make biocontrol fungicide, rice blast and rice sheath blight disease are all had preventive effect.
Technical scheme
A kind of biocontrol strain JH21 that prevents and treats rice sheath blight disease and rice blast be pantoea agglomerans (
Pantoea agglomerans), on June 18th, 2009 was preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, address, and Institute of Microorganism, Academia Sinica, culture presevation number is CGMCC NO.3126.
The method that biocontrol strain JH21 is used to prepare biocontrol fungicide is:
1) this inoculation is cultivated 30 ℃ of 180rpm respectively in the PDA nutrient solution, in Bechtop, take a sample respectively after 16～20 hours and survey the OD value at 600nm place, return to zero with the nutrient solution that does not connect bacterium in the mensuration process;
2) when being cultured to the OD value between 0.5～0.8 the time, to plant daughter bacteria liquid and add in the PDA nutrient solution, 30 ℃ of 180rpm shaking culture 20～24 hours, the centrifugal 10min of 6000rpm then with the 1:100 volume ratio, every 1000ml nutrient solution obtains the wet bacterium of 13～16g, and the wet bacterium of every gram contains 4 * 10
10～10
15Individual thalline is got wet bacterium and preservative fluid for thallus and is made into microbial inoculum by 1 g:40ml, and the viable bacteria total concn is 3 * 10 in the finished product
9～3 * 10
12CFU/ml is used to prevent and treat rice blast and rice sheath blight disease.
Used PDA nutrient solution compound method is: be cut into small pieces to be put in the water after peeling with 200 gram potatoes and boil, boil at least after the boiling more than 20 minutes, then filter with four layers of hospital gauze, filtrate adds 20 gram common sucrose, be settled to 1000 milliliters, the pH value transfers to 7.2～7.4, sterilizes 20 minutes for 121 ℃ then.Preservative fluid for thallus is the 0.01mol/L phosphate buffered saline buffer, and also containing concentration is the Tween20 nonionic surface active agent of 0.001mol/L.
Beneficial effect
Advantage of the present invention and positively effect show: the present invention is special biological prevention and control agent at rice blast and rice sheath blight disease exploitation.Owing to be biotechnological formulation, there are not a series of problems that use brought fully because of chemical pesticide, thereby the ecosystem that can effectively protect the fields, help the nuisanceless production of rice, implement the Sustainable development of agricultural.This biocontrol fungicide preventive effect is good, and price is lower than chemical pesticide, can be the peasant and reduces expenses; This microbial inoculum also has the short volume increase function of giving birth to, and the high-quality pollution-free rice of producing can be the peasant and increases income.
Dull and stereotyped antagonistic effect shows that the JH21 preparation has the obvious suppression effect to rice blast pathogen and rice sheath blight disease pathogenic bacteria.Greenhouse test shows: the JH21 preparation can effectively be controlled Pyricularia oryzae and Rhizoctonia solani Kuhn to the infecting of paddy rice, and suppresses mycelial growth, reaches the effect that disease controlling takes place.Simultaneously, the JH21 preparation can promote the growth of rice plant and improve its resistance.Field experiment result shows: the JH21 microbial inoculum is to being about 65～70% to the rice blast preventive effect, and volume increase can reach 40～45%; To the rice sheath blight disease preventive effect is more than 60～70%, and volume increase can reach 40～60%.
This microbial inoculum (belongs to this laboratory granted patent with wet bacterium and this breadboard preservative fluid for thallus, patent No. ZL02112559.7, publication CN1358838, open day: on July 17th, 2002) be made into microbial inoculum by 1:40 (g/ml), but room temperature preservation 2～3 years.
This mix bacterium agent uses by spraying and prevents and treats disease, also can handle by seed soaking.
Four embodiments
A kind of from paddy rice rhizosphere soil the biocontrol strain JH21 that prevents and treats rice sheath blight disease and rice blast that obtains of screening, this bacterium be general Pseudomonas (
Pantoea agglomerans), 8h produces single bacterium colony on the LB substratum, and bacterium colony is yellow behind the cultivation 24h, and the quality homogeneous is circular to irregular, moistening, central uplift, smooth surface, somatic cells is a direct rod shape, G
-
The preparation method of JH21 microbial inoculum adopts conventional bacterial cultivation:
1) this inoculation is cultivated 30 ℃ of 180rpm respectively in the PDA nutrient solution, in Bechtop, take a sample respectively after 16～20 hours and survey the OD value at 600nm place, return to zero with the nutrient solution that does not connect bacterium in the mensuration process;
2) when the OD value is not between 0.1～1, each bacterium is continued to cultivate, surveyed once every 3 hours, up to each OD value all in this scope. when being cultured to the OD value between 0.5～0.8 the time, to plant daughter bacteria liquid adds in the PDA nutrient solution with the 1:100 volume ratio, 30 ℃ of 180rpm shaking culture 20～24 hours, the centrifugal 10min of 6000rpm then, every 1000ml nutrient solution obtain the wet bacterium of 13～16g, and the wet bacterium of every gram contains 4 * 10
10～10
15Individual thalline;
Get wet bacterium and preservative fluid for thallus and be made into the JH21 biocontrol fungicide by 1 g:40ml, the viable bacteria total concn is 3 * 10 in the finished product
9～3 * 10
12CFU/ml.Preservative fluid for thallus is the 0.01mol/L phosphate buffered saline buffer, and also containing concentration is the Tween20 nonionic surface active agent of 0.001mol/L.
Used PDA nutrient solution compound method is: be cut into small pieces to be put in the water after peeling with 200 gram potatoes and boil, boil at least after the boiling more than 20 minutes, then filter with four layers of hospital gauze, filtrate adds 20 gram common sucrose, be settled to 1000 milliliters, the pH value transfers to 7.2～7.4, sterilizes 20 minutes for 121 ℃ then.
(1) dull and stereotyped antagonism bacteriostatic test
Utilize dull and stereotyped face-off culture method, measure the restraining effect of JH21 bacterial strain Rhizoctonia solani Kuhn and Pyricularia oryzae.
(2) greenhouse test
1. seed treatment:
Rice paddy seed carries out surface sterilization to be handled: sterilized water is washed three times after handling 15 s with 75% alcohol, and sterilized water was washed three times after 5% chlorine bleach liquor handled 15min.It is 10 that the rice paddy seed of surface sterilization is soaked in concentration
8Behind the CFU/mlJH21 microbial inoculum 30min, sow seedling raising after the vernalization of preserving moisture.
2. inoculation of pathogenic bacteria and JH21 microbial inoculum uses
With plastics keg dress rice field soil, transplant 3 caves, 5 rice shoots in every cave for every barrel.Inoculation pathogenic bacteria when rice plant grows to tillering phase.With Rhizoctonia solani Kuhn (
Rhizoctonia solani) AG-1 (IA) for for the examination pathogenic bacteria (Inagaki K. Dispersal of rice sheath blight fungus,
Rhizoctonia solaniAG-1 (IA), and subsequent disease development in paddy fields, from survey of vegetative compatibility groups[J].
Mycoscience1998 (39): 391-397.), adopt toothpick inoculation method (Yi Runhua, Zhu Xiru, Zhou Erxun. the research [J] of Rhizoctonia solani Kuhn artificial inoculation method. Guangzhou University's journal (natural science edition), 2003, (03)) inoculation: will sign the leaf sheath inboard that embeds plant through the wooden brachydont in the length 2 cm left and right sides that connects the bacterium cultivation, preserve moisture.
With Pyricularia oryzae (
Magnaporthe grisea) the 91-17-2 physiological strain is pathogenic bacteria (Mu Changqing; Liu Xue; Lu Qingguang; Jiang is carefully good; Zhu Changxiong. subtilis B-332 bacterial strain reaches the effect [J] of growing surely to the prevention effect of rice blast. plant protection journal, 2007, (02)); inoculate seedling stage: after germ is induced the product spore, inoculate to the foliage-spray spore suspension.
When treating that the state of an illness appears in clear water control group paddy rice, begin to spray the JH21 microbial inoculum and handle.
3. statistics
Statistics JH21 microbial inoculum is to the preventive effect of rice sheath blight disease and rice blast, and to the growth promoting function of paddy rice.
(3) field test
Use the JH21 microbial inoculum to carry out the field test of field control rice blast and rice sheath blight disease at Jiangsu Province's Tongshan County and Anhui Province's Lujiang County in May, 08 to November, added up the JH21 microbial inoculum to the prevention effect of disease with to the short living effect of increasing production of paddy rice.
Experimental result
1. dull and stereotyped bacteriostatic test
According to " planting the disease organon " middle plateform face-off culture experiment method, JH21 is estimated the inhibition effect of Pyricularia oryzae and Rhizoctonia solani Kuhn, statistics is as follows when treating that clear water control treatment group germ mycelia is covered with flat board, JH21 is 74.44% to the Pyricularia oryzae growth inhibition ratio, and the Rhizoctonia solani Kuhn growth inhibition ratio is shown in the 47.78%(table 1).
2. the sick greenhouse test of leaf blast in rice
Behind the inoculation magnaporthe grisea spore, when control group begins to fall ill, spray the JH21 microbial inoculum for the first time, after this sprayed one time the JH21 microbial inoculum every 7-10 days.To rice ear sprouting period, investigation preventive effect and biomass.With 20% tricyclazole WP(Jiangsu bumper harvest farmingization company limited) serve as the contrast medicament.The JH21 microbial inoculum reaches 73.60% to the rice blast preventive effect in greenhouse test, be higher than the tricyclazole treatment group.Simultaneously, use the JH21 microbial inoculum can obviously promote the growth of rice plant, the biomass rate of increase is shown in the 40.83%(table 2).
3. rice sheath blight disease greenhouse test
Behind the inoculation Rhizoctonia solani Kuhn, spray the JH21 microbial inoculum when control group begins to fall ill for the first time, after this sprayed once every 10-15 days.Paddy rice is investigated preventive effect and biomass to milk stage.With the contrast medicament is the very clear SC (Shanghai Nongle Biological Product Co., Ltd.) of 20% line.The JH21 microbial inoculum reaches 81.07% to the rice sheath blight disease preventive effect in greenhouse test, be higher than the agricultural chemicals control group.Simultaneously, the JH21 microbial inoculum can promote the growth of rice plant, and the biomass rate of increase is shown in the 62.41%(table 3).
4. rice blast is prevented and treated field test
Field test is chosen in rice blast and takes place in heavier Anhui Province's Lujiang County and the Jiangsu Province's Tongshan County rice terrace year after year.Establish 3 processing altogether: A:JH21 microbial inoculum 450ml converts water to 40L/667m
2B:20% tricyclazole (WP) 100g converts water to 40L/667m
2C: clear water contrast.Every processing is established 4 sub-districts and is repeated, and every sub-district area is 330m
2, each handles district's group arrangement at random.Isolate with guard rows between the sub-district, rich water quality management is carried out in the experimental plot routinely.Test is forecast medication with reference to the local plant protection unit state of an illness, and the medication for the first time in early stage of tillering prevents and treats medication every 15-17d in conjunction with each field rice disease and pest general status afterwards.The paddy rice milk stage, discontinues medication and adds up the disease severity.Whole rice growing season is prevented and treated 4 times altogether.Application method is: seed soaking is handled before the sowing; Adopt hand sprayer carefully to be sprayed to paddy rice leaf table after transplanting Honda.In order to estimate the ability that the JH21 microbial inoculum promotes paddy growth, carried out biomass mensuration in earlier stage in paddy rice booting, add up output behind the rice harvesting.
Test-results shows, the JH21 microbial inoculum is all obtained effect preferably in the field test of two provinces two places control rice blast, preventive effect difference 61.99% and 64.53% at Anhui Lujiang County and Jiangsu Tongshan County, all be higher than pesticide-treated group JH21 microbial inoculum and can promote the growth of rice plant and increase rice yield, units increased in production be respectively 48.13% and 36.26%(table 4 and 5 shown in).
5. rice sheath blight disease field test
Field test is chosen in rice blast and takes place in heavier Anhui Province's Lujiang County and the Jiangsu Province's Tongshan County rice terrace year after year.Establish 3 processing altogether: A:JH21 microbial inoculum 450ml converts water to 40L/667m
2The very clear SC 75ml of B:20% line converts water to 40L/667m
2C: clear water contrast.Every processing is established 4 sub-districts and is repeated, and every sub-district area is 330m
2, each handles district's group arrangement at random.Isolate with guard rows between the sub-district, rich water quality management is carried out in the experimental plot routinely.Test is with reference to local plant protection unit state of an illness forecast medication, and the medication for the first time in early stage of tillering is afterwards every the each field of 15-17d rice disease and pest general status control medication.The paddy rice milk stage, discontinues medication and adds up the disease severity.Whole rice growing season is prevented and treated 4 times altogether, and application method adopts hand sprayer slightly to be sprayed to rice clump base portion.In order to estimate the promotes growth ability of JH21 microbial inoculum to paddy rice, biomass mensuration has been carried out in paddy rice booting in earlier stage.Add up output behind the rice harvesting.
Test-results shows that the JH21 microbial inoculum is all obtained effect preferably in two provinces two places field control rice sheath blight disease, is respectively 67.81% and 71.57% in the preventive effect of Anhui Lujiang County and Jiangsu Tongshan County, all is higher than the pesticide-treated group.The JH21 microbial inoculum can promote the growth of rice plant and increase rice yield, units increased in production be respectively 59.20% and 56.57%(table 6 and 7 shown in).
Claims (5)
1. biocontrol strain JH21 who prevents and treats rice blast and rice sheath blight disease, for pantoea agglomerans (
Pantoea agglomerans), on June 18th, 2009 was preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, address, and Institute of Microorganism, Academia Sinica, culture presevation number is CGMCC NO.3126.
2. with the control rice blast of the described biocontrol strain JH21 preparation of claim 1 and the biocontrol fungicide of rice sheath blight disease.
3. prepare the method for the described biocontrol fungicide of claim 2, it is characterized in that:
1) the described inoculation of claim 1 30 ℃ of 180rpm in the PDA nutrient solution is cultivated after 16～20 hours in Bechtop the OD value that the 600nm place is surveyed in sampling respectively, return to zero with the nutrient solution that does not connect bacterium in the mensuration process;
2) when being cultured to the OD value between 0.5～0.8 the time, to plant daughter bacteria liquid and add in the PDA nutrient solution, 30 ℃ of 180rpm shaking culture 20～24 hours, the centrifugal 10min of 6000rpm then with the 1:100 volume ratio, every 1000ml nutrient solution obtains the wet bacterium of 13～16g, and the wet bacterium of every gram contains 4 * 10
10～10
15Individual thalline;
3) get wet bacterium and preservative fluid for thallus and be made into microbial inoculum by 1 g:40ml, the viable bacteria total concn is 3 * 10 in the finished product
9～3 * 10
12CFU/ml.
4. method according to claim 2 is characterized in that, used PDA nutrient solution compound method is:
Be cut into small pieces to be put in the water after peeling with 200 gram potatoes and boil, boil at least after the boiling more than 20 minutes, then filter with four layers of hospital gauze, filtrate adds 20 gram common sucrose, is settled to 1000 milliliters, and the pH value transfers to 7.2～7.4, sterilizes 20 minutes for 121 ℃ then; Preservative fluid for thallus is the 0.01mol/L phosphate buffered saline buffer, and also containing concentration is the Tween20 nonionic surface active agent of 0.001mol/L.
5. the application of the described biocontrol fungicide JH21 of claim 2-4 product aspect control rice blast and rice sheath blight disease.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103952338A (en) * | 2014-04-11 | 2014-07-30 | 山西省农业科学院农产品贮藏保鲜研究所 | Pantoea agglomeran new bacterial strain XM2, preparation method for suspension of same, and method for controlling pear black spot |
| CN104694422A (en) * | 2015-02-11 | 2015-06-10 | 齐齐哈尔大学 | Separated pantoea ananatis HYL01 strain and application thereof in promotion of root growth of watermelons |
| CN108192850A (en) * | 2018-03-13 | 2018-06-22 | 长沙艾格里生物肥料技术开发有限公司 | Thiophilic small red oomycetes bacterial strain, thiophilic small red oomycetes microbial inoculum and its application |
| CN110643551A (en) * | 2019-11-18 | 2020-01-03 | 吉林大学 | Stenotrophomonas rhizophila S11 for preventing and treating rice blast and application thereof |
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| CN1358838A (en) * | 2002-01-18 | 2002-07-17 | 南京农业大学 | Preservation method for live bacterial preparation |
| CN1831116A (en) * | 2006-04-03 | 2006-09-13 | 西北农林科技大学 | One strain forming a block type fanjun fungus, fermentation culturing method and application thereof |
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2011
- 2011-03-28 CN CN2011100743369A patent/CN102191198B/en not_active Expired - Fee Related
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| CN1358838A (en) * | 2002-01-18 | 2002-07-17 | 南京农业大学 | Preservation method for live bacterial preparation |
| CN1831116A (en) * | 2006-04-03 | 2006-09-13 | 西北农林科技大学 | One strain forming a block type fanjun fungus, fermentation culturing method and application thereof |
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| 《Appl Microbiol Biotechnol》 20011231 E. Costa等 Production of the biocontrol agent Pantoea agglomerans strain CPA-2 using commercial products and by-products 第367-371页 1-5 第56卷, * |
| 《中国生物防治》 20081231 薛梦林等 拮抗菌B501的鉴定及其对采后冬枣黑斑病的抑制效果 第122-127页 1-5 第24卷, 第2期 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103952338A (en) * | 2014-04-11 | 2014-07-30 | 山西省农业科学院农产品贮藏保鲜研究所 | Pantoea agglomeran new bacterial strain XM2, preparation method for suspension of same, and method for controlling pear black spot |
| CN103952338B (en) * | 2014-04-11 | 2016-08-17 | 山西省农业科学院农产品贮藏保鲜研究所 | Pantoea agglomerans strain X M2 and the preparation method of bacteria suspension thereof and the prevention and controls to Pear black spot |
| CN104694422A (en) * | 2015-02-11 | 2015-06-10 | 齐齐哈尔大学 | Separated pantoea ananatis HYL01 strain and application thereof in promotion of root growth of watermelons |
| CN108192850A (en) * | 2018-03-13 | 2018-06-22 | 长沙艾格里生物肥料技术开发有限公司 | Thiophilic small red oomycetes bacterial strain, thiophilic small red oomycetes microbial inoculum and its application |
| CN110643551A (en) * | 2019-11-18 | 2020-01-03 | 吉林大学 | Stenotrophomonas rhizophila S11 for preventing and treating rice blast and application thereof |
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| Publication number | Publication date |
|---|---|
| CN102191198B (en) | 2012-12-12 |
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