CN102154162B - Bacillus amyloliquefaciens and application thereof - Google Patents
Bacillus amyloliquefaciens and application thereof Download PDFInfo
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- CN102154162B CN102154162B CN2010106149488A CN201010614948A CN102154162B CN 102154162 B CN102154162 B CN 102154162B CN 2010106149488 A CN2010106149488 A CN 2010106149488A CN 201010614948 A CN201010614948 A CN 201010614948A CN 102154162 B CN102154162 B CN 102154162B
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- algae
- bacillus amyloliquefaciens
- cgmcc
- anabena
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Abstract
The invention discloses a bacillus amyloliquefaciens and an application thereof. The bacillus amyloliquefaciens with the China general microbiological culture collection number 4516 (China general microbiological culture collection (CGMCC) NO: 4516) has an obvious inhibiting effect on the growth of algae, especially has a high-efficiency inhibiting effect on the growth of anabaena flos-aquae, and can be applied to algal bloom control or algal removal of water environments such as lakes, urban landscape water and the like which are in eutrophication state.
Description
Technical field
The present invention relates to a kind of bacterium, be specifically related to a kind of molten algae bacterium with efficient effect of algae restraint.
Background technology
In the body eutrophication problem day by day under the serious situation; Economic, convenient, as to have environment friendly biological method (
etc., 1998; Carpenter etc., 1992) become eutrophication water and repair hot research fields.Dissolve the research of algae bacterium control algae by progressively carrying out the sixties in 20th century, come into one's own day by day.The researcher is from marine red tide (Imai etc. both at home and abroad; 2001), eutrophic lake (Rashidan etc.; 2001), reservoir (S.W.Jung etc.; 2008), farmland drainage canal (Jeffrey; 1981), sewage (James etc.; 1969), bottom mud in lake (Choi Hee-jin etc.; 2005) etc. isolate molten algae bacterium in the environment with effect of algae restraint.
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is extensively to be present in natural a kind of non-pathogenic bacteria.At present, domestic it is studied only is confined to Optimizing Conditions of Fermentation, degrading herbicide Butachlor technical 92 (Tan Wenjie etc.; 2005), suppress plant pathogenic fungi and virus (Shen Lili etc.; 2010) etc. at biological control (Wang Yingguo etc.; 2007) Application Areas in, the research in control algae field still is blank.Research shows that bacillus amyloliquefaciens can produce multiple antibacterial substance (Deng Jianliang etc.; 2007), suppress phytopathogen, fungi, virus, nematode (Yoshida etc. through producing microbiotic, antibacterial protein or polypeptide isoreactivity material; 2001), thus play the effect of biological control.
Summary of the invention
The purpose of this invention is to provide a kind of bacillus amyloliquefaciens with efficient effect of algae restraint.
Technical scheme provided by the invention is following:
Scheme 1: a kind of bacillus amyloliquefaciens (Bacillus amyloliquefaciens); Be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on December 28th, 2010 and (be called for short CGMCC; The address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City), preserving number is CGMCC NO:4516.
Scheme 2: be applied to scheme 1 said bacillus amyloliquefaciens CGMCC NO:4516 the algal bloom control of water surrounding or remove algae.
Scheme 3: a special case as scheme 2, it is characterized in that, utilize bacillus amyloliquefaciens CGMCC NO:4516 to suppress algae grows in the water surrounding.
Scheme 4: a special case as scheme 3 is characterized in that said algae is a blue-green algae.
Scheme 5: a special case as scheme 4 is characterized in that said blue-green algae is the wawter bloom anabena.
Beneficial effect of the present invention: bacillus amyloliquefaciens CGMCC NO:4516 provided by the invention can be used for the algal bloom control of water surrounding or removes algae; Algae grows in the water surrounding there is the obvious suppression effect, especially for the very high molten algae effect of having of wawter bloom anabena.
The biomaterial preservation
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens); Be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on December 28th, 2010 and (be called for short CGMCC; The address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City), preserving number is CGMCC NO:4516.
Embodiment
Through a specific embodiment the present invention is further specified below.
(1) separation of bacillus amyloliquefaciens CGMCC NO:4516:
With water sampler in the Dian Chi 1m place, eutrophic lake underwater water sampling, with the top layer water sample of gathering through filter paper filtering, to eliminate protozoic influence and to remove impurity in the water.Getting the aseptic technique of 20mL filtration water sample is injected in 5～7 days the wawter bloom anabena (wawter bloom anabena (Anabaena flos-aquae) algae kind is available from Inst. of Hydrobiology, Chinese Academy of Sciences algae kind storehouse (FACHB)) of the preparatory cultivation of 100mL; Control group injects the autoclaving water sample after 20mL filters; The algae kind is inoculated in the 150mL Erlenmeyer flask; Seal preservation with sealing film, be placed on photosynthetic plant culturing indoor cultivation, photosynthetic culture cycle is 14h:10h, 25 ± 1 ℃ of temperature, illumination 2000lux; Frond inoculation and the process of going down to posterity strictly observe the aseptic technique rule, as above cultivate for 1 week.To contrast normally, the algae liquid that etiolation appears in experimental group is used for next step experiment.This test and Selection wawter bloom anabena is as the main blue-green algae algae kind of being tried.
Collection is cultured to the wawter bloom anabena of logarithmic phase, in normal temperature with the centrifugal 12～15min of 3500～4000r/min.With aseptic BG11 nutrient solution yellow algae liquid is diluted 10,100 and 1000 times.The diluent and the 2mL that get the different gradients of 1mL respectively concentrate wawter bloom anabena liquid, in small test tube, fully be uniformly coated on the BG11 solid medium behind the mixing, and blank is set.These algae plates inversions are placed on the indoor illumination cultivation of photosynthetic plant culturing.After treating plaque to occur on the algae plate, get typical plaque (every plate is got 2～3), be resuspended in the aseptic BG11 liquid nutrient medium of 5mL with the aseptic inoculation ring cutting, with the method for shop algae plate to this plaque purifying three times.More than operation is all carried out in clean room.
Beef-protein medium separates: through cultivating in a few days, the bacterium colony that occurs being scattered around the plaque on the algae plate with aseptic inoculation ring picking colonies typical, is rule on beef-protein medium and is separated, and separates through 3～5 line, obtains the single bacterium colony of bacterium.
Be experimental subjects with wawter bloom anabena, microcystic aeruginosa and synechococcus respectively, in glass orifice plate aperture, add the algae liquid of 3ml logarithmic phase, put into the algae liquid that is cultured to logarithmic phase, illumination cultivation with the part of aseptic inoculation ring picking list bacterium colony.To the single bacterium colony that occurs etiolation in the 7 days growth of ruling once more, purifying repeatedly repeatedly.More than operation is all carried out in clean room.
Dissolving the algae bacterium selects: join wawter bloom anabena, microcystic aeruginosa and synechococcus through purifying molten phycomycete repeatedly repeatedly, dissolve the algae effect test, from various bacteria, choose the molten best bacterial strain DC1 of algae effect.
(2) the biochemical physics and chemistry of bacillus amyloliquefaciens CGMCC NO:4516 is identified
Measure the result through effect of algae restraint preliminary examination experiment and growth curve of bacteria, choose the DC1 bacterial strain that effect of algae restraint optimum, growth adaptation phase are short, the short period of time enrichment culture can obtain the high density thalline.The cellular form of DC1 and physics and chemistry test-results see the following form.
Table 1 cellular form and physicochemical property
Annotate: "+" positive reaction, "-" negative reaction
According to cellular form, Physiology and biochemistry data, 16S rRNA gene order data; The qualification result that dissolves algae bacterium DC1 is bacillus amyloliquefaciens (Bacillus amyloliquefaciens); Be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on December 28th, 2010, preserving number is CGMCC NO:4516.
(3) bacillus amyloliquefaciens CGMCC NO:4516 dissolves the algae effect
Bacillus amyloliquefaciens CGMCC NO:4516 significantly dissolves the algae effect to the wawter bloom anabena to having, and the cell concentration of bacillus amyloliquefaciens CGMCC NO:4516 has very big influence to dissolving the algae effect.Under the identical situation of initial Chl-a concentration, cell concentration is big more, and is big more to the removal efficient of anabena.Initial Chl-a concentration is 0.23mgL
-1Add 1.45 * 10 in the algae liquid
7CellsmL
-1, 3.63 * 10
7CellsmL
-1, 7.26 * 10
7CellsmL
-1, 14.5 * 10
7CellsmL
-1Bacillus amyloliquefaciens CGMCC NO:4516, the average removal rate to anabena behind the illumination cultivation 6d is respectively 43.68%, 66.72%, 82.83%, 95.54%.Under the same experimental conditions, initial Chl-a concentration 0.61mgL
-1The average removal rate of algae liquid is respectively 35.50%, 59.47%, 71.60%, 94.93%; Initial Chl-a concentration 1.06mgL
-1The average removal rate of algae liquid is respectively 31.78%, 51.54%, 60.47%, 91.35%; Initial Chl-a concentration 1.37mgL
-1The average removal rate of algae liquid is respectively 26.09%, 40.32%, 51.59%, 92.26%; Initial Chl-a concentration 1.92mgL
-1The average removal rate of algae liquid is respectively 6.85%, 29.59%, 45.87%, 93.29%.Initial Chl-a concentration is 0.23mgL
-1, 0.61mgL
-1, 1.06mgL
-1, 1.37mgL
-1And 1.92mgL
-1Anabena liquid adds 14.5 * 10
7CellsmL
-1The bacterium amount, behind the illumination cultivation 6d, Chl-a concentration can be reduced to 0.07mgL
-1, 0.18mgL
-1, 0.39mgL
-1, 0.44mgL
-1And 0.4mgL
-1The result shows; Bacillus amyloliquefaciens CGMCC NO:4516 has significant removal effect to anabena; Along with the rising of cell concentration,, prove that the removal of anabena and bacillus amyloliquefaciens CGMCC NO:4516 have positive connection to the also corresponding increase of the clearance of Chl-a.
When initial Chl-a concentration is 0.23mgL
-1, 0.61mgL
-1, 1.06mgL
-1, 1.37mgL
-1, 1.92mgL
-1Gradient raises, and adds 14.5 * 10 in the algae liquid
7CellsmL
-1Bacillus amyloliquefaciens CGMCC NO:4516, the average removal rate to anabena behind the illumination cultivation 6d is respectively 95.54%, 94.93%, 91.35%, 92.26%, 93.29%.This explanation, under bacterium liquid that is fit to and algae liquid concentration, bacillus amyloliquefaciens CGMCC NO:4516 may command algae liquid Chl-a concentration can effectively be controlled the anabena growth in low scope.
Claims (2)
1. a bacillus amyloliquefaciens (Bacillus amyloliquefaciens), its preserving number is CGMCC NO:4516.
2. to algal bloom control or the application that removes algae aspect, said algae is the wawter bloom anabena to the said bacillus amyloliquefaciens of claim 1 in the water body of algal bloom.
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102719383B (en) * | 2012-06-29 | 2014-04-02 | 武汉工业学院 | Bacillus amyloliquefaciens, inoculant and application thereof |
| CN102816718B (en) * | 2012-08-08 | 2014-08-06 | 上海交通大学 | Bacillus and application thereof |
| CN102796685B (en) * | 2012-08-10 | 2013-11-27 | 上海交通大学 | Bacillus SSAL-6 and application thereof in degrading anabaena flos-aquae |
| CN103013851B (en) * | 2012-10-08 | 2014-06-25 | 上海交通大学 | Complex microbial inoculant and applications thereof |
| CN104140963A (en) * | 2014-06-26 | 2014-11-12 | 浙江大学 | Preparation method of compound microbial algaecide |
| CN105265485A (en) * | 2014-07-24 | 2016-01-27 | 天津荣利生物科技发展有限公司 | Microorganism type grass control composition used in aquaculture |
| CN107177538B (en) * | 2017-07-19 | 2019-12-06 | 扬州大学 | Marine bacillus cereus and application thereof in preventing and controlling anabaena flos-aquae |
| CN108949618B (en) * | 2018-06-26 | 2021-05-28 | 武汉市鄂正农科技发展有限公司 | Algae-lysing bacteria and application thereof |
| CN109022303B (en) * | 2018-06-26 | 2021-06-15 | 武汉市鄂正农科技发展有限公司 | Composition with functions of dissolving algae and destroying microalgae air bags and application thereof |
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Non-Patent Citations (3)
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| 史顺玉.溶藻细菌对藻类的生理生态效应及作用机理研究.《工程科技I辑》.2006,(第10期),全文. * |
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Inventor after: Wu Weizhong Inventor after: Li Chao Inventor after: Wu Guanqing Inventor after: Wu Weilong Inventor after: Yang Luhua Inventor after: An Chengcai Inventor before: Wu Weizhong Inventor before: Li Chao Inventor before: Wu Weilong Inventor before: Yang Luhua Inventor before: An Chengcai |
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Free format text: CORRECT: INVENTOR; FROM: WU WEIZHONG LI CHAO WU WEILONG YANG LUHUA AN CHENGCAI TO: WU WEIZHONG LI CHAO WU GUANQING WU WEILONG YANG LUHUA AN CHENGCAI |
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