CN105176933A - Toxic Microcystis cracking cyanophage and separation method and application thereof - Google Patents
Toxic Microcystis cracking cyanophage and separation method and application thereof Download PDFInfo
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Abstract
The invention discloses toxic Microcystis cracking cyanophage and a separation method and application thereof. The toxic Microcystis cracking cyanophage is characterized in that the cyanophage is a MaSSC-P strain, is classified and named as Microcystis aeruginosa single-stranded DNA cyanophage MaSSC-P, collected at the China General Microbiological Culture Collection Center on 25th May, 2015 and numbered as CGMCC No.10598. A purified cyanophage suspension is added into a Microcystis water sample, and the toxic Microcystis cracking cyanophage has specific cracking effect on Microcystis in the water sample and has cracking effect on Microcystis aeruginosa, Microcystis wesenbergii and Microcystis viridis. The toxic Microcystis cracking cyanophage has the advantage of being capable of safely, efficiently and quickly cracking toxic Microcystis in fresh water.
Description
Technical field
The present invention relates to a kind of poisonous Microcystis aeruginosa cracking phycophage, especially relate to a kind of poisonous Microcystis aeruginosa cracking phycophage and separation method thereof and application.
Background technology
In recent years, the economic activity of the mankind increases the weight of day by day, various sanitary sewage and factory effluent enter in the slow flow water bodies such as lake, river mouth, bay, make the nutritive substances such as the nitrogen phosphorus in the water body too much eutrophication become, the algal bloom caused thus, the generation of " wawter bloom " is day by day serious, and wherein blue-green algae becomes the Main Algae causing breakout of water bloom." wawter bloom " is one of the most serious environmental hazard in the world today, wherein China is one of the most serious country of breakout of water bloom, the lake of nearly 75% faces eutrophication pollution in various degree, as Taihu Lake, Chaohu and Dian Chi are very serious with regard to eutrophication, frequent outburst blue-green alga bloom, affect view, and bloom blue algae produces a large amount of toxin, serious reduction drinking water safety.Therefore effective control of blue-green alga bloom and improvement are had important practical significance.
Blue-green algae is the most original, the simplest prokaryotic organism of a class, and the more germy features of tool, are therefore often called again cyanobacteria (Cyanobacterium), is the main primary producer in natural water.Cyanophyta is divided into two guiding principles: chroococcoid guiding principle and hormocystangium guiding principle.Chroococcoid guiding principle algae is unicellular; Hormocystangium guiding principle algae is thread, has hormocystangium.Known blue-green algae about has 2000 kinds, and blue-green algae is widely distributed, various places all over the world, all can grow in seawater and fresh water.Microcystic aeruginosa is a kind of highly toxic blue-green algae, plays an important role in worldwide breakout of water bloom.Microcystic aeruginosa has very large harm to people and domestic animal, and microcystic aeruginosa produces very strong Algae toxins, this Algae toxins can the Eukaryotic PP2A of narrow spectrum suppression (PP2A) activity and cause liver cancer.Wawter bloom can cause Dissolved Oxygen in Water amount to decline, water quality deterioration, and the mortality of fish and other biological also causes a lot of problems in water harnessing.
At present, the technology controlling algae mainly contains Physical, chemical method and biological process.Though Physical control algae instant effect, but need the manpower and materials of at substantial.Chemical method mainly controls algae by chemical agent, and its shortcoming is that chemical agent can pollute in water body.Biological control effect is comparatively obvious, and economic environmental protection, there is huge application prospect.
The virus infecting blue-green algae is called phycophage.First strain phycophage within 1963, to be separated by Saferman with Mor-ris to obtain, it can infect sheath silk algae (Lynbya), seat algae (Phormidi-um) and knitline algae (Plectonema) simultaneously, and this kind of phycophage found the earliest is named as " LPP " phycophage.Phycophage has host specificity.Screening obtains the phycophage of the poisonous Bloom-causing Algal of high, the single-minded infection cracking of lytic activity, cultivate phycophage, form initial stage and outbreak period at blue-green algae algal tufa and add phycophage, allow phycophage cracking algal tufa algae, be the main development approach of Biological control, tool is of great significance.
Summary of the invention
Technical problem to be solved by this invention be to provide a kind of can the safely, efficiently, fastly poisonous Microcystis aeruginosa cracking type phycophage of poisonous Microcystis aeruginosa and separation method thereof and application in cracking fresh water.
The present invention solves the problems of the technologies described above adopted technical scheme: a kind of poisonous Microcystis aeruginosa cracking phycophage, called after microcystic aeruginosa single stranded DNA phycophage (
microcysticaeruginosasingle-strandDNAcyanophage) MaSSC-P strain, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on May 25th, 2015, deposit number is CGMCCNo.10598.
The biological property of this phycophage is as follows: MaSSC-P phycophage has head construction in regular polygon and elongated afterbody, head diameter 85nm, the long 250nm of afterbody; Phycophage can be formed on algae flat board size shape homogeneous bite algae spot, around without halo, edge clear is regular; This phycophage can by the enzymic digestion of DnaseI, S1 single-chain nucleic acid, and can not by RnaseA collagenase treatment.
The biological property of this phycophage is as follows: this phycophage places 1h in 40 ~ 50 DEG C, still keeps infection activity; Then inactivation under more than 60 DEG C of conditions; Phycophage, under the environment of pH3.0 ~ 11.0, keeps infection activity; The resistance to ultraviolet of phycophage, responsive to chloroform, under 4 DEG C or-80 DEG C of temperature can be stored in.
The separation purification method of above-mentioned poisonous Microcystis aeruginosa cracking phycophage, specifically comprises the steps:
(1) separation of phycophage
By top layer, the waters water sample containing poisonous Microcystis aeruginosa cracking phycophage of collection successively through Medium speed filter paper, 0.45 μm and 0.22 μm of nitrocellulose filter filtration; By filtrate and the microcystic aeruginosa being in logarithmic phase by volume the ratio of 1:4 infect and mix, be 2000lx in intensity of illumination, temperature is 25 DEG C, light dark period is cultivate under 12h: 12h condition, the nutrient solution that yellow occurs is carried out continuous passage infection until keep stable to bite algae property, obtain phycophage and infect liquid;
(2) amplification cultivation of phycophage and purifying
After ratio phycophage being infected liquid and microcystic aeruginosa liquid 1:9 by volume mixes, incubated at room temperature 30min on shaking table, then pour into be cooled to 42 DEG C containing quality volume fraction 0.7% agar BG-11 substratum in, pour into after abundant mixing in the culture dish of bottom containing the BG-11 culture medium flat plate of quality volume fraction 1.5% agar, treat that it solidifies, be placed in 2000lx, 25 DEG C, cultivate in the incubator of periodicity of illumination: 12h/12h, cultivate after 5 ~ 7 days, what scraping was formed bites algae spot;
(3) phycophage is further purified
The algae spot of biting above-mentioned steps (2) obtained is suspended in BG-11 substratum, after vortex 4 DEG C spends the night, after centrifuging and taking supernatant, repetition above-mentioned steps (2) is bitten algae spot and is tested 2 times, obtain shape, uniformly bite algae spot, then will bite algae spot is suspended in BG-11 substratum, after 0.22 μm of filter membrane centrifuging, gets filtrate, in filtrate, adding PEG8000 to its quality volume final concentration is 9%, 4 DEG C of sedimentations are spent the night, at 4 DEG C 36,000
× gcentrifugal 2h, with the washing precipitation of BG-11 substratum, obtains the phycophage suspension of purifying.
The phycophage suspension of purifying is added in microcystis waterbloom sample, has Specific lytic effect to the Microcystis aeruginosa in water sample.Its to microcystis kutz (
microcystis) microcystic aeruginosa
m.aeruginosafACHB-924
, M.aeruginosafACHB-925, Hui Shi Microcystis aeruginosa
m.wesenbergiifACHB-908
, M.wesenbergiifACHB-929, Microcystis viridis
m.viridisfACHB-979 all has splitting action.
Compared with prior art, the invention has the advantages that: a kind of poisonous Microcystis aeruginosa cracking phycophage of the present invention and separation method thereof and application, it makes public for the first time microcystic aeruginosa single stranded DNA phycophage, after this cracking performance phycophage infects blue-green algae, can in host fast breeding, and make it cracking.Microcystis aeruginosa cracking performance phycophage can specific infection cracking microcystis kutz, and thus in wawter bloom is administered, Microcystis aeruginosa cracking performance phycophage can control as " the control algae factor " water bloom of water body that Microcystis aeruginosa causes, and is separating obtained from natural water area.This cracking type phycophage can safe, efficient, fast poisonous Microcystis aeruginosa in cracking fresh water, makes the Microcystis aeruginosa (10 of exponential growth
6~ 10
7individual/ml) cracking death is down to 10
3~ 10
4individual/ml.Comparatively strong to Microcystis aeruginosa host specificity, can not harm be produced to environment and the mankind; Culture condition is simple simultaneously, easy enlarged culturing, can be used as a kind of " control algae product " and means of potential novel improvement blue-green alga bloom.
Poisonous Microcystis aeruginosa cracking phycophage, called after microcystic aeruginosa single stranded DNA phycophage (
microcysticaeruginosasingle-strandDNAcyanophage) MaSSC-P strain, China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved on May 25th, 2015, deposit number is CGMCCNo.10598, depositary institution address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, postcode 100101.
Accompanying drawing explanation
Fig. 1 is the Electronic Speculum figure of MaSSC-P phycophage;
Fig. 2 MaSSC-P nucleic acid is agarose gel electrophoresis figure (swimming lane 1: untreated nucleic acid, swimming lane 2:DnaseI ferment treatment, swimming lane 3:RnaseA ferment treatment, swimming lane 4:S1 strand ferment treatment) after DnaseI enzyme, RnaseA enzyme and S1 single-chain nucleic acid ferment treatment;
Fig. 3 is temperature, ultraviolet, chloroform and the pH impact on MaSSC-P phycophage stability;
Fig. 4 is molten algae situation and the one step growth of phycophage.
Embodiment
Below in conjunction with accompanying drawing embodiment, the present invention is described in further detail.
Embodiment 1
The preparation of phycophage MaSSC-P
Water sample picks up from Ningbo, Ningbo City, Zhejiang Province side the Cultural Park; Host algae is microcystic aeruginosa
m.aeruginosafACHB-925, concrete preparation method is as follows:
(1) separation of phycophage
By top layer, the waters water sample containing poisonous Microcystis aeruginosa cracking phycophage of collection successively through Medium speed filter paper, 0.45 μm and 0.22 μm of nitrocellulose filter filtration; By filtrate and the microcystic aeruginosa being in logarithmic phase by volume the ratio of 1:4 infect and mix; Meanwhile, by the water sample (without phycophage particle) after tangential flow 10KD membrane filtration, infect the microcystic aeruginosa of logarithmic phase in contrast in same ratio; Be 2000lx in intensity of illumination, temperature is 25 DEG C, and light dark period is cultivate under 12h: 12h condition; The nutrient solution that yellow occurs is carried out continuous passage infection until keep stable to bite algae property, obtain phycophage and infect liquid;
(2) amplification cultivation of phycophage and purifying
Phycophage is infected liquid (different gradient concentration 10
-1cell/mL, 10
-2cell/mL, 10
-3cell/mL, 10
-4cell/mL, 10
– 5cell/mL, 10
– 6cell/mL, 10
-7cell/mL); With 10
8~ 10
9after the ratio mixing of the algae liquid of cell/mL 1:9 by volume, incubated at room temperature 30min on shaking table, phycophage and frond are fully adsorbed, then pour into be cooled to 42 DEG C containing quality volume fraction 0.7% agar BG-11 substratum in, pour into after abundant mixing bottom containing quality volume fraction 1.5% agar BG-11 culture medium flat plate culture dish in, treat that it solidifies, inversion is placed in 2000lx, 25 DEG C, cultivate in the incubator of periodicity of illumination: 12h/12h, cultivate after 5 ~ 7 days, observe and record and bite algae spot, what scraping was formed bites algae spot;
(3) phycophage is further purified
The algae spot of biting above-mentioned steps (2) obtained is suspended in BG-11 substratum, after vortex 4 DEG C spends the night, after centrifuging and taking supernatant, repeat above-mentioned steps (2) and bite algae spot and test 2 times, obtain shape, uniformly bite algae spot, then will bite algae spot is suspended in BG-11 substratum, after 0.22 μm of filter membrane centrifuging, adding PEG8000 in filtrate to its quality volume final concentration is 9%(w/v), 4 DEG C of sedimentations are spent the night, at 4 DEG C 36,000
× gcentrifugal 2h, with the washing precipitation of BG-11 substratum, obtains the phycophage suspension of purifying.
The formula of BG-11 substratum is as follows:
The phage of this purifying is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is: CGMCCNo.10598, preservation date is on May 25th, 2015, depositary institution address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, postcode 100101.
Embodiment 2
The morphologic observation of microcystic aeruginosa cracking type phycophage
The phycophage of Example 1 separation and purification, in copper mesh, carries out negative staining with 2% uranyl acetate solution (w/v), then observes the form of phycophage under being placed in electron microscope (Hitachi H-7650).
As shown in Figure 1, this phycophage has head construction in regular polygon and elongated afterbody to result, and head diameter is about 85nm, and afterbody is about 250nm.
The phycophage of purifying is infected with the microcystic aeruginosa of exponential growth and mixes, carry out biting the experiment of algae spot, can obtain size shape homogeneous bite algae spot, around without halo, edge clear is regular.
Embodiment 3
Phycophage genomic nucleic acids type identification
The phycophage particle of separation and purification in Example 1, after extracting nucleic acid, adopt DnaseI, RnaseA and S1 single-chain nucleic acid enzyme effect phycophage genomic nucleic acids respectively, effect collection of illustrative plates as shown in Figure 2, (swimming lane 1: untreated nucleic acid is shown in figure, swimming lane 2:DnaseI ferment treatment, swimming lane 3:RnaseA ferment treatment, swimming lane 4:S1 strand ferment treatment) this phycophage can by DnaseI, single-chain nucleic acid enzymic digestion, and by RnaseA collagenase treatment, and then can not show that phycophage MaSSC-P is the virus particle containing single stranded DNA (ssDNA); Be standard according to " virus taxis-ICTV the 8th time report " that ICTV (ICTV) delivers for 2005, this phycophage does not belong to myovirus section, Styloviridae and Podoviridae, should represent new branch.We are its called after
microcysticaeruginosasingle-strandDNAcyanophage(MaSSC-P).
Embodiment 4
Differing temps, ultraviolet (UV), chloroform and pH are on the impact of MaSSC-P phycophage stability
1, thermostability experiment
The phycophage suspension that Example 1 separation and purification obtains 5 parts is placed in test tube, be placed in 40 DEG C respectively, 45 DEG C, 50 DEG C, in 60 DEG C and 80 DEG C of water-baths, action time should be 0min, l0min, 30min and 1h mutually, at room temperature cools after effect terminates, by the phycophage suspension new BG11 substratum dilution (10 processed
-1, 10
-2, 10
-3, 10
-4, 10
– 5, 10
– 6, 10
-7), carry out biting the experiment of algae spot, and arrange 3 parallel group, be 2000lx in intensity of illumination, temperature is 25 DEG C, and light dark period is cultivate 5 ~ 7 days under 12h:12h condition, observes and records the number biting algae spot.
Result shows, as shown in Figure 3, phycophage is at 40 DEG C, and after 45 DEG C of water bath processing 1h, infective activity is almost constant; Algae spot is bitten by 1.1 × 10 after 50 DEG C of water bath processing
7/ ml drops to 3 × 10
4/ ml; After 60 DEG C of water bath processing, infective activity is lost; Illustrate that phycophage is in physical environment, the rising of temperature can not be very large on its impact.
2, ultraviolet is on the impact of phycophage
The phycophage suspension that Example 1 separation and purification obtains, be placed in culture dish, irradiate under ultraviolet, take out phycophage suspension after irradiating 5min, 10min, 30min, lh, 2h respectively to carry out biting the experiment of algae spot, be 2000lx in intensity of illumination, temperature is 25 DEG C, and light dark period is cultivate 5 ~ 7 days under 12h:12h condition, observes and record.
Result shows, as shown in Figure 3, after UV treatment 2h, the infective activity of phycophage is almost constant, illustrates that phycophage has stronger resistance to ultraviolet ability.
3, phycophage is to the susceptibility of chloroform
Get the phycophage suspension that 5ml embodiment 1 separation and purification obtains, add 0.2ml chloroform (1:25), cultivate 30min after mixing on 4 DEG C of shaking tables, centrifuging and taking liquid phase carries out biting the experiment of algae spot, bites algae spot by observed and recorded, judges that phycophage is to the susceptibility of chloroform.
Result shows, as shown in Figure 3, after chloroform process, phycophage loses infection activity completely, then illustrate that separating obtained phycophage is responsive to chloroform.
4, pH is on the impact of phycophage
PH is adjusted to 3,5,9 and 11 by phycophage suspension that Example 1 separation and purification obtains respectively, after process 30min, pH value is recalled to initial value (about pH7), by the microcystic aeruginosa of the phycophage suspension after process and logarithmic phase by volume 1:4 infect and carry out biting algae spot and test, be 2000lx in intensity of illumination, temperature is 25 DEG C, light dark period is cultivate 5 ~ 7 days under 12h:12h condition, observes and record.
Result shows, as shown in Figure 3, after phycophage processes half an hour under pH is respectively 3 and 11 conditions, bites algae spot number by 1.1 × 10
7/ ml reduces to about 7 × 10
5/ ml (pH11) and 6.7 × 10
4/ ml (pH3), and when pH is 5 and 9, infection activity is almost constant.Illustrate that phycophage MaSSC-P has stronger acid and alkali-resistance ability.
Embodiment 5
The host range of phycophage
By the phycophage that screens respectively to the chroococcoid FACHB-193 forming algal tufa, the floating algae FACHB-1166 that quivers of A Shi, nostoc FACHB-596, the floating algae FACHB-920 that quivers of A Shi, swim the algae FACHB-708 that quivers, Anabaena Flos-aquae FACHB-245, aphanizomenon flos aquae FACHB-1039, microcystic aeruginosa FACHB-469, 905, 924, 925, 942, Microcystis aeruginosa FACHB-915, Hui Shi Microcystis aeruginosa FACHB-908, 929, Microcystis viridis FACHB-979, wawter bloom Microcystis aeruginosa FACHB-1028, magnificent Microcystis aeruginosa 916 carries out infection experiment, detect the specificity of phycophage infection host.Result is as shown in table 1, and phycophage MaSSC-P can infect cracking microcystic aeruginosa FACHB-924,925, Hui Shi Microcystis aeruginosa FACHB-908, and 929, Microcystis viridis FACHB-979, there is Specific lytic effect to Microcystis aeruginosa.
The host range experiment of table 1 phycophage
| Kind | Strain | Result of infection |
| Microcystic aeruginosa | FACHB-905 | - |
| FACHB-942 | - | |
| FACHB-469 | - | |
| FACHB-924 | + | |
| FACHB-925 | + | |
| Hui Shi Microcystis aeruginosa | FACHB-908 | + |
| FACHB-929 | + | |
| Microcystis viridis | FACHB-979 | + |
| Wawter bloom Microcystis aeruginosa | FACHB-1028 | - |
| Magnificent Microcystis aeruginosa | FACHB-916 | - |
| Microcystis aeruginosa | FACHB-915 | - |
| Aphanizomenon flos aquae | FACHB-1039 | - |
| Chroococcoid | FACHB-193 | - |
| The floating algae that quivers of A Shi | FACHB-1166 | - |
| FACHB-920 | - | |
| Nostoc | FACHB-596 | - |
| Swim the algae that quivers | FACHB-708 | - |
| Anabaena Flos-aquae | FACHB-245 | - |
Note: "+": positive; "-": negative.
Embodiment 6
The application of phycophage in the model of small-sized wawter bloom waters
By the microcystic aeruginosa of the phycophage suspension of embodiment 1 separation and purification and logarithmic phase by volume the ratio of 1:4 infect and mix; By the mixed solution not containing phycophage in contrast; Be 2000lx in intensity of illumination, temperature is 25 DEG C, and light dark period is cultivate 5 ~ 7 days under 12h:12h condition, observes and record.After yellow, continue enlarged culturing.After obtaining a certain amount of algae lysate, by biting the infection titer of algae spot measuring lysate, and join in the small-sized wawter bloom waters model of simulation.By the yellow of visual inspection algae liquid; Take out part experimental group and control group respectively every 24h, with blood counting chamber, frustule is counted under microscope; Meanwhile, to experimental group bite algae spot experiment, arrange respectively 3 parallel, observe and record and bite algae spot.
As shown in Figure 4, compared with control group in 7 days, the frustule number generation significant difference of experimental group microcystic aeruginosa, by initial 4.05 × 10 for result
6individual/ml is down to 8.25 × 10
4individual/ml, and control group frustule number rises to 10
8individual/about ml; Meanwhile, algae spot number is bitten as shown in Figure 4 from 1.8 × 10
5individual/ml rises to 9.67 × 10
7individual/ml, the change in conjunction with frustule number can show that viral burst size is approximately 24 units/frustule, and then illustrates that MaSSC-P phycophage has stronger splitting action to microcystic aeruginosa.
Above-mentioned explanation is not limitation of the present invention, and the present invention is also not limited to above-mentioned citing.Those skilled in the art are in essential scope of the present invention, and the change made, remodeling, interpolation or replacement, also should belong to protection scope of the present invention, protection scope of the present invention is as the criterion with claims.
Claims (6)
1. a poisonous Microcystis aeruginosa cracking phycophage, it is characterized in that: this phycophage called after microcystic aeruginosa single stranded DNA phycophage MaSSC-P strain, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on May 25th, 2015, deposit number is CGMCCNo.10598.
2. one according to claim 1 poisonous Microcystis aeruginosa cracking phycophage, is characterized in that the biological property of this phycophage is as follows: MaSSC-P phycophage has head construction in regular polygon and elongated afterbody, head diameter 85nm, the long 250nm of afterbody; Phycophage can be formed on algae flat board size shape homogeneous bite algae spot, around without halo, edge clear is regular; This phycophage can by DnaseI, single-chain nucleic acid enzymic digestion, and can not by RnaseA collagenase treatment.
3. one according to claim 1 poisonous Microcystis aeruginosa cracking phycophage, is characterized in that the biological property of this phycophage is as follows: after 40 ~ 50 DEG C of water bath processing 1h, still keep infection activity; Then inactivation under more than 60 DEG C of conditions; Phycophage, under the environment of pH3.0 ~ 11.0, keeps infection activity; The resistance to ultraviolet of phycophage, responsive to chloroform, under 4 DEG C or-80 DEG C of temperature can be stored in.
4. a separation purification method for poisonous Microcystis aeruginosa cracking phycophage according to claim 1, is characterized in that specifically comprising the steps:
(1) separation of phycophage
By top layer, the waters water sample containing poisonous Microcystis aeruginosa cracking phycophage of collection successively through Medium speed filter paper, 0.45 μm and 0.22 μm of nitrocellulose filter filtration; By filtrate and the microcystic aeruginosa being in logarithmic phase by volume the ratio of 1:4 infect and mix, be 2000lx in intensity of illumination, temperature is 25 DEG C, and light dark period is cultivate under 12h: 12h condition; The nutrient solution that yellow occurs is carried out continuous passage infection until keep stable to bite algae property, obtain phycophage and infect liquid;
(2) amplification cultivation of phycophage and purifying
After ratio phycophage being infected liquid and microcystic aeruginosa liquid 1:9 by volume mixes, incubated at room temperature 30min on shaking table, then pour into be cooled to 42 DEG C containing quality volume fraction 0.7% agar BG-11 substratum in, pour into after abundant mixing in the culture dish of bottom containing the BG-11 culture medium flat plate of quality volume fraction 1.5% agar, treat that it solidifies, be placed in 2000lx, 25 DEG C, cultivate in the incubator of periodicity of illumination: 12h/12h, cultivate after 5 ~ 7 days, what scraping was formed bites algae spot;
(3) phycophage is further purified
The algae spot of biting above-mentioned steps (2) obtained is suspended in BG-11 substratum, after vortex 4 DEG C spends the night, after centrifuging and taking supernatant, repeat above-mentioned steps (2) and bite algae spot and test 2 times, obtain shape, uniformly bite algae spot, then will bite algae spot is suspended in BG-11 substratum, after 0.22 μm of filter membrane centrifuging, adding PEG8000 in filtrate to its quality volume final concentration is that 9%, 4 DEG C of sedimentations are spent the night, at 4 DEG C 36,000
× gcentrifugal 2h, with the washing precipitation of BG-11 substratum, obtains the phycophage suspension of purifying.
5. an application for poisonous Microcystis aeruginosa cracking phycophage according to claim 1, is characterized in that the phycophage suspension of purifying to add in microcystis waterbloom sample, has Specific lytic effect to the Microcystis aeruginosa in water sample.
6. the application of a kind of poisonous Microcystis aeruginosa cracking phycophage according to claim 5, is characterized in that its microcystic aeruginosa to microcystis kutz
m.aeruginosafACHB-924
, M.aeruginosafACHB-925, Hui Shi Microcystis aeruginosa
m.wesenbergiifACHB-908
, M.wesenbergiifACHB-929, Microcystis viridis
m.viridisfACHB-979 all has splitting action.
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| CN109097339A (en) * | 2017-06-21 | 2018-12-28 | 宁波大学 | A kind of cracking performance cyanophage MACPNOA1 and its application |
| CN109097339B (en) * | 2017-06-21 | 2021-12-21 | 宁波大学 | Lytic phycophage MACPNOA1 and application thereof |
| CN111269891A (en) * | 2018-12-05 | 2020-06-12 | 宁波大学 | Broad-spectrum virulent cyanophage Me-ZS1 and application thereof |
| CN113736745A (en) * | 2020-05-27 | 2021-12-03 | 宁波大学 | Nostoc high-efficiency lytic phagophytum YongM and application thereof |
| CN113736745B (en) * | 2020-05-27 | 2023-12-29 | 宁波大学 | Nostoc high-efficiency lytic phaeophaga YonggM and application thereof |
| CN113862228A (en) * | 2021-09-28 | 2021-12-31 | 北京化工大学 | Broad-spectrum virulent cyanophage MinS1 and application thereof |
| CN113862228B (en) * | 2021-09-28 | 2023-11-24 | 北京化工大学 | Broad-spectrum virulent phycophage MinS1 and application thereof |
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