Toxic Microcystis cracking cyanophage and separation method and application thereof

A technology of cyanophage and Microcystis, which is applied in the field of pyrolysis and separation of toxic Microcystis cyanophage, can solve the problems of costing a lot of manpower and material resources, and chemical agent pollution, etc. simple effect

Active Publication Date: 2015-12-23
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the physical method is quick to control algae, it needs a lot of manpower and material resources
The chemical method main

Method used

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  • Toxic Microcystis cracking cyanophage and separation method and application thereof
  • Toxic Microcystis cracking cyanophage and separation method and application thereof
  • Toxic Microcystis cracking cyanophage and separation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Preparation of cyanophage MaSSC-P

[0027] The water sample was collected from Ningbo Bang Cultural Park, Ningbo City, Zhejiang Province; the host algae was Microcystis aeruginosa M. aeruginosa FACHB-925, the specific preparation method is as follows:

[0028] (1) Isolation of cyanophages

[0029] The collected surface water samples containing toxic Microcystis lysed cyanophages were filtered through medium-speed filter paper, 0.45 μm and 0.22 μm nitrocellulose filter membranes; the filtrate was mixed with Microcystis aeruginosa in logarithmic growth phase by volume Infection mixture at a ratio of 1:4; at the same time, the water sample (without cyanophage particles) filtered through a tangential flow 10KD membrane was infected with Microcystis aeruginosa in the logarithmic growth phase at the same ratio as a control; The temperature is 2000lx, the temperature is 25°C, and the light-dark cycle is 12h:12h; the yellowed culture solution is continuously subcultured unti...

Embodiment 2

[0038] Morphological Observation of Lysing Cycphages of Microcystis aeruginosa

[0039] Take the cyanophage isolated and purified in Example 1 on the copper grid, negatively stain with 2% uranyl acetate solution (w / v), and then observe the cyanophage under an electron microscope (Hitachi H-7650). form.

[0040] The result is as figure 1 As shown, the cyanophage has a regular polyhedral head structure and a slender tail, the diameter of the head is about 85nm, and the length of the tail is about 250nm.

[0041] The purified cyanophages were infected and mixed with exponentially growing Microcystis aeruginosa, and the algal plaque experiment was carried out, and the algal plaques with uniform size and shape could be obtained, without halos around, and clear and regular edges.

Embodiment 3

[0043] Nucleic acid type identification of cyanophage genome

[0044] Take the cyanophage particles separated and purified in Example 1, and after extracting the nucleic acid, use DnaseI, RnaseA and S1 single-stranded nuclease to act on the cyanophage genome nucleic acid respectively, and the action spectrum is as follows figure 2 As shown in the figure (lane 1: untreated nucleic acid, lane 2: DnaseI enzyme treatment, lane 3: RNaseA enzyme treatment, lane 4: S1 single-chain enzyme treatment) the cyanophage can be digested by DnaseI and single-chain nuclease , and cannot be digested by RNaseA enzyme, and then it is concluded that cyanophage MaSSC-P is a virus particle containing single-stranded DNA (ssDNA); The Eighth Report" is the standard, and the cyanophage does not belong to Myoviridae, Longoviridae and Brachyviridae, and should represent a new subfamily. we named it Microcystica eruginosa single-strand DNA cyanophage (MaSSC-P).

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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 [0001] The invention relates to a toxic Microcystis lytic cyanophage, in particular to a toxic Microcystis lytic cyanophage and its separation method and application. Background technique [0002] In recent years, human economic activities have become more and more serious, and various domestic sewage and production wastewater have been discharged into slow-flowing water bodies such as lakes, estuaries, and bays, causing excessive nutrients such as nitrogen and phosphorus in the water body to become eutrophic. The algae that causes is multiplied in great numbers, and the occurrence of " water bloom " becomes more and more serious, and wherein blue-green algae becomes the main alga that causes water bloom outbreak. "Water bloom" is one of the most serious environmental disasters in the world today. Among them, my country is one of the countries with the most serious outbreak of water bloom. About 75% of the lakes are facing different degrees of eutrophication ...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N7/02C02F3/32
Inventor 刘飞李登峰严小军刘联国吴寒华顾叶华陈梅娟
Owner NINGBO UNIV
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