Skeletonema costatum lytic virus and its separation method and use

A technology for splitting viruses and sclerophytes, which is applied in the field of algae splitting viruses, can solve the problems of undiscovered algae viruses, and achieve the effects of no environmental pollution, good development prospects, and low cost

Active Publication Date: 2016-01-20
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since Nagasaki et al. first discovered the root canal algae virus in 2004, 16 strains of diatom viruses ...

Method used

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  • Skeletonema costatum lytic virus and its separation method and use
  • Skeletonema costatum lytic virus and its separation method and use
  • Skeletonema costatum lytic virus and its separation method and use

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Isolation and Purification of Lytic Virus ScssDNAV from Sclerotina costatum

[0024] The surface water samples were collected from Ninghai Shuangpantu Aquaculture Farm in Ningbo City, Zhejiang Province, and were stored under ice baths. After the water samples were passed through medium-speed filter paper, 0.45 μm, and 0.22 μm filter membranes in turn, they were concentrated into concentrated water samples by an ultrafiltration system. Its concentration factor is 100 times.

[0025] Concentrated water samples were prepared and mixed with Skeletalum costarum NMBguh004-2 in the exponential growth phase at a volume ratio of 1:4, and then cultured in a constant temperature light incubator for 7 days at a temperature of 20°C, light intensity of 2500LX, and light-to-dark ratio (L / D) 12h:12h, obtain the Sclerostetrascens lysate. The virus-free concentrate was co-cultured with Skeletalum costatum in exponential growth phase as a control group. The growth of algae was observed ...

Embodiment 2

[0030] Observation of Morphology of Lysing Virus of Sclerosteina costatum by Light Microscope and Electron Microscope

[0031] The purified virus was used to infect S. costarum NMBguh004-2. After 4 days, the color of the algae liquid became lighter. The algal cells were observed under an optical microscope, and most of the algal cells were found to be lysed.

[0032] Take the isolated and purified Skeletalum costa lysate virus isolated and purified in Example 1, drop it on the copper grid, carry out negative staining with 2% uranyl acetate solution (W / V), and then observe it under an electron microscope (Hitachi H-7650) Morphology of the Skeletalum costa lytic virus.

[0033] The result is as figure 1As shown, the lysed viral particles of Sclerostena costa are spherical, with a diameter of 98±2nm, no flagella and no outer membrane coating.

Embodiment 3

[0035] Effects of different pH, temperature, ultraviolet rays, and freeze-thaw on the stability of Skeletons costa lytic virus ScssDNAV

[0036] (1) Effects of different pH on the stability of ScssDNAV, a lysing virus of Sclerotina costatum

[0037] Take 2 parts of the fresh virus solution obtained by separation and purification in Example 1, measure the original pH value of the virus solution and record it, then use 0.1M NaOH and 0.1M HCl to adjust the corresponding pH=3 and pH=10 of the virus solution and treat them for 30 minutes respectively as the experimental group, Use 0.1M NaOH and 0.1M HCl to adjust the water into pH=3 and pH=10 aqueous solutions as the control group; take 5mL algae liquid to extract chlorophyll a on the 0th, 4th and 10th day respectively, and detect the concentration of chlorophyll a in the experimental group and the control group Variety.

[0038] The results show that if figure 2 As shown, the lysed virus liquid of Sclerotina costarum was treate...

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Abstract

The invention discloses a skeletonema costatum lytic virus and its separation method and use. The skeletonema costatum lytic virus is a skeletonema costatum single chain DNA lytic virus ScssDNAV, is preserved in the China general microbiological culture collection center (CGMCC) Oon May 25, 2015, has a preservation number of CGMCC No. 10599, is used for inhibiting skeletonema costatum growth and killing skeletonema costatum, has strong pyrolysis effects on skeletonema costatum NMBguh004-1 and NMBguh004-2, realizes safe, effective, fast and specific degradation of skeletonema costatum in sea water and has an algae chlorophyll a concentration reduction rate of 76.14%.

Description

technical field [0001] The present invention relates to an algal lytic virus, in particular to a Skeletalum costa lytic virus and its separation method and application. Background technique [0002] Red tide is a recognized marine environmental disaster, which is very destructive to marine fishery production and marine ecological balance. Red tide algae enter the gills of fish through the water flow, causing fish to suffocate and die; some red tide algae also secrete toxins, which will cause fish death after swallowing. In addition, after the death of the algae, a large amount of dissolved oxygen in the water will be consumed during the decomposition process, resulting in the death of a large number of marine organisms due to lack of oxygen, resulting in deterioration of water quality and affecting the rational use of water resources. At present, the algae removal methods at home and abroad are mainly divided into three categories: physical method, chemical method and biolo...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N7/02C02F3/34
Inventor 吴寒华李登峰严小军周成旭刘联国
Owner NINGBO UNIV
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