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Nostoc high-efficiency cracking phycophage YongM and application thereof

A technology of cyanophage and Nostoc, which is applied in the field of high-efficiency and potent cyanophage of Nostoc, can solve the problems of changing the photosynthesis center of the host, reducing the incidence of photosynthesis, and affecting the primary productivity of water bodies, achieving good development prospects and high replication rate, the effect of high infection rate

Pending Publication Date: 2021-12-03
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invasion of cyanophages will also change the photosynthesis center of the host, reduce the incidence of photosynthesis, and indirectly affect the primary productivity of the water body

Method used

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  • Nostoc high-efficiency cracking phycophage YongM and application thereof
  • Nostoc high-efficiency cracking phycophage YongM and application thereof
  • Nostoc high-efficiency cracking phycophage YongM and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Preparation of cyanophage YongM

[0034] (1) Cultivation of Nostoc sp.FACHB-596

[0035] Take 5mL of Nostoc sp.FACHB-596 algae liquid, dilute it into a conical flask filled with 500mL of BG11 liquid medium at a volume ratio of 1:100, and place it at a temperature of 25°C, with a light intensity of 2000lx and a light-dark cycle of In a 12h:12h light incubator, the logarithmic phase FACHB-596 algae liquid can be obtained in about 10 days.

[0036] (2) Enrichment and screening of cyanophages

[0037] Water sample treatment: Collect surface water samples from various places in Dianchi Lake, Yunnan, and bring them back to the laboratory for processing as soon as possible. Water samples were centrifuged at 12,000 g for 20 minutes at 4°C to remove precipitates. Take the supernatant for the experiment.

[0038]Take a number of sterile Erlenmeyer flasks corresponding to the number of samples, and add to each Erlenmeyer flask: 80mL of the above-treated water sample supernatan...

Embodiment 2

[0055] Morphological observation of cyanophage YongM

[0056] Take the cyanophage-algal culture lysate separated and purified in Example 1 and centrifuge at low speed (4°C, 5000g, 5min) to discard the precipitate, distribute the supernatant evenly to two tubes, and inject slowly and gently from the bottom with a syringe 5-10 mL of 20% (w / v) sucrose solution was centrifuged at 35,000 g at 4°C for 1 h, and the supernatant was discarded. Gently add 0.01M PBS, discard PBS, add 0.01M PBS again and place at 4°C overnight to suspend the precipitate. Use a pipette gun to take a drop of cyanophage suspension onto the copper grid, let it stand for 10 minutes, and then use neutral filter paper to absorb excess water from the side. Drop a drop of 3% uranyl acetate on the copper grid, and after staining for 20 seconds, use neutral filter paper to absorb the staining agent from the side. After standing for 10 minutes to dry, the morphology of cyanophages was observed with a transmission e...

Embodiment 3

[0059] Sequence Alignment of DNA Polymerase Genes of Phage YongM

[0060] Genome extraction: Add DNase I and RNase A to the cyanophage YongM suspension to a final concentration of 1 μg / mL, digest overnight at 37°C, and inactivate at 80°C for 15 minutes. Lysis solution (0.5% SDS, 50 μg / mL proteinase K, 20 nM EDTA, both final concentrations) was added to the system, and incubated at 56° C. for 1 h. Add an equal volume of equilibrated phenol, and after gentle shaking, centrifuge at 10000g for 5min at 4°C. The upper layer was collected, and an equal volume of phenol-chloroform-isoamyl alcohol (25:24:1) was added, and after gentle shaking, it was centrifuged at 10,000 g for 5 min at 4°C. Collect the upper layer liquid, add an equal volume of chloroform, mix well, centrifuge at 10000g for 5min, collect the upper layer liquid, and repeat 2 times. Add an equal volume of isopropanol, place at -20°C for at least 30min, centrifuge at 10,000g for 20min at 4°C, and wash the precipitate t...

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Abstract

The invention discloses a nostoc high-efficiency cracking phycophage as well as a separation method and application thereof. The nostoc high-efficiency cracking phycophage is characterized in that the phycophage is a novel Myoviridae phycophage, is named as YongM, is preserved in the General Microbiological Center of Microbiological Culture Collection Center on January 13, 2020, and is numbered as CGMCC NO.18383. The YongM has the characteristics of high efficiency and broad spectrum, can be used for efficiently splitting nostoc and can be used for infecting and splitting various microcystis, Chroococcus, Anabaena aquatica, oscillatoria planktonic and floating hylonema adami. According to the nostoc high-efficiency cracking phycophage, with the application of the YongM, the algae liquid of the nostoc FACHB-596, the algae liquid of the anabaena flos-aquae FACHB-245, the algae liquid of the Chroococcus FACHB-193, the oscillatoria planktonic FACHB-708 can be subjected to cracking clarification within 24 h.

Description

technical field [0001] The invention relates to a cyanophage of cyanobacteria, in particular to YongM, a high-efficiency and potent cyanophage of Nostoc and its application. Background technique [0002] In recent years, with the development of my country's economy and the increase of human activities, the degree of eutrophication of water bodies and algal blooms have become increasingly serious, and they have leapt to the forefront of the world. The prevention and control of algae blooms is very urgent. It can be divided into mechanical removal method, aeration mixing method, nutrient control method, chemical method, hydrodynamic control method and biological control method. Among them, the biological control method using microorganisms to remove algae has broad application prospects. [0003] Cycophages are a group of viruses that specifically infect prokaryotic algae. The cyanophage is a constituent factor of the ecological environment and is closely related to the life a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/02A01N63/40A01P13/02
CPCC12N7/00A01N63/40C12N2795/10121C12N2795/10151
Inventor 秦伟南李登峰许丽华林威童贻刚
Owner NINGBO UNIV
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