Separation culture method of sponge symbiosis blue alga

A technology of sponge symbiotic cyanobacteria, separation and culture, which is applied in the field of cyanobacteria separation to achieve ideal effects, simple separation and purification operations, and high purity.

Active Publication Date: 2014-06-11
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, at present, there is no published research report about obtaining sponge symbiotic cyanobacteria through rapid separation methods at home and abroad, and then carrying out monoculture or pure culture.

Method used

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  • Separation culture method of sponge symbiosis blue alga
  • Separation culture method of sponge symbiosis blue alga

Examples

Experimental program
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Effect test

Embodiment 1

[0022] A method for separating and cultivating sponge symbiotic cyanobacteria, specifically comprising the following steps:

[0023] (1) Screening of sponge samples: screening intact sponge samples with blue-green or yellow color from the ocean;

[0024] (2) Pretreatment of sponge samples: soak the screened sponge samples in boiled and cooled sterilized seawater, then remove debris and wash them, and then cut the sponge samples into 1cm 3 The size of the sponge block is placed in the culture bottle, and at the same time, sterilized seawater containing kanamycin or neomycin is added to the culture bottle until the lower half of the sponge block is submerged in the water, and the upper part is exposed to the water surface, and then Place the culture bottle in an environment with a carbon dioxide volume fraction of 0.05%, and use 200 μmol / (m 2 s) light intensity, pretreatment 36h;

[0025] (3) Sponge sample selection: cut the pretreated sponge block into 0.5cm 3 The size of th...

Embodiment 2

[0033] With above-mentioned embodiment 1, its difference is:

[0034] Sponge sample cut into 0.5cm in step (2) 3 The size of a sponge block; place the culture bottle in an environment with a carbon dioxide volume fraction of 0.035%, and use 150μmol / (m 2 s) light intensity, pretreatment 48h;

[0035] In step (3), the pretreated sponge block is cut into 0.25cm 3 size of sponge block;

[0036] In step (4), put the sponge block in a food grinder, and mash it twice for 20 seconds at a speed of 1200 rpm;

[0037] In step (5), the cell culture plate was placed in the incubator, and the light intensity was 20 μmol / (m 2 s), cultivated for 15 days at an ambient temperature of 20°C;

[0038] In step (6), dilute the sterilized seawater containing cyanobacteria algae by 10 times;

[0039] In step (7), the cell culture plate was placed in a light intensity of 20 μmol / (m 2 · s), the light cycle is 12 / 12 hours, and the water temperature is 20° C. for 15 days.

[0040] The concentration ...

Embodiment 3

[0042] With above-mentioned embodiment 1, its difference is:

[0043] Sponge sample cut into 1.5cm in step (2) 3 The size of a sponge block; the culture bottle is placed in an environment with a carbon dioxide volume fraction of 0.065%, and under the high temperature stress condition of 32°C, 250μmol / (m 2 s) light intensity, pretreatment 24h;

[0044] In step (3), the pretreated sponge block is cut into 0.75cm 3 size of sponge block;

[0045] In step (4), put the sponge block in a food grinder, and mash it 4 times at a speed of 1500 rpm for 10 seconds each time;

[0046] In step (5), the cell culture plate was placed in the incubator, and the light intensity was 50 μmol / (m 2 s), the photoperiod is 12 / 12 hours, and the ambient temperature is 25°C for 10 days;

[0047] In step (6), the sterilized seawater containing cyanobacteria is diluted 20 times;

[0048] In step (7), place the cell culture plate at a light intensity of 50 μmol / (m 2 ·s), cultivated for 10 days under t...

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Abstract

The invention discloses a separation culture method of sponge symbiosis blue algae. The method is characterized by comprising the following steps: selecting a sample with blue-green or yellow sponge from ocean, soaking the sponge sample, subsequently removing the impurities and then cutting into pieces, adding the sponge pieces and sterilized seawater containing antibiotics into a culture bottle till the lower parts of the sponge pieces are soaked in the water and the upper parts are exposed out of the water level, further treating the culture bottle in an carbon dioxide environment under certain temperature and illumination conditions, cutting the sponge pieces into pieces, taking the sponge pieces of dark color, stirring the sponge pieces, then mixing the sponge pieces with the sterilized seawater, putting into a cell culture plate for culturing till blue alga colonies grow out from the cell culture plate, diluting the sterilized seawater with the blue alga colonies, selecting a microsphere with only one algal filament till each hole of the cell culture plate contains the microsphere with one algal filament, and performing temporary rearing on the cell culture plate so as to obtain a product. The method has the advantages that a great deal of symbiosis glue algae can be rapidly separated with good culture effect and operation is convenient.

Description

technical field [0001] The invention relates to a method for isolating cyanobacteria, in particular to a method for isolating and culturing sponge symbiotic cyanobacteria. Background technique [0002] Sponges are important members of benthic communities, widely distributed in temperate, subtropical, tropical and polar regions, and their abundance, diversity and distribution patterns are diverse in different regions. The sponge itself has specific functions, so it has multiple roles in the marine ecosystem, and can also provide a large number of novel biologically active substances. The unique and important ecological functions of sponges, rich biodiversity, and symbiotic relationship with microorganisms can be used as resources for new natural products, thus becoming a research hotspot in marine biology. For example, the invention patent "sponge anti-tumor compound" with patent publication number CN1429203 discloses a new anti-tumor compound isolated from sponges. Publica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
Inventor 何山骆其君严小军张金荣蒋莹
Owner NINGBO UNIV
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