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Mutagenic breeding method of high temperature resistant pasteur Du algae

A Dunaliella pasteuria high-temperature-resistant technology, applied in the field of single-cell green algae, can solve the problem that Dunaliella can not tolerate high temperature, etc., and achieve the effect of prolonging the open-air cultivation time and improving production efficiency

Inactive Publication Date: 2006-04-12
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to aim at the defect that the existing Dunaliella cannot tolerate high temperature, and provide a method for breeding high-temperature-resistant Dunaliella pasteurii by ultraviolet mutagenesis and its identification method

Method used

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  • Mutagenic breeding method of high temperature resistant pasteur Du algae
  • Mutagenic breeding method of high temperature resistant pasteur Du algae
  • Mutagenic breeding method of high temperature resistant pasteur Du algae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] The specific steps for mutagenizing and cultivating high temperature-resistant Dunaliella pasteurii with ultraviolet rays are as follows:

[0062] 1. Preparation of artificial seawater medium for cultivating Dunaliella pasteurii;

[0063] 2. Divide the above-mentioned artificial seawater medium into 100mL Erlenmeyer flasks, each containing 20mL, plug it tightly with a cotton plug, and wrap it with kraft paper;

[0064] 3. Place the packaged above-mentioned culture medium in an autoclave, sterilize at 121°C and a pressure of 0.1mPa for 15 minutes;

[0065] 4. Take out the sterilized culture medium, after cooling to room temperature, inoculate 1mL of Dunaliella pasteurii algae liquid into the sterile medium, shake it gently, and plug it with a cotton plug;

[0066] 5. Place the culture medium in a light incubator, light intensity 1000lux, culture temperature 20°C, daily light time 12h, rotate and shake, 90rpm;

[0067] 6. Take out 0.3mL of algae liquid every day and mix...

Embodiment 2

[0082] Similar to Example 1, the difference is that the artificial seawater culture medium is divided into 250mL Erlenmeyer flasks, each containing 50mL, sealed with 8 layers of gauze, and wrapped with kraft paper. Place the packaged medium above in an autoclave, and sterilize at 121° C. and a pressure of 0.1 mPa for 20 minutes. Take out the sterilized culture medium, cool it down to room temperature, inoculate 5 mL of Dunaliella pasteurella liquid into the sterile culture medium, shake it gently, and seal it with gauze. The culture solution was placed in a light incubator with a light intensity of 4000 lux, a culture temperature of 30°C, and 24 hours of light per day, for static culture, and gently shaken 1 to 3 times a day for 10 to 20 seconds each time.

[0083] The next day, take out 0.3mL of algae liquid and mix with an equal amount of bromophenol blue solution to inactivate Dunaliella pasteurii. Immediately take a sample and calculate the density of Dunaliella pasteurii...

Embodiment 3

[0093] Similar to Example 1, the difference is that the artificial seawater culture medium is divided into 200mL Erlenmeyer flasks, each containing 40mL, sealed with a cotton plug, and wrapped with kraft paper. Place the packaged medium above in an autoclave, and sterilize at 121° C. and a pressure of 0.1 mPa for 15 minutes. Take out the sterilized culture medium, cool it down to room temperature, inoculate 3 mL of Dunaliella pasteurella liquid into the sterile medium, shake it gently, and stuff it with a cotton plug. The culture solution was placed in a light incubator with a light intensity of 3000 lux, a culture temperature of 25°C, and 24 hours of light per day, for static culture, and gently shaken 1 to 3 times a day for 10 to 20 seconds each time.

[0094] Take out 0.2mL of algae liquid every day and mix it with an equal amount of iodine solution to inactivate Dunaliella pasteurii. Immediately take a sample and read it on a hemocytometer to calculate the density of Duna...

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Abstract

A method for mutagenizing and selectively culturing a refractory single-cell green alga includes such steps as sterilizing and cooling culture medium, inoculating alga liquid, culturing, mixing with iodine solution or bromophenol blue, deactivating, calculating alga density in alga liquid, ultraviolet mutagenizing, dark culturing, mixing the cultured alga liquid with fresh culture medium, culturing, high-temp screening, amplifying culture of living alga cells, culturing under light radiation, inoculating yellow alga, culturing, testing its mutagenized effect, measuring the long and short diameters of cell, extracting DNA, random amplifying of polymorphic DNA, calculating genetic similarity coefficient, extracting H-42 and general protein, electrophoresis, dyeing with Coomassic brilliant blue, recording result and observing the electropherogram to obtain result.

Description

technical field [0001] The invention relates to a single-cell green algae, in particular to a method for obtaining high-temperature resistant Dunaliella pasteurii through ultraviolet mutagenesis and an identification method thereof. Background technique [0002] Dunaliella bardawil is a single-celled eukaryotic green alga belonging to the class Chlorophyceae, Volvocales, Dunaliellaceae, and the genus Dunaliella. Dunaliella naturally lacks a cell wall and is one of the few extremely salt-tolerant eukaryotes discovered so far. Dunaliella is rich in β-carotene, glycerol and protein, so it has become one of the main cultured algae for human beings and has high utilization value in commercial development. The optimal growth temperature of Dunaliella is 25-30°C. When the temperature exceeds 35°C, the growth is completely inhibited and the cells gradually die. Our country is hot and hot in summer, especially in the south, so it is difficult to cultivate Dunaliella in the open air...

Claims

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Application Information

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IPC IPC(8): C12N1/12C12N13/00C12Q1/04C12Q1/68C12R1/89
Inventor 刘广发黄瑞芳
Owner XIAMEN UNIV
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