Mutagenesis breeding method for high-fat dunaliella salina

A technology of Dunaliella liposum and Dunaliella, which is applied in the field of Dunaliella, and can solve problems such as the absence of high-fat Dunaliella

Inactive Publication Date: 2008-05-07
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there have been data on the use of ultraviolet light to mutate lower organisms, including Dunaliella, there has been no report on obtaining high-fat Dunaliella through mutagenesis and screening.

Method used

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  • Mutagenesis breeding method for high-fat dunaliella salina

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The specific steps for the ultraviolet mutagenesis of Dunaliella and the breeding of Dunaliella with high lipids are as follows:

[0054] 1. Prepare artificial seawater culture solution for cultivating Dunaliella;

[0055] 2. Divide the above-mentioned artificial seawater culture solution into 100mL Erlenmeyer flasks, each with a volume of 20mL, plug it tightly with a cotton plug, and wrap it with kraft paper;

[0056] 3. Place the packaged above-mentioned culture solution in an autoclave, and sterilize at 121°C and a pressure of 0.1mPa for 15 minutes;

[0057] 4. Take out the sterilized culture medium, cool it to room temperature, inoculate 1mL Dunaliella algae liquid, shake it gently, and plug it with a cotton plug;

[0058] 5. Place the culture medium in a light incubator with a light intensity of 8000 lux, a culture temperature of 28°C, a daily light of 24 hours, and rotate and shake at 90 rpm;

[0059] 6. Take out 3.5mL of algae liquid every day, use the artifici...

Embodiment 2

[0071] Similar to Example 1, the difference is:

[0072] The artificial seawater culture solution was divided into 250mL Erlenmeyer flasks, each containing 50mL, sealed with 8 layers of gauze, and wrapped with kraft paper. Place the packaged above-mentioned culture solution 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 5mL Dunaliella algae liquid into the sterile medium, shake it gently, and seal it with gauze. Place the culture solution in a light incubator with a light intensity of 10,000 lux, a culture temperature of 30°C, and light for 24 hours a day, for static culture, and gently shake 1 to 3 times a day for 10 to 20 seconds each time.

[0073] Dunaliella in exponential growth phase (OD 630 Value is 0.545), evenly spread on the agar medium (the NaCl concentration of the used Dunaliella agar medium is increased to 2.5mol / L, and each plate is coated w...

Embodiment 3

[0082] Similar to Example 1, the difference is:

[0083] The artificial seawater culture solution was divided into 200mL Erlenmeyer flasks, each containing 40mL, sealed with cotton plugs, and wrapped with kraft paper. Place the packaged above-mentioned culture solution in an autoclave, and sterilize at 121° C. and a pressure of 0.1 mPa for 15 minutes. Take out the sterilized culture medium, after cooling to room temperature, inoculate 3 mL of Dunaliella algae liquid into the sterile culture medium, shake it gently, and plug it with a cotton plug. The culture solution was placed in a light incubator with a light intensity of 15000 lux, a culture temperature of 25°C, 12 hours of light per day and 12 hours of darkness, static culture, and gently shaking 1-3 times a day, 10-20s each time.

[0084] Dunaliella algae liquid (OD 630 The value is 0.485) 1mL, injected directly into the sterile plate. Place the plate at 13cm under a 30W ultraviolet lamp in an ultra-clean workbench, an...

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Abstract

The invention relates to a mutation breeding method for high lipid dunaliella, in particular to a mutation breeding method to obtain high lipid content dunaliella through ultraviolet mutation and extraction and screening by ether and other reagent, which belongs to dunaliella field; wherein, the obtained high lipid dunaliella is Danaliella bardawil var. HL. The invention is characterized in that: sterilized culture medium is cooled to inoculate dunaliella, and a dunaliella solution is obtained; the dunaliella solution is light-cultured, and the OD630 value is measured; parent strain is coated on the culture medium, and is cultured after ultraviolet radiation mutation; single colonies after mutation treatment is selected and cultured; the culture medium is subject to centrifugal elutriation, and a fresh dunaliella is obtained; the fresh dunaliella is collected and extracted, and the extract is dried and weighted; a mutational strain is selected, and is inoculated with the parent strain for culture; dunaliella cell is collected through centrifugation, and is dried into dunaliella powder; the total lipid content of the dunaliella powder is measured, and is determined higher than that of the mutational strain of the parent strain; lipid in the dunaliella powder of the high lipid mutational strain is subject to secondary screening, and the mutational train is verified to be a high lipid dunaliella mutational train.

Description

technical field [0001] The invention relates to a dunaliella (or called salina), in particular to a method for obtaining dunaliella (salina) with high lipid content through ultraviolet mutagenesis, extraction and screening with reagents such as ether. Background technique [0002] Dunaliella sp. is a single-celled eukaryotic green alga belonging to the class Chlorophyceae, Volvocales, and Dunaliellaceae. Dunaliella has the characteristics of high photosynthetic efficiency, strong environmental adaptability, short growth cycle and high biological yield. Dunaliella is rich in β-carotene, glycerol and protein, and its total lipid content accounts for about 15% of dry weight (including various fatty acids and their derivatives and compounds). If a mutant strain with a significantly higher total lipid content than the original strain can be obtained by mutagenesis, Dunaliella hyperlipidemia will provide cheap raw materials for the pyrolysis of biomass oil and combustible gas. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/12C12N15/01
Inventor 刘广发陈昱贾立山林锦明
Owner XIAMEN UNIV
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