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Mutagenesis, selective breeding and identification method of cold tolerant Danaliella

A technology of Dunaliella gracilis and low temperature resistance, applied in the field of Dunaliella

Inactive Publication Date: 2007-01-03
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The purpose of the present invention is to solve the problems that existing Dunaliella is difficult to mutate and breed at low temperature, and provide a low-temperature-resistant Dunaliella mutant strain and the technical route and identification of using ultraviolet mutagenesis and breeding of low-temperature-resistant Dunaliella method

Method used

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  • Mutagenesis, selective breeding and identification method of cold tolerant Danaliella
  • Mutagenesis, selective breeding and identification method of cold tolerant Danaliella
  • Mutagenesis, selective breeding and identification method of cold tolerant Danaliella

Examples

Experimental program
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Effect test

Embodiment 1

[0053] Ultraviolet mutagenesis is used to cultivate low-temperature-resistant Dunaliella, and the artificial seawater culture solution for cultivating Dunaliella bardawil is prepared first, and the components are grouped in advance to form a mother solution of 50 to 1000 times, and then diluted according to the needs when used. culture medium. The composition of artificial seawater culture solution for preparing and cultivating Dunaliella is shown in Table 1. The artificial seawater culture solution was divided into 100mL Erlenmeyer flasks, each containing 20mL, tightly plugged 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, cool it to room temperature, inoculate 1mL of Dunaliella pasteurii algae liquid into the sterile culture medium, shake it gently, and plug it with a cotton plug. In an incubator a...

Embodiment 2

[0057] Similar to Example 1, the difference is that the artificial seawater culture solution for the cultivation of Dunaliella salina (Dunaliella salina) is prepared, the artificial seawater culture solution is subpackaged in 250mL Erlenmeyer flasks, each bottle is filled with 50mL, sealed with 8 layers of gauze, Wrap in brown 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 5 mL of Dunaliella salina algae liquid into the sterile culture medium, shake it gently, and seal it with 8 layers of gauze. In an incubator at 30°C, the light is 12h, 2000Lx per day, and the darkness is 12h. Dunaliella salina was cultured continuously for 5 days to obtain synchronous growth of Dunaliella salina, that is, a large number of cell divisions occurred in Dunaliella salina from 6 hours after dark culture to 2 hours afte...

Embodiment 3

[0060] Similar to Example 1, the difference is that the artificial seawater culture solution for preparing and cultivating Dunaliella bardawil (Dunaliella bardawil) is divided into 150mL Erlenmeyer flasks with artificial seawater culture solution, and the amount of each bottle is 30mL, and the cotton plug is tightly plugged. Wrap in brown 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, cool it down to room temperature, inoculate 3 mL of Dunaliella pasteurii algae liquid into the sterile culture medium, shake it gently, and plug it with a cotton plug. In an incubator at 25°C, the light is 12h, 3000Lx per day, and the darkness is 12h. Continuously cultured for 3 days to obtain Dunaliella pasteurii with synchronous growth, that is, a large number of cell divisions occurred in Dunaliella pasteraii from 6 hours after dark culture to 2 hours after l...

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Abstract

The ultraviolet ray mutagenesis, selective breeding and identification method of cold tolerant Dunaliella includes: inoculating Dunaliella to culture medium; lighting and dark inducing for synchronized growth; mixing Dunaliella liquid with iodine solution or bromophenol blue solution to deactivate Dunaliella, injecting Dunaliella liquid to culture dish, mutagenesis under ultraviolet lamp before dark culturing, mixing with fresh culture liquid and painting the mixture to Dunaliella culture medium for low temperature lighting culture; inoculating single Dunaliella colony to the culture liquid and low temperature lighting culturing; sampling detection to comparing the low temperature lighting growth curves of cold tolerant Dunaliella mutant and wild prototype strain; extracting total DNA for RAPD comparison; calculating genetic similarity coefficient; extracting total protein and post-electrophoresis staining while record results; comparing protein electropherogram; and confirming the obtained cold tolerant Dunaliella mutant.

Description

technical field [0001] The present invention relates to a kind of Dunaliella, in particular to a kind of unicellular green alga - two species in Dunaliella genus, Dunaliella bardawil (Dunaliella bardawil) and salina (Dunaliella salina), and the use of ultraviolet light to induce Mutation and selection to obtain these two Dunaliella low-temperature-resistant mutant strains and their identification methods. Background technique [0002] Dunaliella sp., also known as salina, is a type of single-cell eukaryotic green algae, which naturally has no cell wall and is easily digested. The ash content in Dunaliella cells is low, rich in glycerin, protein, β-carotene, carbohydrates and a certain amount of unsaturated fatty acids and vitamins. It is one of the ideal health foods and can also be used as high-quality bait and feed for animals , so it has become one of the main cultured microalgae for humans, and has high utilization value in commercial development. The optimum growth te...

Claims

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Application Information

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IPC IPC(8): C12N1/12C12N13/00
Inventor 刘广发周韬陈昱
Owner XIAMEN UNIV
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