High-growth-rate Dunaliella tertiolecta obtained through ultraviolet mutation breeding

A technology of growth rate and Dunaliella, which is applied in the field of microbial engineering, can solve the problem that the screening research of growth rate is blank and so on

Active Publication Date: 2009-12-09
ENN SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ultraviolet breeding technology has been widely reported in algae. It has been reported that ultraviolet rays play a big role in the mutation breeding of chlorella and spirulina, but there are few reports in Dunaliella. Screening studies were also blank

Method used

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  • High-growth-rate Dunaliella tertiolecta obtained through ultraviolet mutation breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Mutagenesis Breeding of High Production Rate Dunaliella Mutants

[0055] The wild strain of Dunaliella tertiolecta UTEX LB 999 (purchased from the University of Texas) was used as the starting algal strain for ultraviolet mutagenesis, and then isolated and subcultured for preservation. After activation of the above-mentioned mutagenized algae strains, they were screened in an artificial climate box under normal culture conditions (25°C, 4000lux, 12hL / 12hD), and the OD 750 The value is to measure the mutagenized algae strains with high growth rate step by step.

[0056] The composition and content of the culture solution are shown in Table 1. When preparing the culture solution, some components are added to distilled water in solid form, and some components will be made into a mother solution of 50-1000 times in advance, and then diluted as needed when using Prepared as a culture medium.

[0057] Table 1 Composition and content of EM medium

[0058] ...

Embodiment 2

[0072] Identification characteristics of mutant algal strain ENN0001-6 obtained by screening in Example 2

[0073]The algal cells of the mutant algal strain ENN0001-6 were pear-shaped or oval, about 6-10 μM in length, with 2 equal-length flagella, no cellulosic cell wall, and only an outer membrane composed of glycoprotein and neuraminic acid. It can grow in EM, F / 2 and other media. The algae are evenly dispersed in the liquid medium and can swim. It grows well on EM solid medium, and obvious single algae colonies can be formed in 5-7 days. The algae fall into a regular circle, and the algae are green.

Embodiment 3

[0074] Example 3 Comparison of production rate of mutant algal strain ENN0001-6 obtained by screening and wild-type algal strain

[0075] Depend on figure 1 It can be seen that the biomass (OD 750 ) and chlorophyll content (OD 680 ). The normal culture conditions are: 25°C, 4000lux, 12h L / 12h D. The re-screening steps are as follows: the mutagenized algae strains determined by the primary screening are activated by transferring to a solid medium plate once, and a single algal colony is picked and inoculated into a 100ml Erlenmeyer flask added with 20ml EM medium. Cultured for 10 days under normal culture conditions, measured OD 750 value, and adjust the same OD 750 The value is 0.1, inoculated and subcultured in a new 100ml Erlenmeyer flask added with 20ml EM medium, and three parallel samples were set up for each plant. After ten days of culture under normal culture conditions, measure the OD 750 Value, and take 10ml of algae liquid and add 90ml of EM medium (1:10 i...

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Abstract

The invention relates to a method adopting ultraviolet to perform mutagenesis on Dunaliella tertiolecta, which performs screening by using the growth rate of microalgae as an index under a normal culture condition to finally obtain a mutagenic strain ENN0001-6 with higher growth rate compared with a wild strain. After re-screening culture, the biomass (OD750) of the mutagenic strain is increased by 15 percent compared with the wild strain, and the chlorophyll content (OD680) is increased by 1 percent compared with the wild strain.

Description

technical field [0001] The present invention relates to the field of microbial engineering, in particular, the present invention relates to a method for breeding Dunaliella by ultraviolet mutagenesis, and a Dunaliella mutant strain ENN0001-6 with high growth rate obtained by the method. Background technique [0002] Dunaliella tertiolecta belongs to Chlorophyta, Chlorophyceae, Chlamydomonadales, and Dunaliellaceae. It has the characteristics of single cell, no cell wall and small genome. Dunaliella naturally lacks cell walls and is one of the few extremely salt-tolerant eukaryotes discovered so far. Dunaliella is a widely distributed unicellular algae in the genus Dunaliella, with only a thin, elastic outer membrane composed of glycoproteins and neuraminic acid surrounding the protoplasts. Dunaliella contains high beta-carotene, the content of carotenoid is as high as 14% of dry weight, and the production of glycerol increases with the increase of the salinity of the extern...

Claims

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Application Information

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IPC IPC(8): C12N1/12C12N15/01C12N13/00C12P23/00C12P17/04C12P17/06C12P7/64C12R1/89
CPCY02E50/13Y02E50/10
Inventor 吴洪尹顺吉王媛媛邓平蔡忠贞袁普卫
Owner ENN SCI & TECH DEV
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