High-growth-rate Dunaliella tertiolecta obtained through ethylmethane sulfonate mutation breeding

A technology of ethyl methyl sulfonate and growth rate, applied in the field of microbial engineering, can solve the problems of less breeding research, low probability of mutant strains, time-consuming and labor-intensive

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

However, there are few breeding studies on its growth rate.
At present, the mutation breeding of Dunaliella mainly uses ultraviolet mutagenesis, but ultraviolet mutagenesis has the disadvantages of time-consuming and labor-intensive, and the probability of obtaining mutant strains is small.
Ethyl methanesulfonate (EMS), as a strong chemical mutagen, is mainly used in the mutagenesis breeding of Spirulina in algae breeding, but it has not been reported for the growth rate screening of fast-growing algal strains

Method used

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  • High-growth-rate Dunaliella tertiolecta obtained through ethylmethane sulfonate mutation breeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Screening of Dunaliella mutagenic strains with high growth rate

[0059] The wild strain of Dunaliella tertiolecta UTEX LB 999 (purchased from the University of Texas) was used as the starting algal strain, added ethyl methanesulfonate (EMS) (1% final concentration, volume ratio) for mutagenesis, and then isolated and subcultured for preservation. After activation of the above-mentioned mutagenic algae strains, they were screened in multiple steps under normal culture conditions (25°C, 4000lux, 12h L / 12h D) in an artificial climate box, and the OD 750 The value is to measure the mutagenized algae strains with high growth rate step by step.

[0060] The composition and content of the culture medium are shown in Table 1.

[0061] The composition and content of table 1EM medium

[0062] #

Element

quantity

Stock solution concentration

Final concentration at the time of use

1

Artificial Seawater

3L

2 ...

Embodiment 2

[0077] Example 2 The identification characteristics of the mutant algae strain ENN0001-7 obtained by screening

[0078] The algal cells of the mutant algal strain ENN0001-7 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

[0079] Example 3 Mutant algal strain ENN0001-7 obtained from screening and wild-type algal strain UTEX LB 999 Comparison of growth rate and chlorophyll content of

[0080] 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 liqui...

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Abstract

The invention relates to a method adopting ethylmethane sulfonate (EMS) to perform chemomorphosis 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 ENN0001-7 with higher growth rate compared with a wild strain. Compared with the wild strain, the chlorophyll content (OD680) of the mutagenic strain obtained by the method is increased by 8.9 percent, and the biomass (OD750) is increased by 5.9 percent. The strain with stable and high growth rate obtained by the method provides a valuable germ plasm resource for mass production.

Description

technical field [0001] The present invention relates to the field of microbial engineering, in particular, the present invention relates to a method for using ethyl methanesulfonate to induce and breed Dunaliella, and a Dunaliella mutagenic strain ENN0001 with a high growth rate obtained by said method -7. Background technique [0002] Dunaliella is a kind of unicellular eukaryotic green algae without cellulose outer wall. They are mostly found in salt lakes and oceans, although they have been reported to have been found in fresh water. Dunaliella resembles Chlamydomonas, and domestic scholars classify them into the genus Euchlorophyceae, Volvox, Salinaceae, and Dunaliella. Dunaliella cells are very tiny. Judging from the dozen species of Dunaliella that have been reported so far. The largest cell is D. salina, with a volume of about 2600 μm 3 (24~25×12~16μm), the smallest is D.minuta only about 42μm 3 (9×3μm), its size is only a fraction to one hundredth of that of a ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/12C12N15/01C12P23/00C12P17/04C12P17/06C12P7/22C12P7/64C12R1/89
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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