Dunaliella tertiolecta mutant strain with high growth rate and complex mutation breeding method thereof
A compound mutagenesis and Dunaliella technology, applied in the field of microbial engineering, can solve the problems of short screening time, prone to reverse mutation, poor genetic stability of mutant algal strains, etc.
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Embodiment 1
[0062] The screening of the Dunaliella mutagenic strain of embodiment 1 high growth rate
[0063] 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 exposed to high temperature and high light outdoors (temperature 37°C to 45°C, light intensity 10000Lux-50000Lux), and the growth was good. Save after isolation and subculture. After activation of the above-mentioned mutagenic algae strains, they were subjected to multi-step screening in an artificial climate box under normal culture conditions (25°C, 4000lux, 12h L / 12h D), using a WFZ UV-2102PC UV-Vis spectrophotometer (purchased from U.S. Niko Instrument Co., Ltd.) measured the OD value at 750nm of the algae liquid as a measure, and screened out the mutagenized algae strains with high growth rate step by step.
[0064] The composition and content of the culture solution are shown in Table 1. When preparing t...
Embodiment 2
[0080] Identification characteristics of the mutant algae strain ENN0001-1 obtained by screening in embodiment 2
[0081] The algal cells of the mutant algal strain ENN0001-1 are pear-shaped or oval, about 6-10 μM in length, with 2 equal-length flagella, no cellulosic cell wall, and only an outer membrane consisting 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 yellow-green.
Embodiment 3
[0082] The production rate comparison of the mutant algal strain ENN0001-1 obtained by the screening of embodiment 3 and the wild-type algal strain
[0083] 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 o...
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