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Mutagenesis and screening method for improving yield of microalgae oil

A microalgae oil and high oil technology, applied in biochemical equipment and methods, microbial determination/inspection, microorganisms, etc., can solve the problem of lack of high oil production verification, inability to obtain mutant algal strains, lack of analysis of photosynthetic performance of algal strains, etc. problems to avoid omissions

Pending Publication Date: 2022-04-12
XI'AN UNIVERSITY OF ARCHITECTURE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, the current screening methods cannot obtain all the mutant strains with high oil production traits, which is not beneficial for more effective screening of target algal strains
At the same time, the existing technology only measures the macroscopic oil yield and content of high-yield performance, lacks the analysis of photosynthetic performance of algae strains, and the determination of key enzyme activities and gene expression that regulate oil synthesis, and lacks the verification of high-yield oil

Method used

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  • Mutagenesis and screening method for improving yield of microalgae oil
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  • Mutagenesis and screening method for improving yield of microalgae oil

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] This embodiment provides a method for cultivating microalgae liquid to the logarithmic phase, comprising the following steps:

[0060] (1) All the BG11 medium and Erlenmeyer flasks were sterilized at 121°C for 20 minutes;

[0061] (2) Inoculate 100mL of Scenedesmus liquid into a 500mL conical flask containing 120mL of BG11 medium, seal the mouth of the bottle with a sterile film, and place it in a constant temperature light incubator at 25°C with a light intensity of 80μmol·( m 2 s) -1 , continuous culture under the condition of light cycle 12L / 12D, in order to prevent cell attachment from affecting growth during the culture process, shake the flask 3-4 times a day and change the position of the Erlenmeyer flask randomly;

[0062] (3) To be cultured to the logarithmic phase, the cell concentration is 10 6 About one / mL, take 0.1mL of algae liquid and put it in a sterilized plastic cover for mutagenesis;

Embodiment 2

[0064] This embodiment provides a method for utilizing ARTP to mutate microalgae, comprising the steps of:

[0065] (1) Get 0.1 mL of the algae liquid in the logarithmic phase provided in Example 1, apply it on the sterilized plastic cover, and move the plastic cover to the stage below the plasma with tweezers;

[0066] (2) Air is used as the working gas of the plasma, the power of the mutagenometer is adjusted to 120W, the radiation distance between the plastic cover and the outlet of the mutagenometer is adjusted to 2mm, the gas flow rate is 5SLM, and the mutagenesis treatment time is set for mutagenesis For each experimental condition, three parallel experiments were performed.

[0067] Performance analysis: exploring the optimal mutagenesis time

[0068] The optimal mutagenesis time of the method for the ARTP mutagenesis microalga provided by embodiment 2 is explored with reference to the following method:

[0069] (1) Set the mutagenesis treatment time as 0s, 10s, 20s, ...

Embodiment 3

[0078] This embodiment provides a method for screening algae strains using a medium containing malonate after ARTP mutagenesis, comprising the following steps:

[0079] (1) Utilize the ARTP mutagenesis method provided in Example 2 to carry out mutagenesis treatment to the Scenedesmus algae fluid in the logarithmic phase of Example 1, and the mutagenesis time is 90s;

[0080] (2) Transfer the mutagenized algae solution to the protection solution (BG-11 medium containing 5% glycerol), suspend and stabilize it in the light incubator for 3 hours;

[0081] (3) Take 1 mL of the suspended algae solution and add it to 9 mL of the mixed solution (BG-11 medium containing 5% glycerol) for dilution;

[0082] (4) Add 200 μL of diluted algae liquid to 90 mL of solid plate containing BG-11 medium and spread it evenly with a sterilized triangular coating rod, wherein malonic acid is added to BG-11 medium, the concentration If it is 100mg / L, cover it immediately and put it upside down into a ...

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Abstract

The invention relates to a mutagenesis and screening method for improving the yield of microalgae oil. The method comprises the following steps: mutagenizing microalgae by using an ARTP mutagenesis method, and culturing and screening the mutagenized microalgae in a culture medium containing malonic acid. According to the present invention, the malonic acid is added after the ARTP mutagenesis to carry out the directed resistance screening, such that the positive mutation rate of the microalgae strain can be improved so as to obtain the microalgae strain with excellent oil production performance, and the microalgae strain with the product value of the specific growth rate and the relative fluorescence intensity being 40% or more higher than the original microalgae strain is adopted as the positive mutation strain so as to provide the high screening index threshold value, the oil-producing algal strains with more excellent performance can be screened, and screening omission is avoided.

Description

technical field [0001] The disclosure relates to the field of biotechnology, in particular to a method for mutagenesis and screening for improving oil production of microalgae. Background technique [0002] Microalgae has become the most potential raw material for biodiesel production due to its high photosynthetic efficiency and fast growth rate. The main component of microalgae oil, triglyceride, is an important precursor for biodiesel production. Fuel production is very important, therefore, improving the oil production of microalgae is the most important factor to promote the industrialization of microalgae biodiesel. [0003] Due to the poor oil accumulation performance of wild microalgae, the breeding of high oil-yielding algae strains has become a bottleneck for microalgae biodiesel production; Atmospheric room temperature plasma (ARTP) mutagenesis technology, as a new and The field of species improvement has become one of the effective methods to quickly change the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/01C12N13/00C12Q1/06G01N21/64C12Q1/6895
Inventor 孙昕孟令顺
Owner XI'AN UNIVERSITY OF ARCHITECTURE AND TECHNOLOGY
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