Breeding method of new high-lipid-content Botryococcus strain

A technology of botrytis, new strains, applied in microorganism-based methods, biochemical equipment and methods, treatment of microorganisms with electricity/wave energy, etc., can solve the problems of low efficiency, large screening workload, low success rate, etc.

Inactive Publication Date: 2013-06-12
ZHEJIANG UNIV
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Problems solved by technology

[0003] At present, the breeding of new strains of Botrytis with high lipid content mainly faces the following two technical problems: 1. Botrytis is a colony formed by tens to hundreds of cells, and its mutation frequency and mutant The screening efficiency is much lower than that of unicellular microalgae such as Chlorella; 2. The selection pressure suitable for screening mutants

Method used

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Embodiment Construction

[0024] In order to increase the mutagenesis frequency of mutagenesis breeding and the effective screening rate of mutants, it is usually necessary to single-cell the mutagenesis material. In the method, ultrasonic treatment with appropriate power is used to prepare the colony-shaped botrytis into single cells. In order to reduce the mechanical damage of ultrasonic waves to algae cells, this method adds a small amount of Xanthan gum to the algae liquid before ultrasonic treatment, and uses it to form a film on the surface of algal cells, and can make the ultrasonic energy transmission change. To be more smooth and other characteristics, and achieved good results. Ethyl methanesulfonate (EMS) is a highly effective chemical mutagen that can mutate the genetic material DNA of organisms. At the same time, Nile red (9-(diethylamino) benzo[a]phenoxazin-5(5H)-one) is a kind of lipophilic oxazine dyes, which can combine with intracellular lipids and emit specific wavelengths after ent...

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Abstract

The invention relates to development and utilization of Botryococcus, and aims to provide a breeding method of a new high-lipid-content Botryococcus strain. The method comprises the following steps: adding particle Xanthan gum into a Botryococcus solution, putting in an ice water bath, and applying ultrasonic to prepare the Botryococcus colony into unicells; after carrying out chemical mutagen ethylmethane sulfonate mutagenesis and low-melting-point agarose immobilization culture, carrying out NR fluorescence microscopy in-vivo sieving analysis to obtain high-lipid-content Botryococcus candidate mutants; and carrying out amplification culture and lipid content determination to select and breed the new high-lipid-content Botryococcus strain. Compared with the traditional selecting and breeding method, the method provided by the invention is more simple and convenient, has the advantages of higher efficiency and the like, can implement high-flux sieving analysis, and can provide a necessary technique for breeding Botryococcus.

Description

technical field [0001] The invention belongs to the development and utilization technology of energy microalgae-botrytis, in particular to a breeding method of a new strain of botrytis with high lipid content. Background technique [0002] Botryococcus is a kind of energy microalgae widely distributed in the world. The lipid content of this algae is usually 25-35% of its dry cell weight, which is generally higher than other microalgae such as Chlorella, and the lipid composition It is also most similar to fossil crude oil in structure, and is known as "oil algae". The production of high-quality fuels such as aviation fuel has been one of the research and development hotspots in the global bioenergy field in recent years [Metzger & Largeau. Botryococcus braunii: a rich source for hydrocarbons and related ether lipids. Appl Microbiol Biotechnol. 2005, 66: 486–496; Sakamoto, et al. Optimization of light for growth, photosynthesis, and hydrocarbon production by the colonial micr...

Claims

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Application Information

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IPC IPC(8): C12N13/00C12N15/01C12R1/89
Inventor 汪志平于金鑫刘新颖王冬云马丽芳
Owner ZHEJIANG UNIV
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