Method for selecting and breeding new high-lipid-content Botryococcus strain

Abstract

The invention relates to development and utilization of Botryococcus, and aims to provide a method for selecting and breeding 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 ultraviolet radiation 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 technology for the development and utilization of energy microalgae-botrytis, in particular to a method for breeding 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 colo...

AI Technical Summary

Problems solved by technology

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 with high lipid content has not been established, so the algae liquid can only be applied to a flat plate for isolation and culture, and then a large amount of The clones were inoculated one by one into the culture medium to expand the culture, and then the lipid determination was performed, so the screening workload was large, the efficiency was low, and the success rate was low.