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Method of screening high grease algae from natural water body

A natural water body, high-fat technology, applied in the direction of microorganism-based methods, single-cell algae, biochemical equipment and methods, etc., can solve problems such as lack, and achieve the effect of simple screening process

Inactive Publication Date: 2014-02-05
SANYA INST OF DEEP SEA SCI & ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved in the present invention is to provide a method for screening high-fat algae from natural water bodies in order to overcome the technical defect of lack of methods for screening high-fat algae from natural water bodies in the prior art

Method used

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  • Method of screening high grease algae from natural water body
  • Method of screening high grease algae from natural water body
  • Method of screening high grease algae from natural water body

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] For two commonly used lipid-specific fluorescent dyes, different concentrations of dyes were selected and combined with a double-focus microscope to detect the optimal dyeing concentration

[0028] 1. Preparation of materials and reagents:

[0029] 1. Algae species: Chlorella is a microalgae with relatively high oil content. The chlorella in the logarithmic phase cultivated in the laboratory is selected for dyeing to find the best dyeing concentration.

[0030] 2. Dye solution preparation:

[0031] BODIPY505 / 515: Dissolve BODIPY505 / 515 dye with organic solvent dimethyl sulfoxide (DMSO), and make 5mM mother solution, store at -20°C.

[0032] Nile Red: Dissolve Nile Red dye in acetone, prepare a 25 μg / mL stock solution, and store at room temperature in the dark.

[0033] 3. Staining: use different concentrations of BODIPY505 / 515 and Nile Red dyes to stain Chlorella for 15 minutes at room temperature and dark. The dye concentrations are BODIPY 505 / 515: 2 μl / 5ml,...

Embodiment 2

[0035] Flow cytometry combined with fluorescent dyes to detect intracellular lipids in single microalgae

[0036] Algae selected for log phase culture: Phaeodactylum tricornutum Phaeodactylum and chlorella.

[0037] BODIPY505 / 515 dye at a concentration of 20 μl / ml was used to stain a single algae species and a mixture of the two algae at room temperature in the dark for 15 minutes, and the specific fluorescence intensity of the algae before and after staining was measured by flow cytometry (excitation length 488nm, emission wavelength 530nm). Draw 1ml of sample into a dedicated flow analysis test tube, set the filter: forward scattered light, side scattered light, red fluorescence and fluorescent signal. Then put the sample into the sampling system, and adjust the sampling speed at 0.1-1ml / h. 488nm laser excitation produces side scattered light, and the dye also emits fluorescent signals after passing through the laser beam. The scattered light signals and fluorescent s...

Embodiment 3

[0040] Obtained by the groping test of embodiment 1 and embodiment 2: a kind of method for screening high-fat algae from natural water body should comprise the steps:

[0041] S1. Collect water samples containing algae, filter to remove large organisms and impurities;

[0042] S2. Stain the water sample containing algae with a fluorescent dye with a concentration of 20-40 μl / ml (the fluorescent dye is BODIPY505 / 515 or Nile Red) at room temperature in the dark for 15 minutes, and use a flow cytometer with a sorting function to measure before and after staining. Algae fluorescence intensity and signal changes;

[0043] S3. Use a flow cytometer with a sorting function to screen out some water samples with strong intermediate specificity signals to obtain water samples of high-fat algae groups;

[0044] S4. Separating and culturing the water samples of high-fat algae groups to obtain pure microalgae species with high-fat content.

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Abstract

The invention relates to the technical field of algae screening, and specifically discloses a method of screening high grease algae from a natural water body quickly and effectively. The method comprises the following steps: firstly, dyeing a collected algae water sample by a fluorescent dye; then, quickly screening high grease excellent algae groups in the natural water body environment through a flow cytometry screening technology with a sorter; and further separately culturing under a condition of a laboratory to obtain pure micro-algae seeds with high grease content. The method provided by the invention can quickly and conveniently separate algae with high grease content from natural samples in various types, provide a genetic resources center for development of a biological energy, and have prospect and potential of producing algae energy resources on a large scale.

Description

technical field [0001] The invention relates to the technical field of algae screening, in particular to a method capable of quickly and effectively screening high-fat algae from natural water bodies. Separation and screening of algae with high oil content in natural water bodies provides a germplasm resource bank for the development of bioenergy. Background technique [0002] With the rapid development of the economy and the rapid growth of the world population, the energy issue has become the focus of great attention of all countries in the world. At present, oil reserves are decreasing, fossil energy prices are rising, environmental pollution, and energy demand are increasing, so it is urgent to seek a renewable and sustainable clean energy. Biomass energy is the most common clean and renewable energy on the earth. At present, various countries are carrying out the research and production of biofuel and biofuel to replace the use of traditional fuel oil which is expensiv...

Claims

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Application Information

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IPC IPC(8): C12N1/12G01N21/64C12R1/89
Inventor 荆红梅刘红斌郑丽平叶承孙海燕
Owner SANYA INST OF DEEP SEA SCI & ENG
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