Breeding method for high-growing-capability crypthecodium cohnii mutant algal strain

A technology of Cryptodinoflagellate algae and Cryptodinoflagellate, which is applied in the biological field, can solve the problem of no screening of mutant strains of Cryptodinoflagellate, and achieve the effect of low operating cost, simple equipment, high-efficiency and rapid breeding

Active Publication Date: 2017-05-31
KUNMING ZAONENG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] So far, there is no report on the use of sethoxydim for the screening of Cryptidinium mutants

Method used

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  • Breeding method for high-growing-capability crypthecodium cohnii mutant algal strain
  • Breeding method for high-growing-capability crypthecodium cohnii mutant algal strain
  • Breeding method for high-growing-capability crypthecodium cohnii mutant algal strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Crypthecodinium cohnii (Crypthecodinium cohnii) ATCC 30556 was mutagenized using the atmospheric pressure room temperature plasma (ARTP) mutagen developed by Wuxi Yuanqing Tianmu Biotechnology Co., Ltd.

[0056] 1. Adjust the cell concentration of Cryptidinium algae liquid to 10 6 ~10 8 cells / mL, use liquid C9N2 medium for dilution. After the dilution is complete, add glycerol with a final concentration of 5-20%.

[0057] 2. Put the metal slide in the outer flame of an alcohol lamp in the ultra-clean bench and burn it properly. After cooling, put it into a disposable petri dish, take 10-20 μL of algae liquid and evenly spread it on the metal slide.

[0058] 3. Put the petri dish containing the sample slide into the operation chamber of the ARTP mutagenesis instrument, place the slide in the corresponding hole with sterile tweezers, and close the door of the operation chamber.

[0059] 4. The distance between the plasma emission source and the sample is 2mm, the proce...

Embodiment 2

[0069] This example is basically the same as Example 1, except that the initial algae strain used is the Cryptidium mutant strain obtained in Example 1, and the second round of mutagenesis and screening is carried out.

Embodiment 3

[0071] Analysis of the growth characteristics of the mutant strain of Cryptidium. The obtained mutant strain of Cryptidium was cultured in liquid medium with different glucose concentrations. Shaking table setting parameters are rotation speed 100-200rpm, temperature 24-28°C. The steps are: take 5 mL of fresh cells with an OD490nm of 0.8 and add them to 50 mL of culture medium during inoculation, make 3 parallel samples for each group, measure the absorbance value at a wavelength of 490 nm with a UV-1750 spectrophotometer, measure once every 24 hours, and draw a column picture.

[0072] The formula of the cryptodinoflagellate medium of different glucose concentration of table 1

[0073] Media name Glucose (g / L) Yeast extract powder (g / L) Sea salt (g / L) C9N2 9 2 25 C27N6 27 6 25 C33N7.33 33 7.33 25

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Abstract

The invention discloses a crypthecodium cohnii mutant algal strain breeding method taking an inhibitor (including but not limited to sethoxydim) of a key enzyme (Acetyl CoA carboxylase) for fatty acid synthesis as selective pressure. The method comprises atmospheric and room temperature plasma (ARTP) mutation of a crypthecodium cohnii algal strain and mutant algal strain screening under the selective pressure of the inhibitor (including but not limited to sethoxydim) of Acetyl CoA carboxylase, as well as crypthecodium cohnii algal strain acclimation under the selective pressure of the inhibitor (including but not limited to sethoxydim) of Acetyl CoA carboxylase. The method is characterized in that the ARTP mutation technique and the algal strain screening method based on the inhibitor (including but not limited to sethoxydim) of Acetyl CoA carboxylase are combined together, and also the algal strain acclimation method based on the inhibitor (including but not limited to sethoxydim) of Acetyl CoA carboxylase is used. Compared with a conventional crypthecodium cohnii algal strain breeding method, the breeding method provided by the invention has the characteristic that the high-growing-capability algal strain can be efficiently and quickly obtained.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for breeding mutant algae strains of Cryptodinoflagellate with high growth capacity. Background technique [0002] Since polyunsaturated fatty acids play an important role in the physiological functions of the human body, their research and production are increasingly valued by people. Among them, docosahexaenoic acid (DHA) in omega-3 unsaturated fatty acids is an extremely important fatty acid with high nutritional value, which is important for the normal development of infant brain and the normal function of adult brain function. have an important role. In addition, DHA plays an important role in the prevention of human retina and cardiovascular diseases. The human body cannot synthesize DHA by itself and must be ingested from food. [0003] Until now, deep sea fish oil has been the main source of commercially available DHA. But this main source has some fatal problems, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N13/00C12N1/36C12N1/12
CPCC12N1/12C12N1/36C12N13/00
Inventor 张卫文陈磊刘璐刘晶
Owner KUNMING ZAONENG BIOTECH CO LTD
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