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Nannochloropsis sp.OZ-1 mutant strain and heavy ion irradiation selection method for the same

A technology of Nannochloropsis and mutant strains, applied in the field of microbial engineering, can solve the problems of reverse mutation, poor genetic stability of mutant strains, short screening time, etc., and achieve the effect of ensuring stability and high growth rate

Inactive Publication Date: 2013-03-20
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI +1
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  • Description
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  • Application Information

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Problems solved by technology

However, the mutagenesis technology currently used for algae breeding has a short screening time, and usually only passes through one screening, and the mutant strains obtained have poor genetic stability and are prone to reverse mutations.

Method used

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  • Nannochloropsis sp.OZ-1 mutant strain and heavy ion irradiation selection method for the same
  • Nannochloropsis sp.OZ-1 mutant strain and heavy ion irradiation selection method for the same
  • Nannochloropsis sp.OZ-1 mutant strain and heavy ion irradiation selection method for the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Creation of Nannochloropsis heavy ion mutagenesis library and screening of mutant strains with high growth rate

[0029] 1) Take Nannochloropsis sp.OZ-1 as the starting strain; inoculate the strain from the plate into a 250mL Erlenmeyer flask containing 100mL Blue-Green Medium (BG11 for short) culture solution in a light incubator at 25°C, 100μmol / m 2 s, cultivated under continuous light until the logarithmic growth phase.

[0030] The composition and preparation of the BG11 culture solution are shown in Table 1. When preparing the culture solution, add the following components in solid form to distilled water to make a 100-1000 times mother solution, and then dilute it as needed to make a culture solution. Then the BG11 medium was subpackaged and sealed, and sterilized. Take out the sterilized medium, cool to room temperature, and set aside.

[0031] Table 1 Composition and content of BG11 medium

[0032] Element Stock solution concentration ...

Embodiment 2

[0041] Embodiment 2: Biological curve drawing of Nannochloropsis mutant strain HP-1 and wild-type strain

[0042] The high growth rate mutant strain HP-1 and the wild strain obtained above were cultured in a bubbling column photoreactor (outer diameter 4.0 cm, inner diameter 3.8 cm, column height 60 cm) containing 400 mL of BG11 liquid medium until logarithmic growth phase , the culture condition is 25±1℃, the light intensity is 100μmol / m 2 · s, continuous light culture. And continue bubbling ventilation at the bottom of the reactor, the ventilation volume is 0.25vvm, and the gas fed is pure CO 2 Mix with natural air to make the CO in the culture medium 2 The concentration is 2% (volume ratio), and it is sterilized by filtration with a 0.22 μm filter membrane; the above-mentioned mutant strains and wild strains cultivated to the logarithmic growth phase are used as seed liquids and inoculated respectively in bubbling columns containing 400 mL of BG11 liquid medium In the t...

Embodiment 3

[0044] Example 3: Determination of the pigment content of Nannochloropsis mutant strains and wild-type algae strains respectively take 2 mL of the above-mentioned examples and culture them on the 0th day, the 2nd day, the 4th day, the 6th day, the 8th day, and the 8th day respectively. The 10th day, 12th day, 16th day and 18th day mutant algae liquid and wild type algae liquid were centrifuged at 5000rpm for 10min respectively, then added 2mL of methanol, bathed in water at 60℃ for 1h, then centrifuged at 5000rpm for 10min, and finally used Spectrophotometer measures its absorbance respectively at 665, 666, and 470nm, according to the calculation formula C 叶绿素a μg / mg=[13.43A665v / (lV)] / D and C 类胡萝卜素 μg / mg=[(1000A470-44.76A666 / 221)v / (lV)] / D respectively measure the content of chlorophyll a and carotenoids, A665, A470, A666 in the formula are the absorbance values ​​at 665, 666 and 470nm respectively , v is the volume of methanol, l cuvette path length, V is the sample volume. ...

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Abstract

The present invention relates to the field of microbial engineering, specifically to a nannochloropsis sp.OZ-1 mutant strain and a heavy ion irradiation selection method for the nannochloropsis sp.OZ-1 mutant strain. The nannochloropsis sp.OZ-1 mutant strain has a taxonomy name of Nannochloropsis sp.OZ-1HP-1, and is preserved in the China General Microbiological Culture Collection Center (CGMCC) on May 14, 2012, wherein a preservation register number is CGMCC No.6140. According to the present invention, biomass accumulation of the nannochloropsis sp.OZ-1 mutant strain is increased by 19% at a culture final phase compared with the wild-type strain; chlorophyll content is increased by 45% at the fourth day of culture compared with the wild-type strain; lipochrome content is increased by 47% compared with the wild-type strain; biomass of the nannochloropsis sp.OZ-1 mutant strain is increased by 33% at an oil accumulation final phase compared with the wild-type strain, and an oil yield is increased by 28%; fatty acid composition of the nannochloropsis sp.OZ-1 mutant strain does not show significant change compared with fatty acid composition of the wild-type strain, and oil component analysis of the nannochloropsis sp.OZ-1 mutant strain shows that TAG content is increased by 14% compared than the wild-type strain; and the nannochloropsis sp.OZ-1 mutant strain has characteristics of fast growth, high oil yield and high TAG content, and can provide high quality algae species for annochloropsis sp.OZ-1 biofuel industry production.

Description

technical field [0001] The invention relates to the field of microbial engineering, in particular to a Nannochloropsis mutant strain and a heavy ion mutagenic breeding method thereof. Background technique [0002] Energy is the material basis for the survival and development of human society, and efficient and clean energy utilization is an important guarantee for sustainable economic development. With the continuous reduction of conventional non-renewable fossil resources and the increasingly significant global warming, energy, resources and the environment have become important issues facing the sustainable development of human society. Biomass energy, including biodiesel and bioethanol, is considered to be one of the most potential ways to solve the energy crisis due to its renewable and environmentally friendly characteristics. [0003] Microalgae are considered to be the most potential biomass energy raw materials because of their fast biomass accumulation, strong envi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/12C12N13/00C12N15/01C12P7/64C12P17/18C12P23/00C12R1/89
CPCY02E50/13Y02E50/10
Inventor 马玉彬张东远周功克王芝瑶周翠燕
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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