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Method for improving stress resistance of microalgae, increasing biological yield and obtaining high-added-value compounds

A bio-yield, high value-added technology, applied in the biological field, can solve problems such as lack of DXS gene research, and achieve the effect of improving stress resistance, low light resistance, and high value-added compounds

Inactive Publication Date: 2021-01-26
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Compared with higher plants, the current research on DXS genes in algae is relatively lacking

Method used

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  • Method for improving stress resistance of microalgae, increasing biological yield and obtaining high-added-value compounds
  • Method for improving stress resistance of microalgae, increasing biological yield and obtaining high-added-value compounds
  • Method for improving stress resistance of microalgae, increasing biological yield and obtaining high-added-value compounds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment one: the acquisition of AtDXSoe strain

[0040] 1. take Arabidopsis thaliana cDNA as template, adopt SEQ ID NO:1 and SEQ ID NO:2 as primer, adopt conventional PCR technique to amplify, obtain the gene of improving algae and microorganism tolerance and growth rate described in the present invention, its The nucleotide sequence is shown as SEQ ID NO: 3 in the sequence listing.

[0041] Upstream primer (SEQ ID NO: 1): 5'-ATGGCTTCTTCTGCATTTG-3',

[0042] Downstream primer (SEQ ID NO: 2): 5'-AGGGAAGCTCTGTTTTGA-3'.

[0043] The nucleotide sequence shown in SEQ ID NO: 3 was then amplified from the Arabidopsis cDNA by PCR.

[0044] The PCR program is as follows:

[0045] ①95°C for 4min (pre-denaturation)

[0046] ②95℃30s (denaturation)

[0047] ③58℃30s (refolding)

[0048] ④72℃100s (extension)

[0049] ⑤②-④Step cycle 30 times

[0050] ⑥72°C for 5min (final extension)

[0051] ⑦ Store at 4°C.

[0052] Purify the PCR product (see the PCR product purification da...

Embodiment 2

[0071] Embodiment two: AtDXSoe algal strain growth performance comparison

[0072] 1. Select Arabi DXS-OE transformants and WT and culture them in f / 2 until OD750=2.0-3.0.

[0073] 2. Transfer to a columnar photobioreactor (PBR) equipped with fresh f / 2, the initial OD750≈0.10, and cultivate until OD750=2.0~3.0.

[0074] 3. Dilute to the same OD750 value with fresh medium, and distribute into 50mL Erlenmeyer flasks (20mL / bottle), with 4 parallels for each plant.

[0075] 4. After overnight dark adaptation.

[0076] 5. Placed at 50μmol·photons·m -2 ·s -1 Cultivate in an incubator with a certain light intensity (25° C.), measure the OD750 value every 2 days, and measure 5 times in total.

[0077] 6. Comparing the growth rate of wild type and AtDXSoe. Such as figure 2 As shown, the growth rate of AtDXSoe was higher than that of wild type.

Embodiment 3

[0078] Embodiment three: AtDXSoe algae biomass accumulation level comparison

[0079] 1. Select Arabi DXS-OE transformants and WT and culture them in f / 2 until OD750=2.0-3.0.

[0080] 2. Transfer to fresh f / 2 PBR, the initial OD750≈0.10, cultivate to OD750=2.0~3.0.

[0081] 3. Dilute to the same OD750 value with fresh medium, and distribute into 50mL Erlenmeyer flasks (20mL / bottle), with 4 parallels for each plant.

[0082] 4. After overnight dark adaptation.

[0083] 5. Place control culture conditions (50μmol·photons·m -2 ·s -1 , 25℃, f / 2 medium), low light conditions (LL; 10μmol·photons·m -2 ·s -1 ), high light conditions (HL; 200μmol·photons·m-2 ·s -1 ), nitrogen deficiency (f / 2 medium lacking nitrogen source) and trace element deficiency (f / 2 medium lacking trace element).

[0084] 6. After ten days, collect the algae cells and measure the dry weight of the cells, such as image 3 As shown, under all the conditions investigated, the biomass accumulation level of A...

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Abstract

The invention relates to the technical field of biology, in particular to a method for improving the stress resistance of microalgae, increasing the biological yield and increasing high-added-value compounds. The method comprises the step of introducing a gene with a nucleotide sequence shown as SEQ ID NO: 3 or a carrier containing the gene into microalgae. Experiments prove that the growth rate of the microalgae transferred with a DXS gene is obviously improved under the environments of weak light, high light, nitrogen deficiency and trace element deficiency, the biomass and high-value metabolite content in the grown microalgae are also improved, and the DXS gene is proved to be capable of effectively improving the stress resistance in weak light, high light, nitrogen deficiency and traceelement deficiency environments of the microalgae. The method for cultivating the microalgae capable of resisting weak light, high light, nitrogen deficiency and trace element deficiency is simple, convenient and effective, and a new effective choice is provided for improving resistance of plants to weak light, high light, nitrogen deficiency and trace element deficiency and improving the contentof high-value compounds.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for improving stress resistance of microalgae, increasing biological output and high value-added compounds. Background technique [0002] Nannochloropsis is a spherical unicellular eukaryotic microalgae with a diameter of 2-5 μm. Ecologically, Nannochloropsis is located at the beginning of the food chain and plays an important role in the global carbon cycle and the flow of mineral elements. Many Nannochloropsis algae strains have higher oil content, faster growth and high-efficiency synthesis of high-value compounds (such as EPA, algal polysaccharides, chlorophyll, natural vitamins A, B, C, D, E and iron, zinc, magnesium, etc.) , calcium and other minerals, but also contains protein, essential amino acids for the human body, lutein, dietary fiber, nucleic acid, etc.), and has very high nutritional health and economic value. [0003] In plants, the production of isoprene a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/54C12N1/13C12R1/89
CPCC12N15/8206C12N9/0008C12Y202/01007
Inventor 路延笃陈玉婷韩笑李发兰
Owner HAINAN UNIVERSITY
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