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Chroococcidiorella tianjinensis as well as culture method and application thereof

A method for culturing chlorella and a technology for chlorella, which are applied in the field of green algae-Chlorella titanosa and its culturing field, can solve the problems of unknown origin and formation process, unclear genetic background of green algae, errors, etc., so as to improve international competition. strength, good for eye health, clear genetic background

Active Publication Date: 2020-11-13
HEBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003]However, the existing green algae are isolated and collected from the wild environment, and their origin and formation process are unknown, which causes two problems: first, The genetic background of green algae is unclear, making it difficult to accurately determine its unique metabolic properties and potential advantages for targeted and efficient development and application
Second, it is difficult to accurately classify and name, causing confusion and errors in the names of green algae used in the production process, which in turn leads to heterogeneity in composition and function of products, especially foods processed with algal powder, health products, animal feed, and bait sex
Even for common chlorella, the algae species used by different manufacturers are not exactly the same due to different sources, or even completely different due to misclassification. The product quality and nutritional value vary greatly, and it is impossible to meet a unified standard

Method used

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  • Chroococcidiorella tianjinensis as well as culture method and application thereof
  • Chroococcidiorella tianjinensis as well as culture method and application thereof
  • Chroococcidiorella tianjinensis as well as culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Morphology and ultrastructural observation of TDX16-DE

[0042] 1. Morphological observation of TDX16-DE cells

[0043] One inoculation loop of TDX16-DE single colony cells was inserted into 100ml of BBM medium (NaNO 3 , 250mg / l; MgSO 4 .7H 2 O, 75mg / l; NaCl, 25mg / l; K 2 HPO 4 ,75mg / l; KH 2 PO 4 ,175mg / l;CaCl 2 .2H 2 O, 25mg / l; ZnSO 4 .7H 2 O, 8.82 mg / l; MnCl 2 .4H 2 O, 1.44mg / l; MoO 3 ,0.71mg / l;CuSO 4 .5H 2 O, 1.57mg / l; Co(NO 3 ) 2 .6H 2 O, 0.49mg / l; FeSO 4 .7H 2 O, 4.98 mg / l; H 3 BO 3 , 11.4mg / l; KOH, 31mg / l; EDTANa 2 , 50mg / l) in a 200ml conical flask, placed in a light incubator for cultivation, continuous light 60μmol photons m -2 s -1 , 25℃, after culturing for 6 days, take samples to observe the cell morphology and take pictures of the cells under an oil microscope (100×) with a light microscope. like figure 1 As shown, TDX16-DE is green, round or oval, 2.0–3.6 μm in diameter, with 1 adherent chloroplast. Cells proliferate by ...

Embodiment 2

[0048] Example 2 Sequencing of TDX16-DE

[0049] (1) The DNA of TDX16-DE was extracted by CTAB method.

[0050] (2) DNA was amplified with primers RP8 (5'-ACCTGGTTGATCCTGCCAGTAG-3') and RP9 (5'-ACCTTGTTACGACTTCTCCTTCCTC-3') and sequenced. The 18S rRNA sequence of TDX16-DE is shown in SEQ ID NO.1. The blast comparison analysis showed that the 18S rRNA (1687bp) of TDX16-DE had the highest similarity with Chlorella vulgaris, reaching 99.8% (query cover 75%).

[0051] (3) DNA was amplified with primers ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5-TCCTCCGCTTATTGATATGC-3') and sequenced. The ITS sequence of TDX16-DE is shown in SEQ ID NO.2. The results of blast comparison analysis showed that the ITS (761bp) of TDX16-DE had the highest similarity with Chlorella vulgaris, reaching 100% (query cover 99%).

[0052] The 18S rRNA and ITS sequences of TDX16-DE had the highest similarity with Chlorella vulgaris, indicating that TDX16-DE was most similar to Chlorella vulgaris. However, T...

Embodiment 3

[0056] Example 3 Growth of Chlorella tetrazolium Tianjin under different temperature conditions

[0057] An inoculation loop of single algal colony cells of Chlorella tertiaryensis was inserted into a 200ml conical flask containing 100ml of BBM medium, and placed in a light incubator for cultivation at different temperatures, with continuous light of 60μmol photons m -2 s -1 , shake twice a day, after culturing for 10 days, measure the absorbance OD680 of the culture medium. The experimental results showed that Chlorella tianjinensis can grow at 10-40℃, and grow best at 25-35℃.

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Abstract

The invention discloses a first green alga TDX16-DE with a known origin as well as a culture method and application of the green alga TDX16-DE, and belongs to the technical field of microalgae biology. The TDX16-DE is formed by transforming endosymbiotic blue algae (cyanobacteria)-chromonas paramamosain TDX16 after obtaining DNA (deoxyribonucleic acid) of host green algae-haematococcus pluvialis of the endosymbiotic blue algae (cyanobacteria)-chromonas paramamosain TDX16; the TDX16-DE is classified and named as Chroococcidiorella tianjinensis, is preserved in the China Center for Type CultureCollection on August 6, 2020, and has a preservation number of CCTCC AF 2020007 T. The Chroococcidiorella tianjinensis has strong adaptability to the environment; a large amount of carotenoid, such asprotein, polysaccharide, lipid and astaxanthin as well as active substances, bacteriostatic substances and antioxidant substances for promoting cell proliferation and growth can grow and be synthesized rapidly under autotrophic, heterotrophic and mixed nutritional conditions; and thus theChroococcidiorella tianjinensis can be used for producing foods, medicines, health care products, cosmetics, carotenoids, animal feeds, baits, biodiesel and biological fertilizers, and treating waste gas and wastewater.

Description

technical field [0001] The invention belongs to the field of microalgae biotechnology, and particularly relates to the first known origin of green algae - Chlorella tianjinensis and a culture method and application thereof. Background technique [0002] Green algae are eukaryotic algae containing chlorophyll b, with many species and wide distribution, and have unique application and development value in the field of biotechnology. At present, green algae are mainly used for the production of food, health products, pigments, animal feed and biodiesel, as well as the treatment of waste gas and wastewater. [0003] However, the existing green algae are all isolated and collected from the wild environment, and their origin and formation process are unknown, which causes two problems: First, the genetic background of green algae is unclear, so it is difficult to accurately determine its Unique metabolic properties and potential advantages for efficient development and applicatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/12C12P23/00C12R1/89
CPCC12N1/12C12P23/00C12R2001/89C12N1/125Y02A40/80
Inventor 董庆霖邢向英
Owner HEBEI UNIV OF TECH
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