Hermatypic coral symbiotic zooxanthellae algal species as well as separation and purification method and culture method thereof

A technology for separation and purification of corals, applied in the field of reef-building stony coral symbiotic zooxanthellae species and their separation and purification

Pending Publication Date: 2022-05-13
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for the A, B, C, D and other lineages that are widely symbiotic in reef-building corals, by 2014, only one coralzoxanthellae had been cultured in vitro in the world, and it was limited to the laboratory culture stage (Lin et al. , 2014)

Method used

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  • Hermatypic coral symbiotic zooxanthellae algal species as well as separation and purification method and culture method thereof
  • Hermatypic coral symbiotic zooxanthellae algal species as well as separation and purification method and culture method thereof
  • Hermatypic coral symbiotic zooxanthellae algal species as well as separation and purification method and culture method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Take the flat brain coral live on the tray, pay attention to splashing seawater during the experiment, so as not to affect the health of the coral. Light the alcohol lamp and sterilize the mortar, tweezers and other tools. After disinfection, select plump and undamaged coral polyps for extraction, put the removed tissue into a mortar and grind, add buffer solution during the grinding, the buffer solution should not exceed 5ml, and grind until uniform to obtain a tissue suspension. Pour the tissue suspension into a test tube and shake vigorously. Take 1ml of the suspension and inoculate it into 9ml of F / 2 medium, and culture it for 10-15 days under 3000lx light at 28°C to obtain algae liquid in exponential growth phase. After culturing to the exponential growth stage, take the algae liquid and observe it under a microscope. After observing the swimming individuals with flagella, dilute and separate to obtain the single cell algae. Place the unicellular algae in a 1.5ml...

Embodiment 2

[0064] Example 2 Separation, purification and in vitro culture of reef-building coral polyp xanthellae

[0065] a. Burning tweezers, grinding pestle and mortar, set aside;

[0066] Burning uses 95% alcohol to burn the surface, and wait for the item to cool before using it. The above process should be carried out on an ultra-clean workbench;

[0067] b. Use tweezers to pick up the coral tissue, add buffer (EDTA buffer or sterile seawater) into the mortar, and grind evenly to obtain the original suspension A1;

[0068] c. Observe A1 under a microscope to confirm that there are free unicellular algae in the suspension;

[0069] The characteristic of Unicellular algae is that under the optical microscope, the isolated mature cells are round, the nail plate is not obvious, the whole is yellow-green, the size is 10-30 μm, no flagella can be seen, and there is no activity;

[0070] d. Take 1ml of the suspension and inoculate it into 9ml of F / 2 medium, and culture it under 6000lx li...

Embodiment 3

[0077] Example 3 The wide-spectrum temperature-adaptive experiment of reef-building coral zooxanthellae

[0078] Under the culture conditions of 28°C, 3000lx light, and a light cycle of 12 light: 12 dark, set four temperature gradient groups, respectively 18 ± 0.2 ° C, 22 ± 0.2 ° C, 26 ± 0.2 ° C, 30 ± 0.2 ° C, using F / 2 medium, the culture volume is 100ml. The initial concentration of algae cells was 2×10 3 cells / ml, cultivated for 10 days. From the first day of the experiment, take 0.1mL of algae liquid in 0.1mL plankton counting box every 2 days, count under the microscope to obtain the cell density (unit / ml), and take the average of three repetitions. The result is as follows:

[0079] Table 1 Effects of different temperatures on the density of symbiotic algae

[0080]

[0081] The results of the analysis found that the symbiotic algae (zooxanthellae) isolated from the Sinencephalus sinensis in the Wenchang sea area have a wide range of temperature adaptation and ca...

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Abstract

The invention relates to the research field of coral symbiotic algae in marine biology, in particular to a hermatypic coral symbiotic zooxanthellae algal species and a separation and purification method and a culture method thereof. According to the coral and zooxanthellae symbiotic characteristics and the corresponding physiological mechanism, separation, purification and in-vitro culture are performed on zooxanthellae in the coral in a targeted manner, according to the close symbiotic relationship of the zooxanthellae, impurities are not removed in the initial stage of culture, so that the zooxanthellae can gradually adapt to the in-vitro culture environment, and after entering the index growth period, the zooxanthellae can be separated and purified. And separating and purifying to obtain the pure hermatypic coral symbiotic zooxanthellae algal species. The research result of the invention is to promote the physiological function of zooxanthellae and research the symbiotic relationship between coral and zooxanthellae, and provides a feasible thought for protecting the coral reef ecosystem.

Description

technical field [0001] The invention relates to the research field of coral symbiotic algae in marine biology, in particular to reef-building coral symbiotic zooxanthellae species and a separation and purification method and a cultivation method thereof. Background technique [0002] Zooxanthellae are single-celled dinoflagellates that are widely symbiotic in marine invertebrates such as corals, giant clams, and sponges. In the coral reef ecosystem, this mutualistic symbiotic relationship is the basis of the coral reef ecosystem and an important part of the physiological and life activities of corals. The photosynthesis of zooxanthellae can provide corals with oxygen, and at the same time, the fixed carbon is transferred to corals to synthesize various substances needed for life activities. In exchange, corals can provide protection for zooxanthellae and provide the corresponding metabolites of zooxanthellae as raw materials for photosynthesis. This partnership allows cora...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/12C12N1/02C12R1/89
CPCC12N1/12C12N1/02
Inventor 姚雪梅乔立君
Owner HAINAN UNIVERSITY
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