Preparation method for Kappaphycus alvarezii protoplast

A technology of Kappa longicardium and protoplasts, which is applied to plant cells and other directions, and can solve problems such as ineffective effects

Inactive Publication Date: 2014-08-06
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
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Problems solved by technology

But in any case, the preparation of Kappa longcardia protoplasts is the basis and the first step of cultivating seedlings through this method. Due to the limitation of the large amount of κ-carrageenan in the cell wall of this algae, the effect of traditional enzymatic hydrolysis technology is not obvious

Method used

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  • Preparation method for Kappaphycus alvarezii protoplast
  • Preparation method for Kappaphycus alvarezii protoplast
  • Preparation method for Kappaphycus alvarezii protoplast

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Embodiment 1

[0018] Required solution preparation: prepare 50mM sodium phosphate buffer solution with pH 6.0, take 4.4ml of 1mol / L sodium dihydrogen phosphate and 0.6ml of 1mol / L disodium hydrogen phosphate, mix well, dilute the mixed solution to 100ml with distilled water, Store in a 4-degree refrigerator as a stock solution for later use.

[0019] Preparation of sterile seawater solution: Take 50mg of chloramphenicol and add it to 100ml of sterile seawater.

[0020] Preparation of eluent: Take 109.3g of mannitol (0.6M) and 554.9mg of calcium chloride (5mM) and dissolve them in turn into an Erlenmeyer flask (2L ), then put it into a sterilizer, sterilize at high temperature (121°C) and high pressure (102.9kPa) for 25 minutes, transfer it to a storage bottle after cooling, and wait for use.

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Abstract

The invention specifically relates to a method for preparing Kappaphycus alvarezii protoplast by using extract of internal organs of Siganus Concatenalu, which belongs to the field of biotechnology. The method comprises the following steps: feeding omnivorous tropical Siganus Concatenalu with Kappaphycus alvarezii in advance so as to induce Siganus Concatenalu to produce enzyme digesting carragheenan; then preparing mixed enzyme liquid with the internal organs of Siganus Concatenalu as a raw material; and degrading cell walls and intercellular substances of Kappaphycus alvarezii by using the mixed enzyme liquid so as to obtain free protoplast. The method provided by the invention can realize artificial cultivation of a novel strain of Kappaphycus alvarezii in virtue of traditional protoplast cell fusion and callus induction and differentiation and provide a novel approach and related supporting technology for seedling growing and breeding.

Description

technical field [0001] The invention belongs to the field of biological technology, and specifically relates to a method for preparing protoplasts of Kappa longcardia by using the viscera extract of basket fish. Background technique [0002] Kappa longcardia is an important tropical red alga resource that produces κ-carrageenan in the world. κ-carrageenan is a viscous polysaccharide of seaweed cells, composed of macromolecules composed of D-galactose and its derivatives, and has broad market development and application prospects. Carrageenan has the characteristics of gel and water retention, and has a certain hardness. Therefore, it is an important processing raw material supporting food manufacturing industries such as dairy products, ham, and ice cream, and has a very wide application prospect in food, chemical and other fields. Carrageenan also has good application prospects in medicine. Studies have shown that subcutaneous injection of carrageenan can stimulate the rev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
Inventor 刘建国李俊鹏庞通李虎林伟
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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