Method for preparing protoplast of Eucheuma denticulatum from extract of internal organs of Siganus Concatenalu

A technology of Eucheuma fine-toothed and protoplast, applied in the direction of plant cells, etc., can solve the problems of no major breakthrough, slow growth, germplasm degradation, etc., and achieve the effect of wide distribution of fish species, high feeding speed, and complete results

Active Publication Date: 2014-08-06
南通中国科学院海洋研究所海洋科学与技术研究发展中心 +1
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Problems solved by technology

In the past, the large-scale cultivation of this algae mainly relied on vegetative reproduction, but the spore reproduction and sexual reproduction methods of this algae have been seriously degraded, resulting in germplas

Method used

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  • Method for preparing protoplast of Eucheuma denticulatum from extract of internal organs of Siganus Concatenalu
  • Method for preparing protoplast of Eucheuma denticulatum from extract of internal organs of Siganus Concatenalu
  • Method for preparing protoplast of Eucheuma denticulatum from extract of internal organs of Siganus Concatenalu

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Embodiment 1

[0029] 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 them evenly, and dilute it to 100ml with distilled water as The stock solution is ready for use.

[0030] Preparation of disinfection medium: take 50 mg of chloramphenicol and add it to 100 ml of sterile sea water.

[0031] Preparation of eluent: Take 109.3g mannitol (0.6M) and 554.9mg calcium chloride (5mM), respectively dissolve them into conical flasks (2L) containing 1 liter of F / 2 medium, and put them in a sterilizing pot , sterilized under high temperature and high pressure for 25 minutes, and transferred to a storage bottle after cooling for later use.

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Abstract

The invention discloses a method for preparing protoplast of Eucheuma denticulatum from the extract of internal organs of Siganus Concatenalu. The method comprises the following steps: (1) feeding Siganus Concatenalu with Eucheuma denticulatum; (2) acquiring crude enzyme extract from the internal organs of Siganus Concatenalu; and (3) subjecting Eucheuma denticulatum to enzymatic hydrolysis with the crude enzyme extract so as to obtain the protoplast. The method provided by the invention uses easily available raw materials and has complete and stable results; with the method, the protoplast with a complete structure and without cell walls can be obtained, and a good base is laid for subsequent germplasm seedling cultivation through protoplast healing and tissue culture.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing protoplasts. Background technique [0002] Eucheuma serrata is an important ι-carrageenan-producing red algae with broad market development and application prospects. In my country, Eucheuma fine-toothed is except producing iota carrageenan and is used in food and pharmaceutical processing and manufacturing, and can also be directly used to be processed into cold and dressed with sauce instant food products, and only one item of cold and dressed with sauce instant food products consumes about 2000 tons of dry products every year in my country. [0003] Internationally, the cultivation of Eucheuma serrata is mostly carried out in tropical and subtropical regions, and Eucheuma serrata is mainly produced in Southeast Asian countries and regions such as Indonesia and the Philippines. In the past, the large-scale cultivation of this algae mainly relied on vegetativ...

Claims

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Application Information

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IPC IPC(8): C12N5/04
Inventor 刘建国李俊鹏庞通李虎林伟
Owner 南通中国科学院海洋研究所海洋科学与技术研究发展中心
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