阅读说明：本技术 一种用有机合成高分子固定化微藻保种技术 (Microalgae seed conservation technology by organic synthesis polymer immobilization ) 是由 邓敏丽 于 2018-06-23 设计创作，主要内容包括：本发明提供了微藻保种技术邻域内的一种用有机合成高分子固定化微藻保种技术,其先定量称取有机合成高分子固定剂、氯化钠、藻液,然后将有机合成高分子固定剂和氯化钠分别加入蒸馏水,配成一定浓度的凝胶溶液,将有机合成高分子固定剂凝胶溶液煮沸消毒,再将有机合成高分子固定剂凝胶溶液与藻液按一定比例充分混匀,注射器均匀的速率滴入CaCl<Sub>2</Sub>溶液中,边滴边摇动,一定时间后形成固化的微藻胶珠,形成的微藻酸钙胶珠用消毒蒸馏水洗涤数次,得到固定藻细胞,保存培养,最后定时取样进行复活试验,室温下培养10天,先目测,后镜检,以判断是否存活；本发明的备方法简单,操作简单,不易污染,保种时间较长。(The invention provides a microalgae seed-preserving technology by immobilizing microalgae with an organic synthetic polymer, which comprises the steps of weighing an organic synthetic polymer fixing agent, sodium chloride and algae liquid in a certain amount, then adding the organic synthetic polymer fixing agent and the sodium chloride into distilled water respectively to prepare a gel solution with a certain concentration, boiling the organic synthetic polymer fixing agent gel solution for sterilization, then fully and uniformly mixing the organic synthetic polymer fixing agent gel solution and the algae liquid according to a certain proportion, and dropping CaCl into a syringe at a uniform rate 2 Dripping while shaking in the solution, forming solidified microalgae colloidal beads after a certain time, washing the formed microalgae calcium colloidal beads with sterilized distilled water for several times to obtain fixed algae cells, preserving and culturing, sampling at regular time, performing a revival test, culturing at room temperature for 10 days, firstly performing visual inspection, and then performing microscopic inspection to judge whether the microalgae cells survive; the preparation method is simple, the operation is simple, the pollution is not easy to occur, and the seed preservation time is longer.)

1. A microalgae seed conservation technology by immobilizing organic synthetic polymer is characterized in that the preparation method comprises the following steps:

(A1) weighing and liquid preparation: quantitatively weighing an organic synthetic polymer fixing agent, sodium chloride and algae liquid, and respectively adding the organic synthetic polymer fixing agent and the sodium chloride into distilled water to prepare a gel solution;

(A2) fixing: sterilizing the organic synthetic polymer fixative gel solution, mixing the organic synthetic polymer fixative gel solution and algae solution at a certain ratio, and dripping 2% CaCl with an injector at a rate of 6mL/min₂Dripping while shaking in the solution, and forming solidified microalgae colloidal beads after a certain time to obtain fixed algae cells;

(A3) and (4) preservation: washing the formed microalgae calcium colloid beads with sterilized distilled water for several times, and preserving and culturing;

(A4) and (3) detection: sampling at regular time for a revival test, culturing at room temperature for 10 days, and performing visual inspection and microscopic inspection to judge whether the plant survives.

2. The method according to claim 1, wherein in step (A1), the organic polymer fixative comprises one of polyacrylamide, polyvinyl alcohol, porous silica gel and polyurethane.

3. The method according to claim 1, wherein in step (A1), the concentration of the organic polymer fixative is 1-5%.

4. The method according to claim 1, wherein in step (A2), the organic synthetic polymer fixative is mixed with the algae solution at a ratio of 1:1 to 4: 1.

Background

The microalgae is rich in nutrition and has wide application in life and production practice of people, a seed preservation technology is very important in culture and application of the microalgae, the large-scale microalgae industry is not enough only by culturing the microalgae, and the algae seeds must be preserved for later use to meet the requirements, but the microalgae is small and difficult to separate, and the variation of unicellular pure species of the algae seeds along with the change of the environment and the loss of the physiological activity and genetic characteristics of the unicellular pure species of the algae seeds are easy to occur in the seed preservation process, so the algae seed preservation is particularly important.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to overcome the defects in the prior art and provide a microalgae seed preservation technology immobilized by organic synthetic polymers, which has the advantages of simple preparation method, simple operation, difficult pollution and longer seed preservation time.

The purpose of the invention is realized by the following technical scheme: a microalgae seed conservation technology by using organic synthetic polymer immobilization comprises the following steps:

(A1) weighing and liquid preparation: quantitatively weighing an organic synthetic polymer fixing agent, sodium chloride and algae liquid, and respectively adding the organic synthetic polymer fixing agent and the sodium chloride into distilled water to prepare a gel solution;

(A2) fixing: sterilizing the organic synthetic polymer fixative gel solution, mixing the organic synthetic polymer fixative gel solution and algae solution at a certain ratio, and dripping 2% CaCl with an injector at a rate of 6ml/min₂Dripping while shaking in the solution, and forming solidified microalgae colloidal beads after a certain time to obtain fixed algae cells;

(A3) and (4) preservation: washing the formed microalgae calcium colloid beads with sterilized distilled water for several times, and preserving and culturing;

(A4) and (3) detection: sampling at regular time for a revival test, culturing at room temperature for 10 days, and performing visual inspection and microscopic inspection to judge whether the plant survives.

Selecting an organic synthetic polymer fixing agent, weighing the organic synthetic polymer fixing agent, sodium chloride and algae liquid quantitatively by a balance, adding the organic synthetic polymer fixing agent and the sodium chloride into distilled water respectively to prepare a gel solution with a certain concentration, boiling the organic synthetic polymer fixing agent gel solution for sterilization for later use, then fully and uniformly mixing the organic synthetic polymer fixing agent gel solution and the algae liquid according to a certain proportion, and dripping CaCl into the CaCl at a uniform rate by an injector₂Dripping while shaking to embed free microalgae cells in a net support made of polysaccharide or polymer, forming solidified microalgae gel beads after a certain time, washing the formed microalgae calcium gel beads with distilled water for several times to obtain fixed microalgae cells, storing and culturing, sampling at regular time to perform a revival test, culturing at room temperature for 10 days, performing visual inspection and microscopic inspection to determine whether to perform a revival testSurvival; compared with the prior art, the invention has the beneficial effects that: the method has the advantages of simple preparation method, simple operation, difficult pollution and long seed preservation time.

As a further improvement of the invention, the organic synthetic polymer fixing agent comprises one of polyacrylamide, polyvinyl alcohol, porous silica gel and polyurethane.

As a further improvement of the invention, the concentration of the organic synthetic polymer fixing agent is 1-5%.

As a further improvement of the invention, the mixing ratio of the organic synthetic polymer fixing agent to the algae liquid is 1: 1-4: 1.

Detailed Description