Fabrication method of seagrass seedling protoplast

Abstract

The invention discloses a fabrication method of seagrass seedling protoplast, comprising the following steps: temporarily raise the seagrass seedlings in double-anti sea water, select the seedlings that just sprout and sequentially put them into alcohol with the concentration of 75 percent, sodium hypochlorite solution with the concentration of 1 percent and potassium iodide solution with the concentration of 1.5 percent for disinfection, then wash, soak and restore the seedlings by sterile seawater. Then, the seagrass seedlings are shredded and placed into centrifugal tube with enzyme solution for enzymolysis. After the enzymolysis, glucose liquid is added before filtration. Wash the seedlings thrice with hypertonic sea water and obtain the precipitate after centrifugation. Add medium to the precipitate and obtain the protoplast through centrifugation. The invention can avoid pollution and obtain sterile seagrass seedling protoplast and has laid a solid foundation for the protoplast fusion, genetic transformation, plant regeneration as well as new breeds breeding. Moreover, the invention can also provide technical reserves for seagrass propagation of cell engineering, rapid propagation of fine lines seagrass and improving seagrass genetic engineering.

Description

Technical field [0001] The present invention relates to a method for preparing protoplasts, in particular to a method for preparing sterile Euphyllum protoplasts. Background technique [0002] Protoplasts are spherical cells exposed after removing the cell wall of plant cells. They are totipotent and can reproduce rapidly in large numbers under artificial control conditions. They can also be induced to fuse to form hybrid cells, providing experimental materials for somatic cell hybridization. There are mechanical, microbiological, and enzymatic methods for isolating protoplasts from seaweeds. The general enzymatic method is a more ideal and commonly used method. The protoplasts obtained by mechanical methods suffer greater damage, which is not conducive to subsequent experiments. Microbial methods can easily cause contamination by miscellaneous bacteria, which can cause contamination to subsequent callus formation and seedling growth. Zostera marina L., commonly known as ...

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Problems solved by technology

However, using the above-mentioned conventional disinfection methods and enzymatic hydrolysis conditions can not obtain the protoplasts of Z.

According to the analysis, the plant Zorophyllum does not belong to seaweed, but belongs to an angiosperm living in seawater. It is precisely because of the physiological characteristics of Zinariae, that there is no relevant report on the preparation method of Zinariae protoplasts.

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