A kind of preparation method of eelgrass seedling protoplast

Abstract

The invention discloses a method for preparing protoplasts of aseptic eelgrass seedlings. The specific steps are: put the eelgrass seedlings into double-resistant seawater for temporary cultivation for 2 days; select newly germinated eelgrass seedlings; Disinfection with 75% alcohol, 1% sodium hypochlorite solution, and 1.5% potassium iodide solution; rinse, soak and restore with sterile seawater; add enzyme solution to the centrifuge tube and cut it into small pieces, and then carry out enzymatic hydrolysis ; Add glucose liquid after enzymatic hydrolysis and filter; wash 3 times with hypertonic sea water and centrifuge to obtain a precipitate; add culture medium to the precipitate and centrifuge to obtain protoplasts. The present invention can overcome the pollution, obtain the protoplasts of the aseptic eelgrass seedlings, and lay the foundation for protoplast fusion, genetic transformation, plant regeneration, new variety cultivation, etc. In addition, the present invention can also be used for the establishment of eelgrass cell engineering propagation technology , rapid breeding of excellent strains of eel algae, and improved eel algae genetic engineering to provide technical reserves.

Description

technical field [0001] The invention relates to a method for preparing protoplasts, in particular to a method for preparing the protoplasts of aseptic eosinoplastus. Background technique [0002] Protoplasts are spherical cells exposed after removal of 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 methods, microbial methods, and enzymatic methods for isolating protoplasts from algae. The general enzymatic method is an ideal and commonly used method. The protoplasts obtained by the mechanical method suffered greater damage, which was not conducive to the subsequent experiments. The microbial method is easy to cause contamination by bacteria, which will cause pollution to the subsequent callus formation and seedling growth. Zostera marina L., commonly k...

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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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