Fabrication method of seagrass seedling protoplast

A technology of protoplasts and eelgrass, which is applied to plant cells, etc., can solve the problems of not being able to obtain eelgrass protoplasts, achieve the effect of overcoming pollution and prolonging the time of enzymatic hydrolysis

Inactive Publication Date: 2010-07-07
SHANDONG ORIENTAL OCEAN SCI TECH
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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 sea

Method used

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  • Fabrication method of seagrass seedling protoplast
  • Fabrication method of seagrass seedling protoplast
  • Fabrication method of seagrass seedling protoplast

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Embodiment Construction

[0028] Below in conjunction with accompanying drawing and embodiment describe in detail:

[0029] like figure 1 Shown, a kind of preparation method of aseptic eosinopsis seedling protoplast, its concrete steps are:

[0030] (1) Put the eel algae seedlings into the double-resistant seawater for temporary cultivation, and temporarily raise them in the double-resistant seawater at a temperature of 4 degrees Celsius for 2 days. Eelweed seedlings can be grown naturally, or cultured or cultivated artificially. For one method of cultivating eelgrass seedlings, see the applicant's invention patent application "A method for rapid germination of eelweed seeds", the application number being 200910223107.1.

[0031] The double-antibody seawater formula of this embodiment is: the concentration of penicillin is 100 units per milliliter (100 units / ml), and the concentration of streptomycin is 100 units per milliliter (100 units / ml), that is, 1 liter of seawater needs 0.48 g (800,000 units) ...

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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 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...

Claims

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Application Information

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IPC IPC(8): C12N5/04
Inventor 钱冠兰李晓捷罗世菊张壮志刘延岭王宏梅李志凌赵楠赛珊宋少峰
Owner SHANDONG ORIENTAL OCEAN SCI TECH
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