A method for improving the direct seedling ratio of Chinese cabbage embryoid body by using liquid culture technology
A technology for liquid culture and embryoid body, which is applied in horticultural methods, botanical equipment and methods, applications, etc., can solve the problems of labor and time consumption, low efficiency, etc.
Active Publication Date: 2021-03-16
沈阳静冶生物科技有限公司
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Problems solved by technology
The direct seedling formation of microspore embryoid bodies is affected by many factors. The conventional inoculation of embryoid bodies in this field uses containers such as triangular flasks, canned bottles, and tissue culture bottles to make seedlings. The culture method adopts solid culture, which is inefficient and consumes a lot of time. Manpower and time, so it is very important to establish an efficient method for direct seedling formation of microspores
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[0014] 1. The present invention selects the embryoid body that three different cabbage varieties produce through microspore culture, carries out solid culture and liquid culture respectively, and the solid culture mode is: select for use containing sucrose 25~30g / L, 6-BA 0.2~0.3mg / L, agar 6.5~7.5g / L, B5 medium with pH 5.8~6.0, divided into tissue culture bottles (7.5cm in diameter, 11cm in height) to make the thickness reach 4cm, and sterilized by 121℃ Transfer the cabbage embryoids to the surface of the culture medium in the tissue culture bottle in the bacterial operation bench, and culture them for 20-30 days at a temperature of 25±3°C, a light intensity of 3000-4000 lx, and a photoperiod of 16h / 8h. The liquid culture method is as follows: choose B5 medium containing 25-30g / L sucrose, 0.2-0.3mg / L 6-BA, 0.1-0.15mg / L Amp, pH 5.8-6.0, and pack them into tissue culture devices (diameter 7.5 cm, height 11cm, and a plastic net 4cm away from the bottom) to make the liquid level r...
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The invention provides a method for improving the direct seedling formation rate of Chinese cabbage embryoids by using a liquid culture technique. In the method, a special device applicable for a liquid culture medium is used; the used liquid culture medium is a B5 culture medium, with the pH of 5.8 to 6.0, containing 25 to 30 g / L of saccharose, 0.2 to 0.3 mg / L of 6-BA and 0.1 to 0.15 mg / L of penbritin (Amp); in the culture process, aeration needs to be carried out in the culture medium at an interval of 15 min, the aeration time lasts for 2 to 3 min, the liquid level is raised until the embryoids are incompletely submerged in the aeration process, the quantity of seedlings can be counted after cultivation for 20 to 30 d under the conditions that the temperature is 22-28 DEG C, the light intensity is 3000 to 4000 lx and the light period is 16h / 8h, and the seedlings are subcultured in the next step. Through counting and calculation, the direct seedling formation rate of the embryoids can be 80% to 90% through cultivation by adopting the method, and is 1.5 to 2.0 times of a seedling formation rate in a traditional culture mode.
Description
technical field [0001] The invention relates to the technical field of cell breeding, in particular to a method for improving the direct seedling ratio of Chinese cabbage embryoid bodies by using liquid culture technology. Background technique [0002] Microspore culture is a technique that gives fresh free microspores appropriate medium and culture conditions to develop into embryoid bodies, and then develop into regenerated plants. Double haploid pure lines can be quickly obtained as parents. It is a practical cell breeding technology that is applied to dominant hybrid breeding and shortens the breeding period. [0003] Microspore culture technology includes two processes of microspore embryo formation and embryoid body seedling formation. The mature microspore culture technology should have a high rate of microspore embryo formation and high embryoid body direct seedling formation rate. The direct seedling formation of microspore embryoid bodies is affected by many facto...
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IPC IPC(8): A01H4/00
CPCA01H4/008
Inventor 戴速航
Owner 沈阳静冶生物科技有限公司