Supersonic wave wall breaking culture method for Chinese terrestrial orchid progenitor cell embryo

Abstract

The invention provides a supersonic wave wall breaking culture method for Chinese terrestrial orchid progenitor cell embryo. The method comprises the following steps: disinfecting non-cracking orchid fruit with 75% alcohol and taking out seeds; immersing for 5-10 min with 10% sodium hypochlorite; taking out and flushing with sterile water three times for surface disinfection; cutting the fruit on an aseptic bench; pouring progenitor cell embryo powder particle into sterile water; rotating on a rotation shaking table to promote affinity with water; pouring into aseptic supersonic wave oscillation container and oscillating repeatedly until plume seed coat is observed to be ruptured by supersonic wave energy by a stereo microscope; sucking into a prepared medium for aseptic culture. An oscillation mode is as below: oscillating for 1 min and standing for 3 min; then oscillating for 1 min and standing for 3 min; and repeating the process. The invention can start progenitor cell embryo differentiation germination of Chinese terrestrial orchids, such as cymbidium, cymbidium kanran makino, green orchid, qiuzhi, cymbidium sinense and torchlily to culture complete plant, and has great significance for realizing artificial breeding cultivation of rare orchids and saving endangered species.

Description

technical field [0001] The invention belongs to a cell tissue culture method of Chinese terrestrial orchids, in particular to a pretreatment method before initiation of the development of the cultured orchid primordial cell embryos, and a method for ultrasonically breaking the wall of the Chinese terrestrial orchid primordial cell embryos. Background technique [0002] The seeds of orchids grown in China are very thin, as fine as dust. There is only an incomplete embryo in the seed, without germ and endosperm, without the basic structure of the seed, and without nutrients for its germination, so it cannot be called a seed. It can only be called the primary cell embryo, and there is a layer of winged and feathery seed coat, which is not easy to absorb water. For example, the seeds of the lotus petal orchid can not absorb water and nutrients provided by the outside after being soaked in water for more than three years. The required fungi are different, and people have not be...

AI Technical Summary

Problems solved by technology

The existing technology to break through the seed coat is to use sodium hydroxide solution or acid solution to soak and corrode the seed coat. After the skin is broken, the sodium hydroxide solution or acid solution also corrodes the cell embryo and makes it inactivated. It is difficult to control it just right and successfully. Very low rate

At present, it is only a theoretical approach and cannot achieve practical purposes.

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