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

A primary cell embryo, ultrasonic technology, applied in plant regeneration, horticultural methods, botanical equipment and methods, etc., can solve problems such as difficulty in control, low success rate, inactivation, etc.

Inactive Publication Date: 2012-03-21
张笑逸
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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 solutio

Method used

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

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Embodiment Construction

[0013] Further illustrate positive effect of the present invention with example below.

[0014] Beginning in 2007, successively to spring orchid, cold orchid, green orchid, autumn branch, black orchid, sword leaf orchid six kinds of Chinese ground orchid species fruit, experiment with the method of the present invention, cultivate with common culture method after processing, obtained success. The processing procedure of each orchid seed is as follows:

[0015] Select uncracked orchid fruits, sterilize the surface with 75% alcohol, take out the seeds, soak them in 10% sodium hypochlorite for 5-10 minutes, take them out and rinse them with sterile water for 3 times to complete the surface disinfection, and put them on the sterile operating table Cut the fruit, pour the primary cell embryo powder into 100ml of sterile water, drop in 0.5ml of surfactant Tween 80, and process (rotate) on a rotary shaker for 1 minute to promote its affinity with water, then Pour it into a steriliz...

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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...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 张笑逸
Owner 张笑逸
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