Tissue culture method of dendrobium stem seedlings

Abstract

The invention discloses a tissue culture method of dendrobium stem seedlings. The method comprises serial steps of building an intermittent immersion solution culture system, wherein the steps comprise a first step of preparing and sterilizing an MS culture solution, wherein components and use amounts thereof of the MS culture solution are as follows: MS, 0.5 to 5.0 mg / L of NAA, 50 to 200 g / L of potato juice and 30 g / L of saccharose; adjusting a pH value to be 5.70 to 6.00; after the MS culture solution is prepared, placing the MS culture solution into a reagent bottle; transferring the reagent bottle into a moist heat high-pressure sterilization pot; sterilizing the reagent bottle by using a moist heat sterilization method for future use; a second step of sterilizing an intermittent immersion type bioreactor by using the moist heat sterilization method; a third step of inoculating explants into the intermittent immersion type bioreactor containing the MS culture solution; a fourth step of setting the relative parameters of the intermittent immersion type bioreactor. The method disclosed by the invention is used for breeding the dendrobium stem seedlings, and has the beneficial technical effects of high proliferation rate, high degree of automation, small amount of labor, good tissue culture seedling quality and high adaptability.

Description

technical field [0001] The invention relates to the technical field of medicinal plant tissue culture, in particular to a method for tissue culture and propagation of dendrobium nobile seedlings. Background technique [0002] Dendrobium nobile is a perennial herbaceous plant of Orchidaceae, also known as flat nobile, flat yellow grass, flat grass, yellow grass, etc. The seeds of Dendrobium nobile are tiny, the embryo is not fully developed, and it is extremely difficult to germinate under natural conditions; if it is propagated by ramets or axillary buds, it takes a long time and has a low reproduction coefficient. The natural environment on which Dendrobium nobile depends has changed, and wild resources have declined sharply , In addition to long-term excavation, the germplasm resources are scarce and are on the verge of extinction, which can no longer meet the market demand. [0003] The application of plant tissue culture method has solved the shortage of Dendrobium nobi...

AI Technical Summary

Problems solved by technology

However, the traditional method of plant tissue culture using agar as a support for solid or semi-solid culture has become increasingly prominent that it cannot meet the reality and development needs of the industry calling for low consumption and high efficiency.

The traditional tissue culture method has a single nutrient component in the culture medium, and requires a large amount of agar for cultivation. It also needs to deal with a large number of containers when filling and cleaning, and also requires a lot of labor. It has the disadvantages of high labor costs, high energy consumption, and high materials.

At the same time, the traditional tissue culture method of Dendrobium nobile seedlings generally can grow up to 30 plants in one bottle, and the yield is low. When transplanting, the solid medium of the roots needs to be gently cleaned by hand, which is a lot of work and is easy to damage the young roots; When extracting effective secondary metabolites, they must be extracted from agar, which requires a lot of work

These unfavorable factors limit the industrial application of traditional tissue culture technology, and it is urgent to upgrade the existing mainstream tissue culture methods and devices in an economical and automated way.

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