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Method for culturing dendrobium candidum protocorms

A technology of dendrobium candidum and protocorms, applied in horticultural methods, botanical equipment and methods, horticulture, etc., to achieve the effects of saving cultivation costs, reducing production costs, and reducing labor

Active Publication Date: 2015-06-17
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] In view of the fact that the cultivation of protocorms of Dendrobium officinale by using the intermittent immersion culture method has not been reported and publicly applied, it is necessary to develop a method for the industrial expansion of protocorms of Dendrobium candidum by using intermittent immersion bioreactors

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  • Method for culturing dendrobium candidum protocorms

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: as figure 1 Shown, the method for cultivating the protocorm of dendrobium officinale comprises the following steps:

[0038] (1) Preliminary preparation:

[0039] ① Preparation of solid medium: the formula of solid medium is 1 / 2MS basic medium, add NAA 0.5mg / L; After high-pressure moist heat sterilization, it is ready for use;

[0040]②Preparation of liquid proliferation medium: the formula of liquid proliferation medium is 1 / 2MS basic medium, add NAA0.6mg / L; 6-BA 6.0 mg / L; sucrose concentration 30g / L; banana homogenate 60g / L, adjust The pH value is 5.8. After the preparation of the culture solution is completed, it is boiled, subpackaged, sterilized by high temperature and high pressure, and then set aside;

[0041] ③Sterilization of bioreactor: Assemble the bioreactor, then bandage and sterilize it; after high temperature and high pressure damp heat sterilization, it is ready for use;

[0042] (2) Germination and proliferation of protocorms:

[00...

Embodiment 2

[0050] The method for cultivating the protocorm of Dendrobium officinale, the specific steps are as shown in Example 1, the difference is that: in the assembly process, the glass beads are put into the bioreactor, and the glass beads of 4 mm are selected to be tiled on the bioreactor. On the reaction plate; the formula of the selected liquid proliferation medium is 1 / 2MS+0.5 mg / L NAA+4.0 mg / L 6-BA+30 g / L sucrose+50g / L banana homogenate, adjust the pH value to 6.0; The inoculation density in the liquid medium was 45g / L. After 4 days of observation and cultivation, they were sterile and then placed in the batch immersion culture bioreactor; the immersion frequency was 7 min / 6 h; the culture time was 50 days.

[0051] The cultured protocorm of Dendrobium officinale has a proliferation coefficient of 26.43, a light yellow color, good quality, few and large bud heads, and no obvious differentiation.

Embodiment 3

[0053] The method for cultivating the protocorm of Dendrobium officinale, the specific steps are as shown in Example 1, the difference is that: in the assembly process, the glass beads are put into the bioreactor, and the glass beads of 6 mm are selected to be tiled on the bioreactor. On the reaction plate; the formula of the selected liquid proliferation medium is 1 / 2MS+0.4 mg / L NAA+2.0 mg / L 6-BA+30 g / L sucrose+40 g / L banana homogenate, adjust the pH value to 5.8 ; The inoculation density of inoculation into liquid medium was 30 g / L. After 3 days of observation and cultivation, they were sterile and then placed in a batch immersion culture bioreactor with an immersion frequency of 5 min / 4 h; the culture time was 40 days.

[0054] The cultured protocorm of Dendrobium officinale has a proliferation coefficient of 28.46, a light yellow color, high quality, few and small bud heads, and no obvious differentiation.

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Abstract

The invention belongs to the field of plant biotechnology, relates to medicinal plant cultivation technology and particularly relates to a method for culturing dendrobium candidum protocorms. The method comprises the following production steps in sequence: preparation, protocorm germination and proliferation, transitional observation and culture, and bioreactor production. The method for producing the dendrobium candidum protocorms by using an intermittent immersion bioreactor is carried out under sterile conditions, the sterilization treatment method is damp and hot sterilization at high temperature and high pressure, in the technical scheme, the intermittent immersion bioreactor is used for culturing the dendrobium candidum protocorms, so that a high proliferation rate is obtained within a short time, the culture period of the dendrobium candidum protocorms is shortened and the quality of the cultured dendrobium candidum protocorms is high; the method is simple to operate and is automatic, thereby saving time and labor; meanwhile, the use of culture dishes is reduced to save the manpower cost and the culture cost and achieve a low-cost culture method, and thus the method is suitable for industrial promotion.

Description

technical field [0001] The invention belongs to the technical field of medicinal plant tissue culture, and in particular relates to a method for industrialized propagation of protocorms of Dendrobium officinale by using an intermittent immersion bioreactor. Background technique [0002] Dendrobium officinale is a rare Chinese herbal medicine in my country. It is one of the "Nine Chinese Immortal Herbs", also known as "Qianjin Grass". "Shen Nong's Materia Medica" records that Dendrobium "mainly injures, eliminates numbness, lowers qi, nourishes five viscera, strengthens yin, and thickens the stomach after long-term use. ". Modern pharmacological studies have shown that Dendrobium officinale contains a variety of active ingredients such as alkaloids, polysaccharides, and rich trace elements. Dendrobium officinale plants have low survival rate, slow growth rate, low reproduction rate, low yield and high price, which cannot meet the market demand at all. Replacing the orig...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 陈集双蒋海侠张本厚宋莲
Owner NANJING UNIV OF TECH
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