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Tissue culture method for club moss

A technology of tissue culture and male stone pine, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of sensitive fungicide response, no authoritative mature technology for tissue culture, and no detection, etc., and achieve a high survival rate High, solve the effect of poor natural fertility and strong root vitality

Inactive Publication Date: 2016-07-20
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The fleshy stems are rich in endophytic bacteria, are sensitive to fungicides, and are easy to change color. There is no authoritative and mature technology for tissue culture, and no reports on tissue culture of male stone pine have been found.

Method used

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  • Tissue culture method for club moss
  • Tissue culture method for club moss
  • Tissue culture method for club moss

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] A male stone pine tissue culture method, comprising the following:

[0018] (1) Take the wild male stone pine as explants and cultivate them after 2 days of clean sand, remove the stems of the roots and leaves, wash the surface, soak in 1wt.% sodium hypochlorite solution for 15 minutes, rinse with tap water, put in 75% Sterilize in v / v alcohol for 30 seconds, then sterilize with 0.1wt.% mercuric chloride for 7 minutes, rinse with sterile water 4 times;

[0019] (2) Under sterile conditions, cut the stems into 1.5cm-long stem segments, inoculate them on the induction medium, continuously light for 8 hours a day, light intensity 1500lx, culture temperature 23±1℃, and cultivate for 150 days;

[0020] (3) Subculture; the stem buds formed on the induction medium were transplanted to the growth medium under sterile conditions, continuously illuminated for 12 hours a day, the light intensity was 2000lx, the culture temperature was 25±1°C, and cultured for 130 days. Complete r...

Embodiment 2

[0024] A male stone pine tissue culture method, comprising the following:

[0025] (1) Use wild male stone pine as an explant and cultivate it after 3 days of clean sand, remove the stems of the roots and leaves, after cleaning the surface, soak in 1wt.% sodium hypochlorite solution for 15 minutes, rinse with tap water, put in 75% Sterilize in v / v alcohol for 30 seconds, then sterilize with 0.1wt.% mercuric chloride for 10 minutes, rinse with sterile water 5 times;

[0026] (2) Under sterile conditions, cut the stems into 1.5cm long stem segments, inoculate them on the induction medium, continuously light for 8 hours a day, light intensity 2000lx, culture temperature 23±1℃, and cultivate for 150 days;

[0027] (3) Subculture; the stem buds formed on the induction medium were transplanted to the growth medium under sterile conditions, continuously illuminated for 12 hours a day, the light intensity was 2500lx, the culture temperature was 25±1°C, and cultured for 130 days. Comp...

Embodiment 3

[0031] A male stone pine tissue culture method, comprising the following:

[0032] (1) Use wild male stone pine as an explant and cultivate it after 3 days of clean sand, remove the stems of the roots and leaves, after cleaning the surface, soak in 1wt.% sodium hypochlorite solution for 15 minutes, rinse with tap water, put in 75% Sterilize in v / v alcohol for 30 seconds, then sterilize with 0.1wt.% mercuric chloride for 8 minutes, rinse with sterile water 5 times;

[0033] (2) Under sterile conditions, cut the stems into 1.5cm-long stem segments, inoculate them on the induction medium, continuously light for 8 hours a day, light intensity 1700lx, culture temperature 23±1℃, and cultivate for 150 days;

[0034] (3) Subculture; the stem buds formed on the induction medium were transplanted to the growth medium under aseptic conditions, continuously illuminated for 12 hours a day, the light intensity was 2300lx, the culture temperature was 25±1°C, and cultured for 130 days. Compl...

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Abstract

The invention provides a tissue culture method for club moss. Wild club moss is taken as an explant for establishing a sterile system, and then inductive culture and subculture are performed. The technical system overcomes the difficulties of more endophyte of succulent stems and high probability of discoloring. Seedlings formed in the stage of subculture are robust, the differentiation of roots, stems and leaves is relatively complete, and the roots are strong in vigor; the rooting is quickly realized in a planting process, the survival rate is high (90% and above), the growth is quick and the color is excellent. According to the tissue culture method for club moss, the rapid propagation of seedlings under an aseptic condition is realized, so that the problem of shortage in natural wild resources caused by poor natural reproductive capacity of club moss is solved; the technical system is beneficial to the development and utilization of the Chinese herbal medicine with long application history and exact efficacy and fills up the blank of tissue culture for club moss.

Description

technical field [0001] The invention belongs to biological cell tissue cloning technology, utilizes modern biotechnology to rapidly propagate male stone pine, and specifically relates to a male stone pine tissue culture method. Background technique [0002] Gongshisong: Ludisia discolor (Ker-Gawl.) A.Rich., a plant of the Orchidaceae family, also known as Heterophyllum heterocolor, Cadison silkworm, Shishang lotus root, Stone face lotus, etc., is a folk herbal medicine with local characteristics in southern Fujian. The medicine has a long history of application, has definite efficacy and is widely used. It is sweet, slightly astringent, and cool in nature. It has the functions of clearing heat and cooling blood, promoting body fluid and reducing fire, promoting water and dredging collaterals, and relieving cough. "Chinese Materia Medica" records its efficacy: "moisten the lungs, invigorate the spleen, calm the nerves, and treat mainly tuberculosis and hemoptysis, neurasthen...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 林成辉廖琳琳
Owner FUJIAN AGRI & FORESTRY UNIV
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