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A kind of male stone pine tissue culture method

A technology of tissue culture and male stone pine, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of tissue culture without more authoritative mature technology, sensitive fungicide response, and undetected, etc., to achieve a natural solution The effect of poor fecundity, high survival rate and strong root vitality

Inactive Publication Date: 2018-01-12
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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  • A kind of male stone pine tissue culture method
  • A kind of male stone pine tissue culture method
  • A kind of male stone pine tissue culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0018] (1) Use wild male stone pine as an explant and cultivate it 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, the stems were cut into 1.5cm long stem segments, inoculated on the induction medium, continuously illuminated for 8 hours a day, the light intensity was 1500 lx, the culture temperature was 23±1 ℃, and cultivated 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 ...

Embodiment 2

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

[0025] (1) The wild male stone pine was used as explants after 3 days of clean sand cultivation, the stems of the roots and leaves were removed, the surface was cleaned, soaked in 1wt.% sodium hypochlorite solution for 15 minutes, rinsed with tap water, and 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, the stems were cut into 1.5 cm long stem segments, inoculated on the induction medium, and continuously illuminated for 8 h every day, with a light intensity of 2 000 lx, and a culture temperature of 23±1 °C for 150 days. ;

[0027] (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, with a light intensity of 2500 lx, and a culture temperature of 2...

Embodiment 3

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

[0032] (1) The wild male stone pine was used as explants after 3 days of clean sand cultivation, the stems of the roots and leaves were removed, the surface was cleaned, soaked in 1wt.% sodium hypochlorite solution for 15 minutes, rinsed with tap water, and 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 every day, light intensity 1700 lx, 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 2300 lx, the culture temperature was 25±1°C, and cultured f...

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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, is also known as Heterophyllum discolor, Stone silkworm, Shishang lotus root, Stone face lotus, etc. It is a 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, neurasthenia...

Claims

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Application Information

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