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A method for rapid propagation of mountain turtle tissue culture

A technology of tissue culture and mountain tortoise, applied in gardening methods, botanical equipment and methods, plant regeneration, etc., can solve the problems of small seeds, poor reproductive ability, population and individual number reduction, etc.

Active Publication Date: 2021-12-14
CHENGDU JIANJIANG PHARMA FACTORY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, its medicinal value has been continuously discovered, and the market demand has increased. However, due to its dioeciousness, small seeds, and hard seed shells, it is difficult to germinate under natural conditions and its reproductive ability is extremely poor. Consumption, the deterioration of the environment, the population and the number of individuals have decreased sharply, and the traditional methods of sowing and dividing can no longer meet the growing market demand.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] (1) Collect the young stems with leaves and buds of the mountain turtle, soak them in a saturated solution of washing powder for 30 minutes, and gently brush the surface with a brush, then rinse them with running water for 30 minutes; place them on an ultra-clean workbench, and use a volume fraction of 75%. sterilized with alcohol for 30 seconds, rinsed twice with sterile water, then sterilized with 0.1% mercuric chloride for 6 minutes, rinsed with sterile water for 5 times, and cut off 2 mm at both ends of the stem with sterile surgical scissors to obtain sterile Stem section, set aside;

[0025] (2) Inoculate the aseptic stem segment obtained in step (1) into MS+6-BA 1.0mg / L+NAA 0.1mg / L, agar 5.8g / L, sucrose 30g / L, pH 5.8 induction medium , at 25°C, light intensity 2300l x, light duration 12h / d, culture for 15 days, induce adventitious buds;

[0026] (3) Cut the adventitious buds obtained in step (2) into 1.5cm segments and inoculate MS+6-BA0.4mg / L+IBA 0.2mg / L in a v...

Embodiment 2

[0031] (1) Collect the tender stems with leaves and buds of the mountain tortoise, soak them in a saturated solution of washing powder for 30 minutes, and gently brush the surface with a brush, then rinse them with running water for 30 minutes; Disinfect with alcohol for 30 seconds, rinse with sterile water for 2 times, then disinfect with 0.1% mercury chloride for 6 minutes, rinse with sterile water for 5 times, use sterile surgical scissors to cut off 2 mm at both ends of the stem to obtain sterile stems segment, to be used;

[0032] (2) Inoculate the aseptic stem segment obtained in step (1) into MS+6-BA 1.0mg / L+NAA 0.1mg / L, agar 5.8g / L, sucrose 30g / L, pH 5.8 induction medium , at 25°C, light intensity 2300l x, light duration 12h / d, culture for 15 days, induce adventitious buds;

[0033] (3) Cut the adventitious buds obtained in step (2) into 1.5cm segments and inoculate them horizontally with MS+6-BA0.4mg / L+IBA 0.2mg / L, agar 5.8g / L, sucrose 30g / L , a proliferation medium...

Embodiment 3

[0037] (1) Collect the young stems with leaves and buds of the mountain turtle, soak them in a saturated solution of washing powder for 30 minutes, and gently brush the surface with a brush, and then rinse them with running water for 30 minutes; % alcohol for 30 seconds, rinse twice with sterile water, then disinfect with 0.1% mercuric chloride for 6 minutes, rinse with sterile water for 5 times, and dry the surface with sterile filter paper on the ultra-clean workbench , use aseptic surgical scissors to cut off each 2mm of the stem section at both ends to obtain a sterile stem section for use;

[0038] (2) Inoculate the aseptic stem section obtained in step (1) into MS+6-BA 1.5mg / L+NAA 0.2mg / L, agar 5.8g / L, sucrose 30g / L, pH 5.8 induction medium , at 25°C, light intensity 2500lx, light duration 12h / d, culture for 14 days, induce adventitious buds;

[0039] (3) Cut the adventitious buds that step (2) obtains into 1cm segments, inoculate vertically to MS+6-BA 0.6mg / L+IBA0.3mg / ...

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Abstract

The invention belongs to the technical field of tissue culture of Chinese herbal medicine plants, and in particular relates to a method for rapid propagation of mountain turtle tissue culture, comprising the following steps: collecting young and budded stem sections of mountain turtle, removing leaves, soaking in a saturated solution of washing powder for 30 minutes, and using Gently brush the surface with a brush, rinse with running water for 30 minutes, place it on an ultra-clean workbench, disinfect with 75% alcohol by volume for 30 seconds, rinse twice with sterile water, and then disinfect with 0.1% mercuric chloride 6min, washed 5 times with sterile water, and then blotted the surface moisture with sterile filter paper, then cut off 2 mm at both ends of the stem section with sterile surgical scissors to obtain sterile stem sections, and put them on the induction medium, proliferation medium, The strong seedling medium and the rooting medium can be cultivated to obtain strong rooted seedlings; the method of the invention can obtain a large number of complete strong aseptic seedlings, which reduces the production cost and improves the planting efficiency. And it has a remarkable effect on the induction rate of adventitious buds, value-added coefficient, and rooting rate.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture of traditional Chinese medicinal materials, and in particular relates to a method for rapid propagation of mountain turtle tissue culture. Background technique [0002] Mountain tortoise is a kind of plant of the genus Stephania whose root tubers are rich in various alkaloids, and it is a traditional Chinese medicinal material in my country. The isolation of a variety of active alkaloids, flavonoids, sterols and sugars from its roots is of great significance in clinical, agricultural and other fields. At present, commercial mountain tortoises are still mainly collected in the wild. In recent years, its medicinal value has been continuously explored, and the market demand has increased. However, due to its dioeciousness, small seeds, and hard seed shells, it is difficult to germinate under natural conditions and its reproductive ability is extremely poor. Consumption, the deteriorati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 林良科王倩芸
Owner CHENGDU JIANJIANG PHARMA FACTORY