Method for reducing tissue culture rate of anoectochilus roxburghii and culture medium formula thereof

A technology of medium formula and pollution rate, which is applied in the field of plant tissue culture, can solve the problems of polluted clematis seedling breeding, genetic variation of culture materials, and difficulty in transplanting tissue culture seedlings, etc., and achieves good sterilization effect and repeatability Good, easy-to-grasp effects

Inactive Publication Date: 2013-08-21
ZHEJIANG FORESTRY UNIVERSITY +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of tissue culture production, the pollution problem caused by microorganisms such as bacteria and fungi (the pollution rate is generally more than 20%) has always been the main obstacle restricting the breeding of golden clematis seedlings.
In the early stage of pollution, the proliferation efficiency will be reduced, and the growth of culture materials will be delayed; in the later stage of pollution, it will cause difficulty in transplanting and death of tissue culture seedlings, and some will even cause genetic variation of culture materials

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] 1) Selection of explants: Naturally grown clematis plants were used as explants, washed with tap water, rinsed with 70% ethanol for 30 seconds, sterilized with 0.1% mercuric chloride for 10 minutes, sterile water Rinse 5-6 times repeatedly, drain the water, cut into 1 noded stem segment, and set aside.

[0028] 2) Medium preparation: use 1 / 2MS as the basic medium, add banana 60g / L, activated carbon 1.0g / L, NAA1.0mg / L, 6-BA0.6mg / L, sucrose 26g / L, agar powder 6.0 g / L, onion 50g / L, and crab shell powder 15g / L are mixed to adjust the pH value of the culture medium to 6.0. Heat the prepared medium until the agar is completely dissolved, dispense it into clean culture bottles while hot, and cover them.

[0029] 3) Sterilization of culture medium and inoculation equipment: Use high-pressure steam to sterilize the culture medium and inoculation equipment. Keep the temperature of the autoclave at 121°C, 105kPa, 20min. Metal utensils such as knives, scissors, and tweezers were...

Embodiment 2

[0034] 1) Selection of explants: Aseptic tissue-cultured seedlings of Clematis clematis were used as explants, the leaves of Clematis clematis were removed, and the stems were cut into segments with 1 node for later use.

[0035] 2) Medium preparation: use 1 / 2MS as the basic medium, add banana 50g / L, activated carbon 0.8g / L, NAA1.2mg / L, 6-BA0.4mg / L, sucrose 22g / L, agar powder 5.5 g / L, onion 60g / L, and crab shell powder 20g / L are mixed to adjust the pH value of the culture medium to 5.8. Heat the prepared medium until the agar is completely dissolved, dispense it into clean culture bottles while hot, and cover them.

[0036] 3) Sterilization of culture medium and inoculation equipment: Use high-pressure steam to sterilize the culture medium and inoculation equipment. Keep the temperature of the autoclave at 121°C, 105kPa, 20min. Metal utensils such as knives, scissors, and tweezers were wiped with 75% alcohol cotton balls before inoculation, and then sterilized at high temper...

Embodiment 3

[0041] 1) Selection of explants: Aseptic tissue-cultured seedlings of A. clematis were used as explants, and the terminal buds of 1 node of A. clematis were cut for later use.

[0042] 2) Medium preparation: use 1 / 2MS as the basic medium, add banana 50g / L, activated carbon 1.2g / L, NAA0.8mg / L, 6-BA0.8mg / L, sucrose 30g / L, agar powder 6.5 g / L, onion 60g / L, and crab shell powder 20g / L are mixed to adjust the pH value of the culture medium to 5.8. Heat the prepared medium until the agar is completely dissolved, dispense it into clean culture bottles while hot, and cover them.

[0043] 3) Sterilization of culture medium and inoculation equipment: Use high-pressure steam to sterilize the culture medium and inoculation equipment. Keep the temperature of the autoclave at 121°C, 105kPa, 20min. Metal utensils such as knives, scissors, and tweezers were wiped with 75% alcohol cotton balls before inoculation, and then sterilized at high temperature in a sterilizer with quartz sand for 8 ...

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PUM

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Abstract

The invention relates to a method for reducing the tissue culture rate of anoectochilus roxburghii and a culture medium formula thereof. The technical scheme is as follows: the culture medium formula is characterized by comprising the following components: 1/2 of MS, 40-60g/L of banana, 0.8-1.2g/L of active carbon, 0.8-1.2g/L of NAA, 0.4-0.8mg/L of 6-BA, 22-30g/L of cane sugar, 5.5-6.5g/L of agar, 40-60g/L of onion and 10-20g/L of crab shell powder. The method is characterized by comprising the following steps of: 1) disinfection of an inoculation chamber; 2) disinfection of a culture chamber; 3) preparation of culture medium; 4) sterilization of the culture medium and inoculation instruments; 5) inoculation of explants; and 6) culture of tissue culture seedlings.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for reducing the pollution rate of tissue culture of Auranthus aurantiacum and its medium formula. Background technique [0002] Anoectochilus roxburghii (Anoectochilus roxburghii), also known as Golden Silk Grass, is a perennial herb of the family Orchidaceae. It is a traditional traditional Chinese medicine in my country. It enjoys the laudatory title of "King of Medicine" among the people. In southern Zhejiang, Fujian, Taiwan, and Guangdong and Guangxi, clematis has been used to boil water, make tea, and make soup for a long time for physical fitness, nourishing and recuperation. Modern research shows that clematis contains polysaccharides, flavonoids, cardiac glycosides and other components, which can enhance human immunity, improve body resistance, prevent disease invasion, enhance and improve physical fitness and other effects. [0003] Fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 邵清松胡润淮王莹季宗富易浩君
Owner ZHEJIANG FORESTRY UNIVERSITY
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