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Ligusticum wallichii tissue cultivation fast-propagating method with young leaflet tablet as explant

A tissue culture and explant technology, applied in the fields of botanical equipment and methods, horticultural methods, cultivation, etc., can solve the problems of "ling seed" easy aging, unstable germplasm resources, long breeding time, etc., and achieve robust growth. , Not easy to be infected with viruses, strong practical effect

Inactive Publication Date: 2008-08-20
CHENGDU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a fast tissue culture of Rhizoma Chuanxiong with young leaves as explants in view of the problems in the prior art that the breeding time is long, the germplasm resources are unstable, "Lingzhong" itself is easy to age and carry bacteria. This method can quickly reproduce a large number of Ligusticum chuanxiong tissue culture seedlings with fast growth, high quality and low bacterial infection rate.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Disinfection of explants:

[0018] Put the young leaves of Ligusticum chuanxiong as explants into washing powder solution for 2 minutes, rinse them with running water for 2.5 hours, and then disinfect them with 70% ethanol at a concentration of 0.1% for 10 seconds under sterile conditions. mercuric chloride for 3 minutes, and finally washed 4 times with sterile water;

[0019] (2) Culture of callus:

[0020] Cut the sterilized leaves into appropriate size, inoculate them on the callus medium MS+6-BA0.5mg / L+2,4-D2mg / L+sucrose 30g / L+agar 7.0g / L, and inoculate at 24~26 ℃, 12 hours of light per day, and light intensity of 1500-2000Lx for cultivation, the statistical induction rate is 65% after 45 days;

[0021] (3) Differentiation culture:

[0022] After 45 days, the callus was divided into appropriate sizes, transferred to the differentiation medium MS+6-BA0.5mg / L+NAA0.5mg / L+sucrose 30g / L+agar 7.0g / L, and kept at 24~26℃, Illuminated for 12 hours a day, and the ligh...

Embodiment 2

[0028] The callus culture medium adopts MS+2,4-D 2mg / L+sucrose 30g / L+agar 7.0g / L, and other steps are the same as in Example 1. The callus culture medium was used to induce callus, and the induction rate was 67.5% after 45 days, which was slightly higher than that in Example 1. The resulting callus grows faster, and it is very beneficial to the cell suspension culture of Ligusticum chuanxiong, but it is not conducive to the differentiation of adventitious buds in the later stage. Although the callus culture medium adopted in Example 1 has a slightly lower induction rate and a slightly slower callus growth rate, it is beneficial to the differentiation of adventitious buds in the later stage.

Embodiment 3

[0030] The callus culture medium adopts MS+6-BA0.5mg / L+2,4-D3mg / L+sucrose 30g / L+agar 7.0g / L, and other steps are the same as in Example 1. The callus culture medium was used to induce callus, and the induction rate was 55% after 45 days, which was lower than Example 1 and Example 2, but not obvious. However, when the concentration of 2,4-D was 4 mg / L, the induction rate was only 25% after 45 days, showing a significant decline.

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PUM

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Abstract

The invention discloses a ligustcum chuanxiong horn tissue culture fast propagation method by taking tender leaves as explants, comprising the steps of the sterilization of explants, the culture of callus, differentiation culture, rooting culture, seedling hardening and transplanting. The invention is characterized in that, the adopted callus culture medium is MS+6-BA0 to 0.5mg / L+2, 4-D1 to 3mg / L+sucrose30g / L+agar6.5 to 7.0g / L; the differentiation culture medium is MS+6-BA0.1 to 0.5mg / L+NAA0.1 to 0.8mg / L+sucrose30g / L+agar6.5 to 7.0g / L; the rooting culture medium is 1 / 2MS+ NAA0.1 to 0.5mg / L+IBA0.1 to 0.5mg / L+sucrose15g / L+agar6.5 to 7.0g / L. The method has the advantages that, ligustcum chuanxiong horn tissue culture seedlings with fast growth rate, good quality, and low pollution rate can be rapidly propagated in large amount, and the requirement to produce ligustcum chuanxiong horn on a large scale is satisfied.

Description

technical field [0001] The invention relates to a plant regeneration method through tissue culture technology, in particular to a rapid propagation method of Rhizoma Chuanxiong tissue culture with young leaves as explants. Background technique [0002] Chuanxiong is one of the most common Chinese herbal medicines in my country, and it is also a best-selling herbal medicine in the international market in recent years. It is an important raw material for many health care products and medicines. Because the medicinal material is limited by geographical conditions, its planting area is limited, and it is mainly distributed in Dujiangyan, Sichuan Province, my country. Ligusticum chuanxiong is a biennial plant. Due to the difficulty in fruiting of Ligusticum chuanxiong, the breeding method of chuanxiong has always been the traditional vegetative propagation and cultivation of stem nodes. This method needs to be cultivated in mountainous areas and planted in dam areas during the br...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01G31/00C12N5/04
Inventor 王跃华孙雁霞马丹炜吴洁邬晓勇徐文俊张海强刘畅
Owner CHENGDU UNIV
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