Callus induction method taking paris polyphylla coleoptiles as explants

A callus induction and callus technology, applied in the field of callus induction, can solve the problem of no obvious breakthrough in explants, and achieve the effects of solving the shortage of medicinal materials, reducing production costs, and realizing mass production.

Inactive Publication Date: 2012-01-04
CHENGDU UNIV
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AI Technical Summary

Problems solved by technology

At present, the tissue culture of Paris polyphylla has not achieved satisfactory results, the main reason

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] (1) Select the white crypto-buds that will germinate on the rhizomes of P. chinensis, are free from diseases and insect pests, and have a diameter of about 2 cm. Rinse the crypto-buds with tap water first, then dry them in the shade, and then sterilize them with 70% alcohol on a super-clean bench 30s, followed by disinfection with 0.1% mercuric chloride for 8 minutes, and finally washed 5 times with aseptic water shock, after washing, use sterilized filter paper to repeatedly absorb the moisture on the surface of cryptobuds;

[0015] (2) Inoculate the sterilized cryptobuds in medium MS+6-BA 2.0 mg·L -1 + coconut milk 80mg·L -1 + sucrose 30g·L -1 +Agar 6.0g·L -1 The pre-cultivation was carried out in the medium, the pH value used was 5.8, the culture temperature was 20°C, the light was 8 hours a day, and the light intensity was 1200lx;

[0016] (3) Select the buds that are obviously enlarged after pre-cultivation for 25 days, peel off the outer layer of the buds on th...

Embodiment 2

[0018] Change the pre-medium in Example 1 step (2) to MS+6-BA2.0mg L -1 + sucrose 30g·L -1 +Agar 6.0g·L -1 , the callus induction medium in step (3) was changed to MS+6-BA1.0mg L -1 +NAA3.0mg·L -1 + Coconut juice 30mg·L -1 + sucrose 30g·L -1 +Agar 6.0g·L -1 , other steps were the same as in Example 1, and the callus induction rate was 18.14% after induction culture for 55 days.

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Abstract

The invention discloses a callus induction method taking paris polyphylla coleoptiles as explants. The method comprises the following steps: firstly sterilizing and pre-culturing latent buds to be germinated on a paris polyphylla rootstock; and selecting the white inner-layer coleoptiles of the buds the volume of which is obviously increased after preculture as the explants, inoculating the explants into a callus induction culture medium with the concentration of MS+6-BA of being 0.5-2.0mg/L<-1>, the concentration of NAA of being 0.5-4mg/L<-1>, the concentration of coconut juice of being 20-80mg/L<-1>, the concentration of sucrose of being 30-50g/L<-1> and the concentration of agar of being 6.0-7.0g/L<-1>, and culturing for 50-60 days to obtain white or yellow-white compact calluses formed on the surfaces of the explants. The method disclosed by the invention better has the beneficial effects of better solving the problems of the traditional paris polyphylla production method such as long growth time, high cost and the like, and effectively alleviating the shortage phenomenon of the current paris polyphylla medicinal material.

Description

technical field [0001] The present invention relates to a method for culturing the pachyrhiza tissue, in particular to a method for inducing callus with the bud sheath of the polyphylla as an explant. Background technique [0002] Zhonglou is one of the precious Chinese medicinal materials. There are 24 kinds in the world, and 19 kinds in my country. Among them, the types and resources of the provinces in Southwest China are particularly rich, such as Yunnan, Sichuan, Guizhou and other places. The main active ingredient of Chonglou is steroidal saponins, and its aglycones are mainly diosgenin and pinanogenin of isospirostanols, and contain compounds such as amino acids, sterones, ecdysones, and flavonoid glycosides. Modern pharmacological research shows that Chonglou has anti-tumor, hemostasis, cough and asthma, antibacterial, antiviral and other effects, and its good curative effect has been used by many pharmaceutical industries. The main ingredient of important Chinese p...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 王跃华田梦良蒋亭亭李文光付伟杨军张珏张红玉陈燕王勇
Owner CHENGDU UNIV
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