Method for induction and subculture of taxus chinensis leaf callus and culture medium used by method

A technology for inducing culture medium and callus, applied in the directions of botanical equipment and methods, horticultural methods, applications, etc., can solve the problem that the amount of yew resources cannot meet the demand for paclitaxel production, and achieves low pollution rate and induction rate. High, fast growth effect

Inactive Publication Date: 2013-10-16
SHANGHAI INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, with the continuous strengthening of protection measures for wild yew resources in the world, it is impossible to use wild resources to produce paclitaxel, and the amount of artificially planted yew resources at this stage is far from meeting the demand for paclitaxel production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A method for inducing a yew leaf callus, specifically comprising the steps of:

[0029] (1) Disinfection of explants

[0030] Take the young leaves of Taxus chinensis in the same year, wash them with running water for 4 hours, disinfect them with 70% alcohol for 30 seconds, disinfect them with 1% mercury chloride for 8 minutes, and rinse them with sterile water;

[0031] (2), callus induction culture

[0032] Scratch the leaves of the sterilized yew explants obtained in step (1) with a blade and inoculate them on the induction medium, and culture them in the dark at a controlled temperature of 20-22°C for 20 days to obtain callus after induction ;

[0033] The induction medium, calculated per liter, consists of 400mg NH 4 N0 3 , 556mg Ca(NO 3 ) 2 .4H 2 O, 990 mg K 2 SO 4 , 96 mg CaCl 2 .2H 2 O, 170 mg KH 2 PO 4 , 0.25 mg Na 2 MoO 4 .2H 2 O, 370 mg MgSO 4 .7H 2 O, 22.4mg MnSO 4 .H 2 O, 8.6mg ZnSO 4 .7H 2 O, 0.25mg CuSO 4 .5H 2 O, 27.8 mg FeSO 4 ....

Embodiment 2

[0038] A method for inducing a yew leaf callus, specifically comprising the steps of:

[0039] (1) Disinfection of explants

[0040] Take the young leaves of Taxus chinensis in the same year, wash them with running water for 4 hours, disinfect them with 70% alcohol for 30 seconds, disinfect them with 1% mercury chloride for 8 minutes, and rinse them with sterile water;

[0041] (2), callus induction culture

[0042] Scratch the leaves of the sterilized yew explants obtained in step (1) with a blade and inoculate them on the induction medium. After 20 days of dark culture under the control temperature of 20-22°C, the callus after induction culture is obtained. ;

[0043] The induction medium, calculated per liter, consists of 400mg NH 4 N0 3 , 556mg Ca(NO 3 ) 2 .4H 2 O, 990 mg K 2 SO 4 , 96 mg CaCl 2 .2H 2 O, 170 mg KH 2 PO 4 , 0.25 mg Na 2 MoO 4 .2H 2 O, 370 mg MgSO 4 .7H 2 O, 22.4mg MnSO 4 .H 2 O, 8.6mg ZnSO 4 .7H 2 O, 0.25mg CuSO 4 .5H 2 O, 27.8 mg Fe...

Embodiment 3

[0048] A method for inducing a yew leaf callus, specifically comprising the steps of:

[0049] (1) Disinfection of explants

[0050] Take the young leaves of Taxus chinensis in the same year, wash them with running water for 4 hours, disinfect them with 70% alcohol for 30 seconds, disinfect them with 1% mercury chloride for 8 minutes, and rinse them with sterile water;

[0051] (2), callus induction culture

[0052] Scratch the leaves of the sterilized yew explants obtained in step (1) with a blade and inoculate them on the induction medium, and culture them in the dark at a controlled temperature of 20-22°C for 20 days to obtain callus after induction ;

[0053] The induction medium is calculated per liter, consisting of 1650mg NH 4 NO 3 , 1900mg KNO 3 , 440mg CaCl 2 2H 2 O, 370mg MgSO 4 ·7H 2 O, 170mg KH 2 PO 4 , 0.83mg KI, 6.2 mgH 3 BO 3 , 22.3mg, MnSO 4 4H 2 O, 8.6mg ZnSO 4 ·7H 2 O, 0.25mg Na 2 MnO 4 2H 2 O, 0.025mg CuSO 4 ·5H 2 O, 0.025mg CoCl 2 ·6H...

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Abstract

The invention discloses a method for induction and subculture of taxus chinensis leaf callus. The method comprises the following steps of: disinfecting tender leaves (which are taken as an explants) of south taxus chinensis growing in the current year, inoculating to a specific induced culture medium, selecting loosen and flaxen callus after culturing for 20 days in dark at the temperature of 20-22 DEG C, transferring into a specific subculture medium, and subculturing 20 days at the temperature of 20-22 DEG C, thus obtaining the taxus chinensis leaf callus. The method for the induction and subculture of the taxus chinensis leaf callus provided by the invention has the characteristics of high inductivity, short callus time, rapid growth rate, low pollution rate and the like; and the method achieves the purpose that the taxus chinensis callus is cultured on large scale through a plant cell culture technology.

Description

technical field [0001] The invention relates to a method for induction and subculture of yew leaf callus and the used culture medium. Background technique [0002] Paclitaxel (Taxo1) is from Taxaceae Taxus ( Taxus spp. ) A natural anti-cancer taxane compound extracted from the bark of the plant, it is currently recognized as the best broad-spectrum, strong active natural anti-cancer drug in the world. Due to the slow growth of Taxus plants, the content of paclitaxel is low, accounting for only 0.01% - 0.02% of the dry mass of the bark, and about one hundred thousandth of the dry mass of the branches and leaves. Therefore, with the continuous strengthening of protection measures for wild yew resources in the world, it is impossible to use wild resources to produce paclitaxel, and the amount of artificially planted yew resources at this stage is far from meeting the demand for paclitaxel production. [0003] Large-scale cultivation of Taxus chinensis calli by plant cell cult...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 宋丽莉赵华强姜育龙侯梅芳张妤萍
Owner SHANGHAI INST OF TECH
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