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High-content camptothecine camplotheca acuminata callus culture method

A technology of callus and culture method, applied in the field of camptothecin callus culture with high content of camptothecin

Inactive Publication Date: 2003-11-12
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the induction of callus containing camptothecin on MS as the basic medium

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] (1) Explant preparation

[0030] Material disinfection: Soak the removed philodendron leaves or petioles in 70% alcohol for 10-30 seconds on the ultra-clean workbench, rinse with sterile water twice for 5 minutes each time, and insert 0.1% HgCl 2 Sterilize in the solution for 3 to 7 minutes, rinse with sterile water for 3 to 5 times, each time for 5 minutes, and blot dry on sterile filter paper.

[0031] (2) Callus induction

[0032] (1) The treated leaves of Campylodontia chinensis were cut into 0.5cm×0.5cm pieces, and the stalks of Campylodontia cypress leaves were cut into 0.5cm long sections and inserted into callus induction medium to induce callus. In this embodiment, the culture medium for inducing callus is prepared by adding the following raw materials and their proportions to 1 liter of MS basic medium:

[0033] Sucrose 30g

[0034] Agar powder 6.5g

[0035] 2,4-Dichlorophenoxyacetic acid 2.0mg

[0036] 6-(Benzylamino)purine 1.0mg

[0037...

Embodiment 2

[0039] In the callus induction culture step of the present embodiment, the callus induction medium used is prepared by adding the following raw materials and their proportions to 1 liter of MS basic medium:

[0040] Sucrose 20g

[0041] Agar powder 5.4g

[0042] 2,4-Dichlorophenoxyacetic acid 0.25mg

[0043] 6-(Benzylamino)purine 0.5mg

[0044] In this example, other cultivation steps are the same as in Example 1.

Embodiment 3

[0046] In the callus induction culture step of the present embodiment, the callus induction medium used is prepared by adding the following raw materials and their proportions to 1 liter of MS basic medium:

[0047] Sucrose 40g

[0048] Agar powder 7.2g

[0049] 2,4-Dichlorophenoxyacetic acid 3.0mg

[0050] 6-(Benzylamino)purine 3.0mg

[0051] In this example, other cultivation steps are the same as in Example 1.

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Abstract

The invention is a high-content camptothecin callus cultivating method of camptotheca acuminata, its steps including: (1) prepare outer plant body; (2) induce the callus; cut the happy-tree laminae into blocks of 0.5X0.5-1.0 cm2, and the leafstalk cut into 0.5-1.0 cm segments inserted in the callus-inducing culture medium for inducing. The above culture medium is prepared by adding sucrose, 20-40g, agar powder, 5.4-7.2g, 2,4-dichlorophenoxyacetic acid, 0.25-3.0 mg, and 6-(benzyl amino) purine, 0.5-3.0 mg in 1 litre of MS basic culture medium, pH 6.0. Hold the prepared culture medium in the nutritious bottles, 25ml / bottle, then sterilize for 25 minutes under a 0.1-0.15 MPa pressure, then place the bottles in the hothouse for cultivating, temperature 22-26 deg.C and luminous intensity 2000-3000 lx.

Description

technical field [0001] The invention belongs to the plant regeneration of plant tissue culture technology and the technical field of undifferentiated human, animal or plant cells, and specifically relates to a method for culturing camptothecin callus with high content of camptothecin. Background technique [0002] Camptothecin (CPT) is an alkaloid contained in the unique tree species Camptotheca acuminate Decne in my country. It is another promising plant anticancer drug after paclitaxel. The only anticancer drug that exerts cytotoxicity exclusively by inhibiting topoisomerase I. [0003] The traditional camptothecin extraction method uses the root bark, fruit and leaves of camptothecin to extract camptothecin. The root bark as raw material needs to be cut down a lot of camptothecin, and the fruit as raw material is subject to seasonal restrictions, and the consumption is large and the output is small , which is not conducive to the effective utilization of camphor tree reso...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 王喆之强小利田宇红李发荣
Owner SHAANXI NORMAL UNIV
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