Induction method for Camptotheca acuminata Decne polyploid

A technology of polyploidy and tetraploidy, applied in horticultural methods, botany equipment and methods, plant cells, etc., can solve the problem of low content of camptothecin, achieve high content of active ingredients, improved genetics, and high efficiency Effect

Inactive Publication Date: 2011-02-09
CHENGDU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The object of the present invention is to solve the problem of low content of camptothecin in existing camptothecin. Higher than normal diploids, polyploid plants also have the characteristics of organ gigantism and provide a method for inducing camptothecin polyploids, which can breed camptothecins with high content of active ingredients (i.e. camptothecin) Ploid tissue cultured seedlings to meet the growing demand for camptothecin anticancer drugs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Processing of explants: collect the young and tender stems of Campylodon chinensis as explants, disinfect them with 70% alcohol for 30 seconds, rinse them with sterile water for 3 times, and then use 0.1% mercuric chloride After 30 minutes of disinfection, rinse with sterile water 5 times, and then dry the water with sterile filter paper;

[0018] (2) Callus culture: cut the sterilized stems into 2cm lengths and inoculate them in B 5 +NAAlmg / L+2,4-D0.5mg / L+KT0.5mg / L callus culture medium;

[0019] (3) Polyploid induction: Soak the callus in the colchicine solution that has been filtered and sterilized for 40 hours, the solution concentration is 400 mg / L, then rinse with sterile water for 3 to 4 times, and then transfer to a place without Colchicine MS+6-BA 1mg / L+NAA0.2mg / L bud differentiation medium cultured in the dark for 7 days, then transferred to light culture, the light intensity was 1500-2000Lx, and the light was 12 hours a day;

[0020] (4) Identification ...

Embodiment 2

[0025] (1) Treatment of explants: collect the young and tender leaves of Campylodon chinensis as explants, disinfect them with 75% alcohol for 5 seconds, rinse them with sterile water for 3 times, and then use 0.1% mercuric chloride After 5 minutes of disinfection, rinse with sterile water 5 times, and then dry the water with sterile filter paper;

[0026] (2) Callus culture: Cut the sterilized young leaves in half and inoculate them in B 5 +2,4-D1.5mg / L+KT0.5mg / L callus culture medium;

[0027] (3) Polyploid induction: Soak the callus in the colchicine solution that has been filtered and sterilized for 30 hours. Colchicine MS+6-BA0.5mg / L+NAA0.2mg / L bud differentiation medium cultured in the dark for 7 days, then transferred to light culture, the light intensity was 1500-2000Lx, and the light was 12 hours a day;

[0028] (4) Polyploid identification: the adventitious buds after 50 days of growth were pretreated in a 0.2% colchicine solution for 2.3 hours, washed with water, ...

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Abstract

The invention discloses an induction method for camptotheca acuminate polyploidy, comprising the steps that: 1 the stem or leaves of camptotheca acuminate are used as explant and disinfection treatment is performed; 2 callus is cultivated; 3 the polyploidy is induced and bud differentiation culture is performed to the induced callus; 4 the polyploidy is identified; 5 the polyploidy seedlings are cultivated; 6 the seedlings are harden and transplanted. The induction method has the advantages that, through the polyploidy induction to camptotheca acuminate plants, the camptotheca acuminate tetraploid tissue culture seedlings having high content of camptothecin can be cultivated; therefore the great demand of human to anticancer camptothecin derivatives is satisfied.

Description

technical field [0001] The invention relates to a method for inducing camptophyll polyploid. Background technique [0002] Campylodontaceae belongs to the Eclipta genus in the Davidiaceae family. It is a unique economic tree species in China. In August 1998, it was approved as the first batch of national key II protected wild plants. Camptothecin contained in camptothecin has significant anti-tumor activity and unique anti-cancer mechanism. Studies have confirmed that camptothecin prevents the synthesis of cellular DNA and RNA by inhibiting the activity of DNA topoisomerase I and the replication of retroviruses. Phytoactive ingredients. Pre-clinical trials have shown that camptothecin and its derivatives have varying degrees of curative effect on more than 30 tumors such as bladder cancer, brain cancer, breast cancer, cervical cancer, colon cancer, leukemia, lung cancer, lymphoma and liver cancer, which has caused people to The widespread attention of camptophylla has bec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H1/08C12N5/04A01G31/00G01N33/48A01H4/00
Inventor 王跃华孙雁霞张海强徐作英邬晓勇刘碧崇
Owner CHENGDU UNIV
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