Method for rapidly breeding transgene abrotanum using micro adventive bud technique

A technology of transgenic and Artemisia annua, which is applied in the biological field, can solve the problems of long-term cultivation of pure varieties, restrictions on large-scale promotion of transgenic Artemisia annua, lack of genetic stability, etc., and achieve the effect of increasing the production of artemisinin

Inactive Publication Date: 2008-05-14
SUZHOU TANGJI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, transgenic Artemisia annua is generally a heterozygote, which does not have genetic stability, and the genes in the offspring will segregate, which is not conducive to agricultural production
It takes a long time to breed pure varieties from transgenic Artemisia annua, and consumes a lot of manpower and material resources, which limits the large-scale promotion of transgenic Artemisia annua
Have not yet found relevant reports on the rapid propagation of transgenic Artemisia annua using micro-adventitious bud technology mentioned in the subject of the present invention

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Disinfection of Transgenic Artemisia annua Explants and Induction of Adventitious Buds

[0022] Young stem segments (1 cm long) containing growth points were cut from transgenic Artemisia annua plants as explants. Disinfect with 75% alcohol for 20 seconds, wash once with sterile water, 1.5% HgCl 2 Disinfect for 6 minutes and wash with sterile water 5 times. The sterilized transgenic Artemisia annua explants were transferred to the adventitious bud induction medium, which was MS medium+0.5mg / L 6-benzyladenine (6-BA)+0.05 mg / L naphthaleneacetic acid (NAA)+3% sucrose+0.65% agar powder. The culture temperature is 25±1° C., light is 16 hours a day, subculture is performed every 14 days, and a large number of microadventitious buds (length 1.5 cm) grow around the explant after 1 subculture. The induction rate of adventitious buds reached 91%.

[0023] Elongation and rooting of transgenic adventitious shoots of Artemisia annua

[0024] The explants with a large number of ...

Embodiment 2

[0028] Disinfection of Transgenic Artemisia annua Explants and Induction of Adventitious Buds

[0029] Young stem segments (1.5 cm long) containing growth points were cut from transgenic Artemisia annua plants as explants. Disinfect with 75% alcohol for 20 seconds, wash once with sterile water, 1.5% HgCl 2 Disinfect for 6 minutes and wash with sterile water 5 times. The sterilized transgenic Artemisia annua explants were transferred to the adventitious bud induction medium, which was MS medium+0.5mg / L 6-benzyladenine (6-BA)+0.05 mg / L naphthaleneacetic acid (NAA)+3% sucrose+0.65% agar powder. The culture temperature is 25±1° C., light is 16 hours a day, subculture is performed every 14 days, and a large number of microadventitious buds (length 1.5 cm) grow around the explant after 1 subculture. The induction rate of adventitious buds reached 94%.

[0030] Elongation and rooting of transgenic adventitious shoots of Artemisia annua

[0031] The explants with a large number o...

Embodiment 3

[0035] Disinfection of Transgenic Artemisia annua Explants and Induction of Adventitious Buds

[0036] Young stem segments (2 cm long) containing growth points were cut from transgenic Artemisia annua plants as explants. Disinfect with 75% alcohol for 20 seconds, wash once with sterile water, 1.5% HgCl 2Disinfect for 6 minutes and wash with sterile water 5 times. The sterilized transgenic Artemisia annua explants were transferred to the adventitious bud induction medium, which was MS medium+0.5mg / L 6-benzyladenine (6-BA)+0.05 mg / L naphthaleneacetic acid (NAA)+3% sucrose+0.65% agar powder. The culture temperature was 25±1° C., and the light was 16 hours a day. Subculture was performed every 20 days. After one subculture, a large number of microadventitious buds (1.5 cm in length) grew around the explants. The induction rate of adventitious buds reached 98%.

[0037] Elongation and rooting of transgenic adventitious shoots of Artemisia annua

[0038] The explants with a lar...

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Abstract

The invention relates to a method for rapidly propagating transgenic Artemisia annua by microadventitious bud technology in the field of biotechnology. In the present invention, after the transgenic Artemisia annua explants are sterilized, they are placed on the medium for inducing adventitious buds, cultivated under light, and a large number of adventitious buds grow around the explants; the explants with a large number of adventitious buds are transferred to Micro-adventitious buds are elongated in the medium for elongation of adventitious buds; the elongated adventitious buds are cut and transferred to the rooting medium of adventitious buds to produce adventitious roots; the complete plants that grow robustly are transplanted into a medium containing transplanting A large number of normal-growing transgenic Artemisia annua plants were obtained through domestication. The present invention screens out a medium suitable for induction and rooting of transgenic Artemisia annua explants, the induction rate of adventitious buds of transgenic Artemisia annua explants exceeds 90%, the rooting rate of adventitious buds is 100%, and the survival rate of transplanted regenerated plants is 100%. , The reproduction coefficient reaches 80-100.

Description

technical field [0001] The invention relates to a method in the field of biotechnology, in particular to a method for rapidly propagating transgenic Artemisia annua using microadventitious bud technology. Background technique [0002] Artemisia annua L. is an annual herb belonging to the family Asteraceae. Artemisinin (artemisinin) is a sesquiterpene lactone compound containing a peroxide bridge structure isolated from its aerial part. It is currently the most effective drug for treating malaria in the world, especially for cerebral malaria and anti-malaria Chloroquine malaria has the characteristics of quick action and low toxicity. Currently, the most effective treatment for malaria recommended by the World Health Organization is artemisinin combination therapy (ACTs). In addition, with the deepening of pharmacological research on artemisinin, scientists have found that artemisinin and its derivatives also have anti-inflammatory, anti-schistosomiasis, anti-tumor and immu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C12N5/04
Inventor 唐克轩张凌景福远王国丰
Owner SUZHOU TANGJI BIOTECH CO LTD
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