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Inducing culture method of Aconitum vilmorinianum Kom. embryoid

A cultivation method and technology of embryoid bodies, which are applied in horticultural methods, botanical equipment and methods, plant regeneration, etc., and can solve problems such as shortage of seedlings

Active Publication Date: 2020-10-23
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The object of the present invention is to provide a method for inducing and culturing the Aconitum aconitifolia embryoid, which utilizes the Aconitum aconitifolia bud to induce and cultivate the Aconitum aconitifolia embryoid, which has the characteristics of an embryo. Under certain conditions, it can quickly grow into regenerated disease-free plants, which can effectively solve the problem of shortage of seedlings in the production of Aconitum japonica, and realize the rapid propagation of high-quality Aconitum japonica seedlings

Method used

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  • Inducing culture method of Aconitum vilmorinianum Kom. embryoid
  • Inducing culture method of Aconitum vilmorinianum Kom. embryoid
  • Inducing culture method of Aconitum vilmorinianum Kom. embryoid

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Embodiment 1

[0021] Embodiment 1: The method for culturing the embryoid body of Aconitum aconitifolia provided by this embodiment comprises the following steps:

[0022] (1) Selection of bulbils: Select Aconitum aconitifolia plants that grow robustly, and cut young bulbils between the leaf axils as explants ( figure 1 A);

[0023] (2) Bulb disinfection: Sterilize the Aconitum japonica selected in step (1) in 75% ethanol for 30 seconds, wash it with sterile water for 5 times, and then put it in 0.1% HgCl 2 Disinfect in the solution for 10 minutes, then wash 5 times with sterile water, and dry the water with sterile paper;

[0024] (3) Callus induction: such as figure 1 As shown in B, insert the Aconitum aconitum sterilized in step (2) into solid medium (2,4-dichlorophenoxyacetic acid 2.0mg / L, KT 0.5mg / L, PVP 1.5g are added to MS medium / L, agar 5.0g / L, sucrose 20g / L, medium pH value 5.8), cultured under the conditions of culture temperature 25℃, light 12h per day, and light intensity 3...

Embodiment 2

[0027] Embodiment 2: The method for culturing the embryoid body of Aconitum aconitifolia provided by this embodiment comprises the following steps:

[0028] (1) Selection of bulbils: Select the vigorously growing plants of Aconiti japonica, and cut the tender bulbils between the leaf axils as explants;

[0029] (2) Bulb disinfection: Sterilize the Aconitum japonica selected in step (1) in 75% ethanol for 60 seconds, wash it with sterile water for 6 times, and then put it in 0.1% HgCl 2 Disinfect in the solution for 15 minutes, then wash 5 times with sterile water, and dry the water with sterile paper;

[0030](3) Callus induction: insert the sterilized Aconitum aconitifolia buds in step (2) into solid medium (add 2,4-D 1.5mg / L, KT 0.3mg / L, PVP 1.5g to MS medium / L, agar 5.5g / L, sucrose 30g / L, medium pH value 5.8), cultured under conditions of culture temperature 25℃, light 12h per day, and light intensity 3500lx. rate of 0%, browning rate of 0%, callus induction rate of 87.2...

Embodiment 3

[0033] Embodiment 3: The method for culturing the embryoid body of Aconitum aconitifolia provided by this embodiment comprises the following steps:

[0034] (1) Selection of bulbils: Select the vigorously growing plants of Aconiti japonica, and cut the tender bulbils between the leaf axils as explants;

[0035] (2) Bulb disinfection: Sterilize the Aconitum japonica selected in step (1) in 75% ethanol for 50 seconds, wash it with sterile water for 4 times, and then put it in 0.1% HgCl 2 Disinfect in the solution for 12 minutes, then wash 6 times with sterile water, and dry the water with sterile paper;

[0036] (3) Callus induction: insert the sterilized Aconitum aconitifolia buds in step (2) into solid medium (add 2,4-D 1.0mg / L, KT 0.1mg / L, PVP 1.5g to MS medium / L, agar 6.0g / L, sucrose 35g / L, medium pH value 5.8), cultured under conditions of culture temperature 23℃, light 10h per day, and light intensity 3000lx. rate of 0%, browning rate of 0%, callus induction rate of 62....

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Abstract

The invention discloses an inducing culture method of an Aconitum vilmorinianum Kom. embryoid. The inducing culture method includes the steps that bulbils of Aconitum vilmorinianum Kom. are used as explants, after the bulbils are disinfected, callus is first induced, and embryonic callus with the color of light yellow with green and vigorous division is selected from the callus for inducing culture, and obtain the Aconitum vilmorinianum Kom. embryoid is obtained. A new way is provided for the mass cultivation and rapid propagation of high-quality seedlings of the Aconitum vilmorinianum Kom., and the problems of the shortage of plant resources and seedling resources of the Aconitum vilmorinianum Kom. as well as serious diseases and the like can be effectively solved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for inducing and culturing the embryoid body of Aconiti fruticosa. Background technique [0002] Huangcaowu ( Aconitum vilmorinianum Kom.) is a perennial herbaceous vine of the genus Aconitum in the family Ranunculaceae. Its roots are highly toxic and have the functions of expelling wind and cold, dehumidification and pain relief, detoxification and swelling, etc. It is often used to treat bruises, rheumatism, cold-damp arthralgia, Cold hands and feet, sores and other diseases. As an important traditional Chinese medicine, Huangcaowu has been directly harvesting wild resources for medicinal purposes for a long time, but it has been over-harvested and its quantity has decreased sharply. In recent years, artificial planting has begun to solve the problem of resource scarcity, mainly using methods such as seed propagation, bulbil propagation and small root propagation. Howe...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/005A01H4/001
Inventor 李昆志李一果
Owner KUNMING UNIV OF SCI & TECH
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