A kind of tissue culture and in vitro regeneration method of Ting taiqing

A technology of in vitro regeneration and tissue culture, which is applied in the field of bioengineering to achieve the effect of shortening the culture time, saving the workload, and improving the quality of the seedlings

Active Publication Date: 2019-08-09
SOUTHWEST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] According to the relevant literature, there is no report on the method of realizing rapid propagation of Ting taiqing through tissue culture. Therefore, it is urgent to explore a method that can realize rapid propagation to meet the needs of modern production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A method for tissue culture and in vitro regeneration of Ting taiqing, the steps comprising:

[0023] (1) Disinfection of explants: select the leaves of Ting taiqing seedlings as explants, wash the soil with tap water, put it into a beaker, add water and a small amount of washing powder, make a solution for soaking, shake gently for 5min, Rinse with tap water for 2 hours, and rinse the washing powder; finally transfer to the ultra-clean workbench, disinfect with 75% ethanol solution for 3 seconds, wash with sterile water twice, and use 0.1% mercuric chloride solution containing Tween (2 drops) Disinfect for 4 minutes, wash 6 times with sterile water, and wash for later use.

[0024] (2) Induction of callus and differentiation of adventitious buds: under aseptic operation, cut the sterilized leaves into 1cm 2 Leaves of different sizes were inoculated into callus induction medium (WPM medium+TDZ2.0mg / L+NAA 0.2mg / L+agar 4.5g / L+sucrose 20g / L, pH=5.8) During the process, m...

Embodiment 2

[0028] A method for tissue culture and in vitro regeneration of Ting taiqing, the steps comprising:

[0029] (1) Disinfection of explants: select the leaves of Ting taiqing seedlings as explants, wash the soil with tap water, put it into a beaker, add water and a small amount of washing powder, make a solution for soaking, shake gently for 5min, Rinse again with tap water for 2 hours, and rinse off the washing powder; finally transfer to the ultra-clean workbench, disinfect with 75% ethanol solution for 5 seconds, wash with sterile water twice, and use 0.1% mercuric chloride solution containing Tween (2 drops) Disinfect for 5 minutes, wash with sterile water for 6 times and set aside;

[0030] (2) Induction of callus and differentiation of adventitious buds: under aseptic operation, cut the sterilized leaves into 1.2cm 2size, inoculated into the callus induction medium (MS medium + TDZ 1.5mg / L + NAA0.1mg / L + agar 4.5g / L + sucrose 20g / L, pH = 5.8) in the way that the leaves ar...

Embodiment 3

[0034] A method for tissue culture and in vitro regeneration of Ting taiqing, the steps comprising:

[0035] (1) Disinfection of explants: select the leaves of Ting taiqing seedlings as explants, wash the soil with tap water, put it into a beaker, add water and a small amount of washing powder, make a solution for soaking, shake gently for 5min, Rinse again with tap water for 2 hours, and rinse off the washing powder; finally transfer to the ultra-clean workbench, disinfect with 75% ethanol solution for 3 seconds, wash with sterile water twice, and disinfect with 0.1% mercuric chloride solution containing Tween (2 drops) 4min, wash 6 times with sterile water, wash and set aside;

[0036] (2) Induction of callus and differentiation of adventitious buds: under aseptic operation, cut the sterilized leaves into 1.5cm 2 Size, inoculated into callus induction medium (1 / 2MS medium + TDZ 2.5mg / L + NAA 0.3mg / L + agar 4.5g / L + sucrose 20g / L, pH = 5.8) In the medium, make the incision ...

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Abstract

The invention relates to a tissue culture and regeneration in vitro method for clerodendrum subscaposum. The method comprises the steps of selecting and disinfecting an explant, inducing a callus tissue, performing differentiation induction on an adventitious bud, cultivating a strong seedling, rooting and the like. A seedling leaf of the clerodendrum subscaposum is taken as the explant, a tissueculture system of the clerodendrum subscaposum is explored, the ideal culture conditions for acquiring an asepsis material, inducing the callus tissue, performing differentiation induction on the adventitious bud, cultivating the strong seedling, rooting and the like are determined, a vegetative propagation system of the clerodendrum subscaposum is finally successfully established, and certain theoretical and technical foundation is provided for the full use and genetic improvement of the resource of the clerodendrum subscaposum medicine as well as culture of clonal seedlings. The culture method has the advantages that inducing the callus tissue and differentiating the adventitious bud are finished in one step, the operation is simple, the culture time is greatly shortened while the workload is saved, a large quantity of tissue culture seedlings are produced within a short period of time, the reproductive rate is high, the quality of the seedling is high, and the method is suitable forindustrialized production.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for tissue culture and in-vitro regeneration of Singing chinensis. Background technique [0002] Clerodendrum subscaposum is a perennial deciduous or evergreen shrub of Verbenaceae Daqing. The whole plant is used as medicine. Its flowers can calm the nerves and stop bleeding. It is mainly used to treat palpitations, insomnia, and bleeding hemorrhoids; Indications for migraine, bruises, bruises, carbuncle and sores; roots and stems can clear lung heat, facilitate urination, cool blood and stop bleeding. Rheumatic Ostealgia Treatment. [0003] Stems erect, unbranched or less branched, branchlets quadrangular. The leaves are alternately opposite, thin leathery, with long stalks, heart-shaped or oval leaves, sharp or acuminate at the top, heart-shaped at the base, fine teeth on the edges, and small coarse hairs scattered on the leaves. Racemes terminal, oblique to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 邓国宾王连春杨燕张媛蔡光辉胡鹏飞周炳江
Owner SOUTHWEST FORESTRY UNIVERSITY
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