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Method for producing anoectochilus formosanus by tissue culture

A technology of tissue culture and culture medium, applied in the field of plant tissue culture, can solve the problems affecting the in-depth research and application of Taiwan Aurea clematis, and achieve the effects of short cycle, large output and increased output

Inactive Publication Date: 2010-06-02
烟台汇鹏生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The 2005 edition of the Chinese Pharmacopoeia has not yet recorded Clematis
Have not found a practical and effective culture technology to solve the doubts and difficulties in the induction and proliferation of A. taiwanensis, which has affected the in-depth research and application of A. taiwanensis

Method used

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  • Method for producing anoectochilus formosanus by tissue culture
  • Method for producing anoectochilus formosanus by tissue culture
  • Method for producing anoectochilus formosanus by tissue culture

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1: a kind of method of producing Aureus taiwanensis by tissue culture, it comprises the steps:

[0026] a. Plant selection and aseptic treatment:

[0027] a) select the wild plants of Taiwan golden clematis (Anoectochilus formosanus Hayata) grown in July, rinse with tap water to remove the roots and leaves, then rinse with distilled water, and put them in the ultra-clean bench; b) remove the roots and leaves Soak the stem plant body in 75% alcohol for 30 seconds to sterilize the surface, and then clean it with sterile water; c) Soak the stem plant body sterilized by alcohol in 0.5% sodium hypochlorite solution for 5 minutes. Rinse with water for 5 times, then blot the sterile water with filter paper; d) under aseptic conditions, cut the stem section sterilized by sodium chlorate into small sections of clematis taiwanensis with stem nodes of about 1 cm;

[0028] b. Cluster bud induction culture:

[0029] The aseptically treated small segment of A. taiwanensi...

Embodiment 2

[0032] Embodiment 2: a kind of method of producing Taiwan clematis by tissue culture, it comprises the steps:

[0033] a. Plant selection and aseptic treatment:

[0034] a) Select the wild plants of Taiwan golden clematis (Anoectochilus formosanus Hayata) grown in August, rinse with tap water to remove the roots and leaves, then rinse with distilled water, and put them in the ultra-clean workbench; b) remove the roots and leaves Soak the stem section plant body with 78% alcohol for 28 seconds to carry out surface sterilization, and then clean it with sterile water; c) soak and disinfect the stem section plant body through alcohol sterilization with 0.6% sodium hypochlorite solution for 4 minutes, and then use sterile water to clean the plant body. Rinse with water for 4 times, then blot dry the sterile water with filter paper; d) under aseptic conditions, cut the stem section sterilized by sodium chlorate into small sections of clematis taiwanensis with stem nodes of about 1 c...

Embodiment 3

[0039] Embodiment 3: detect the total flavonoids of the regenerated plant of A. taiwanensis cultivated in wild A. taiwanensis, embodiment 1 and embodiment 2. The specific detection method includes the following steps:

[0040] (1) Preparation of reference substance solution

[0041] Precisely weigh 9.9mg of rutin reference substance dried to constant weight, put it in a 5mL volumetric flask, add ethanol to dissolve it and adjust the volume to the mark, and shake well to obtain a concentration of 1.98mg·mL -1The rutin reference substance solution, set aside. Precisely measure 0, 0.1, 0.2, 0.3, 0.4, 0.6, 0.8, 1.0mL of the reference substance solution, put it in a 25mL graduated test tube, add water to make up to 5mL; add 1.0mL of 5% sodium nitrite solution, mix well, and place for 6 Minutes; add 1.0mL of 10% aluminum nitrate solution, and let stand for 6 minutes; add 10.0mL of 4% sodium hydroxide solution, add water to make up to the mark, mix well, let stand for 15 minutes. A...

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Abstract

The invention discloses a method for producing anoectochilus formosanus by tissue culture, comprising the following steps: frond selection and asepsis, cluster bud inducing culture and large-scale culture. The key to the technique is characterized in that two different culture media are applied to the inducing culture and the large-scale culture, wherein the inducing culture medium is as follows: H culture medium+ 10-50g / L of cane sugar + 0.1-3.0mg / L of alpha-naphthylacetic acid+ 0.1-5.0mg / L of 6-benzyladenine, and pH value is 5.0-7.0; the large-scale culture medium is as follows: B5 culture medium+ 20-40g / L of cane sugar+ 6-8g / L of agar+ 2.0-3.0g / L of active carbon+ 1.0-2.0mg / L of alpha-naphthylacetic acid+ 2.0-3.0mg / L of benzyladenine, and pH value is 5.5-6.5. The method is easy to operate, features low production cost, high output, high quality and no pollution to the environment and can realize batch production.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for producing Aureus taiwanensis through tissue culture. Background technique [0002] Anoectochilus formosanus Hayata is an endangered and rare medicinal plant. Medicinal whole herb, known as the "King of Medicine", is a traditional precious medicinal material in our country. It is flat in nature and sweet in taste, has the effects of clearing heat and cooling blood, dispelling wind and dampness, and is mainly used for hemoptysis, bronchitis, nephritis, cystitis, diabetes, hematuria, rheumatoid arthritis, convulsions in children, snake bites, etc. The 2005 edition of the Chinese Pharmacopoeia has not yet recorded Clematis clematis. [0003] Clematis taiwanensis is a promising traditional Chinese medicine immune enhancer, and has anti-tumor, hypoglycemic, anti-oxidation, anti-mutagenic effects, and has been highly valued by scholars at home and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 贾景明
Owner 烟台汇鹏生物科技有限公司
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