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Method for promoting synthesis of secondary metabolites in common bletilla pseudobulb suspension cultured cells

A technology for secondary metabolites, suspension culture cells, applied in biochemical equipment and methods, microorganisms, plant cells, etc.

Active Publication Date: 2019-09-27
ZUNYI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] At present, it has been reported in the literature that the liquid suspension culture system of leukoplakia cells has been successfully established, but there are few research reports on how to promote the accumulation of secondary metabolites in leukariae suspension culture cells by regulating the performance of suspension culture

Method used

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  • Method for promoting synthesis of secondary metabolites in common bletilla pseudobulb suspension cultured cells
  • Method for promoting synthesis of secondary metabolites in common bletilla pseudobulb suspension cultured cells
  • Method for promoting synthesis of secondary metabolites in common bletilla pseudobulb suspension cultured cells

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Experimental program
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Embodiment 1

[0029] Embodiment 1: A method for promoting the synthesis of secondary metabolites in white and suspension culture cells, comprising the following steps:

[0030] Step 1. Take the seeds of Baiji and induce culture on the induction medium of MS+1.0mg / L 6-BA+3.0mg / L 2,4-D+30.0g / L sucrose for 30d, then in MS+0.1mg / L The loose callus obtained by subculture for 2 generations for 40 days on the subculture medium of L 6-BA+2.0mg / L 2,4-D+30.0g / L sucrose;

[0031] Step 2: Accurately weigh the bletilla callus obtained in step 1, add it to the liquid medium of MS+1.0mg / L 6-BA+3.0mg / L 2,4-D, and add it to the medium Add sucrose to suspension culture for 35 days;

[0032] Regulate the pH value of liquid medium in the scope of 5.85~6.05, the volume of liquid medium is 21.74mL, the addition amount of white and callus is 0.92g, the concentration of sucrose in the culture liquid medium is 28.77g / L, The culture temperature is 20°C-25°C, the culture environment is dark culture, and the suspens...

Embodiment 2

[0033] Embodiment 2: A method for promoting the synthesis of secondary metabolites in white and suspension culture cells, comprising the following steps:

[0034] Step 1. Take the seeds of Baiji and induce culture on the induction medium of MS+1.0mg / L 6-BA+3.0mg / L 2,4-D+30.0g / L sucrose for 25d, then in MS+0.1mg / L The loose callus obtained by subculture for 2 generations for 40 days on the subculture medium of L 6-BA+2.0mg / L 2,4-D+30.0g / L sucrose;

[0035] Step 2: Accurately weigh the bletilla callus obtained in step 1, add it to the liquid medium of MS+1.0mg / L 6-BA+3.0mg / L 2,4-D, and add it to the medium Add sucrose to suspension culture;

[0036] Adjust the pH value of the liquid medium in the range of 5.85 to 6.05, the volume of the liquid medium is 40.13mL, the amount of white and callus is 0.82g, the concentration of sucrose in the liquid medium is 30.73g / L, and the culture temperature is 20°C-25°C, the culture environment is dark culture, and the suspension culture time...

Embodiment 3

[0037] Embodiment 3: A method for promoting the synthesis of secondary metabolites in white and suspension culture cells, comprising the following steps:

[0038] Step 1. Take the seeds of Baiji and induce culture on the induction medium of MS+1.0mg / L 6-BA+3.0mg / L 2,4-D+30.0g / L sucrose for 35d, then in MS+0.1mg / L The loose callus obtained by subculture for 2 generations on the subculture medium of L 6-BA+2.0mg / L 2,4-D+30.0g / L sucrose for 20 days;

[0039] Step 2: Accurately weigh the bletilla callus obtained in step 1, add it to the liquid medium of MS+1.0mg / L 6-BA+3.0mg / L 2,4-D, and add it to the medium Add sucrose to suspension culture;

[0040] Adjust the pH value of the liquid medium in the range of 5.85 to 6.05, the volume of the liquid medium is 38.35mL, the amount of white and callus added is 1.40g, the concentration of sucrose in the liquid medium is 38.32g / L, and the culture temperature The temperature is 20°C-25°C, the culture environment is dark culture, and the s...

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Abstract

The invention discloses a method for promoting synthesis of secondary metabolites in common bletilla pseudobulb suspension cultured cells, and belongs to the technical field of plant cell culture. The method comprises the following steps: accurately weighing common bletilla pseudobulb calli, adding the common bletilla pseudobulb calli in a liquid culture medium of MS+ 1.0 mg / L 6-BA+ 3.0 mg / L 2,4-D, and then adding saccharose in the culture medium for suspension culture. The pH value of the liquid culture medium is 5.85-6.05, the volume of the liquid culture medium is 21.74-40.13 mL, the adding amount of the common bletilla pseudobulb calli is 0.82-1.40 g, the concentration of the saccharose in the liquid culture medium is 28.77-38.32 g / L, the culture temperature is 20-25 DEG C, a culture environment is dark culture, and the time of suspension culture lasts for 30-40 d. In common bletilla pseudobulb suspension culture cells obtained by culture under culture conditions of the invention, the accumulation amounts of p-hydroxybenzyl alcohol, dactylorhin A, militarine and coelonin can be obviously increased.

Description

technical field [0001] The invention relates to the technical field of plant cell culture, in particular to a method for promoting the synthesis of secondary metabolites in bletilla suspension culture cells. Background technique [0002] Baiji (Bletilla striata (Thunb.) Rchb.f.), also known as Baiji, is a traditional Chinese herbal medicine. Baiji is used as medicine with dried tubers: bitter, sweet, astringent, slightly cold; astringent to stop bleeding, reduce swelling and promote muscle growth; it can be used for hemoptysis and hematemesis, traumatic bleeding, tuberculosis hemoptysis, ulcer bleeding, and is a good medicine for hemostasis. In recent years, its chemical composition and pharmacological research have shown that it is a variety of secondary metabolites in medicinal materials, such as bibenzyls, dihydrophenanthrenes, phenanthrenes, biphenanthrenes, quinone derivatives and other compounds. The specific components Including p-hydroxybenzyl alcohol, 1,4-bis[4-(gl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04C12N5/02
CPCC12N5/0025
Inventor 徐德林潘胤池肖世基李林闻伟锷上官艳妮刘厚伯黄恻隐李梦竹
Owner ZUNYI MEDICAL UNIVERSITY