Separation and culture method using ginseng cambium stem cells

A cultivation method and cambium technology, applied in the field of plant stem cell separation and cultivation, can solve the problems of weak division ability, degeneration of cell lines, limited division ability, etc., and achieve the effects of strong stress resistance, fast growth speed and reduced production cost

Inactive Publication Date: 2014-01-15
鹭港生物药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the essence of this kind of cells is derived from differentiated somatic cells, so the ability to divide is limited and the ability to resist stress is not strong
In the process of industrial mass production, cell lines often degenerate and have weak division ability, making it difficult to culture in large volumes

Method used

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  • Separation and culture method using ginseng cambium stem cells
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  • Separation and culture method using ginseng cambium stem cells

Examples

Experimental program
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Effect test

Embodiment 1

[0021] (1) Firstly, the root tuber of Northeastern ginseng with a diameter of 2 cm was sterilized with 0.1% mercury chloride for 10 minutes, and then cut into 0.2 cm thick semicircular slices including periderm, phloem and cambium, see figure 1 .

[0022] (2) Place the above semicircular slices in a solution containing 1.0 mg.L -1 Picloram and 0.5mg.L -1 2,4-dichlorophenoxyacetic acid in WPM medium, cultured at 25°C in the dark. Due to the rapid division of cambium stem cells, a cluster of cambium stem cells accumulates significantly on the explant to form a bulge, see figure 2 , this stem cell mass has a dense texture and a smoother surface.

[0023] (3) Two weeks later, the explants whose cambium proliferated significantly were taken out, and the cambium stem cells were separated and transferred to the subculture medium for culture, see image 3 . The subculture medium contains 0.5mg.L -1 Picloram and 0.5mg.L -1 The WPM medium of 2,4-dichlorophenoxyacetic acid is sub...

Embodiment 2

[0025] (1) Firstly, the 2-cm-diameter roots of Northeastern ginseng were sterilized with 0.1% mercury chloride for 10 minutes, and then cut transversely into 0.2-cm-thick semicircular slices including the periderm, phloem and cambium.

[0026] (2) Place the semicircular slices in a solution containing 0.1 mg.L -1 Picloram and 0.5mg.L -1 2,4-dichlorophenoxyacetic acid in WPM medium, cultured at 25°C in the dark.

[0027] (3) Two weeks later, the explants whose cambium proliferated significantly were taken out, and the cambium stem cells were separated and transferred to the subculture medium for culture; the subculture medium contained 1.0mg.L -1 Picloram and 2.0mg.L -1 The WPM medium of 2,4-dichlorophenoxyacetic acid is subcultured every two weeks, and a large number of stem cells can be obtained in a short period of time.

[0028] The values ​​in Table 1 below are the concentrations of picloram and 2,4-dichlorophenoxyacetic acid in the WPM medium and subculture medium used...

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Abstract

The invention discloses a separation and culture method using ginseng cambium stem cells. The separation and culture method using ginseng cambium stem cells is characterized by comprising the following steps: (1) firstly performing disinfection treatment on the root tubers with the diameter of 2 cm of northeast ginseng by 0.1% mercury bichloride for 10 minutes, and then transversely cutting the northeast ginseng into semicircular slices with the thickness of 0.2 cm and containing periderms, phloems and cambiums; (2) placing the semicircular slices on a WPM (woody plant medium) containing 0.1-4.0 mg / L of picloram and 0.1-4.0 mg / L of 2,4-dichlorphenoxyacetic acid, and culturing at 25 DEG C in the dark; (3) taking out the obviously-multiplied explants of the cambiums after two weeks, separating the cambium stem cells, transferring the cambium stem cells on a subculture medium and culturing, wherein the subculture medium is a WPM containing 0.1-4.0 mg / L of picloram and 0.1-4.0 mg / L of 2,4-dichlorphenoxyacetic acid, and subculture is performed every two weeks. According to the method, separation and culture for ginseng stem cells are performed by using the unsplit cambium stem cells, and the cambium stem cells are infinitely multiplied, so that production can be effectively expanded.

Description

technical field [0001] The invention relates to a plant stem cell separation and cultivation method, in particular to a ginseng cambium stem cell separation and cultivation method. Background technique [0002] Ginseng is a famous and precious Chinese medicinal material, which is deeply loved by consumers and has a great market demand. The resources of wild ginseng are limited, and the method of artificially planting ginseng is currently used in production to meet market demand. [0003] However, due to the long growth cycle of ginseng, the occupation of cultivated land, and the production process is easily affected by environmental conditions, pests and heavy metal pollution, the cost of this traditional production technology is relatively high and the quality is difficult to control. [0004] In order to solve the above-mentioned problems, a technology of producing medicinal components of ginseng by in vitro suspension culture of ginseng cells has been developed. This tec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
Inventor 亓琴
Owner 鹭港生物药业有限公司
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