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Plant stem cells for angelica plant storage root cambia as well as preparation method and culture method thereof

A plant stem cell and angelica technology, applied in plant cells and other directions, can solve the problem of inability to isolate the cambium stem cell line and so on

Active Publication Date: 2015-04-22
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The shortcomings of the prior art exist in the following aspects. Through actual application, imitating the experiment of the patent on the storage root cambium cell separation method, it was found that in some Angelica plants such as Angelica sinensis (Angelica sinensis (Oliv.) Diels), Angelica dahurica (Fisch. ex Hoffm.) Benth. et Hook. f. ex Franch. et Sav), Cambial stem cell lines cannot be isolated from plants such as Angelica decursiva (Miq.) Franch. et Savat)

Method used

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  • Plant stem cells for angelica plant storage root cambia as well as preparation method and culture method thereof
  • Plant stem cells for angelica plant storage root cambia as well as preparation method and culture method thereof
  • Plant stem cells for angelica plant storage root cambia as well as preparation method and culture method thereof

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preparation example Construction

[0046] Preparation method of plant stem cells of storage root cambium of Angelica plant , including the following steps:

[0047] 1) Selection and sterilization of explants: take the storage roots of Angelica genus plants as explants, clean them, and then perform sterilization treatment to make the surface and interior of the explants free of bacteria;

[0048] 2) Slicing of explants: Slicing the above-mentioned sterilized explants under aseptic conditions to obtain explant slices;

[0049] 3) Screening culture: attach the slices to the screening medium; treat at 4°C-30°C for 16-72 hours, so that the cells in the non-cambium part of the slices do not divide;

[0050] 4) Separation culture: Take out the explant slices after the above-mentioned screening culture and put them into the slice liquid for recovery, and then put the slices into the separation medium for culture, so as to promote the division of the cambium cells and prevent them from dedifferentiation , while other...

Embodiment 1

[0071] Embodiment 1: the preparation method of the plant stem cell of angelica plant storage root cambium

[0072] (1) Selection and sterilization of explants

[0073] Select the complete root of the annual angelica seedling with a diameter of about 0.8 cm, remove the surface soil, and rinse with tap water for 30 minutes to obtain a clean explant. Soak the explants in 75% ethanol for 30 seconds in a sterile operating table, and wash them twice with sterile water. Then soak in 1-2% sodium hypochlorite and shake for 4 minutes, then wash twice with sterile water, and finally soak and shake with 0.1% mercury chloride for 4 minutes, and wash with sterile water for 4-5 times to obtain a completely sterilized surface. Explant.

[0074] Put the explants that have been sterilized on the surface into the anti-browning deep layer sterilization solution and shake for 30 minutes. The anti-browning deep layer sterilization solution contains 3% sucrose, 200mg / L penicillin, 300mg / L strep...

Embodiment 2

[0113] Example 2: Method for culturing plant stem cells in storage root cambium of Angelica plants

[0114] Method for Suspension Culture of Plant Stem Cells in Storage Root Cambium of Angelica Plant

[0115]In the application of plant cell engineering, it is necessary to establish a suspension system to facilitate further research. The specific suspension culture operation is as follows: inoculate the cambium stem cell line obtained in Example 1 in a liquid medium with a wet weight of 40 g / L, and culture it at 100-120 rpm, 23°C-27°C, and the suspension medium formula As shown in Table 5. After 15 days of suspension culture, it was observed that the cambium stem cell line was white and uniform cells (such as Figure 6 shown), while the callus cells have the characteristics of browning, and the size of tissue clumps in the callus is different (such as Figure 7 shown).

[0116] Table 6 Suspension Medium Components

[0117]

[0118] The relevant functions of the camb...

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Abstract

The invention discloses plant stem cells for angelica plant storage root cambia as well as a preparation method and a culture method thereof. The preparation method comprises the following steps: slicing sterilized storage roots, placing sliced storage roots on a screening culture medium, processing so that the non-cambial cells in the root slices partially die or are damaged or stop growing; placing the slices into a separation culture medium for culturing so that the division of cambial cells is promoted and no dedifferentiation is caused, and preliminarily separating to obtain the cambial cells; transferring the cambial cells into a subculture medium for carrying out subculture, transferring the subcultured cambial cells into the separation culture medium for alternately and repeatedly culturing until a stable cell containing a large amount of small vacuoles is obtained. The method for culturing the cambial stem cells comprises a suspension culture method and a fermenter culture method. By adopting the preparation method, the separation of the angelica cambial stem cells is achieved, a new method is provided for the expanding culture of the angelica cambial stem cells and the preparation method lays the foundation for a large amount of industrial production of angelica plant storage root cambia stem cells and also provides a new source of cells for angelica plant storage root cambia stem cells.

Description

technical field [0001] The invention relates to a plant stem cell of storage root cambium of Angelica plant and its preparation and culture method. Background technique [0002] At present, plant biotechnology is researched by inducing callus, cultivating plant organs or isolating protoplasts, but the method of separating and culturing cambium has only recently appeared. [0003] Callus is the parenchyma tissue formed after dedifferentiation of plants, but after dedifferentiation, the amount and type of secondary metabolites will decrease, and it also contains a central large vacuole, which will become larger in the expanded culture (such as fermenter culture, etc.). Vacuoles are prone to fragmentation leading to cell death, and the culture environment is harsh. [0004] The organ culture of plants generally includes the cultivation of plant roots or buds, which maintains some states of plant cells. Compared with callus, the content of secondary metabolites is more, but bec...

Claims

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Application Information

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IPC IPC(8): C12N5/04
Inventor 严春艳丁楠韩丽娟
Owner GUANGDONG PHARMA UNIV
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