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Method for reducing plant cell browning risk through absorption of quinone substances with silica gel

A silica gel adsorption and plant cell technology, which is applied in the biological field of plant cells, can solve the problems of plant cell culture system instability, production capacity decline, etc., and achieve high adsorption target specificity, improved capacity, improved stability and efficiency. Effect

Inactive Publication Date: 2014-07-16
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to provide a method for reducing the risk of browning of plant cells by using silica gel to absorb quinones to solve the problem of unstable plant cell (especially yew cells) culture system and decreased production capacity caused by browning

Method used

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  • Method for reducing plant cell browning risk through absorption of quinone substances with silica gel
  • Method for reducing plant cell browning risk through absorption of quinone substances with silica gel
  • Method for reducing plant cell browning risk through absorption of quinone substances with silica gel

Examples

Experimental program
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Effect test

Embodiment 1

[0028] This example relates to a method for inhibiting the browning of yew cells by using sponge-like silica gel to absorb quinones. Including the following steps:

[0029] (1) Preparation of sponge-like silica gel blocks: Take a food-grade sponge silica gel plate, cut into small pieces of 1 cm square, wash with ultrapure water, sterilize at 121 degrees for 20 minutes, and set aside.

[0030] (2) Establishment of the yew culture system: Take a 250ml Erlenmeyer flask, add 75ml of B5 medium, add 25g / L of sucrose, 4mg / L of plant hormone naphthalene acetic acid, and 0.15mg / L of 6-benzylbenzylaminoadenine , adjust the pH to 5.8, and seal it with a breathable sterile film. Sterilized for later use. After being lowered to room temperature, the yew cells were inoculated, and the inoculation amount was 12g / L (calculated according to the fresh weight of the cells). There were 20 groups in the experiment, and 10 bottles were the experimental group. After the inoculation, a piece of st...

Embodiment 2

[0036] In this example, the regenerated silica gel block used in Example 1 was used as an adsorbent, and the effect of inhibiting the browning of Taxus chinensis cells by adsorbing quinones on the silica gel block after elution and regeneration was investigated.

[0037] After the regenerated silica gel blocks were cleaned with ultrapure water, they were sterilized for use. All the implementation steps are exactly the same as in Example 1 except that the silica gel block used is for elution regeneration.

[0038] During the cultivation process, it was observed that: 5 bottles in the control group had obvious browning, and three of them were severely browned, and the cultures were dark reddish brown. The browning rate was 50%. The silica gel adsorption group was all normal without browning.

[0039] After the culture, samples were taken to measure the cell density and paclitaxel content. It was found that the cell density of the control group was 22±1.9g / L, while the cell den...

Embodiment 3

[0042] This example is a method of using rod-shaped silica gel to absorb quinones in a cell reactor to inhibit the browning of Taxus chinensis cells.

[0043] (1) Rod-shaped silica gel preparation: Take a medical-grade silica gel rod with a diameter of 1 cm, cut it into a length of 20 cm, clean it with ultrapure water, and install it on a 2-liter NBS cell culture reactor according to the electrode installation method (such as figure 2 as shown, figure 2 It is a schematic diagram of the application of rod-shaped silica gel to adsorb quinones in the yew cell culture in the reactor; the thick rod extending into the yew culture at the lower end on the right represents the rod-shaped silica gel adsorption material, and the thorny ball represents the yew cell mass).

[0044] (2) Establishment of yew culture system: prepare 1 liter of B5 medium, which contains 4 mg / L of plant hormone naphthalene acetic acid, 0.15 mg / L of 6-benzylbenzylaminoadenine, and 25 g / L of sucrose, and adjust...

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Abstract

The present invention discloses a method for reducing plant cell browning risk through absorption of quinone substances with silica gel. The method comprises: preparing a silica gel absorption material having a specific shape, and sterilizing so as to be spare; during a cell culture process, completely contacting the silica gel absorption material and a plant cell culture, wherein the color of the silica gel absorption material is changed into red-brown after absorbing quinone substances; and taking out the silica gel absorption material after the absorption achieves a saturation state or the culture is completed. According to the present invention, the silica gel is selected as the absorption agent for the cell browning related quinone substances so as to avoid absorption on cell nutrients in the culture medium, reduce the content of the quinone substances in the cell microenvironment, avoid plant (Taxus chinensis) cell browning death, and easily increase stability and cell activity of plant (Taxus chinensis) cell culture; and browning generally causes the decreased cell metabolism level, such that the secondary metabolite having drug activity in the cells can be increased by using silica gel absorption to avoid browning, wherein the yield of the anticancer drug paclitaxel in the Taxus chinensis cells can be increased.

Description

technical field [0001] The invention belongs to the biological field of plant cells, and in particular relates to a method for reducing the risk of browning of plant cells by using silica gel to absorb quinone substances. Background technique [0002] Most plant secondary metabolites have the ability to regulate cell physiology and are important drugs or drug precursors, such as the anticancer drug paclitaxel in yew. However, due to the scarcity of yew resources, it is a national protected plant, and it is forbidden to cut down natural forest trees. In natural or artificially planted yew plants, the content of paclitaxel is very low, less than 1 / 10,000. Therefore, separating yew cells and culturing them under artificial conditions can precisely regulate the cell microenvironment, facilitate stress induction, and paclitaxel production The capacity is much higher than that of yew planting, and it has become an efficient, environmentally friendly and sustainable commercial pro...

Claims

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Application Information

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IPC IPC(8): C12N5/04
Inventor 苗辰飞朱闪烁苗志奇
Owner SHANGHAI JIAO TONG UNIV
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