Cryopreservation method of tobacco leaf callus cell

A callus, cryopreservation technology, applied in the preservation of plants, plant cells, botanical equipment and methods, etc., to achieve the effect of preventing freezing denaturation, protecting freezing damage, and preventing denaturation

Inactive Publication Date: 2015-04-15
WUHAN SHANGCHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The cryopreservation method of tobacco callus has not been reported yet, so it is very important to find a simple and efficient cryopreservation method

Method used

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  • Cryopreservation method of tobacco leaf callus cell

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Embodiment Construction

[0024] The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0025] In this example, the callus cells of tobacco leaves were first cultured in the culture medium for 5 days to obtain a cell suspension. They were divided into three groups for cultivation, and the three groups were cultured in 3%, 4% and 8% sucrose liquid medium respectively.

[0026] Then, the cell suspension was centrifuged at 800 rpm for 5 minutes at 4°C and the supernatant was removed to obtain the cell liquid, and 2 mL of the cell liquid was transferred to a cryopreservation tube, and 0.858 mL of cryoprotectant was added. Tighten the cap to seal after dispensing. The cryoprotectant is a mixture of sterilized sorbitol and dimethyl sulfoxide, wherein the volume ratio of the two is 2:1.

[0027] Cool down the cryopreservation tube to 4°C and let it stand for 30 minutes at this condition, then continue to cool down to -20°C and let it...

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Abstract

The present invention provides a cryopreservation method of tobacco leaf callus cells, comprising the steps of: culturing the tobacco leaf callus cells in a medium to prepare a cell suspension; centrifuging the cell suspension and removing supernatant to get a cell solution, transferring the cell solution to a cryopreservation tube, adding a cryoprotectant therein, then sealing, cooling the cryopreservation tube to 4 DEG C and standing at the condition for 20-40 minutes, continue cooling to -20 DEG C and standing for 30-40 minutes at the temperature, continue cooling to -40 DEG C and standing for 3-4 hours at the temperature, and last storing the cryopreservation tube at -80 DEG C. The method provided by the invention is simple and easy, the cryopreserved cells have high survival rate, and the method does not consume a large amount of manpower and resources and is low in cost.

Description

technical field [0001] The invention provides a method for cell preservation, in particular to a method for cryopreservation of tobacco leaf callus cells, and belongs to the field of biotechnology. Background technique [0002] Tobacco (tobacco) is an annual herbaceous plant belonging to the Solanaceae (Solanaceae) genus Nicotiana tabacum, and there are about 60 species. Tobacco is characterized by being easy to carry out tissue culture and easy to obtain transformed plants to become a typical model plant. It is also known as the "fruit fly" of the plant kingdom along with Arabidopsis thaliana. In technical applications, it can be used as a bioreactor to produce antibiotics. The gene products of cancer are then extracted using biotechnology for medical treatment. In addition, many potential uses of tobacco in the development of food and drug resources will continue to be discovered and utilized. Therefore, obtaining tobacco cells through tissue culture has important medica...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04A01N3/00
Inventor 余光辉向文豪覃永华
Owner WUHAN SHANGCHENG BIOTECH
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