Simple cryopreservation method of plant BY-2 cell

A BY-2, cryopreservation technology, used in the field of plant cell cryopreservation and recovery, to reduce the chance of ice crystals, reduce salt precipitation, and achieve good protection effects

Inactive Publication Date: 2014-11-26
SOUTH CENTRAL UNIVERSITY FOR NATIONALITIES
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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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  • Simple cryopreservation method of plant BY-2 cell
  • Simple cryopreservation method of plant BY-2 cell
  • Simple cryopreservation method of plant BY-2 cell

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Embodiment Construction

[0027] The preferred embodiments of the present invention will be described in detail below; it should be understood that the preferred embodiments are only for illustrating the present invention, rather than limiting the protection scope of the present invention.

[0028] The cryopreservation method of tobacco callus comprises the steps:

[0029] Pre-cultivation: Pre-cultivate the calli of tobacco leaves in liquid medium containing 8% sucrose for 5 days; pre-treatment with cryoprotectant: centrifuge the pre-cultured cell suspension at 800 rpm for 5 min at 4°C, and suck off the supernatant Finally, keep 2ml of cell fluid and transfer it to a cryopreservation tube, then add cryoprotectant to the preserved cell fluid and seal it;

[0030] The method of adding cryoprotectant: 0.572ml of 50% sorbitol, 0.286ml of dimethyl sulfoxide, and 2ml of cell suspension, wherein, sorbitol is sterilized by high-pressure steam, and dimethyl sulfoxide is sterilized by ultraviolet lamp;

[0031]...

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Abstract

The invention discloses a simple cryopreservation method of a plant BY-2 cell. The cryopreservation method comprises the steps of pre-culturing the BY-2 cell in 8% sucrose liquid culture medium, then adopting cooling and freezing steps in the presence of cryoprotectants (sorbitol and dimethyl sulfoxide), and finally storing at -80 DEG C for later use after carrying out cooling treatment in stages at 4 DEG C, -20 DEG C and -40 DEG C. Frozen cell sap is melted by adopting a rapid melting method when the cell revives; and a fresh culture medium is used to culture after liquid supernatant is centrifugally removed, and then the cell can be used. The method is simple and feasible; loss of cryopreservation of liquid nitrogen is reduced; the frozen cell has high survival rate; the problems of consumption of manpower and material resources, reduction of cell quality, and the like in cultivation and use of the cell in a laboratory can be solved; and the method disclosed by the invention can be used as a general technology of a biology laboratory.

Description

technical field [0001] The invention belongs to the technical field of cryopreservation and recovery of plant cells, and in particular relates to a method for ultra-low temperature cryopreservation of callus of model plant tobacco. 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 t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04
Inventor 余光辉
Owner SOUTH CENTRAL UNIVERSITY FOR NATIONALITIES
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