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A method to optimize the cryopreservation effect of vitrification of Arabidopsis seedlings

A technology of vitrification ultra-low temperature and Arabidopsis thaliana, applied in horticultural methods, plant preservation, botanical equipment and methods, etc., to achieve the effects of improving preservation effects, reducing damage, and changing the formation status

Inactive Publication Date: 2016-04-20
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Adding certain exogenous substances in the existing technology is beneficial to improve the survival rate of cryopreservation, but it is unknown which exogenous substances to choose for specific plants and tissues to improve their preservation effect

Method used

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  • A method to optimize the cryopreservation effect of vitrification of Arabidopsis seedlings
  • A method to optimize the cryopreservation effect of vitrification of Arabidopsis seedlings
  • A method to optimize the cryopreservation effect of vitrification of Arabidopsis seedlings

Examples

Experimental program
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Effect test

Embodiment 1

[0053] 1) Acquisition of Arabidopsis seedlings: Disinfect Arabidopsis seeds with 70% ethanol for 15 seconds, rinse them with sterile water for 4 times, then disinfect them with an aqueous solution containing 20 wt% NaClO and 0.01 wt% Tween 20 for 10 min, rinse them with sterile water 6 times; then the sterilized seeds were vernalized at 4°C for 48 hours and cultured on MS solid medium. 2 , cultivated at a temperature of 25° C.; then cultivated in the dark for 16 h at a culture temperature of 20° C., and cultured the seeds for a total of 60 h to obtain Arabidopsis seedlings with a seedling age of 60 h.

[0054] 2) Divide the 60h Arabidopsis seedlings into the experimental group and the control group, and set up 3 parallel experiments in each group, put 50 seedlings into a 2mL cryovial containing 1mL loading solution, and treat them at room temperature for 20min; Remove by suction, add vitrification solution, and treat at 0°C for 50 minutes; put the cryovial into liquid nitrogen...

Embodiment 2

[0066] This example is mainly to test and analyze the parameters of Arabidopsis thaliana seedlings at various stages of cryopreservation, so as to demonstrate the regulation of exogenous graphene quantum dots on membrane damage and oxidative stress during 60h cryopreservation of Arabidopsis seedlings.

[0067] 1) Experimental material: the material in group 1 is: the seedling of Arabidopsis thaliana germinated according to the method described in Example 1 of the present invention for 60h

[0068] Materials in group 2 are: seedlings treated with vitrification solution in Example 1;

[0069] Group 3 is: the seedlings after thawing after vitrification cryopreservation according to the method in Example 1;

[0070] Group 4 is the seedling after washing with washing liquid in embodiment 1;

[0071] Group 5 is the seedlings after recovery culture treatment in Example 1.

[0072] 1) Experimental materials: Arabidopsis thaliana seedlings germinated for 60 hours and seedlings after ...

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Abstract

The invention discloses a method for optimizing a vitrification ultra-low temperature preservation effect of arabidopsis seedlings. According to the invention, the arabidopsis seedlings are processed by adopting a vitrification solution which contains a carbon nanometer material so as to improve the preservation effect of the arabidopsis seedlings. The method specifically comprises the steps of loading liquid treatment, vitrification solution treatment and liquid nitrogen preservation, wherein the vitrification solution is the MS culture solution containing 0.1-0.5 g / L of grapheme quantum dots. The method disclosed by the invention optimizes the preservation effect of the arabidopsis seedlings remarkably; as the grapheme quantum dots are added to serve as an allogenic material, the vitrification ultra-low temperature preservation of plants is facilitated.

Description

technical field [0001] The invention relates to the field of preservation of plants or parts thereof, in particular to a method for optimizing the effect of vitrification cryopreservation of Arabidopsis seedlings. Background technique [0002] Cryopreservation is a modern in vitro preservation technology for germplasm resources developed in the 1970s. Usually stored in liquid nitrogen, the substance metabolism and growth activities in the preserved material cells are almost completely stopped, and they are in a relatively stable biological state, achieving the purpose of long-term preservation of germplasm. Ultra-low temperature preservation is currently the only method that does not require continuous subculture. Medium and long-term preservation method. Vitrification cryopreservation is to place cells or tissues in a vitrification solution composed of a certain proportion of permeable and non-permeable protective agents, so that the material and its vitrification solution...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N3/00A01H4/00
Inventor 任丽陈冠群申晓辉张荻
Owner SHANGHAI JIAOTONG UNIV
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