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A vitrification ultra-low temperature preservation method for effectively preserving tartary buckwheat callus

A technology of vitrification of ultra-low temperature and callus, which is applied in the field of ultra-low temperature preservation of plant germplasm resources, can solve the problems of low cell survival rate and poor preservation effect of tartary buckwheat callus, and achieve the goal of improving survival rate and alleviating Damage, reduction of residual effects

Inactive Publication Date: 2011-11-30
CHENGDU UNIV
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Problems solved by technology

The traditional washing method often uses MS + sucrose 1.2mol / L solution to wash the preserved materials, usually in three steps, each step is 10 minutes, but a large number of experimental research results show that this method is not effective for the preservation of tartary buckwheat callus. The effect is not good, the survival rate of the cells after storage is not high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Pre-cultivation: get the callus of tartary buckwheat, place it in a freezing tube equipped with culture medium, and pre-culture it for 1 day at normal temperature. Alcohol 0.4mol;

[0018] (2) Loading: Take the callus pre-cultured for 1 day and place it in a cryovial containing a loading solution, and load it at 4°C for 20 minutes. The loading solution is composed of 0.5 mol / L sucrose solution and vitrified The solution is prepared at a volume ratio of 40:60, and the vitrification solution contains per liter: 1.0 mol of sucrose + 0.2 mol of sorbitol + 100 g of polyethylene glycol + 100 mL of 1,2-propylene glycol solution;

[0019] (3) Dehydration: Take the loaded callus and place it in a cryotube equipped with a vitrification solution, and dehydrate it at 0°C for 60 minutes. The vitrification solution contains per liter: 1.0mol of sucrose+0.2mol of sorbitol+ Polyethylene glycol 100g+1,2-propylene glycol solution 100mL;

[0020] (4) cryopreservation: put the above-...

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Abstract

The invention discloses a vitrification ultra-low temperature preserving method for effectively preserving bitter buckwheat callus. In the method, a vitrification ultra-low temperature preserving technology is applied to the preservation of the bitter buckwheat callus for the first time, a novel vitrification protectant (counted by liter) which comprises 1.0 mol of cane sugar, 0.2 mol of sorbic alcohol, 100 grams of polyethylene glycol and 100 mL of 1,2-dihydroxypropane solution is provided, a washing way is improved, and three-step gradient elution is applied to vitrification ultra-low temperature preservation of plants for the first time, wherein the adopted three-step gradient washing solutions (counted by lifter) comprise 30 grams of bovine serum albumin, 1.0 mol of cane sugar and 0.5mol of sorbic alcohol, 30 grams of bovine serum albumin, 0.5 mol of cane surgar and 1.0 mol of sorbic alcohol, and 30 grams of bovine serum albumin and 0.5 mol of cane sugar. Due to the adoption of the method, the survival rate of a material preserved at ultra-low temperature can be increased remarkably, and is up to 90.02 percent in maximum.

Description

technical field [0001] The invention relates to a cryopreservation technology of plant germplasm resources, in particular to a vitrification cryopreservation method for effectively preserving tartary buckwheat callus. Background technique [0002] Tartary buckwheat is a natural functional food integrating nutrition, health care and medical treatment, and its economic benefits are very high after processing. At present, the planting of tartary buckwheat is mainly planted after the germination of tartary buckwheat seeds. However, long-term research has found that there is a problem of seed aging during the storage of tartary buckwheat seeds, which is manifested by a decrease in germination rate year by year, which is very unfavorable for tartary buckwheat breeding. , is also quite detrimental to the preservation of its germplasm resources. [0003] Cryopreservation is a main content of modern biotechnology, and it is also an ideal method in plant germplasm preservation techno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N3/00
Inventor 赵钢王跃华刘益丽林抗雪孙雁霞邬晓勇马良良张珏
Owner CHENGDU UNIV
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