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A method for overcoming the vitrification of regenerated seedlings after cryopreservation of Jerusalem artichoke stem tips

A technology for ultra-low temperature preservation and regeneration of seedlings, which is applied in the direction of plant cells, can solve the problems of less research on vitrification, achieve the effects of avoiding vitrification, low cost, and overcoming vitrification

Active Publication Date: 2015-11-04
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on overcoming the vitrification phenomenon reported in the literature in the past mostly focused on overcoming or eliminating the vitrification phenomenon of the test-tube plantlets in the tissue culture process, and there were few studies on overcoming the vitrification phenomenon after cryopreservation

Method used

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  • A method for overcoming the vitrification of regenerated seedlings after cryopreservation of Jerusalem artichoke stem tips
  • A method for overcoming the vitrification of regenerated seedlings after cryopreservation of Jerusalem artichoke stem tips
  • A method for overcoming the vitrification of regenerated seedlings after cryopreservation of Jerusalem artichoke stem tips

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Experimental program
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Effect test

Embodiment 1

[0029] Example 1 The phenomenon of vitrification of Jerusalem artichoke stem tips during ultra-low temperature storage

[0030] 1. Reagent formulations

[0031] Pre-culture medium: MS liquid medium containing 0.4mol / L sucrose.

[0032] Loading solution: MS liquid medium containing 2.0mol / L glycerol and 0.4mol / L sucrose.

[0033] The preparation method of PVS2 vitrification protective agent: thoroughly mix 3.26mol glycerin, 2.42mol ethylene glycol, 1.9mol dimethyl sulfoxide and 0.4mol sucrose with MS liquid medium to 1L.

[0034] Unloading solution: MS liquid medium containing 1.20mol / L sucrose.

[0035] 2. Ultra-low temperature preservation of Jerusalem artichoke stem tip

[0036] Get the Jerusalem artichoke tissue culture seedling ( figure 1 ), peel off the outer leaves layer by layer until the remaining 1-2 leaf primordia surround the shoot apex meristem, and cut off the shoot tip of 1.5-2mm ( figure 2 ), soaked in the pre-culture solution at 25°C for 3 days in the dar...

Embodiment 2

[0040] Embodiment 2 The overcoming of the vitrification phenomenon of Jerusalem artichoke stem tip cryopreservation

[0041] 1. Reagent formulations

[0042] Pre-culture solution containing fructose: MS liquid medium containing 0.4mol / L fructose (substitute sucrose with fructose).

[0043] Mannitol-containing pre-culture solution: MS liquid medium containing 0.4mol / L mannitol (replacing sucrose with mannitol).

[0044] Loading solution containing fructose: MS liquid medium containing 2.0 mol / L glycerol and 0.4 mol / L fructose (substitute sucrose with fructose).

[0045] Loading solution containing mannitol: MS liquid medium containing 2.0mol / L glycerol and 0.4mol / L mannitol (mannitol is used instead of sucrose).

[0046] Preparation method of fructose-containing vitrification agent: 3.26mol glycerin, 2.42mol ethylene glycol, 1.9mol dimethyl sulfoxide and 0.4mol fructose were thoroughly mixed with MS liquid medium to 1L (sucrose was replaced by fructose).

[0047] The prepara...

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Abstract

The invention provides a method for preventing vitrification of regenerated seedlings of jerusalem artichoke stem tips subjected to ultralow-temperature storage. In reagents used in a pre-cultivation stage, a loading stage, a PVS2 vitrification protection stage and a washing and unloading stage, fructose replaces saccharose and ultralow-temperature storage treatment is executed. According to the method, the phenomenon of vitrification of the regenerated seedlings of the jerusalem artichoke stem tips subjected to ultralow-temperature storage can be effectively prevented; the direct regeneration rate of plants which are not subjected to healing can be over 40 percent; the method is easy to operate and suitable for ultralow-temperature storage of the jerusalem artichoke stem tips.

Description

technical field [0001] The invention relates to a droplet vitrification ultra-low temperature preservation technology of plant germplasm resources, in particular to a method for overcoming the vitrification of regenerated seedlings after ultra-low temperature preservation of Jerusalem artichoke stem tips. Background technique [0002] Jerusalem artichoke (Helianthus tuberous L.) is a perennial herbaceous plant belonging to the genus Sunflower in the Asteraceae family. It is an ecological and economical plant with high economic value, medicinal value, ornamental value and ecological value. It gets its name because it looks like a chrysanthemum on the ground and a taro on the ground. It is native to North America and widely distributed in the United States, Canada, Mexico and other countries. It was introduced into my country through Europe in the 17th century, and is distributed in the three northeastern provinces of my country, Inner Mongolia, Hebei, Henan and other provinc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04
Inventor 张金梅陈晓玲韩丽卢新雄辛霞尹广鹍何娟娟
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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