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Ultralow-temperature preservation method and restoration culture method for carnation

A technology of ultra-low temperature preservation and preservation method, which is applied in the ultra-low temperature preservation method and the field of re-cultivation of carnation, and achieves the effect of restoring a good growth condition

Inactive Publication Date: 2012-03-21
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since then, the small droplet vitrification method based on the conventional vitrification method, and the practice of preserving the stem tip of carnation has not been reported so far.

Method used

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  • Ultralow-temperature preservation method and restoration culture method for carnation
  • Ultralow-temperature preservation method and restoration culture method for carnation
  • Ultralow-temperature preservation method and restoration culture method for carnation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The cryopreservation of embodiment 1 carnation

[0032] Material:

[0033] Carnation variety: safflower variety, purchased from Shanghai Flower Market. The flower stems are cut to propagate the test-tube plantlets, and the flower stem clones are obtained.

[0034] Pre-culture medium: containing 0.4mol L -1 Sucrose MS solid medium;

[0035] Pretreatment solution: containing 2mol L -1 glycerol and 0.4mol L-1 MS liquid medium of sucrose;

[0036] Vitrification protective agent: containing 3.26mol L -1 Glycerin, 2.42mol·L -1 Ethylene glycol, 1.9mol·L -1 DMSO and 0.4mol L -1 A solution of sucrose.

[0037] method:

[0038] Select robust tissue-cultured seedlings of the above carnation species, cut out the shoot tips (2-3mm), and culture them in the dark at 4°C for 5 days in the pre-culture medium, soak the shoot tips in the pretreatment solution at room temperature for 30 minutes, and then transfer to Treat in vitrification agent at 4°C for 60-80 minutes, then A: ...

Embodiment 2

[0042] The cryopreservation of carnation of embodiment 2---the influence of sucrose concentration and pretreatment time on preservation effect in the pre-cultivation medium

[0043] The physiological state of the experimental material has a significant impact on the survival rate after freezing. The shoot tip is pre-cultured with high-concentration sucrose for a short period of time, so that the tissue can lose part of the water more gently, and at the same time promote the cells to secrete protective substances, which is conducive to survival after freezing. The usual method of operation is to cut out larger shoot tip tissues for pre-culture, and then cut out appropriate size shoot tips for cryopreservation. Carnation stem tip tissue is easy to brown after cutting, and then pre-cultivated with high-concentration sucrose, it will cause the shoot tip tissue to become soft and water-soaked, which will cause great inconvenience to the subsequent stripping operation, and the stem ...

Embodiment 3

[0057] The ultra-low temperature preservation of carnation of embodiment 3---the impact of vitrification treatment time on preservation effect

[0058] The vitrification reagent can dehydrate the tissue and gently infiltrate the tissue, enter the vitrified state during cryopreservation, and effectively protect the cell structure. However, since the protective agent contains toxic and variable components (such as dimethyl sulfoxide), it needs to be strictly controlled. Under the premise of ensuring a sufficiently high survival rate, the processing time should be shortened as much as possible.

[0059] Material:

[0060] Carnation variety: Carnation genus Carnation of Caryophyllaceae safflower variety, purchased from Shanghai Flower Market. Cut the flower stems to propagate the test-tube plantlets, and obtain the receptacle cloning lines. Cut out the receptacle to propagate the test-tube plantlet, and obtain the flower stem cloning line.

[0061] Pre-culture medium: containi...

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Abstract

The invention relates to an ultralow-temperature preservation method for carnations by using a droplet vitrification method. Pre-culture, pretreatment and vitrification protection treatment are conducted to the stem tips of the carnations and finally the carnations are put into liquid nitrogen for ultralow-temperature preservation. The invention additionally provides a restoration culture method for the carnations preserved at ultralow temperature. The ultralow-temperature preservation method for the carnations provided by the invention is simple, convenient, feasible, stable and reliable, the restoration growth condition of the preserved carnations is good and the regeneration rate of plants highly reaches more than 90 percent.

Description

technical field [0001] The invention relates to a method for ultra-low temperature preservation of plants, in particular to a method for ultra-low temperature preservation of carnation and a recultivation method. Background technique [0002] Carnation (Diarahus caryophfllus L.) is an evergreen subshrub perennial flower of the family Caryophyllaceae. It is one of the four famous cut flowers in the world. The effective preservation of its cultivated species and wild species is very important for the localization of carnation. Carnation resources are usually subcultured regularly through cultivar cuttings and tissue culture. However, if Carnation is infected with viruses, long-term asexual reproduction will easily lead to continuous accumulation of viruses in the body. It is difficult to effectively preserve the germplasm for a long time. It is necessary to find the most suitable preservation method. [0003] In a broad sense, there are two types of conservation of germplasm r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N3/00A01H4/00
Inventor 林田刘灶长杨华李天菲陈海荣罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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