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Cryopreservation method for chrysanthemum

A technology of ultra-low temperature and chrysanthemum, which is applied in the field of preservation of chrysanthemum germplasm, and can solve the problems that the ultra-low temperature preservation technology has not yet been established

Inactive Publication Date: 2009-04-08
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the cryopreservation technology for chrysanthemum germplasm resources has not been established

Method used

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  • Cryopreservation method for chrysanthemum
  • Cryopreservation method for chrysanthemum
  • Cryopreservation method for chrysanthemum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The cryopreservation of embodiment 1 chrysanthemum

[0023] Material:

[0024] Chrysanthemum varieties: 4 varieties of chrysanthemums of the genus Dendranthema molifolium (Dendranthema molifolium), purple chrysanthemums, cut chrysanthemums 'Shenma' (only one chrysanthemum), and the related dahlia (Dahliapinnata Cav.), Buy from Shanghai and Hangzhou flower markets. Cut out the receptacle to propagate the test-tube plantlets, and obtain the receptacle cloning line.

[0025] Pre-culture medium: containing 0.4mol L -1 MS solid medium with sucrose

[0026] Pretreatment solution: containing 2mol L -1 glycerol and 0.4mol L -1 Sucrose MS Broth

[0027] 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 sucrose

[0028] method:

[0029] Select robust tissue-cultured seedlings of the above-mentioned chrysanthemum varieties, cut out the shoot tips (1.5-2mm), and culture them in the dark at 4°C ...

Embodiment 2

[0033] The cryopreservation of embodiment 2 chrysanthemum---the influence of sucrose concentration in the pre-cultivation medium on preservation effect

[0034] 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 general operation is to cut larger shoot apex tissue for pre-culture, and then cut it to an appropriate size for cryopreservation. In the experiment of this patent, the selected materials are easy to brown after being cut, and then pre-cultivated with high-concentration sucrose, which will cause the shoot tip tissue to become soft and water-soaked, which will cause great inconvenience to the subsequent stripping operation. It is also hig...

Embodiment 3

[0045] The cryopreservation of embodiment 3 chrysanthemum---the impact of pretreatment time on preservation effect

[0046] During the cryopreservation process, chrysanthemum shoot tips were treated with vitrification reagents for 20 minutes under ice bath conditions to obtain viable shoot tips, but treatment time longer than 20 minutes was not conducive to the survival and regeneration of shoot tips. Since the dehydration time of the vitrification reagent on the shoot tip is only 20 minutes, the dehydration step is very important for the successful cryopreservation of chrysanthemum. In this process, for shoot tips in different physiological states, some shoot tips may have been dehydrated too much within 20 minutes, while others may not be dehydrated enough, especially shoot tips with high water content, which will eventually cause the shoot tips to die. Therefore, before vitrification, appropriately prolonging the pretreatment time and strengthening the preliminary dehydrati...

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Abstract

The invention provides a method for preserving chrysanthemum at ultra-low temperature, which comprises the following steps: stem tip of sterile vaccine lymph is picked up and pre-cultured, is put into a pre-treatment solution for performing treatment for 20 to 40 minutes at room temperature, and is then transferred into a vetrification protection agent for performing treatment for 10 to 20 minutes at 4 DEG C or at room temperature; and the stem tip is transferred to a cryovial provided with a fresh vetrification protection agent pre-cooled in an ice bath, and is put into liquid nitrogen for preservation. The stem tip is subjected to recovery culture, and can directly grow into a plantlet. The method is convenient, feasible, stable and reliable; after being preserved, the chrysanthemum has good recovery and growth conditions; and the plantlet breed with the highest regeneration rate can reach more than 85 percent.

Description

technical field [0001] The invention relates to a method for preserving chrysanthemum germplasm, in particular to a method for ultra-low temperature preservation of chrysanthemum. Background technique [0002] Chrysanthemum (chrysanthemum) is one of the four famous fresh cut flowers in the world. my country has abundant germplasm resources of Chrysanthemum and its close relatives. These species contain many excellent genes that cultivated chrysanthemum lacks, such as insect resistance, Drought resistance, waterlogging resistance, etc., can be used for cultivating chrysanthemum germplasm innovation, and breeding high-quality and high-resistance new varieties. Secondly, Chrysanthemum and its close relatives have many species and varieties, which can be cultivated as potted flowers, cut flowers or ground covers, and are widely used for beautification and decoration of homes, conference venues, streets, squares, enterprises and institutions, etc., and have a high market value. F...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N3/02
Inventor 刘灶长刘艳霞林田李天菲罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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