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Method for cryopreservation and plant regeneration of Dianthus caryophyllus shoot tip

A technology of ultra-low temperature preservation and carnation, applied in the field of plant cell engineering, can solve the problems of changing the operation of the tissue culture preservation method, the field preservation method being susceptible to diseases and insect pests, being cumbersome and time-consuming, etc. restore good growth

Inactive Publication Date: 2010-09-22
FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problems that the field preservation method is vulnerable to the invasion of diseases and insect pests, the tissue culture preservation method is easy to cause the change of some genetic traits, and the traditional cryopreservation method is cumbersome and time-consuming, the present invention provides a carnation stem tip cryopreservation and plant regeneration method. method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1, the acquisition of carnation tissue culture seedlings

[0021] Take the stems of two buds of Carnation, disinfect them with alcohol with a volume concentration of 70% for 30 seconds, then disinfect them with 0.2% mercury chloride for 25 minutes, wash them with sterile water for 3 times, and inoculate the stems in the following induction medium Upper: MS+2.0mg / L BA+0.5mol / L NAA, cultured for 20 days under light intensity of 2000lx, light time of 12 hours / day, temperature of 25°C, after the buds survived, transferred to the above induction medium After 20 days of culture, transfer to MS+0.3mg / LBA+0.5mg / LNAA subculture medium and culture for 30 days to obtain carnation aseptic tissue culture plantlets.

Embodiment 2

[0022] Embodiment 2, carnation stem tip cryopreservation and plant regeneration

[0023] A. Cut the carnation tissue culture seedling stem section 10mm of embodiment 1, be placed on the following pre-medium: the sucrose+volume percentage of MS+0.3mol / L is the agar of 5% DMSO+7g / L, under light intensity For: 1500lx, light time: 12 hours / day, temperature: 22°C, culture for 3 days;

[0024] B. Take out the pre-cultured stem segment in A, cut off the 2mm long stem tip, and put it into the following loading solution: MS+2mol / L glycerol+0.4mol / L sucrose, load for 40 minutes;

[0025] C. Transfer the shoot tips from the loading solution to the following vitrification solution PVS2: MS+0.4mol / L sucrose+30% glycerol+15% ethylene glycol+15% by volume DMSO, dehydrated at 0°C for 40 minutes;

[0026] D. Transfer the stem tip in C to the same PVS2 solution as above C, and store it in liquid nitrogen;

[0027] E. Take the shoot tip out of the liquid nitrogen and thaw it quickly in a wate...

Embodiment 3

[0029] Embodiment 3, carnation stem tip cryopreservation and plant regeneration

[0030] A. cut the carnation tissue culture seedling stem section 15mm of embodiment 1, place on the following pre-medium: the sucrose+volume percentage of MS+0.5mol / L is the agar of 5% DMSO+7g / L, under light intensity For: 2000lx, light time: 8 hours / day, temperature: 28°C, culture for 1 day;

[0031] B. Take out the pre-cultured stem segment in A, cut off the 4mm long stem tip, and put it into the following loading solution: MS+2mol / L glycerol+0.4mol / L sucrose, and load for 30 minutes;

[0032] C. Transfer the shoot tips from the loading solution to the following vitrification solution PVS2: MS+0.4mol / L sucrose+30% glycerol+15% ethylene glycol+15% by volume DMSO, dehydrated at 0°C for 60 minutes;

[0033] D. Transfer the stem tip in C to the same PVS2 solution as above C, and store it in liquid nitrogen;

[0034] E. Take the shoot tip out of liquid nitrogen and thaw it quickly in a water bath...

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Abstract

The invention provides a method for cryopreservation and plant regeneration of Dianthus caryophyllus shoot tips. Growth-restored regenerated Dianthus caryophyllus plants are obtained through preculture, loading, dehydration, freezing, thawing, restoration culture and the like. By adopting the method provided by the invention, the regeneration rate of Dianthus caryophyllus can reach 52.4 percent, the device for the vitrification cryopreservation of the Dianthus caryophyllus shoot tips is simple, the process is simplified, the operation is easy, the freezing effect is good, the restoration growth of the preserved Dianthus caryophyllus is good, the hereditary variation probability is small and the method is an effective way for the in vitro preservation of Dianthus caryophyllus germ plasm resources.

Description

technical field [0001] The invention relates to a method for preserving plant germplasm resources, in particular to a method for ultra-low temperature preservation of carnation stem tip vitrification, and belongs to the technical field of plant cell engineering. Background technique [0002] Carnation (Dianthus caryophyllus), also known as carnation, is a perennial herbaceous plant of the genus Caryophyllum in the family Caryophyllaceae. It is one of the world-renowned fresh cut flowers and one of the most important bulk export flowers in Yunnan Province. [0003] At present, the preservation of carnation germplasm resources mainly adopts the field preservation method and the tissue culture preservation method. The field preservation method is vulnerable to the invasion of diseases and insect pests and the threat of harsh environment. Avoidance of changes in certain genetic traits. [0004] Cryopreservation is an ideal way to preserve plant germplasm. Since Nag and Street f...

Claims

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Application Information

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IPC IPC(8): A01H4/00A01N3/00
Inventor 周旭红莫锡君桂敏王继华张颢欧阳德爱何艳李绅崇曹桦李金泽
Owner FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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