Cryopreservation and plant regeneration method of vegetative propagation flower narcissus

A technology of ultra-low temperature preservation and vegetative reproduction, applied in the fields of plant regeneration, botanical equipment and methods, horticultural methods, etc., can solve the problems of loss of ornamental value and commerciality, degeneration of species, degeneration, etc., and achieve the simple and feasible method of plant regeneration. , restore good growth condition

Inactive Publication Date: 2013-06-12
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the narcissus virus disease in my country is very harmful. Under the continuous cultivation of the bulbs used as the provenance for many years, the virus accumulation is becoming more and more serious, resulting in a shrinking yield year by ...

Method used

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  • Cryopreservation and plant regeneration method of vegetative propagation flower narcissus
  • Cryopreservation and plant regeneration method of vegetative propagation flower narcissus
  • Cryopreservation and plant regeneration method of vegetative propagation flower narcissus

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1, Induction of Small Bulbs of Chongming Narcissus

[0023] Such as figure 2 Shown and illustrated is the flow chart of the Chongming narcissus small bulb induction operation flow chart of the present invention. The operation steps are: a', select 3-4 year old Chongming narcissus bulbs; b', sterilize the bulbs of step a'with 70% alcohol 1 min, sterilize with 0.1% mercury for 10-20 min, then rinse with sterile water; c', inoculate the bulb cut pieces of step b'with 1 mg·L -1 BA and 1 mg·L -1 Small bulbs were induced on NAA's MS medium, where the induction culture temperature on the medium was 25±2℃, and the light was 12 h·d -1 ,Light intensity 36 μmol m -2 s -1 , After 3 months of induction, small bulbs with a diameter of 0.5 to 1 cm are obtained.

Embodiment 2

[0024] Example 2, Chongming narcissus pre-training

[0025] Such as figure 1 As shown, the description is the operation flow chart of the preservation method of the present invention. The operation method of the preservation method is: a. Induce the bulbs of vegetatively propagated flower narcissus, and cut the stem tips of 2~5mm in size; b. The shoot tips of root primordium and bud primordium are pre-cultured, and then placed in the pretreatment solution for 15-30 minutes at room temperature; c. Transfer the shoot tips treated in the pretreatment solution to the vitrification protectant for ice bath Treat for 60-120min; d. Transfer the ice-bath treated stem tip to a freezing tube containing fresh pre-cooled vitrification protectant; e. Put the above freezing tube into liquid nitrogen quickly for storage.

Embodiment 3

[0026] Example 3. The effect of stem tip size on the survival rate of Chongming Narcissus under cryopreservation

[0027] The sucrose concentration of 1~1.5 mm, 2~3 mm, 4~5 mm stem tips is 0.3 mol·L -1 After pre-cultured in MS medium for 5 days, it was treated with pretreatment solution for 20 min, and then treated with vitrification protectant ice bath for 60-120 min. The preferred treatment time was 80 min. The result of resuming culture after freezing storage showed: The survival rate of 2~3mm diameter is the highest, up to 80%. The survival rate of stem tips less than 2 mm is only 40%, and the survival rate of stem tips greater than 4 mm is only 10%.

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Abstract

The invention relates to a cryopreservation and plant regeneration method of vegetative propagation flower narcissus. The method comprises the following steps of: inducing bulbs of the narcissus, and cutting stem tips with root primordia and bud primordia for preculture; processing the stem tips in a pre-treatment solution at room temperature, performing ice bath treatment on the stem tips in a vitrification protecting agent, transferring the stem tips into a freezing pipe containing pre-cooled fresh vitrification protecting agent, and rapidly feeding the freezing pipe into liquid nitrogen to store; and when the stem tips are in use, taking out the frozen freezing pipe from the liquid nitrogen, immediately putting the frozen freezing pipe into water batch at 39-41 DEG C to rapidly unfreeze, and then cultivating the stem tips in the freezing pipe in a culture medium. The cryopreservation method of the vegetative propagation flower narcissus recorded by the invention is simple, feasible, stable and reliable, can ensure good restoring growth conditions of the stored narcissus, and has a good reference value to cryopreservation of genetic resources of bulb and ball perennial plants.

Description

technical field [0001] The present invention relates to a method for preserving plant germplasm resources and regenerating plants, in particular to a method for ultra-low temperature preservation and plant regeneration of vegetatively propagated flowers, specifically to a method for ultra-low temperature preservation and plant regeneration of Chongming narcissus , belongs to the technical field of plant cell engineering. Background technique [0002] Chongming Narcissus ( Narcissus tazetta var. chinensis ) is one of the characteristic flower varieties in Shanghai, which was cultivated as early as the Ming Dynasty. It has elegant fragrance, long flowering period, and short plant type. It is suitable for planting on the edge of grassland or arranging flower beds and flower borders in gardens. It has wide application prospects and has been sold in international markets in recent years. During the routine production and preservation process, due to serious viral diseases and p...

Claims

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Application Information

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IPC IPC(8): A01N3/02A01H4/00
Inventor 林田杨华龙萍朱天生刘灶长李天菲罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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