Method for in-vitro conservation and growth recovery after conservation of germplasm resource of cymbidium goeringii

An in-vitro preservation and resource technology, applied in the field of plant tissue culture, can solve the problems of large manpower and material resources, cross-contamination, genetic variation, etc., and achieve the effects of prolonging the preservation time, strong regeneration ability, and stable genetic genes.

Inactive Publication Date: 2017-12-01
SHANDONG CROP GERMPLASM CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Frequent in vitro subculture preservation technology may cause cross-contamination during each subcu...

Method used

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  • Method for in-vitro conservation and growth recovery after conservation of germplasm resource of cymbidium goeringii
  • Method for in-vitro conservation and growth recovery after conservation of germplasm resource of cymbidium goeringii

Examples

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Embodiment 1

[0026] A method for in vitro preservation of Chunlan germplasm resources and recovery of growth after preservation, the specific steps are as follows:

[0027] (1) The shoot tip was stripped from the newly grown pseudobulb of Chunlan, and placed on the surface of the protocorm induction medium supplemented with 0.5 mg / L 6-benzyl adenine in VW medium to cultivate protocorms.

[0028] Among them, the stripping method of the stem tip is as follows: select a pseudobulb with a length of 5 cm, wash the dust on the surface with tap water, peel off the outermost leaf, scrub it with alcohol with a volume concentration of 75% for 3 seconds, and put it in a concentration of 0.1 % mercuric chloride solution for 20 minutes, rinsed with sterile water for 3 times, dried in the air or blotted the surface moisture with filter paper, and stripped the shoot tips with a size of about 0.5 mm under a microscope.

[0029] Place the shoot tip on the surface of the protocorm induction medium in the te...

Embodiment 2

[0043] A method for in vitro preservation of Chunlan germplasm resources and recovery of growth after preservation, the specific steps are as follows:

[0044] (1) The shoot tip was stripped from the newly grown pseudobulb of Chunlan, and placed on the surface of the protocorm induction medium supplemented with VW medium supplemented with 1 mg / L 6-benzyl adenine to cultivate protocorms.

[0045] Among them, the stripping method of the stem tip is as follows: select a pseudobulb with a length of 7 cm, clean the surface dust with tap water, peel off the two outermost leaves, scrub with alcohol with a volume concentration of 75% for 4 seconds, and put in a mass concentration of 0.1 % mercuric chloride solution for 20 minutes, rinsed with sterile water for 4 times, dried in the air or blotted the surface moisture with filter paper, and stripped the shoot tips with a size of about 0.5 mm under a microscope.

[0046] Place the shoot tip on the surface of the protocorm induction medi...

Embodiment 3

[0058] A method for in vitro preservation of Chunlan germplasm resources and recovery of growth after preservation, the specific steps are as follows:

[0059] (1) The shoot tip was stripped from the newly grown pseudobulb of Chunlan, and placed on the surface of the protocorm induction medium supplemented with 0.8mg / L 6-benzyl adenine in VW medium to cultivate protocorms.

[0060] Among them, the stripping method of the stem tip is as follows: select a pseudobulb with a length of 6 cm, clean the surface dust with tap water, peel off one leaf of the outermost layer, scrub it with alcohol with a volume concentration of 75% for 4 seconds, and put it in a mass concentration of 0.1 % mercuric chloride solution for 20 minutes, rinsed with sterile water for 3 times, dried in the air or blotted the surface moisture with filter paper, and stripped the shoot tips with a size of about 0.5 mm under a microscope.

[0061] Place the shoot tip on the surface of the protocorm induction mediu...

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Abstract

The invention discloses a method for in-vitro conservation and growth recovery after conservation of a germplasm resource of cymbidium goeringii. The method comprises the steps: picking off stem tips from newly grown pseudobulbs of cymbidium goeringii, culturing the stem tips on the surface of a protocorm induction culture medium to form protocorms, and carrying out repeated cutting and secondary culture on the protocorms to reproduce a required amount of protocorms; then, culturing the protocorms in an in-vitro conservation culture medium, and carrying out dark culture on the protocorms under the condition of 0-5 DEG C after the protocorms enter a development and growth stage; and transferring the protocorms in dark culture to a differential culture medium, and differentiating the protocorms to produce young seedlings. The protocorms are induced by using a cymbidium goeringii stem tip culture technology and are then differentiated to obtain the young seedlings of cymbidium goeringii. Due to the combination of conserving the protocorms at low temperature and adding a proper amount of paclobutrazol, the conservation time can be greatly prolonged, the secondary culture frequency can be reduced, and the hereditary character of the germplasm resource can be effectively remained. If the germplasm resource is cultured under normal temperature and light conditions after being conserved for a certain time, growth can be rapidly recovered, and the young seedlings can be differentiated.

Description

technical field [0001] The invention relates to a method for in vitro preservation of Chunlan germplasm resources and recovery of growth after preservation. It belongs to the technical field of plant tissue culture. Background technique [0002] Chunlan (scientific name: Cymbidium goeringii (Rchb. f.) Rchb. f.) is a ground plant of the genus Orchid, also known as Duolan, Pudilan, Youlan, Duoduoxiang, and Grass Orchid. One of the oldest and most popular varieties. The plants are generally smaller, and the pseudobulbs are smaller and ovoid. Leaves band-shaped, margin toothless or serrate. The flower colors are mostly green and light brown yellow, and the flowers are fragrant. The propagation method is generally branch propagation, which can only be carried out once every 3 to 4 years, and the propagation speed is relatively slow. Chunlan seeds are very small, powdery, without endosperm, only a simple embryo, surrounded by a loose, transparent, impermeable seed coat, the e...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005A01H4/008
Inventor 颜廷进李群戴双邓翠霞田茜
Owner SHANDONG CROP GERMPLASM CENT
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