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Method for preserving germplasm resource of Chinese cymbidium in vitro and restoring growth after preservation

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

Inactive Publication Date: 2017-08-29
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 subculture process, and multiple subcultures will also cause genetic variation. manpower and material resources

Method used

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  • Method for preserving germplasm resource of Chinese cymbidium in vitro and restoring growth after preservation
  • Method for preserving germplasm resource of Chinese cymbidium in vitro and restoring growth after preservation

Examples

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Effect test

Embodiment 1

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

[0027] (1) Peel off the shoot tips from the newly grown pseudobulbs of Molan, and place them on the surface of protocorm induction medium supplemented with 0.5 mg / L 6-benzyl adenine in VW medium to cultivate protocorms.

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

[0029] Place the shoot tip on the surface of the protocorm induction medium in the test tube or culture bottle, an...

Embodiment 2

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

[0044] (1) Peel off the shoot tips from the newly grown pseudobulbs of Molan, and place them on the surface of the protocorm induction medium supplemented with 1 mg / L 6-benzyl adenine in VW medium to cultivate into protocorms.

[0045] Among them, the peeling method of the stem tip is as follows: select a pseudobulb with a length of 7cm, wash 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, air-dried or blotted with filter paper to dry the surface water, and peeled off 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 medium in the test tube or culture...

Embodiment 3

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

[0059] (1) Peel off the shoot tips from the newly grown pseudobulbs of Molan, and place them on the surface of the protocorm induction medium supplemented with 0.8 mg / L 6-benzyl adenine in VW medium to cultivate protocorms.

[0060] Among them, the peeling method of the stem tip is as follows: select a pseudobulb with a length of 6cm, wash the surface dust with tap water, peel off the outermost leaf, scrub 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, air-dried or blotted with filter paper to dry the surface water, and peeled off the shoot tips with a size of about 0.5mm under a microscope.

[0061] Place the shoot tip on the surface of the protocorm induction medium in the test tube or culture bottle...

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Abstract

The invention discloses a method for preserving the germplasm resource of Chinese cymbidium in vitro and restoring growth after preservation. The method comprises the following steps: peeling stem apexes from newly-growing pseudobulbs of Chinese cymbidium, putting the stem apexes into a protocorm induction culture medium for conducting surface culture on the stem apexes into protocorms, conducting repeated cutting and subculture on the protocorms, thereby reproducing a required quantity of protocorms, then conducting culture by adopting in vitro preservation of the culture medium, and after the protocorms enter the development and growth stages, conducting dark culture in a condition of 0-5DEG C; and transferring the protocorms in dark culture onto a differential medium, and producing seedlings by differentiation of the protocorms. The method uses a Chinese cymbidium stem apex culture technique for inducing the protocorms, and then Chinese cymbidium seedlings are obtained through differentiation of the protocorms. The protocorms are preserved at low temperature, a proper amount of paclobutrazol is added, so the preserving time can be greatly prolonged, the number of times of subculture is reduced, and the hereditary characters of the germplasm resource are effectively maintained. After preservation for a certain period of time, when cultured at normal temperature in an illumination condition, the protocorms can quickly restore growth and differentiate seedlings.

Description

technical field [0001] The invention relates to a method for in vitro preservation and recovery of growth after preservation of Morchia germplasm resources. It belongs to the technical field of plant tissue culture. Background technique [0002] Molan (scientific name: Cymbidium sinense (Jackson ex Andr.) Willd.), also known as the old orchid, is a terrestrial plant of the genus Orchidaceae. Pseudobulb ovoid, enclosed in leaf base. Leaf band-shaped, nearly thin leathery, dark green. Scape from the base of the pseudobulb, erect, thicker, generally slightly longer than the leaves; the color of the flowers varies greatly, usually dark purple or purple-brown with light-colored lips, generally with a strong aroma. The propagation method is generally division propagation, which can only be carried out once every 3 to 4 years, and the propagation speed is relatively slow. Molan seeds are very small and powdery, without endosperm, only a simple embryo, which is covered with loos...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 丁汉凤贾文斌辛富刚王存娥颜廷进
Owner SHANDONG CROP GERMPLASM CENT
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