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Method for in-vitro preservation and growth recovery after preservation of oncidium germplasm resources

A new technology for in vitro preservation and oncidium, applied in the field of plant tissue culture, can solve the problems of cross-contamination, large manpower and material resources, genetic variation, etc., and achieve the effects of strong regeneration ability, extended storage time, and stable genetic genes

Inactive Publication Date: 2017-06-23
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 in-vitro preservation and growth recovery after preservation of oncidium germplasm resources
  • Method for in-vitro preservation and growth recovery after preservation of oncidium germplasm resources

Examples

Experimental program
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Effect test

Embodiment 1

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

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

Embodiment 2

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

[0044] (1) The shoot tip was stripped from the newly grown pseudobulb of Oncidium, 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 medium in the t...

Embodiment 3

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

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

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Abstract

The invention discloses a method for in-vitro preservation and growth recovery after preservation of oncidium germplasm resources. The method comprises the following steps: peeling stem tips from newly grown false bulbs of oncidium, putting the stem tips onto the surface of a protocorm induction medium, culturing so as to obtain protocorm, and performing repeated cutting and subculture on the protocorm so as to breed protocorm of an expected number; culturing by using an in-vitro preservation medium; after the protocorm enters into a development and growth period, performing dark culture at 0-5 DEG C; transferring the dark cultured protocorm onto a differential medium, and performing differentiation on the protocorm, thereby generating seedlings. By adopting an oncidium stem tip culture technique, the protocorm is induced, the protocorm is further differentiated, and thus oncidium seedlings can be generated. The protocorm is preserved at low temperature, and with a proper amount of paclobutrazol, the preservation time can be greatly prolonged, the subculture times can be reduced, and the hereditary characters of the germplasm resources can be effectively maintained. After being preserved for a certain time, the protocorm can be recovered to grow rapidly when being cultured under normal temperature and lighting conditions, and seedlings can be differentiated.

Description

technical field [0001] The invention relates to a method for in vitro preservation of oncidium germplasm resources and recovery of growth after preservation. It belongs to the technical field of plant tissue culture. Background technique [0002] Oncidium, a traditional famous flower in China, is rich in germplasm resources, widely distributed, and has high ornamental value. 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. Oncidium seeds are very small and powdery, without endosperm, only a simple embryo, surrounded by a loose, transparent, impermeable seed coat. The embryo contains very little nutrients, most of which are lipids. Under natural conditions, it is difficult for the seeds of xinlan to germinate. Therefore, traditional seed preservation techniques are not suitable for the preservation of oncidium germplasm resources. [0003] Most of the cultivated var...

Claims

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Application Information

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IPC IPC(8): A01H4/00A01N3/00
CPCA01H4/001A01H4/005A01H4/008A01N3/00
Inventor 丁汉凤王存娥贾文斌辛富刚颜廷进
Owner SHANDONG CROP GERMPLASM CENT
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