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Method for in vitro preservation of cybidium ensifolium germplasm resources and recovery growth after preservation

An in vitro preservation and germplasm 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 prolonged storage time, strong regeneration ability, and stable genetic genes

Inactive Publication Date: 2017-08-22
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 of cybidium ensifolium germplasm resources and recovery growth after preservation
  • Method for in vitro preservation of cybidium ensifolium germplasm resources and recovery growth after preservation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] A method for in vitro preservation of Jianlan 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 Jianlan, and placed on the surface of the protocorm induction medium supplemented with 0.5 mg / L 6-benzyladenine 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 tes...

Embodiment 2

[0043] A method for in vitro preservation of Jianlan 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 Jianlan, and placed on the surface of the protocorm induction medium supplemented with VW medium supplemented with 1mg / 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 mediu...

Embodiment 3

[0058] A method for in vitro preservation of Jianlan 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 Jianlan, 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 ...

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Abstract

The invention discloses a method for in vitro preservation of cybidium ensifolium germplasm resources and recovery growth after preservation. The method comprises the following steps: stripping stem tips from newly-grown false bulbs of cybidium ensifolium, and putting the steam tips on the surface of a protocorm-induced culture medium to form protocorms; carrying out repeated cutting and subculture on the protocorms to breed out the protocorms with required quantity; then culturing by adopting an in vitro preservation culture medium; after the protocorms enter a development and growth stage, carrying out dark culture under the condition of 0 to 5 DEG C; transferring the protocorms in dark culture to a differential culture medium, and differentiating the protocorms to generate seedlings. According to the method disclosed by the invention, the protocorms are induced by using a cybidium ensifolium stem tip culture technology; then the protocorms are differentiated to obtain cybidium ensifolium seedlings; the processes of preserving the protocorms at low temperature and adding a proper amount of paclobutrazol are combined, so that the preservation time can be greatly prolonged, subculture times are reduced, and hereditary characters of the germplasm resources are effectively retained; after the protocorms are preserved for a certain time, the protocorms are putted under the conditions of normal temperature and illumination for culturing, further the growth can be quickly recovered, and the seedlings are differentiated.

Description

technical field [0001] The invention relates to a method for in vitro preservation of Jianlan germplasm resources and recovery of growth after preservation. It belongs to the technical field of plant tissue culture. Background technique [0002] Jianlan (scientific name: Cymbidium ensifolium (L. ) Sw.) is a terrestrial plant, also known as Xionglan, Junhelan, Jianhui, etc., also known as Qiulan. Formation is usually in the golden autumn season, the flowers often have fragrance, and the color changes greatly, usually light yellow-green with purple spots. 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. Jianlan 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. Natural conditions Xia Jianlan seeds are difficult to germ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01N3/00
CPCA01H4/001A01H4/008A01N3/00
Inventor 颜廷进田茜邓翠霞戴双李群
Owner SHANDONG CROP GERMPLASM CENT
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