Daphne genkwa stem explant primary tissue culture method

A tissue culture and explant technology, which is applied in the directions of botanical equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems such as no report on the tissue culture and rapid propagation technology of stem explants, and achieves convenient and easy experimental operation. The effect of strong tolerance and high success rate

Active Publication Date: 2017-06-13
JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the research on Daphne genkwa mainly focuses on its chemical components and pharmacological effects, as well as the research on biological pesticides. There are few

Method used

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  • Daphne genkwa stem explant primary tissue culture method
  • Daphne genkwa stem explant primary tissue culture method
  • Daphne genkwa stem explant primary tissue culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Primary tissue culture medium: MS+0.1mg·L -1 NAA+1.0mg·L -1 ZT, add 30g / L sucrose and 7g / L agar to adjust the pH to pH=5.8.

[0037] Training method:

[0038] (1) Selection of Daphne genkwa explants: collection of robust growth, full axillary buds, no pests and diseases of Daphne genkwa explants in the same year, remove all leaves, cut into 10cm stems with axillary buds as explants;

[0039] (2) Treatment of Daphne genkwa explants: soak the explants in a 0.5% washing powder aqueous solution for 20 minutes with a mass-volume ratio, then rinse them with running water for 12 hours, and place the explants under the aseptic environment of the ultra-clean workbench in the inoculation room. The body was cut into 4cm stem segments, sterilized with 75% ethanol aqueous solution for 30s, and then 1g·L -1Soak in a mixed solution of mercuric chloride and 5 drops of Tween 80 for 12 minutes for disinfection; rinse with sterile water after disinfection, cut the stem section into a l...

Embodiment 2

[0042] Primary tissue culture medium: MS+0.1mg·L -1 NAA+1.5mg·L -1 ZT, add 30g / L sucrose and 7g / L agar to adjust the pH to pH=5.8.

[0043] Training method:

[0044] (1) Selection of Daphne genkwa explants: collection of robust growth, full axillary buds, no pests and diseases of Daphne genkwa explants in the same year, remove all leaves, cut into 10cm stems with axillary buds as explants;

[0045] (2) Treatment of Daphne genkwa explants: soak the explants in 0.5% washing powder aqueous solution for 20 minutes, then rinse them with running water for 12 hours, and place the explants in the aseptic environment of the ultra-clean workbench in the inoculation room. Cut into 4cm stem segments, sterilize with 75% ethanol aqueous solution for 30s, and then use 1g·L -1 Soak in a mixed solution of mercuric chloride and 5 drops of Tween 80 for 12 minutes for disinfection; rinse with sterile water after disinfection, cut the stem section into a length of 2 cm, the upper cut of the ste...

Embodiment 3

[0048] Primary tissue culture medium: MS+0.1mg·L -1 NAA+1.5mg·L -1 ZT, add 25g / L sucrose and 6.5g / L agar to adjust the pH to pH=5.7.

[0049] Training method:

[0050] (1) Selection of explants of Daphne genkwa: collection of strong growth, full axillary buds, no pests and diseases of Daphne genkwa explants in the same year, remove all leaves, cut into 8cm stems with axillary buds as explants;

[0051] (2) Treatment of Daphne genkwa explants: Soak the explants in a 0.2% washing powder aqueous solution for 20 minutes, then rinse them with running water for 12 hours, and inoculate the explants under the aseptic environment of the ultra-clean workbench in the inoculation room. The body was cut into 5cm stem segments, sterilized with 70% ethanol aqueous solution for 45s, and then 1g·L -1 Soak in a mixed solution of mercuric chloride and 3 drops of Tween 80 for 10 minutes for disinfection; rinse with sterile water after disinfection, cut the stem section into a length of 2.5 cm,...

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Abstract

The invention discloses a daphne genkwa stem explant primary tissue culture method which comprises the following steps: (1) selecting a daphne genkwa explant: acquiring a daphne genkwa annual branch, removing all leaves and cutting the branch into axillary bud stems as the explant, (2) treating the daphne genkwa explant: soaking and washing the explant with a washing powder solution, rinsing the explant with running water, performing disinfection treatment and then shifting the explant into a culture medium of daphne genkwa stem explant primary tissue culture, and (3) culturing the daphne genkwa explant: performing irradiation culture on the explant shifted into the primary culture medium in a culture rack of a culture room until inducted clustered buds germinate. According to the method, the stem explant induced clustered buds are adopted, so that a daphne genkwa tissue culture seedling can be simply and quickly obtained, a culture period is obviously shortened, a culture success rate is obviously increased, a germination rate is high, and a vitrification rate is low.

Description

technical field [0001] The invention relates to plant primary tissue culture, in particular to a method for primary tissue culture of Daphne genkwa stem segment explants. Background technique [0002] Genkwa (Daphne genkwa Sieb.et Zucc.) is a deciduous shrub of Daphneaceae Daphne. Its flowers are clustered and colorful, and have high ornamental value. It is an excellent wild flower resource in my country. It is also an important Chinese herbal medicine and biological pesticide in my country. [0003] Since the 1960s, especially since the 1990s, the habitat of Daphne genkwa has been severely damaged, the distribution range of the population has shrunk, the number of populations and individuals has declined sharply, and its wild resources have gradually decreased. It is an urgent requirement to protect and utilize this plant resource to study the reproduction technology of Daphne genkwa. [0004] Daphne genkwa is generally propagated by sowing, but the traits of the offsprin...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 张虎许建民潘静霞宋微
Owner JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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