Tea tree virus-free tissue culture method

A technology of tissue culture and tea tree, which is applied in the field of plant tissue culture, can solve problems such as deformed embryos, influence of experimental results, pollution, etc., and achieve the effects of improving quality and quality, expanding planting scale, and optimal culture conditions

Inactive Publication Date: 2018-11-16
山东亚龙生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Browning, yellowing, glass seedlings, deformed embryos, pollution and many other problems often occur in the process of plant tissue culture, which affect the experimental results and even lead to the failure of the experiment. loss, urgent need to address
[0004] Chinese patent CN 2016101044044 discloses a method for tissue culture of tea tree young leaves. The method is to inoculate the sterilized tea tree young leaf explants on agar-free liquid medium. The 0.5-1cm place is not cut off, and then the explants are inoculated in the liquid medium for dark culture for 3 days and then transferred to the solid medium for dark culture for 11-13 days. The browning rate of the explants processed by this method is 32%, and the germination rate is as high as 68%; the present invention uses the method of combining liquid-solid co-cultivation and dark culture to significantly reduce the browning rate of explants in the tea tree tissue culture process, and significantly improve the germination rate and efficiency, but it cannot completely solve the yellowing , deformity and other issues

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] A tea tree detoxification tissue culture method, comprising:

[0050] (1) material selection: select no rot and no disease in spring, strong 15-20cm branches of the year, and cut the branches into strong sections with 1 axillary bud as explants;

[0051] (2) Surface treatment: rinse the collected explants with running water, soak them in 5% Tween-80 for 20 minutes, and rinse them with running water;

[0052] (3) Disinfection: according to the degree of tenderness of the explants, the old explants were sterilized with 75% alcohol for 5 minutes, 0.1% HgCl solution for 15 minutes, and rinsed with sterile water; the tender explants were sterilized with 75% alcohol Disinfect for 1 min, 0.1% HgCl solution for 5 min, rinse with sterile water;

[0053] (4) cutting: renewing the sterilized explant incision, removing the large bracts;

[0054] (5) Primary culture: inoculate the treated explants on medium Ⅰ, place them at a temperature of 26°C, light for 10 hours a day, light in...

Embodiment 2

[0060] A tea tree detoxification tissue culture method, comprising:

[0061] (1) material selection: select no rot and no disease in spring, strong 15-20cm branches of the year, and cut the branches into strong sections with 1 axillary bud as explants;

[0062] (2) Surface treatment: rinse the collected explants with running water, soak them in 5% Tween-80 for 20 minutes, and rinse them with running water;

[0063] (3) Disinfection: According to the tenderness of the explants, the old explants were sterilized with 75% alcohol for 3 minutes, 0.1% HgCl solution for 20 minutes, and rinsed with sterile water; the tender explants were sterilized with 75% alcohol Disinfect for 5 minutes, 0.1% HgCl solution for 4 minutes, rinse with sterile water;

[0064] (4) cutting: renewing the sterilized explant incision, removing the large bracts;

[0065] (5) Primary culture: inoculate the treated explants on medium Ⅰ, place them at a temperature of 25°C, light for 16 hours a day, light inte...

Embodiment 3

[0071] A tea tree detoxification tissue culture method, comprising:

[0072] (1) material selection: select no rot and no disease in spring, strong 15-20cm branches of the year, and cut the branches into strong sections with 1 axillary bud as explants;

[0073] (2) Surface treatment: rinse the collected explants with running water, soak them in 5% Tween-80 for 20 minutes, and rinse them with running water;

[0074] (3) Disinfection: according to the degree of tenderness of the explants, the old explants were sterilized with 75% alcohol for 5 minutes, 0.1% HgCl solution for 15 minutes, and rinsed with sterile water; the tender explants were sterilized with 75% alcohol Disinfect for 1 min, 0.1% HgCl solution for 5 min, rinse with sterile water;

[0075] (4) cutting: renewing the sterilized explant incision, removing the large bracts;

[0076] (5) Primary culture: inoculate the treated explants on medium Ⅰ, place them at a temperature of 26°C, light for 10 hours a day, light in...

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PUM

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Abstract

The invention discloses a tea tree virus-free tissue culture method which comprises the following steps: (1) material selection: selecting the current-year branches which are robust without decay or diseases in spring; (2) surface treatment: cleaning and disinfecting the bud explant of the tea tree; (3) disinfecting; (4) cutting; (5) primary culture: inoculating to a culture medium I and culturing; (6) multiplication subculture: inoculating an aseptic seedling to a culture medium II, and performing multiplication culture; (7) rooting culture; (8) domestication and seedling hardening. The method disclosed by the invention has the following beneficial effects: (1) through the tea tree tissue culture using the method, the culture medium components controlling differentiation, subculture growth, rooting and the like in tissue culture are determined; the research determines the best optimization selection for the type, concentration and culture conditions of the growth regulation substances, breaks through the technical barrier during the tea tree tissue culture, solves the technical problems of difficult in-vitro regeneration of the tea tree and low genetic transformation efficiency, and realizes a perfect culture effect.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a tea tree detoxification tissue culture method. Background technique [0002] Tea tree is a perennial woody plant, which has the characteristics of long growth cycle, low self-compatibility, short flowering period, difficult to separate linkage traits, and low beneficial mutation rate, which restricts traditional breeding methods. Therefore, genetic transformation is used to develop tea tree varieties. Improvement is increasingly favored by researchers, and tissue culture can provide an ideal recipient for a large number of gene transfers, and has become a common material for studying transgenic plants. However, there are very few successful cases of tea tree genetic transformation research at present, the main reason being that tea tree regeneration in vitro is difficult and the transformation efficiency is extremely low. [0003] The growth rate of te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 金红梅李亚周宇洋
Owner 山东亚龙生物科技有限公司
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