Method for reducing explant pollution in tea tree tissue culture process

A technique of tissue culture and explants, applied in botany equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems of restricting the development of tea tree tissue culture and explant pollution, so as to improve the success rate, reduce pollution, Good antibacterial effect

Active Publication Date: 2014-03-05
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Tea tree is a kind of plant with more endophytic bacteria. Therefore, in the process of tissue culture, the explants are seriously polluted, which seriously restricts the development of tea tree tissue culture.

Method used

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  • Method for reducing explant pollution in tea tree tissue culture process

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Embodiment 1

[0020] A method for reducing explant contamination in the tea tree tissue culture process, the concrete steps are as follows:

[0021] (1) Take tea tree stems that grow vigorously in spring and have full axillary buds as explants;

[0022] (2) Cut the explants into small pieces, disinfect them with 75% alcohol for 20 seconds, rinse them twice with sterile water, disinfect them with 0.1% mercuric acid for 4 minutes, rinse them twice with sterile water, and then use Disinfect with mercury chloride with a mass fraction of 0.1% for 5 minutes, and rinse with sterile water for 5 times;

[0023] (3) Cut off the discolored part of the explant wound due to disinfection, and then cut it into a 1-1.5cm long stem segment with an axillary bud for later use;

[0024] (4) Prepare medium with MS + 1.5 mg / L 6-BA + 0.3mg / L NAA + 30g sucrose + 6.5g agar powder, adjust the pH value to 5.6, and distribute the prepared medium into 500mL wide-mouth Erlenmeyer flasks Medium, high temperature steril...

Embodiment 2

[0028] A method for reducing explant contamination in the tea tree tissue culture process, the concrete steps are as follows:

[0029] (1) Take tea tree stems that grow vigorously in spring and have full axillary buds as explants;

[0030] (2) Cut the explants into small pieces, disinfect them with 75% alcohol for 23 seconds, rinse them twice with sterile water, disinfect them with 0.1% mercuric acid for 4 minutes, rinse them twice with sterile water, and then use Disinfect with mercuric chloride with a mass fraction of 0.1% for 5 minutes, and rinse with sterile water for 4 times;

[0031] (3) Cut off the discolored part of the wound of the explant, and then cut it into a 1-1.5cm long stem segment with an axillary bud for later use;

[0032] (4) Prepare medium with MS + 1.5 mg / L 6-BA + 0.3mg / L NAA + 30g sucrose + 6.5g agar powder, adjust the pH value to 5.7, and distribute the prepared medium into 500mL wide-mouth Erlenmeyer flasks Medium, high temperature sterilization for ...

Embodiment 3

[0036] A method for reducing explant contamination in the tea tree tissue culture process, the concrete steps are as follows:

[0037] (1) Take tea tree stems that grow vigorously in spring and have full axillary buds as explants;

[0038] (2) Cut the explants into small pieces, disinfect them with 75% alcohol for 25 seconds, rinse them twice with sterile water, disinfect them with 0.1% mercuric acid for 4 minutes, rinse them twice with sterile water, and then use Sterilize with mercuric chloride with a mass fraction of 0.1% for 5 minutes, and rinse with sterile water for 3 times;

[0039] (3) Cut off the discolored part of the wound of the explant, and then cut it into a 1-1.5cm long stem segment with an axillary bud for later use;

[0040] (4) Prepare medium with MS + 1.5 mg / L 6-BA + 0.3mg / L NAA + 30g sucrose + 6.5g agar powder, adjust the pH value to 5.8, and distribute the prepared medium in 500mL wide-mouth Erlenmeyer flasks Medium, high temperature sterilization for 20...

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Abstract

The invention discloses a method for reducing explant pollution in tea tree tissue culture process. The treatment method comprises the following steps: sterilizing a selected explant, cutting off the discolored cut part of the explant due to sterilization, cutting the part to a 1-1.5 cm long stem section with an axillary bud, inoculating the axillary bud on a culture medium containing antibiotic, inoculating 1-2 stem sections in each bottle, and placing the stem sections in a culture room for culture after being inoculated. The method disclosed by the invention combines ampicillin with streptomycin sulfate to reduce the pollution rate of the explant in the tea tree tissue culture process so as to improve the survival rate.

Description

technical field [0001] The invention belongs to the technical field of tea tree tissue culture, and in particular relates to a method for reducing explant contamination in the tea tree tissue culture process. Background technique [0002] tea tree( Camellia sinensis L. Kuntze.) belongs to Theaceae, Camellia, is a perennial woody, evergreen plant. Tea tree tissue culture began in the 1960s. At present, tea tree tissue culture has obtained complete regenerated plants by culturing the seeds, stem segments, axillary buds, anthers, etc. of tea trees. Tea tree is a kind of plant with more endophytic bacteria. Therefore, the explants are seriously polluted in the process of tissue culture, which seriously restricts the development of tea tree tissue culture. Therefore, researching a method for reducing the contamination of explants in the tea tree tissue culture process has become an urgent technical problem in this field. Contents of the invention [0003] The object of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 孙威江吴婉婉
Owner FUJIAN AGRI & FORESTRY UNIV
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