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Method for sterilizing turbinaria conoides rhizoid explant

A technology of Tritonella lobularis and a disinfection method, which is applied in the fields of botanical equipment and methods, horticultural methods, plant regeneration, etc., can solve the problem that it is difficult to kill bacteria or fungi that enter the interior of seaweed cells, and it is impossible to obtain a large number of sterile explants. In order to avoid drug resistance, avoid irreversible damage, and reduce the use time

Inactive Publication Date: 2014-03-26
QINGDAO WINCHANCE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The tissue culture technology of marine plants is relatively backward. The main reason is that it is impossible to obtain a large number of sterile and viable explants. Seaweed is mainly composed of cells. Although the structure of large seaweed is relatively complex, the type and dosage of disinfectants The effect of explants is very significant, high doses are easy to kill explants, and low doses are not easy to kill bacteria or fungi that enter the interior of seaweed cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] After the Trumpetella pseudophyllum was collected from the sea, it was cleaned with sea water and transported to the operation room in an ice box. The fronds of the Trumpetella pseudophyllum were removed and the rhizomes were retained as explants for tissue culture. The explants were removed with a brush. After removing the algae and other attachments attached to the body, soak the explants in 1% penicillin for 5 hours, then take out the explants, soak them in 0.5% potassium iodide solution for 2 minutes, and then Take out the explants, sterilize them in 1% povidone-iodine for 1 min, and finally rinse them with cooled filtered high-pressure sterilized sea water; cut the above-mentioned rhizoid explants into sections with a length of 0.2-0.5 cm And inoculated into the liquid PES culture medium, inoculated 1 section in each culture bottle, cultured on a shaker, the speed of the shaker was 100r / min, the culture conditions were light intensity 5000Lx, light time 12h, tempera...

Embodiment 2

[0017] After the Trumpetella pseudophyllum was collected from the sea, it was cleaned with sea water and transported to the operation room in an ice box. The fronds of the Trumpetella pseudophyllum were removed and the rhizomes were retained as explants for tissue culture. The explants were removed with a brush. After removing the algae and other attachments attached to the body, soak the explants in oxytetracycline with a concentration of 5% for 8 hours, then take out the explants and soak them in a potassium iodide solution with a concentration of 1% for 5 minutes , then the explants were taken out, sterilized in 5% povidone-iodine for 3 minutes, and finally rinsed with cooled filtered high-pressure sterilized sea water; the above-mentioned rhizoid explants were cut into lengths of 0.2-0.5cm Cut the sections and inoculate them into the liquid PES culture medium, inoculate one section in each culture bottle, culture on a shaker, the speed of the shaker is 100r / min, the culture...

Embodiment 3

[0019] After the Trumpetella pseudophyllum was collected from the sea, it was cleaned with sea water and transported to the operation room in an ice box. The fronds of the Trumpetella pseudophyllum were removed and the rhizomes were retained as explants for tissue culture. The explants were removed with a brush. After removing the algae and other attachments attached to the body, soak the explants in 2% streptomycin for 6 hours, then take out the explants and soak them in 0.7% potassium iodide solution for 3 minutes , then the explants were taken out, sterilized in 2% povidone-iodine for 2 minutes, and finally rinsed with cooled filtered high-pressure sterilized sea water; the above-mentioned rhizoid explants were cut into lengths of 0.2-0.5cm Cut the sections and inoculate them into the liquid PES culture medium, inoculate one section in each culture bottle, culture on a shaker, the speed of the shaker is 100r / min, the culture conditions are light intensity 5000Lx, light time ...

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Abstract

The invention discloses a method for sterilizing a turbinaria conoides rhizoid explant. The method is characterized by comprising the steps of clearing attachment such as impurity algae on the explant by utilizing a brush and adopting the turbinaria conoides rhizoid as an explant for the tissue culture, placing the explant into 1% to 5% antibiotic to be soaked for 5h to 8h, then taking out the explant, immersing the explant in a potassium iodide solution for 2min to 5min, taking out the explant, sterilizing the explant in povidone iodine for 1min to 3min, and finally washing the explant by utilizing the cooled and filtered high-pressure sterilized seawater. By adopting the method, the application concentration and the application time of the antibiotic can be reduced, and the drug resistance of the explant can be avoided; meanwhile, through the sterilizing effect of the potassium iodide and povidone iodine, not only can the bacteria or fungus on the surface and inside the explant be killed, but also the unrecoverable damage of the sterilizing agent to the explant can be avoided, and 85% bacteria-free feasible rhizoid explant can be obtained through the method.

Description

technical field [0001] The invention relates to the field of disinfection of plant tissue culture technology, in particular to a disinfection method for rhizoid explants of Trumpetella pseudophyllum. Background technique [0002] Pseudomonas microphyllum ( Turbinaria conoides ) belongs to the Phaeophyta, Fucusaceae, Sargassumaceae, and Trumpetella genus, and is a common large brown algae in my country. Trumpetella microphylla is rich in marine steroids such as marine sterols. Compared with sterols in terrestrial plants, marine sterols have more abundant skeletons and branched chains. This unique structure endows it with anti-tumor and anti-viral properties. , anti-inflammatory and bactericidal special effects, and can be used to prepare enzyme inhibitors, therefore, marine steroids have become a class of ingredients that attract attention in the medical and pharmaceutical industries. [0003] At present, the wild resources of Trumpetella pseudophylla are mainly used, but t...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 苏建丽
Owner QINGDAO WINCHANCE TECH
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