Method for carrying out tissue cultivation propagation of anoectochilus roxburghii by intermittent submerged bioreactor

A bioreactor, tissue culture technology, applied in the directions of botanical equipment and methods, horticultural methods, applications, etc., to achieve the effects of strong environmental adaptability, reduced production costs, and strong adaptability

Inactive Publication Date: 2014-05-07
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of the fact that the production of clematis seedlings has not been reported and publicly applied by using the batch immersion culture method, it is necessary to develop a method for rapid propagation of clematis tissue culture using batch immersion bioreactors

Method used

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  • Method for carrying out tissue cultivation propagation of anoectochilus roxburghii by intermittent submerged bioreactor

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] (1) According to the formula MS+NAA1.0mg / L+6-BA1.5mg / L+sucrose 30g / L+Anoectochilus 0.5g / L, adjust the pH value to 5.8, prepare the culture solution, and wrap the reactor, high temperature, high pressure, humidity and heat Standby after sterilization;

[0041] (2) Using wild Anoectochilus stems as explants, sterile treatment:

[0042] Cleaning: Choose healthy and disease-free plants, remove the roots, soak in potassium permanganate solution for 10 minutes, and then wash with running water for 60 minutes;

[0043] Disinfection: In a sterile environment, remove the leaves from the cleaned plants, soak the stems with 75% alcohol for 30 seconds, wash twice with sterile water, then soak with 0.1% mercury liters for 20 minutes, and wash with sterile water for 5~ 7 times.

[0044] (3) Remove the leaves from the aseptically treated stems and cut them into several stem segments each containing one stem node. Firstly, they are inoculated into MS solid medium. After 3 to 5 days of observa...

Embodiment 2

[0049] (1) According to the formula MS+NAA0.7mg / L+6-BA1.2mg / L+sucrose 30g / L+Anoectochilus fresh 0.3g / L, adjust the pH value to 6.0, prepare the culture solution, and wrap the reactor, high temperature and high pressure Use moist heat to sterilize for later use.

[0050] (2) Using wild Anoectochilus stems as explants, sterile treatment:

[0051] Cleaning: Choose healthy and disease-free plants, remove the roots, soak in potassium permanganate solution for 7 minutes, and then wash with running water for 45 minutes;

[0052] Disinfection: In a sterile environment, remove the leaves of the cleaned plants, soak the stems with 75% alcohol for 20 seconds, wash twice with sterile water, then soak with 0.1% mercury for 15 minutes, and wash with sterile water for 5~ 7 times.

[0053] (3) Remove the leaves from the aseptically treated stems and cut them into several stem segments each containing one stem node. Firstly, they are inoculated into MS solid medium. After 3 to 5 days of observation a...

Embodiment 3

[0057] (1) According to the formula MS+NAA0.5mg / L+6-BA1.0mg / L+ sucrose 30g / L+ Anoectochilus 0.2g / L, adjust the pH to 6.2, prepare the culture solution, and wrap the reactor, high temperature and high pressure Use moist heat to sterilize for later use.

[0058] (2) Using wild Anoectochilus stems as explants, sterile treatment:

[0059] Cleaning: Choose healthy and disease-free plants, remove the roots, soak in potassium permanganate solution for 5 minutes, and then wash with running water for 30 minutes;

[0060] Disinfection: In a sterile environment, remove the leaves from the cleaned plants, soak the stems with 75% alcohol for 15 seconds, wash twice with sterile water, then soak with 0.1% mercury for 10 minutes, and wash with sterile water for 5~ 7 times.

[0061] (3) Remove the leaves from the aseptically treated stems and cut them into several stem segments each containing one stem node. Firstly, they are inoculated into MS solid medium. After 3 to 5 days of observation and culti...

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Abstract

The invention belongs to the field of a traditional Chinese medicinal material, and particularly relates to a method for carrying out tissue cultivation propagation of anoectochilus roxburghii by an intermittent submerged bioreactor. The method comprises the steps of early-stage preparation, explant selection, and aseptic treatment, inoculation, cultivation of the bioreactor, and the like. Multiplication and rooting and strong seedling cultivation are finished one time in the reactor; the method has the characteristics of being short in period, high in tissue culture seedling quality and the like; and a method with low cost and high efficiency is provided for rapid propagation of tissue cultivation of the anoectochilus roxburghii.

Description

technical field [0001] The invention belongs to the technical field of tissue culture of medicinal plants, and in particular relates to a method for rapidly multiplying Aurea clematis tissue culture by using an intermittent immersion bioreactor to obtain high-quality seedlings. Background technique [0002] Anoecthilus roxburghii (Anoecthilus roxburghii) is a perennial herbaceous plant of the genus Orchidaceae, also known as Anoecthilus, Anoecthilus, Anoecthilus, and Anoecthilus. It usually grows in the fertile leaf-rotted soil under the shady broad-leaved forest. It is a traditional precious medicinal material in my country. It has the effects of cooling and detoxifying, nourishing yin and reducing fire, reducing inflammation and relieving pain, and has no toxic and side effects. It is known as the "King of Medicine" among the people. reputation. The plant height of clematis is 8~20cm, the plant type is small and exquisite, the leaf shape is beautiful, the leaf veins are go...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 陈集双张本厚贾明良金磊磊
Owner NANJING TECH UNIV
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