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
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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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