Method for reducing tissue culture rate of anoectochilus roxburghii and culture medium formula thereof
A technology of medium formula and pollution rate, which is applied in the field of plant tissue culture, can solve the problems of polluted clematis seedling breeding, genetic variation of culture materials, and difficulty in transplanting tissue culture seedlings, etc., and achieves good sterilization effect and repeatability Good, easy-to-grasp effects
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Embodiment 1
[0027] 1) Selection of explants: Naturally grown clematis plants were used as explants, washed with tap water, rinsed with 70% ethanol for 30 seconds, sterilized with 0.1% mercuric chloride for 10 minutes, sterile water Rinse 5-6 times repeatedly, drain the water, cut into 1 noded stem segment, and set aside.
[0028] 2) Medium preparation: use 1 / 2MS as the basic medium, add banana 60g / L, activated carbon 1.0g / L, NAA1.0mg / L, 6-BA0.6mg / L, sucrose 26g / L, agar powder 6.0 g / L, onion 50g / L, and crab shell powder 15g / L are mixed to adjust the pH value of the culture medium to 6.0. Heat the prepared medium until the agar is completely dissolved, dispense it into clean culture bottles while hot, and cover them.
[0029] 3) Sterilization of culture medium and inoculation equipment: Use high-pressure steam to sterilize the culture medium and inoculation equipment. Keep the temperature of the autoclave at 121°C, 105kPa, 20min. Metal utensils such as knives, scissors, and tweezers were...
Embodiment 2
[0034] 1) Selection of explants: Aseptic tissue-cultured seedlings of Clematis clematis were used as explants, the leaves of Clematis clematis were removed, and the stems were cut into segments with 1 node for later use.
[0035] 2) Medium preparation: use 1 / 2MS as the basic medium, add banana 50g / L, activated carbon 0.8g / L, NAA1.2mg / L, 6-BA0.4mg / L, sucrose 22g / L, agar powder 5.5 g / L, onion 60g / L, and crab shell powder 20g / L are mixed to adjust the pH value of the culture medium to 5.8. Heat the prepared medium until the agar is completely dissolved, dispense it into clean culture bottles while hot, and cover them.
[0036] 3) Sterilization of culture medium and inoculation equipment: Use high-pressure steam to sterilize the culture medium and inoculation equipment. Keep the temperature of the autoclave at 121°C, 105kPa, 20min. Metal utensils such as knives, scissors, and tweezers were wiped with 75% alcohol cotton balls before inoculation, and then sterilized at high temper...
Embodiment 3
[0041] 1) Selection of explants: Aseptic tissue-cultured seedlings of A. clematis were used as explants, and the terminal buds of 1 node of A. clematis were cut for later use.
[0042] 2) Medium preparation: use 1 / 2MS as the basic medium, add banana 50g / L, activated carbon 1.2g / L, NAA0.8mg / L, 6-BA0.8mg / L, sucrose 30g / L, agar powder 6.5 g / L, onion 60g / L, and crab shell powder 20g / L are mixed to adjust the pH value of the culture medium to 5.8. Heat the prepared medium until the agar is completely dissolved, dispense it into clean culture bottles while hot, and cover them.
[0043] 3) Sterilization of culture medium and inoculation equipment: Use high-pressure steam to sterilize the culture medium and inoculation equipment. Keep the temperature of the autoclave at 121°C, 105kPa, 20min. Metal utensils such as knives, scissors, and tweezers were wiped with 75% alcohol cotton balls before inoculation, and then sterilized at high temperature in a sterilizer with quartz sand for 8 ...
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