A kind of method for tissue culture and rapid propagation of plug field bubble
A technology of tissue culture and field soaking, which is applied in horticultural methods, botany equipment and methods, horticulture, etc., can solve the problems that there are no reports on the tissue culture and rapid propagation of Rubus erythroides, and achieve the benefits of industrial production Effect
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Embodiment 1
[0023] A method for culturing and rapidly propagating field bubble tissue, comprising the following steps in turn:
[0024] (1) Disinfection and inoculation of soaked stems in fields
[0025] Take the new stems that were extracted in the same year and cut them into stems with axillary buds. Select the stems with a moderate degree of lignification, first use a toothbrush to gently scrub under tap water, and add an appropriate amount of detergent solution to soak for 2-4 minutes. After rinsing with running water for 0.5h, turn the stem section into a 70% alcohol solution and soak for 2min in an ultra-clean bench, rinse with sterile water for 3 times, and immerse in 0.1% HgCl with 5 drops of Tween-80 added dropwise. 2 Soak and sterilize in the solution for 13-14min, then fully soak with sterile water for 3 times, inoculate in normal polarity direction into WPM basic medium supplemented with TDZ0.5mg / L, 6-BA2.0mg / L, the The amount of sucrose added to the medium was 30 g / L, the am...
Embodiment 2
[0044] The difference between this example and Example 1 lies in step (1): Disinfection and inoculation of the soaked stems in the field: Take the new stems of the soaked in the field that were extracted in the same year, cut them into stems with axillary buds, and divide them into mild and mild ones according to the degree of lignification. , medium and high, first use a toothbrush to gently scrub under tap water, add an appropriate amount of detergent solution to soak for 2-4 minutes, rinse with running water for 0.5 hours, and then transfer the stems to 70% alcohol solution in an ultra-clean bench Soak for 2 minutes, rinse with sterile water for 3 times, immerse in 0.1% HgCl with 5 drops of Tween-80 2 Soak and sterilize in the solution for 11-17min, then fully soak with sterile water for 3 times, inoculate in normal polarity direction into WPM basic medium supplemented with TDZ0.5mg / L, 6-BA2.0mg / L, the The amount of sucrose added to the medium was 30 g / L, the amount of agar...
Embodiment 3
[0048] The difference between this example and Example 1 lies in step (2): Induction of clustered buds: select moderately lignified stem segments from the new stems extracted in the same year, first use a toothbrush to gently scrub under tap water, and add an appropriate amount of detergent Soak in the solution for 2-4min, rinse with running water for 0.5h, then put the stem section into 70% alcohol solution and soak for 2min in the ultra-clean bench, rinse with sterile water for 3 times, add 5 drops of Tween-80 0.1%HgCl 2 Soak and sterilize in the solution for 13-14min, then fully soak with sterile water for 3 times, and inoculate in the direction of normal polarity to add TDZ0-2.0mg / L, 6-BA0-5.0mg / L, IAA0-5.0mg / L L, CH0-1.0mg / L WPM basic medium, the amount of sucrose added in the medium is 30g / L, the amount of agar added is 7g / L, the pH is 5.8-6.0, and the medium is placed at 121°C at high temperature Autoclave for 20min. The culture medium of the explants was first placed...
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