A kind of tissue culture method of Populus tomentosa seedling
A technology of tissue culture and Populus tomentosa, applied in horticultural methods, botany equipment and methods, plant regeneration, etc., can solve the problems of no tissue culture method of Populus tomentosa, and achieve the effect of shortening the reproduction cycle
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Embodiment 1
[0052] Cultivation of Maoxin Yang by in-bottle rooting method:
[0053] (1) Sterilization of explants:
[0054] After sterilizing and sterilizing the young stems of Maoxin Yang in alcohol with a mass concentration of 75% for 40 to 60 seconds, transfer them to a mercuric solution with a mass concentration of 0.2% for sterilization for 10 to 15 minutes, and then use sterile Rinse with water for 3 to 5 times to obtain detoxified explants.
[0055] (2) Adventitious bud culture:
[0056] In a sterile working environment, cut off both ends of the sterilized stem section in contact with the disinfectant, then cut the stem section into sections with a length of 0.5 cm to 1 cm and 1 to 2 growth points, and inoculate them into induced In the medium: the induction medium is based on MS medium, and is obtained by adding 6-BA, IBA, sucrose and agar. The concentration of 6-BA in the induction medium is 0.3 mg / L, and the concentration of IBA is 0.2 mg / L. L, the concentration of sucrose is...
Embodiment 1-1 to Embodiment 1-5
[0064] Change the type and dosage of reagents in the subculture medium in step (3), and observe the effects of different hormone combinations on the reproductive coefficient of adventitious buds of P. Wherein, multiplication factor=total proliferation of explant growth points / total number of differentiated explants, calculated by the method of averaging.
[0065] Table 1
[0066]
[0067] It can be seen from Table 1 that, when the 6-BA concentration is 0.2 mg / L, the plant height and multiplication multiple of P. rhizoma are the best in Comparative Examples 1-1 to 1-3. However, it can be seen from Examples 1-3 to 1-5 that when the concentration of IBA is 0.1 mg / L and the concentration of 6-BA is 0.2 mg / L, the plant height and proliferation effects are the most obvious, especially when NAA is not used, The multiplication factor is above 3.8. Therefore, for the subculture step, the most suitable subculture medium formulation is MS+0.2mg / L of 6-BA+0.1mg / L of IBA.
Embodiment 2
[0069] In the subculture in step (3), the concentration of 6-BA in the subculture medium is 0.2 mg / L, the concentration of IBA is 0.1 mg / L, the concentration of sucrose is 30 g / L, and the concentration of agar is 6 g / L L, NAA is not used, and the pH of the subculture medium is 5.8-6.0.
[0070] In the rooting culture of step (4), the concentration of NAA in the rooting medium is 0.1-0.3 mg / L, the concentration of IBA is 0.1-0.5 mg / L, the concentration of sucrose is 30 g / L, and the concentration of agar is 6 g / L L, and the pH value of the rooting medium is 5.8 to 6.0. Other steps and operation modes are the same as those in Example 1.
PUM
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