In-vitro rapid propagation method of rare and endangered plant ammopiptanthus mongolicus

[0036] (1) Obtaining Ilex mongolica explants: Inoculate Ilex mongolica seeds sterilized by conventional methods into MS medium containing 2.0 mg / L 6-BA, culture in the dark for 7-11 days, and take germinated seedlings , the thickness of the epicotyl is 3-5mm, keep the meristem part of the cotyledon near the hypocotyl 2-5mm and the epicotyl 2-4mm, cut off the rest, and then cut longitudinally along the hypocotyl in the middle of the two cotyledons, Remove the axillary buds, cut 2-4 wounds at the junction of cotyledons and hypocotyls with a scalpel, and obtain two cotyledon node explants.

[0037] (2) Induction culture of clustered buds: with the adaxial side of the cotyledon nodes facing upwards, insert them into B containing 1.0 mg / L 6-BA at an angle of 45°C. 5 In the culture medium, dark induction was performed for 5-7 days, the culture temperature was 22-26°C during the day, and 17-21°C at night, and then artificially assisted with light induction for 15 days. The induction

[0042] The difference from Example 1 is that in the step (2) of the induction culture of clustered buds, light induction was assisted for 20 days.

[0044] The difference from Example 1 is that step (3) assisted light induction for 20 days in the elongation culture of clustered buds.

[0045] With the adaxial side of the cotyledon node facing up, insert it obliquely at 45°C into B containing 1.0 mg / L 6-BA 5 In the culture medium, the dark induction is 5-7 days, the culture temperature is 22-26°C during the day, and 17-21°C at night, then the induction rate of artificial cluster buds is 78.3%, and the average number of buds is 2.6.