In-vitro tissue culture and rapid propagation method for viburnum burejaeticum
A viburnum and tissue culture technology is applied in the field of rapid propagation of pruning viburnum in vitro tissue culture, and can solve the problems of low survival rate and high pollution rate
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Embodiment 1
[0031] Example 1 Disinfection and primary culture of explants
[0032] Choose pruned viburnum young twigs or sprouting strips that are robust and free from diseases and insect pests as explants, take materials from the field, and bring them back to the laboratory for pre-treatment of explants: first soak in detergent for 30 minutes, and cut Remove the leaves on the young branches, leave the petiole 0.3~0.5cm, and then rinse continuously with tap water for 2h, and then enter the sterile operation room for sterilization: sterilize with 0.1% mercury for 10min, rinse with sterile water 4 times to obtain sterility Explants: Cut the germ-bearing stem segments of the sterile explants into about 2cm and inoculate them in the primary medium (the primary medium is to add 6-BA (1.0mg / L), IBA ( 0.1mg / L) sucrose (30g / L), agar powder (8~10g / L), pH value adjusted to 5.8~6.0) for primary culture to make the axillary buds germinate and grow. The culture conditions are: keep light for 14h ( Every...
Embodiment 2
[0037] Example 2 Tissue culture seedling multiplication culture
[0038] Transfer the viable shoots from the primary culture to a proliferation medium for subsequent proliferation and culture, with 14 hours of light per day (from 8:00 am to 22:00 every day), the light intensity is 2000Lx, and the culture temperature is about 25℃. . The suitable proliferation medium formula for tissue culture seedlings is to add 6-BA (2.0mg / L), IBA (0.2mg / L), GA at an appropriate concentration in MS medium. 3 (0.4mg / L), agar powder (8-10g / L), sucrose (30g / L), pH value adjusted to 5.8-6.0. After many subcultures, cluster sprouts are formed, such as figure 2 As shown, the succession cycle is about 30 days. See Table 2 for the effects of different basic media on the proliferation and growth of tissue cultured seedlings. The effects of different types and concentrations of auxin on the proliferation and growth of tissue cultured seedlings are shown in Table 3.
[0039] Table 2 The effect of differen...
Embodiment 3
[0047] Example 3 Rooting culture in bottle
[0048] Inoculate the rootless tissue culture seedlings with a height of about 2~3cm and strong growth on the rooting medium for rooting culture. The culture temperature is controlled at about 25℃, the light intensity is about 2000Lx, and the light time is from 8:00 to evening every day. Tissue cultured seedlings were induced to take root at 22:00 (14h). The best rooting medium is to add appropriate concentration of IBA (2.0mg / L), sucrose (30g / L) and agar powder (8-10g / L) to 1 / 2MS medium, and adjust the pH value to 5.8-6.0. After 15-20 days of cultivation, rooting will begin, and 3-8 thick adventitious roots can be produced within 30 days, such as image 3 As shown, the rooting rate is over 90%. The effects of different types and concentrations of hormones on the rooting of tissue cultured seedlings are shown in Table 4.
[0049] Table 4 The effects of different types and concentrations of hormones on the rooting of tissue cultured seed...
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