Tissue culture bud propagation method formertensia maritime
A technology of oyster leaves and tissue culture, applied in the field of cultivation and reproduction of oyster leaves, can solve the problems of artificial planting, difficulty in planting and reproduction of oyster leaves, and low germination rate, and achieves rapid seedling raising, overcoming difficulty in reproduction and long-term vegetative growth of plants. Aging, to achieve the effect of annual cyclical production
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Embodiment 1
[0021] Embodiment 1: the culture of oyster leaf aseptic bud, and explant is carried out optimization,
[0022] (1) Orthogonal experiment with four factors and three levels was carried out on explants, culture medium, 6-BA and NAA.
[0023] Four factors: explant, medium, 6-BA, NAA.
[0024] Three levels: explant selection 1 (tiller bud), 2 (leaf), 3 (seed) 3 different parts; medium selection 1 (MS), 2 (improved MS), 3 (B5) 3 different parts Medium; 6-BA chooses 1 (0.5mg / L), 2 (1.0mg / L), 3 (1.5mg / L) 3 different concentrations; NAA chooses 1 (0.1mg / L), 2 (0.2mg / L) L), 3 (0.5mg / L) 3 different concentrations. Preliminary screening of suitable explants, medium, 6-BA, NAA was carried out through the above orthogonal design. One treatment of 3 bottles, repeated 3 times.
[0025] (2) Rinse the oyster leaf seeds, leaves, and tiller buds, place them in 75% alcohol for 45 seconds, soak in 0.1% mercuric solution for 45 seconds, and in sterile water for 60 seconds, then inoculate them i...
Embodiment 2
[0030] Embodiment 2: Bud Proliferation Culture
[0031] (1) Get the aseptic buds of oyster leaves obtained by test group 7 in Example 1, carry out branching processing, and then inoculate in different medium formulas+7g / L agar+30g / L sucrose respectively, and 1 tissue culture bottle is connected with 1 See Table 3 for the specific experimental design, cultivate in a culture room at 18°C, light intensity 2000lx, and photoperiod 12h / d, and observe the growth situation regularly.
[0032] Table 2, Bud Proliferation Culture Conditions
[0033] test group Nutrient solution 6-BAmg / L NAAmg / L Culture temperature ℃ YB1 B5 0.5 0.05 24 YB2 B5 0.5 0.1 24 YB3 B5 0.5 0 24 YB4 B5 0.5 0.05 18
[0034] (2) Experimental results
[0035] After 30 days of inoculation, count the number of proliferating buds in each test group, and carry out division processing, and count the multiplication multiple and effective bud number of each test gro...
Embodiment 3
[0039] Embodiment 3: rooting culture
[0040] Table 4, rooting culture treatment table.
[0041] test group Nutrient solution IBA(mg / L) 1 1 / 2B5 0.2 2 1 / 2B5 0.5 3 1 / 2B5 1.0 4 1 / 4B5 0.2 5 1 / 4B5 0.5 6 1 / 4B5 1.0 7 B5 0.2 8 B5 0.5 9 B5 1.0
[0042] Get the aseptic seedling that YB4 test group obtains among the embodiment 2, carry out branching processing in ultra-clean workbench, then inoculate respectively in different culture medium formula+7g / L agar+30g / L sucrose, 1 tissue culture bottle connects 1 bud, the specific experimental design is shown in Table 4, cultivated in a culture room with a light intensity of 2000lx, a photoperiod of 12h / d, and a temperature of 18°C, and the growth conditions were observed regularly.
[0043] The experimental results are shown in Table 5. In terms of inducing the rooting of oyster leaf buds, considering the excessive growth of buds, root length and rooting rate, a better m...
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