Tissue culture and rapid propagation method of new variety 'Shirley' of vertical branch type chionanthus retusus
A technology for tissue culture, rapid propagation and new varieties, which is applied in the field of plant cell engineering to achieve the effects of high multiplication multiple, low mutation rate, and improved reproduction efficiency.
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Embodiment 1
[0042] Based on WPM, add 30g / L sucrose and 6g / L agar, and add different concentration gradients of KT (0.1, 0.5, 1.0mg / L), 6-BA (1.0, 1.5, 2.0mg / L) Two-factor random block design, experimental design shown in Table 1. 30 flasks were inoculated with each medium, with 2 explants per flask. The growth height of stem segments, the condition of new leaves and the proliferation of adventitious buds were observed every week (proliferation coefficient=number of new shoots multiplied / inoculated quantity). Culture conditions: The culture temperature is 24°C±2°C, the light intensity is 2500-3000lx, the light time is 12h / d, and the pH of the medium is 5.8-6.2.
[0043] Table 1: Effects of different growth regulators on stem segment differentiation of 'Xueli' (observation results at 35 days)
[0044]
[0045] After culturing for 35 days, it was observed that the combination of growth regulators in group 5 had the best effect on promoting the differentiation of 'Xueli' stem segments, t...
Embodiment 2
[0047] Based on WPM, add 30g / L sucrose, 6g / L agar, 1.5mg / L 6-BA, 0.5mg / L KT, by adding different concentrations of GA 3 (0.5-4mg / L) to carry out single factor experiment, experimental design is shown in Table 2. Each medium was used to inoculate 30 bottles, and each bottle had 2 stem segments cultured with the medium that induced stem segment differentiation. Observe and record the elongation height of stem segment and seedling height every week. Culture conditions: culture temperature is 24°C±2°C, light intensity is 2500-3000lx, light time is 12h / d, medium pH is 5.8-6.2.
[0048] After culturing for 15 days, it was observed that group 3 (i.e. adding 2.0mg / L of GA 3 ) had the best effect on promoting the elongation of the stem section of 'Xueli', and the stem section of tassel tree could grow by 3.01cm (Table 2 and image 3 ).
[0049] Table 2: GA of stem segments of ‘Xueli’ 3 single factor elongation test
[0050]
Embodiment 3
[0052] With WPM as the basic medium, add 30g / L sucrose and 6g / L agar, add different concentration gradients of IBA (0.1-1.5mg / L), NAA (0.1-1.5mg / L) to carry out two-factor random block design , see Table 3 for the experimental design. Observe and record rooting performance every week. Culture conditions: culture temperature is 24°C±2°C, light intensity is 2500-3000lx, light time is 12h / d, medium pH 5.8-6.2.
[0053] Table 3: Effects of different plant growth regulators on rooting of 'Xueli'
[0054]
[0055] It was observed that the combination of growth regulators in group 8 had the best effect on promoting rooting, and the root primordium was formed after about 15 days of culture. After 35 days of culture, the root length reached 5.1 cm, and the rooting rate reached 62.79%.
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