One-step seedling and efficient in-vitro propagation method with gynura bicolor leaves
A technique for in vitro propagation of G. violaceum, applied in horticultural methods, botanical equipment and methods, horticulture and other directions, can solve the problems of long seedling breeding cycle, easy mutation of germplasm, complicated procedures, etc., and achieves good growth and seedlings. The effect of clump growth is robust
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Embodiment 1
[0031] (1) Medium preparation: MS was used as the basic medium, 0.25 mg of 6-BA, 0.4 mg of NAA, 0.2 mg of IBA, 7 g of carrageenan, and 30 g of sucrose were added per liter, and the pH of the medium was adjusted to 6.0 after mixing ; Dispense each bottle with about 30 mL, put it in an autoclave with sterilization conditions set at 121°C and 105 Kpa for 15 minutes, take out the culture bottle, and cool it for later use;
[0032] (2) Explant treatment and inoculation: According to the conventional inoculation method, the leaves of aseptic seedlings of G. purple-backed seedlings with a leaf diameter of about 0.8-1.5 cm were cut as explants, and the leaves were directly inoculated on the above-mentioned medium (MS+ 0.25 mg / L 6-BA+0.4 mg / L NAA+0.2 mg / L IBA+30 g / L sucrose+7 g / L carrageenan), inoculate 2 explants in each bottle, and aseptically seal the mouth of the bottle;
[0033] (3) Cultivation: Move the inoculated culture bottle into the culture room for culture, the culture cond...
Embodiment 2
[0036] Other operations are the same as in Example 1, except that in step (1), the formula of the medium is (MS+0.25 mg / L 6-BA+0.6 mg / L NAA+0.2 mg / L IBA+30 g / L sucrose + 7 g / L carrageenan); in step ((3), after 15 days of culture, the survival rate of the explants was 100%, the leaves were obviously enlarged, the leaf margins were upturned, and the induction rate of callus cells (or tissues) 100%. After cultivating for 35 days, the leaves of the explants transformed into massive bud clusters, the differentiation rate was 100%, the height of the bud clusters was about 0-7 mm, the growth rate of the bud clusters was faster, and a small amount of complete leaves were formed. Meanwhile, the bud clusters The inside of the clump tissue emits white fluff, and then the fluff grows rapidly into fine roots densely covered with white fluff, and the root tissue is relatively rich. After 45 days of cultivation, the seedling clump begins to form, with bulbs appearing at the base, and the lea...
Embodiment 3
[0038] Other operations are the same as in Example 2, except that in step (1), the formula of the medium is (MS+0.25 mg / L 6-BA+0.8 mg / L NAA+0.2 mg / L IBA+30 g / L sucrose + 7 g / L carrageenan); in step ((3), after 15 days of culture, the survival rate of the explants was 100%, the leaves were significantly enlarged and arched, the leaf margins were upturned, and the callus induction rate was 100%. After 35 days of culture, the leaves of the explants had rapidly transformed into massive bud clusters, the differentiation rate was 100%, the height of the bud clusters was about 2-7 mm, the bud clusters grew vigorously, and a large number of complete leaves were formed. At the same time, the base of the bud clusters had A large number of white fluffy fine roots are formed, and the root system is rich. After 40 days of cultivation, the seedling clusters begin to form, with obvious bulbs at the base, and the leaves are stretched and green. After 65 days of cultivation, the seedling clust...
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