High efficiency quick breeding method for cichorium intybus test tube seedling
A chicory and high-efficiency technology, applied in horticultural methods, botanical equipment and methods, plant cells, etc., can solve the problems of long cultivation period, complicated procedures, and low frequency of plant regeneration, and achieve the effect of reducing cultivation cost and increasing reproduction coefficient.
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Embodiment 1
[0047]Embodiment 1: choose the plump chicory seeds, rinse with tap water, soak and shake for 30s with 70% ethanol, 0.1% HgCl 2 The surface was disinfected for 12 minutes, rinsed with sterile water 6 times, inoculated on 1 / 2 MS basic medium, illuminated for 16 hours a day, the light intensity was 20001x, and the culture temperature was 25±2°C.
[0048] Cut the base of chicory aseptic seedlings cultured for 20 days and inoculate them with 2.0mg / L6-BA, 0.5mg / LNAA, 10mg / L AgNO 3 , 100mg / LV B1 , 300mg / L proline and 4% sucrose MS medium, 25±2°C, 2500lux light culture for 7 days, the surface of the explant and the incision will gradually produce green adventitious buds, after 15 days, the buds develop Clustered adventitious buds, the number of adventitious buds formed by each leaf is at most 44, with an average of 36.08 (such as figure 1 shown). Cut off the adventitious buds more than 2cm, transfer to the MS medium that adds 0.5mg / L IBA, produce adventitious roots after 7 days, ca...
Embodiment 2
[0050] Embodiment 2: choose the plump chicory seeds of grain, wash with tap water, soak and shake 30s with 70% ethanol, 0.1%HgCl 2 Disinfect the surface for 12 minutes, rinse with sterile water 6 times, inoculate on 1 / 2 MS basic medium, light for 16 hours a day, light intensity 20001x, and culture temperature 25±2°C.
[0051] Cut the middle and upper part of the leaves of chicory aseptic seedlings cultured for 30 days, and inoculate them in the supplementary 1.5mg / L6-BA, 0.5mg / L NAA, 5mg / L AgNO 3 , 50mg / L V B1 , 300mg / L proline and 4% sucrose MS medium, 25±2°C, 2500lux light culture for 7 days, the surface of the explant and the incision will produce green adventitious buds one after another, about 15 days, the buds develop Clustered adventitious buds, the number of adventitious buds formed by each leaf is at most 35, with an average of 29.06 (such as figure 2 shown). Cut out adventitious buds over 2cm, transfer them to 1 / 2MS medium supplemented with 0.5mg / L IBA, 5-7d will...
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