Isolated culture method of unfertilized ovary of lilium

[0036] The flower buds of embodiment 1 different periods, different genotypes, the in vitro culture contrast of different culture medium:

[0037] Lily unfertilized ovary culture method in vitro, comprises the following steps:

[0038] (1) Select flower buds, place them in a sterile bottle under aseptic conditions, first sterilize the surface with 75% alcohol for 30 seconds, then sterilize with 10% NaClO for 10 minutes, and then rinse with sterile water 3 times, 3 minutes each time;

[0039] (2) In the ultra-clean workbench, peel off the sterile flower buds to take out the ovary, and cut the ovary into slices with a thickness of 1-2 mm;

[0040] (3) Inoculate ovary slices in Erlenmeyer flasks or petri dishes containing different induction media, and culture them under the conditions of 1600lx light, 14h / 10h (day / night) and (25±1)°C for 20-25d , to induce embryo formation within the ovule;

[0041] (4) The number of ovules containing embryos (embryogenic ovules) was counted

[0052] The character of each culture stage of embodiment 2

[0053] (1) Take flower buds (about 10 cm) 1 day before flowering in the greenhouse, put them in a sterile bottle under sterile conditions, first sterilize the surface with 75% alcohol for 30 seconds, then sterilize with 10% NaClO for 10 minutes, and then use Rinse with sterile water 3 times, 3 minutes each time;

[0054] (2) Peel off the sterilized flower buds, take out the ovary, cut it transversely into slices with a thickness of 1-2 mm with a scalpel, and place the slices horizontally on the induction medium for induction culture ( figure 1 Middle A), the induction medium is: BDS+2,4-D2 mg / L+6-BA2 mg / L+sucrose 100g.L -1 + agar 6g.L -1 , pH6.0. The culture conditions are: light 1600lx, 14h / 10h (day / night), (25±1)°C. Cultivate for 20-25 days.

[0055] (3) After 20-25 days of induction culture, the ovules expanded, and the sliced ​​ovaries also expanded ( figure 1 Middle B), the embryo inside the enlarged