Method for cultivating male sterile tobacco haploids from unfertilized ovules
A technology for male sterility and ovule cultivation, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of un-pollinated ovary cultivation, yield and quality trait deterioration, and breeding problems of male sterile hybrids that have not been seen. Use limited value and other issues to achieve the effect of inhibiting the occurrence of glass seedlings, promoting clustering, and increasing the frequency of induction and plant regeneration
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Embodiment 1
[0038] Embodiment 1: After removing the ovary wall from the ovary of the flower buds in the five stages of K1, K2, K3, K4, K5, put into the embryoid induction medium prepared with H1 medium as the basic medium and cultivate with H2. The embryoid body induction test was carried out in the embryoid body induction medium prepared with the basic medium.
[0039] Wherein, the specific operation process of embryoid induction culture with the embryoid induction medium prepared with H1 medium as the basic medium is as follows:
[0040] (1) Disinfection treatment and inoculation of the material, put the collected flower buds in the above five periods into 0.1% mercuric solution for 7-9min disinfection, then rinse with sterile water 3-4 times, then under sterile conditions Cut off the base of the flower buds of the five stages after disinfection, squeeze out the ovary with tweezers respectively, and remove the ovary wall, and put the ovules of the five stages into five cells containing ...
Embodiment 2
[0045] Example 2: The ovaries of the flower buds in the five stages of K1, K2, K3, K4 and K5 were directly stripped of the ovules, and put into the embryoid induction medium prepared with the H1 medium as the basic medium and the H2 medium respectively. The embryoid body induction test was carried out in the embryoid body induction medium prepared for the basic medium.
Embodiment 3
[0046] Embodiment three: the ovary of above-mentioned K1, K2, K3, K4, K5 five stage flower buds is put into respectively the embryoid body induction medium prepared with H1 medium as basic medium and the H2 medium after circumcision. The embryoid body induction test was carried out in the embryoid body induction medium prepared for the basic medium. After one week of inoculation and culture of the circumcised ovary, the exposed ovules were obviously enlarged, and the ovary walls of the explants with ovary walls turned green. Thickening, no rupture of the ovary wall was observed, and there was basically no sign of enlargement of the inner ovule. At this time, the ovary wall was removed, and the ovule was still browned and died during the culture process.
[0047] In the above-mentioned three embodiments, the ovaries of the five stages of K1 to K5, when the ovary wall is removed or the ovule is directly peeled off for culture, the ovules all have swelling phenomenon, but K1 to K5...
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