Breeding method of male sterile cytoplasm NAU7A of rapeseed
A technology of male sterility and male sterile lines, applied in the field of genetic engineering and plant breeding, can solve the problems of unstable fertility, single and single sterile cytoplasm, etc., and achieve the effect of wide application
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Embodiment 1
[0038] Protoplasts were extracted from leaves of Descurainian sophia and cotyledons of Brassica napus, PEG-high pH, high calcium plus DMSO fusion method, and cultured in a shallow liquid layer to study the effects of Descurainian sophia and Brassica napus 19 regenerated plants were obtained by protoplast fusion factors of Brassica napus. The results showed that: 4% cellulase + 0.5% isolated enzyme + 5 mmol / L MES enzymatic hydrolysis for 10 hours and 1% cellulase + 0.2% isolated enzyme + 3 mmol / L MES enzymatic hydrolysis for 14 hours were suitable for obtaining high-yield Artemisia annua and rape respectively Protoplasts; 30% PEG+0.3mol / L glucose+50mmol / L CaCl2 2H2O+15%DMSO obtained a 10% fusion rate; the protoplast culture effect of the improved B5 medium was better; different fresh cultures were added each time The addition of MES has a good effect on preventing browning; the suitable differentiation medium is MS+4.0mg / L 6-BA+0.3mg / L NAA+5.0mg / L AgNO3; the suitable rooting ...
Embodiment 2
[0041] Sterile plants appeared in fusion hybrids obtained by using protoplast fusion technology. The sterile line was used to cross with NAU7B, and the completely sterile line was further selected from cross F1 to backcross with NAU7A. After 5 generations of backcross, the stable sterile line NAU7A was obtained. The sterility of NAU7A showed stable sterility among years. Through the test cross, the restorer line has been found, and the three-line matching has been realized, which provides a new pollination control system for the utilization of heterosis. The study pioneered the cytoplasmic method, and through this method, cytoplasmic recombination was found from the fusion hybrid to the first generation of backcrossing; the study of gene cloning showed that the cytoplasmic mtDNA remained stable after the first generation of backcrossing, and no recombination occurred.
[0042] NAU7A is one of a series of recombinant cytoplasms obtained by protoplast fusion. In the present in...
Embodiment 3
[0044] Using the NAU7A sterile cytoplasm to cross the Chinese cabbage (Brassica napus) variety "Shanghai Qing", the F1 generation is still sterile, but after the NAP cytoplasmic Brassica napus is crossed with Shanghai Qing, the pollen amount of the F1 is basically normal, indicating that NAU7A sterile cytoplasm can be transferred to close relatives of rapeseed, providing a new method for the utilization of heterosis.
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