Method for widening tetraploid wheat genetic variation
A technique of genetic variation and tetraploidy, applied in botany equipment and methods, application, plant gene improvement, etc., can solve the problems of low frequency of beneficial mutations, high cost of chemical and physical mutations, environmental pollution, etc., to achieve The effects of high mutagenesis efficiency, rich mutagenesis types, and high mutation frequency
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Embodiment 1
[0036] A method for widening the genetic variation of tetraploid wheat, comprising the steps of:
[0037] (1) The genetically stable parental tetraploid wheat AABB was sexually crossed with the genetically stable diploid DD and bagged, and triploid immature embryos ABD were obtained 12-15 days after pollination;
[0038] (2) The triploid immature embryos ABD obtained in step (1) are cultured in vitro in the laboratory using tissue culture technology to obtain ABD plant seedlings;
[0039] Wheat immature embryos are cultured in vitro using MS solid medium. The specific formula of MS solid medium is as follows:
[0040] 1) 500ml of 100× iron salt mother liquor (1.39g of FeSO 4 ·7H 2 O, 1.86 g Na 2 EDTA·2H 2 O);
[0041] 2) 500ml of 20×macroelement mother liquor (16.5g of NH 4 NO 3 , 19 g KNO 3 , 1.7 g KH 2 PO 4 , 3.7 g MgSO 4 ·7H2 O, 3.32 g anhydrous CaCl 2 );
[0042] 3) 100×trace element mother solution 200ml (16.6 mg KI, 124 mg H 3 BO 3 , 338 mg MnSO 4 ·H 2 O...
Embodiment 2
[0062] The difference with embodiment 1 is:
[0063] In step (3), the method for performing chromosome doubling treatment on ABD seedlings is: soaking ABD seedlings with a mixed solution of colchicine and dimethyl sulfoxide (the preparation method of the mixed solution of colchicine and dimethyl sulfoxide is: First prepare a colchicine solution with a mass percentage concentration of 0.06%, and then prepare a volume percentage concentration of 1.2% dimethyl sulfoxide solution; finally, mix the two solutions evenly, which is a mixture of colchicine and dimethyl sulfoxide solution); at the same time, use an oxygen pump to maintain an oxygenated state at the bottom of the mixed solution (it is enough to keep the mixed solution with continuous bubbles during the entire experiment); under the environmental conditions of a temperature of 20 ° C and light (fluorescent lamp irradiation), treat for 18 hours , after flushing for 2.2 hours, the chromosome doubling treatment of ABD seedli...
Embodiment 3
[0066] The difference with embodiment 1 is:
[0067] In step (3), the method for performing chromosome doubling treatment on ABD seedlings is: soaking ABD seedlings with a mixed solution of colchicine and dimethyl sulfoxide (the preparation method of the mixed solution of colchicine and dimethyl sulfoxide is: First prepare a colchicine solution with a mass percentage concentration of 0.04%, and then prepare a volume percentage concentration of 1.8% dimethyl sulfoxide solution; finally, mix the two solutions evenly, which is a mixture of colchicine and dimethyl sulfoxide solution); at the same time, use an oxygen pump to maintain an oxygenated state at the bottom of the mixed solution (it is enough to keep the mixed solution with continuous bubbles during the entire experiment); under the environmental conditions of a temperature of 20 ° C and light (fluorescent lamp irradiation), treat for 22 hours , after flushing for 1.8 hours, the chromosome doubling treatment of ABD seedli...
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