Mutation type PSII D1 or psbA D1 protein, and coding nucleic acid and application thereof
A mutant, protein amino acid technology, applied in the field of agricultural genetic engineering
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Embodiment 1
[0060] In the present invention, a large number of seeds are obtained by planting the mutated rice seeds, and then a large number of second-generation rice seedlings are obtained by sowing, spraying PSII herbicides, sequencing and analyzing the surviving seedlings, the specific steps are as follows:
[0061]Purchase 100 kilograms of Huaidao No. 5 and 100 kilograms of Huanghuazhan seeds from the market, soak them in water for 22 hours in an artificial climate chamber at 28°C, remove and dry the floating water, and then use 0.7% ethyl methanesulfonate (Ethyl methanesulfonate, EMS) Soak for 12 hours. Stir once an hour during this period, so that the ethyl methanesulfonate can be fully and evenly absorbed by the seeds. In addition, part of the seeds were taken, soaked in water again for 12 hours, and treated without adding ethyl methanesulfonate as a control. After 12 hours, pick up the seeds, discard the ethyl methanesulfonate aqueous solution, and rinse repeatedly with a large ...
Embodiment 2
[0066] The resistant rice seedlings of the present invention are cultivated and seeded, and crossed with the wild type. It is found that the mutant type is used as the female parent, and the wild type pollen is used to pollinate the mutant type female parent, and the new seeds obtained all inherit the PSII inhibitor. Drug resistance; conversely, the wild type is used as the female parent, and the pollen of the mutant plant is used to pollinate the wild type, and all the resulting plants are killed. The specific method is as follows:
[0067] According to the sequencing results of rice seedlings, two sets of hybridization experiments were carried out directly using contemporary surviving plants and wild-type plants. One group is Huaidao 5 containing the S264G mutation from atrazine screening and the wild type Huaidao 5, and the other group is Huang Huazhan and the wild type Huang Huazhan containing the V219I mutation selected from Diuron. Fifty healthy rice plants with similar...
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