Herbicide resistance mutant and application thereof
A herbicide and resistance technology, applied in herbicide-resistant mutants and its application in plant breeding, can solve the problems of large environmental pollution, heavy phytotoxicity, and large residues of sulfonylurea herbicides
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Embodiment 1
[0044] Embodiment 1, mutagenesis obtains the mutant resistant to clethodim herbicide
[0045] Seeds of rice Huanghuazhan are sown. Before sowing, the seeds of Huanghuazhan are soaked in water for 22 hours in an incubator at 25°C, then the seeds are taken out and the water is controlled to dry. Then, the rice seeds were soaked in 0.7% (w / w) ethyl methanesulfonate (EMS) for 12 hours, during which the seeds were shaken every 1 hour. After 12 hours, discard the EMS solution, change the tap water to soak the seeds, discard the tap water, repeat the soaking 5 times, soak for 5 minutes each time, then rinse the seeds with tap water for 2 hours, and turn the seeds during the washing process to wash off the EMS. Take a small amount of seeds and place them on the surface of wet paper, put them in an incubator at 28°C, and measure the germination rate to 60-70%, and sow the remaining seeds in the field. After the plants are mature, they are harvested in plots, each plot has an area of ...
Embodiment 2
[0046] Example 2. Genome sequence analysis of clethodim herbicide-resistant mutants
[0047] Take the leaves of the above-mentioned mutant rice plants, together with the leaves of the wild-type Huanghuazhan rice plants, extract their genomic DNA respectively, and entrust Shenzhen Huada Gene Research Institute to perform sequencing. The sequence comparison shows that: compared with the wild-type Huanghuazhan, the acetyl Coenzyme A carboxylase (ACCase) is mutated at position 5372, from A to G, resulting in the change of asparagine to serine at position 1791 of the corresponding encoded protein. The nucleotide sequence of the mutated ACCase gene As shown in SEQ ID NO: 4, the amino acid sequence of the ACCase protein encoded by it is shown in SEQ ID NO: 5. It was backcrossed and purified, and a stable strain KX2-28 was obtained after self-propagation; the other three mutant plants Acetyl-CoA carboxylase (ACCase) has mutated at the same point. The specific mutation is that the 5374...
Embodiment 3
[0048] The seedling spray experiment of embodiment 3, mutant KX2-2, KX2-28 and KX3-89
[0049] The obtained rice mutant lines KX2-2, KX2-28, KX3-89 resistant to ACCase inhibitor herbicides and wild type Huanghuazhan seeds (WT) were sown respectively. Wild-type rice seeds (WT) and mutant KX2-2 were sprayed with 0, 35, 70, 105, 140, 175, 210 g a.i. / hm at the 4-leaf stage, respectively 2 The dose of clethodim was used to verify the resistance, and the experimental results were as follows figure 2 shown at 35g a.i. / hm 2 Under the treatment of clethodim at a certain dose, most of the wild-type Huanghua died, and the growth and development were greatly affected, while KX2-2 was not affected by clethodim and showed strong resistance to clethodim; when clethodim was applied Dose up to 105g a.i. / hm 2 At the same time, the wild-type Huanghua accounted for all the deaths, while about 40% of the resistant mutant strain KX2-2 survived.
[0050] Wild-type rice seeds (WT) and mutant lin...
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