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Herbicide-resistant mutants and their uses

A herbicide and resistance technology, applied in herbicide-resistant mutants and its application in plant breeding, can solve the problems of large residues of sulfonylurea herbicides, low drug efficacy, and large environmental pollution

Active Publication Date: 2022-05-27
深圳洁田模式生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, amide herbicides have low drug efficacy and cause great environmental pollution; sulfonylurea herbicides have large residues and serious phytotoxicity

Method used

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  • Herbicide-resistant mutants and their uses
  • Herbicide-resistant mutants and their uses
  • Herbicide-resistant mutants and their uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1. Mutation to obtain a mutant resistant to clethodim herbicides

[0046] Sow the seeds of Huanghuazhan rice. Before sowing, soak the seeds of Huanghuazhan in water in a 25°C incubator for 22 hours, then remove the seeds and control the moisture. Then, the rice seeds were soaked with 0.7% (w / w) ethyl methanesulfonate (EMS) for 12 hours, during which time 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, and repeat the soaking for a total of 5 times, soaking for 5 minutes each time, and then rinse the seeds with tap water for 2 hours, turning the seeds during the rinsing process to wash the EMS. Take a small amount of seeds and put them on the surface of wet paper, put them in a 28°C incubator, measure the germination rate of 60-70%, and sown the remaining seeds in the field. After the plants are mature, they are harvested in plots, each plot area of ​​30 square meters, mi...

Embodiment 2

[0047] Example 2. Genome sequence analysis of clethodim-resistant herbicide mutants

[0048] Take the leaves of the above-mentioned rice mutant plants, together with the leaves of wild-type Huang Huazhan, extract their genomic DNA respectively, and entrust Shenzhen BGI Research Institute for sequencing. Coenzyme A carboxylase (ACCase) is mutated at the 5372nd position, from A to G, resulting in the 1791st position of the corresponding encoded protein from asparagine to serine. 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. After backcross purification, a stable line KX2-28 was obtained after self-cross propagation; the other three mutant plants had Acetyl-CoA carboxylase (ACCase) has been mutated at the same point. The specific mutation is that the 5374th position of the ACCase gene is mutated, changing from A to T, resulting in the 1792th position of the corresp...

Embodiment 3

[0049] Example 3. Seedling stage spraying experiment of mutants KX2-2, KX2-28 and KX3-89

[0050] The obtained mutant lines KX2-2, KX2-28, KX3-89 and wild-type Huanghuazhan seeds (WT) of rice resistant to ACCase inhibitor herbicides were sown separately. 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 applied dose of clethodim confirms resistance, and the experimental results are as follows figure 2 shown at 35g a.i. / hm 2 Under the treatment of clethodim, the wild-type Huanghua accounted for most of the death, and the growth and development were greatly affected, while KX2-2 was not affected by clethodim and showed strong clethodim resistance; when clethodim was applied Doses up to 105g a.i. / hm 2 At the time, the wild type Huanghua accounted for all deaths, while the resistant mutant line KX2-2 still survived about 40%.

[0051] Wild-type rice seeds (WT) and mutant line KX2-28 wer...

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Abstract

The invention discloses a herbicide-resistant mutant and its application, and belongs to the field of plant biotechnology. Specifically, the invention relates to a protein-nucleic acid mutant sequence capable of causing plants to produce herbicide resistance and the herbicide-resistant mutant sequence. Sex mutants and their use in plant breeding. The present invention uses clethodim herbicide to find six mutants with resistance to ACCase inhibitor herbicides in the rice mutant library, and the resistant mutants can tolerate ACCase inhibitor herbicides, especially ene oxalone. The present invention also develops the application of the protein with ACCase inhibitor herbicide resistance and its coding gene in transgenic or non-transgenic plant breeding, and the herbicide resistance gene and protein can be used to breed plants with herbicide resistance , especially gramineous crops, such as rice, corn, wheat, barley, sorghum, millet, millet, etc., have great application value in plant breeding and agricultural production.

Description

technical field [0001] The present invention belongs to the field of plant biotechnology, in particular, the present invention relates to a protein nucleic acid mutant sequence capable of making plants resistant to herbicides and the application of the herbicide resistant mutant in plant breeding. [0002] technical background [0003] With the large-scale application of rice direct seeding cultivation technology in agricultural production, the problem of farmland weeds in rice direct seeding production has become an important factor affecting crop yield. On the one hand, manual weeding consumes high human resources and costs, and it is not convenient for intensive agricultural production, which seriously restricts the development process of crop planting towards high yield, high quality and low cost. On the other hand, grass weeds in paddy fields are difficult to control, and herbicides that can be used to kill grass weeds can usually kill rice, so the use of herbicides is e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00C12N15/52C12N15/82A01H5/00A01H6/46
CPCC12N9/93C12N15/01C12N15/8274C12Y604/01002
Inventor 唐晓艳邓兴旺周君莉陈竹锋王承旭陈雷许纯珏金名捺谢刚李早霞
Owner 深圳洁田模式生物科技有限公司
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