Rice herbicide resistant protein and application thereof in plant bleeding

A herbicide, plant technology, applied in the field of plant protein

Active Publication Date: 2012-07-18
SHENZHEN XINGWANG BIOLOGICAL SEED IND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, plants with herbicide resistance and their decisive substances (such as proteins, genes, etc.) have not yet been theoretically designed, and can only be obtained through the hard work of scientific researchers, especially long-term screening, and relying on some luck to get

Method used

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  • Rice herbicide resistant protein and application thereof in plant bleeding
  • Rice herbicide resistant protein and application thereof in plant bleeding
  • Rice herbicide resistant protein and application thereof in plant bleeding

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Take 60 kg of seeds of Huanghuazhan and 30 kg of seeds of Huangsizhan, which are commercially available, respectively, soak them in water for 22 hours in an incubator at 28° C., then fish out the seeds and dry them out. Then, soak the seeds of Huanghuazhan with 0.7% (w / w) ethyl methanesulfonate (EMS) and soak the seeds of Huangsizhan with 0.8% (w / w) ethyl methanesulfonate (EMS) for 12 hours each, during which Shake the seeds every 1 hour. After 12 hours, discard the EMS solution, replace the tap water to soak the seeds, discard the tap water, and repeat the soaking 5 times in total, soaking for 5 minutes each time, then rinse the seeds with tap water for 2 hours, and turn the seeds during the rinsing process to wash off the EMS. Then put the seeds on the surface of wet paper, put them in an incubator at 28°C, the germination rate is 60-70%, and then sow. Two weeks after sowing, the rate of white seedlings was detected, among which Huanghua accounted for 3.5%, and Huang...

Embodiment 2

[0040] Refer to the method of Fan ZJ et al. (Specific activity determination of acetolactate synthase from maize (Zea mays L.). See Han WN et al. The Proceedings of the 18 th Asia-Pacific Weed Science Society Conference . 515 - 522 . May 28 -June 2, 2001. Beijing: Standards Press of China.), extracted the ALS enzymes of the wild-type and mutant plants of Huangsizhan and Huanghuazhan mentioned above, and determined the rate of inhibition of the corresponding enzyme activity by imidazolinone herbicides. In brief, 5 g of each plant seedling was taken and chopped, and 10 mL of 50 mmol / L K at pH 7 was added. 2 HPO 4 -KH 2 PO 4 Buffer (which contains 1mmol / L sodium pyruvate, 0.5mmol / L MgCl 2 , 0.5mmol / L TPP, 10μmol / L FAD), ground and pulverized with quartz sand, filtered with 8 layers of gauze, and the filtrate was centrifuged at 4°C and 20,000rpm for 30 minutes. Take the supernatant, add ammonium sulfate to make it reach 50% saturation, then place it at 0°C for 2 hours, then ce...

Embodiment 3

[0045] Beijing Weiming Kaituo Agricultural Biotechnology Co., Ltd. was entrusted to use the conventional Agrobacterium-mediated method to transfer the above-mentioned mutant ALS enzyme coding genes of Huangsizhan and Huanghuazhan into wild-type Arabidopsis plants. In brief, the forward primer (5'-GGTTCATAGTTGACACCTTAG-3') and the reverse primer (5'-GTCATCCTATCCCCACCGACAT-3') were used to PCR amplify the genomic DNA of the above-mentioned Huangsizhan and Huanghuazhan mutant plants, respectively. Amplify the mutant ALS gene, after the sequence is correct, clone the ALS gene whose nucleotide sequence is shown in SEQ ID NO: 1 and 3 into the pCAMBIA1303 plasmid (purchased from Cambia Company), select the positive clone and transform it into Agrobacterium AGL0, and culture the bacteria and transformed Arabidopsis thaliana. The transgenic seeds of the T1 generation were obtained after harvesting the positive Arabidopsis thaliana by PCR. When planting, they were sprayed with the imidaz...

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Abstract

The invention discloses rice herbicide resistant protein and application thereof in plant bleeding. The ALS (acetolactate synthase) protein provided by the invention realizes that a plant has the herbicide resistance, and is derived from rice varieties Huangsizhan and Huanghuazhan, and carries Trp548, Ala96 or Ser627mutation. Furthermore, the invention also provides corresponding nucleic acid, expression cassettes, carriers, cells, plants, and method for obtaining the plants with herbicide resistance and application of the plants, as well as a method for identifying the plants in the invention.

Description

technical field [0001] The invention belongs to the field of plant proteins, in particular, the invention relates to a protein capable of imparting herbicide resistance (tolerance) to plants (especially rice) and its application in plant breeding. Background technique [0002] Weeds in farmland are one of the most important factors affecting crop yields, but manual weeding consumes high human resources and costs, and it is not convenient for intensive agricultural production, which seriously restricts crop planting in the direction of high yield, high quality and low cost development process. Therefore, herbicides have emerged as the times require, and their use has played a significant role in solving farmland weed damage, promoting innovation in cultivation methods, and increasing production. [0003] Common herbicides, including sulfonylureas, imidazolinones, pyrimidotriazoles, pyrimidine salicylate and other herbicides, have been widely used commercially. However, sinc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/88C12N15/60C12N15/63C12N1/21C12N1/19C12N5/10A01H5/00A01H1/02A01H4/00C12N15/84C12Q1/68C12Q1/527
Inventor 陈竹锋王承旭柳威唐晓艳邓兴旺
Owner SHENZHEN XINGWANG BIOLOGICAL SEED IND
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