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Method for identifying glyphosate-resistant paddy rice

A glyphosate-resistant, transgenic rice technology, applied in biochemical equipment and methods, microbial assay/inspection, DNA/RNA fragments, etc., can solve the problems of low efficiency, time-consuming and laborious, and high input cost

Active Publication Date: 2019-06-25
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the yield and quality of direct-seeded rice in dry land are directly threatened by weeds in paddy fields. Traditional weeding methods mainly rely on manual pulling or weeding, which is time-consuming, laborious, inefficient and high in input costs. The promotion and use of chemical weeding methods not only reduces the The production cost has also greatly improved the production efficiency of rice

Method used

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  • Method for identifying glyphosate-resistant paddy rice
  • Method for identifying glyphosate-resistant paddy rice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Example 1, Discovery of Molecular Marker Sequences and Design of Primers

[0094] 1. Obtain multi-generation seeds

[0095] 1. At the flowering stage, the transgenic rice En-12 is used as the male parent and the transgenic rice Ec-8 is used as the female parent to perform hybridization, and the obtained seeds are F1 generation seeds.

[0096] 2. The plants grown from the F1 generation seeds are F1 generation plants, and the F1 generation plants are self-crossed, and the obtained seeds are F2 generation seeds.

[0097] 3. The plants grown from F2 generation seeds are F2 generation plants.

[0098] 2. Identification of glyphosate resistance

[0099] The plants to be tested are: transgenic rice En-12, transgenic rice Ec-8, and F2 generation plants.

[0100] The method of detecting glyphosate resistance: when the plant is growing at the 3-5 leaf stage, spray 2000ppm glyphosate solution, observe after 14 days, if the plant is not dead, the plant is a glyphosate resistant ...

Embodiment 2

[0113] Embodiment 2, establishment of method

[0114] 1. Obtain plants of each generation

[0115] 1. At the flowering stage, the transgenic rice En-12 is used as the male parent and the transgenic rice Ec-8 is used as the female parent to perform hybridization, and the obtained seeds are F1 generation seeds.

[0116]2. The plants grown from the F1 generation seeds are F1 generation plants, and the F1 generation plants are self-crossed, and the obtained seeds are F2 generation seeds.

[0117] 3. The plants grown from the F2 generation seeds are F2 generation plants, the F2 generation plants are self-crossed, and the obtained seeds are F3 generation seeds.

[0118] 4. Plants grown from F3 generation seeds are F3 generation plants.

[0119] 2. Detection of plant genotype

[0120] The plants to be tested are: 20 transgenic rice En-12, 20 transgenic rice Ec-8, 20 F1 generation plants, 390 F2 generation plants, and 500 F3 generation plants for sampling. The rice variety Zhonghu...

Embodiment 3

[0138] Embodiment 3, sensitivity detection

[0139] 1. Take the En / Ec plants (F2 generation plants) obtained in Example 2, and extract genomic DNA. This genomic DNA serves as the target DNA.

[0140] 2. Take Zhonghua 11 and extract genomic DNA. This genomic DNA served as background DNA.

[0141] 3. Mix the target DNA and the background DNA to obtain a template solution. The total DNA content in 1 μl template solution is about 100ng, and the target DNA content is 100 copies, 50 copies, 25 copies, 15 copies or 10 copies.

[0142] 4. Take the template solution obtained in step 3, use the primers designed in Example 1 to perform PCR amplification on A, and perform 1% agarose gel on the PCR amplification product.

[0143] The reaction system of PCR amplification and the reaction procedure of PCR amplification are the same as step 2 of Example 2.

[0144] The results showed that when the content of the target DNA in the system was more than 15 copies, the target band of 521bp c...

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Abstract

The invention discloses a method for identifying glyphosate-resistant paddy rice. The invention provides a primer combination composed of a primer pair A and a primer pair B; the primer pair A is composed of En-F and En-R; the primer pair B is composed of Ec-F and Ec-R; the En-F is shown in a sequence 3; the En-R is shown in a sequence 4; the Ec-F is shown in a sequence 5; the Ec-R is shown in a sequence 6. The invention also provides a method for screening glyphosate-resistant plants in the offspring with transgenic paddy rice En-12 and transgenic paddy rice Ec-8 as parents, wherein the method comprises the steps: the genomic DNA of the offspring plants with the transgenic paddy rice En-12 and the transgenic paddy rice Ec-8 as the parents is used as a template, the primer pair A and the primer pair B are subjected to PCR amplification respectively, a 521 bp product is obtained if the primer pair A is adopted, and a 550 bp product is obtained if the primer pair B is adopted, wherein the offspring plants are glyphosate-resistant plants. The primer combination provided by the invention can be used for identifying the glyphosate-resistant paddy rice, and has the advantages of high accuracy, specificity and sensitivity.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for identifying glyphosate-resistant rice. Background technique [0002] Rice is one of the most important food crops in the world and occupies a pivotal position in my country's food production. However, the production of rice is plagued by problems such as the reduction of farmland area, the shortage of rural labor force, and the shortage of water resources. In order to solve these problems, the mode of rice production is changing from the model of rice transplanting in paddy fields to direct-seeding rice in dry land. However, the yield and quality of direct-seeded rice in dry land are directly threatened by weeds in paddy fields. Traditional weeding methods mainly rely on manual pulling or weeding, which is time-consuming, laborious, inefficient and high in input costs. The promotion and use of chemical weeding methods not only reduces the The cost of product...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCY02P60/20
Inventor 王旭静王志兴董玉凤
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI