Application of osrbr2 Gene in Creating Male Sterile Lines of Rice

A male-sterile line, male-sterile technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve problems such as threats to food security, and achieve the elimination of artificial emasculation, high seed purity, and high outcross seed setting rate. Effect

Active Publication Date: 2021-07-02
FUJIAN AGRI & FORESTRY UNIV
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

At present, the genetic background of the male sterile lines facing the production of hybrid rice in my country is becoming more and more obvious. Most of the male sterile lines of three-line hybrid indica rice have the same or similar genetic relationship with the wild-type cytoplasm. Disease outbreak will seriously threaten my country's food security

Method used

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  • Application of osrbr2 Gene in Creating Male Sterile Lines of Rice
  • Application of osrbr2 Gene in Creating Male Sterile Lines of Rice
  • Application of osrbr2 Gene in Creating Male Sterile Lines of Rice

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1: creation of rice male sterile line

[0026] According to the principle of site-directed knockout of CRISPR / Cas9 editing system, the target site of gene site-directed editing is determined, and the primers shown in SEQ ID NO: 3 and SEQ ID NO: 4 are designed. The synthesized single nucleotide sequence forms a dimer structure by annealing reaction and ligates into such as figure 1 The indicated vector VK005-01 (purchased from Beijing Weishang Lide Biotechnology Co., Ltd., article number: 20160601S), was constructed to contain OsRBR2 Plasmids for gene target sequences OsRBR2 -VK005-01. After the plasmid was transformed into Escherichia coli DH5α competent cells, positive single clones were picked, and the plasmid was extracted after shaking the bacteria, and confirmed by sequencing. will build the OsRBR2 -VK005-01 plasmid was transferred into Agrobacterium rhizogenes EHA105 for genetic transformation of rice.

[0027] SEQ ID NO: 3: 5'-CAGGTTCCTCCGGCGTTAG...

Embodiment 2

[0035] Embodiment 2: Utilization of rice male sterile lines

[0036] Will Osrbr2-1 or Osrbr2-2 The mutants were crossed with indica rice varieties such as Minghui 77, Minghui 63, and 9311, and the F1 plants were normally fertile, but male sterile plants appeared in the F2 generation, and they were fertile: the sterility conformed to the segregation rule of 3:1, proving that OsRBR2 The male sterility characteristic caused by nucleotide sequence changes in other rice varieties can be stably inherited without being affected by genetic background.

[0037] Will Osrbr2-1 or Osrbr2-2 The mutant is used as a male parent to cross with the sterile parent in the three-line or two-line hybrid combination, and self-cross for one generation to obtain the F2 generation; from the F2 generation, select plants with both male sterility and sterility characteristics, and The maintainer line of the original sterile parent is crossed and selfed for one generation to obtain the hybrid F2 ge...

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Abstract

The invention provides OsRBR2 Application of genes in creating male sterile lines of rice. Targeted knockout using CRISPR / Cas9 editing system OsRBR2 Gene, so that the function of the gene is completely lost, and a male sterile line of rice is obtained. said OsRBR2 The nucleotide sequence of the gene is shown in the sequence listing SEQ ID NO.1, and the encoded amino acid sequence is shown in the sequence listing SEQ ID NO.2. The vegetative growth of the rice male sterile line prepared by the invention is completely normal, there is no obvious ear wrapping at the heading stage, the anthers have no pollen, and the outcrossing seed setting rate is high. The male sterile line of rice is used in hybrid rice breeding, and has the advantages of eliminating artificial emasculation, no need to spray plant exogenous hormones, and high seed purity. It can greatly improve production efficiency, reduce production costs, and reduce environmental pollution. Therefore, it is of great importance. Application prospect.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, it involves OsRBR2 The application of genes in the creation of rice male sterile lines, targeted knockout by using CRISPR / Cas9 editing method OsRBR2 Gene, create a new male sterile line of rice, and apply hybrid rice production. Background technique [0002] Rice is an important food crop in the world, and about half of the world's population depends on rice as the staple food. Therefore, rice plays an important role in ensuring global food security. Hybrid rice production technology is an important achievement of modern agriculture, which has a significant effect of increasing yield. my country is the first country in the world to promote hybrid rice, which has made important contributions to ensuring my country's food security and also played a key role in maintaining world food security. my country's hybrid rice production technology has been at the international lea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H5/02A01H6/46
CPCC07K14/415C12N15/8213C12N15/8289
Inventor 段远霖吴为人潘美珍蔡正正
Owner FUJIAN AGRI & FORESTRY UNIV
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