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Application of OsRBR2 gene to development of rice male sterile line

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: 2019-02-22
FUJIAN AGRI & FORESTRY UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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 to development of rice male sterile line
  • Application of OsRBR2 gene to development of rice male sterile line
  • Application of OsRBR2 gene to development of rice male sterile line

Examples

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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 Minghui 77, Minghui 63, 9311 and other indica rice varieties, and the F1 generation plants were normally fertile, but male sterile plants appeared in the F2 generation and 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 the 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, the plants with both male sterility and sterility characteristics are screened out, and the The maintainer line of the original sterile parent is crossed and selfed for one generation to ...

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Abstract

The invention provides an application of an OsRBR2 gene to development of a rice male sterile line. The OsRBR2 gene is knocked out at a fixed point by a CRISPR / Cas9 editing system, the gene loses functions completely, and the rice male sterile line is obtained. The nucleotide sequence of the OsRBR2 gene is shown as SEQ ID NO.1 in a sequence table, and a gene coding amino acid sequence is shown asSEQ ID NO.2 in the sequence table. The prepared rice male sterile line is completely normal in nutrition and growth, has no obvious panicle enclosure in heading stage and is high in outcrossing rate,and anther is pollen-free. The rice male sterile line has the advantages of avoiding artificial castration, avoiding spraying of plant exogenous hormones, being high in seed purity and the like when applied to hybrid rice breeding, can increase production efficiency greatly and reduce production cost and environmental pollution, thereby having important 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 Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/46
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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