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
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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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