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Application of rice OsAPI5 gene in fertility control

A gene and rice technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problem of no function, etc., and achieve the effect of rapid and far-reaching practical application significance of cloning genes

Inactive Publication Date: 2012-05-30
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Compared with the study of the API5 gene in animals, after completing the whole genome sequencing of rice and Arabidopsis, it was shown that there are one and two homologues of the API5 gene in rice and Arabidopsis, respectively, but without any correlation of their functions to report

Method used

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  • Application of rice OsAPI5 gene in fertility control
  • Application of rice OsAPI5 gene in fertility control
  • Application of rice OsAPI5 gene in fertility control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Embodiment 1: Isolation and cloning of OsAPI5 gene

[0070] 1. Obtaining of osapi5-1 mutant

[0071] The rice T-DNA insertion mutant library used was provided by the State Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University using the vector pFX-E24.2-15R (see Figure 7 ) (the creation method of the mutant library has been published, see Wu et al., Development of enhancer trap lines for functional analysis of the rice genome. 2003, Plant J, 35: 418-427; Zhang et al., Non-random distribution of T-DNA insertions at various levels of the genome hierarchy as revealed by analyzing 13,804 T-DNA flanking sequences from an enhancer-trap mutant library. 2007, Plant J, 49: 947-959). In 2005, from the T0 generation mutant library (see http: / / rmd.ncpgr.cn / , Zhang et al., RMD: a rice mutant database for functional analysis of the rice genome. 2006. Nucl Acids Res, 34: D745-D748) selected 2,000 rice transgenic varieties "Zhonghua 11" (a common rice variety...

Embodiment 2

[0078] Example 2: Observation of semi-thin sections of anthers of wild type and mutants at different stages

[0079] Anthers derived from wild type and mutants at different developmental stages were taken in 50% FAA fixative solution (50% dH 2 O, 40% ethanol, 10% acetic acid) fixed for 24 hours (anther development is divided into 8 stages, for details, refer to Feng, et al. 2001. Pollen development and its stages in rice (Oryzasativa L.). Chinese J. Rice Sci 15 ( 1): 21-28.). The fixed material was dehydrated in a series of alcohols (70% alcohol, 90% alcohol, 95% alcohol, and absolute alcohol were placed in sequence for 30 minutes). The dehydrated material was embedded with the embedding agent Technovit 7100 resin (purchased from Heraeus Kulzer Company, Germany) (refer to the reagent manual for detailed methods), and after being placed at 37°C for 3-5 days, the embedding block was made into semi-thin sections (1 micron, slicer model The number is LEICA EM UC6, and the method...

Embodiment 3

[0081] Example 3: Identification of an allelic mutant osapi5-2 of osapi5-1

[0082] The T-DNA insertion site of an allelic mutant osapi5-2 of Osapi5-1 is located in the 13th exon of the gene ( figure 2 A). In 2008, 21 osapi5-2 mutant T1 plants were planted in the experimental field of Huazhong Agricultural University in Shizishan District, Hongshan District, Hubei Province, China. Among them, the 17th and 18th single plants had a mutant phenotype, that is, extremely low fertility. The principle of co-segregation detection is as follows: figure 2 A, The primers used are P4 (5'-GCCCATAAGGTTTGATGACAGA-3'), P5 (5'-CATGAGAAGCCCAAGCAGAG-3') and vector primer P6 (5'-GTTACGTCCTGTAGAAACCCCAA-3'). Depend on Figure 4 B shows that the mutant phenotype of the osapi5-2 family is co-segregated with the T-DNA insertion in the OsAPI5 gene. This further indicated that the mutant phenotype of osapi5-2 was also caused by the insertion mutation of OsAPI5 gene.

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Abstract

The invention belongs to the technical field of plant gene engineering and discloses a rice fertility control gene OsAPI5 and an application thereof, in particular relates to isolation and cloning, functional verification and an application of a gene OsAPI5 for regulating and controlling rice fertility. The nucleotide sequence of the OsAPI5 gene is shown in the sequence table SEQ ID NO:1, and the amino acid sequence coded by the OsAPI5 gene is shown in the sequence table SEQ ID NO:2. In the invention, the cloned gene and the coding protein thereof can be used for regulating and controlling rice fertility and breeding hybrid rice sterile lines and have important application value.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. It specifically relates to the cloning, functional verification and application of a gene OsAPI5 controlling rice fertility. The present invention utilizes T-DNA labeling technology to obtain a gene OsAPI5 that controls rice fertility from a rice T-DNA insertion mutant library through forward genetics methods, and proves it through transgene complementation experiments and allelic mutant analysis Function of the OsAPI5 gene. The invention also relates to genetically improving the fertility control of rice by using the gene through genetic engineering technology. Background technique [0002] Rice is one of the most important food crops in the world. It feeds nearly half of the world's population. How to increase rice production is an important issue related to the national economy and people's livelihood. At present, the vast majority of rice in my country is produced by hybrid rice, a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415A01H5/00
Inventor 吴昌银李兴旺张启发
Owner HUAZHONG AGRI UNIV
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