Application of rice OsRPA1a gene in fertility control

A gene and rice technology, applied in the field of rice hybrid seed production and fertility control, can solve problems such as non-detection, and achieve the effect of improving product quality

Inactive Publication Date: 2009-10-28
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

OsRPA2-1 also had the same expression trend in the elongation zone of stem nodes under the same treatment, but OsRPAla was not detected in this region

Method used

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  • Application of rice OsRPA1a gene in fertility control
  • Application of rice OsRPA1a gene in fertility control
  • Application of rice OsRPA1a gene in fertility control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1: Screening and Character Identification of Mutants

[0062] mutant library T 0 After the generation plants were harvested separately, the next generation, namely T, was planted under normal cultivation conditions. 1 In the second generation, each material was planted in 2 rows, with 10 plants in each row, and a total of 20 plants were planted at a planting density of 15×24 cm. The phenotypes of the mutants were recorded in the field during the entire growth period of rice, and the phenotypes of the mutant plants in the same family were consistent and conformed to the typical 3:1 segregation. The DNA of each individual plant was extracted in time for PCR positive detection. Families with positive PCR results for all mutants were used as materials for follow-up studies. The primers for positive detection by PCR are located on the T-DNA segment. A pair of primers are GV-F: 5-ggc atc ggt aaa cat ctg ct-3, GV-R: 5-gcc tca aga agc tca agt gc-3, The size of the PC...

Embodiment 2

[0067] Example 2: Isolation of mutant T-DNA flanking rice genome sequence

[0068] For the mutation family obtained in Example 1, this example utilizes the TAIL-PCR method (refer to 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 method and steps) isolated the flanking sequence of the mutant T-DNA insertion site.

[0069] Through this example, the flanking sequence of the insertion site of the family's T-DNA was obtained, and the balast search tool provided on the Rice Genome Annotation Project website ( http: / / rice.plantbiology.msu.edu / blast.shtml ), the isolated flanking sequence was located in the intron of the Os02g53680 gene of rice chromosome 2. Such as figure 2 As shown in a, on the KOME website ( http: / / cdna01.dna.affrc.go.jp / cDNA / ) has a full-length cDNA J033031B02 corresponding to it. The...

Embodiment 3

[0072] Example 3: Co-segregation verification of T-DNA and mutant traits

[0073] According to the mutant Osrpala family T-DNA localization result that embodiment 2 obtains, design genome primer O 1 (5'-agc gag tac gtc atcaac ga-3') and O 2 (5'-ccg aaa caa gaa ctt cca gg-3') with primer LBT on the left border of T-DNA 3 (5'-cca gta cta aaa tcc aga tccccc gaa t-3,) paired amplification to verify the genotypes of each individual plant in the T1 and T2 generations, such as figure 2 as shown in c. Corresponding genotype and phenotype to see if co-segregation is consistent. Primer O 1 +O 2 The product size is about 1.1kb, primer O 2 +LBT 3 The size of the product is about 0.9kb. Since the fragment of T-DNA transferred into the genome is about 10kb, when the T-DNA is inserted into the O 1 with O 2 Between, it cannot be amplified because the fragment is too large. Therefore, when a single plant is respectively used with primer O 1 +O 2 and primer O 2 +LBT 3 There are t...

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Abstract

The invention pertains to the technical field of plant genetic engineering, in particular relates to an application of an OsRPA1a gene, a rice fertility control gene, in rice fertility control. The invention is characterized in that: the OsRPA1a gene is one of the following nucleotide sequences: (1) the DNA sequence showed in sequence table SEQ NO:1; or (2) a DNA sequence of a protein with the coding the same as that of the protein in (1). The invention discloses the isolation cloning, biological function verification and genetic transformation steps of the gene and the cloned gene is important for the genetic improvement of rice.

Description

technical field [0001] The invention relates to the field of plant genetic engineering. It specifically involves the use of rice T-DNA random insertion mutant library to isolate and clone a gene OsRPAla that controls rice pollen fertility and embryo sac fertility by participating in rice meiosis, the biological function verification of the gene, genetic transformation and Application as a sterility gene in rice hybrid seed production and fertility control. Background technique [0002] In the life history of eukaryotes, there are diploid sporophyte era and haploid gametophyte era. During the sporophyte era, eukaryotes undergo meiosis to form haploid spores; next, fusion of gametes from the parents reverts the offspring to the diploid level of their parents. Therefore, meiosis plays an important role in the alternation of eukaryotic generations. [0003] In the past ten years, yeast has been used as a model organism to study meiosis, and many genes controlling meiosis have...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/29
Inventor 吴昌银常玉晓张启发
Owner HUAZHONG AGRI UNIV
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