Unlock instant, AI-driven research and patent intelligence for your innovation.

Clone of gene OsRPA2c for controlling rice sterility and application of clone

A gene, rice technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problems of no RPA2 gene fertility control, no function involved, etc., and achieve the effect of rapid isolation and cloning of genes

Inactive Publication Date: 2014-07-02
HUAZHONG AGRI UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although some laboratories have done some research on the RPA gene in rice and Arabidopsis (see below), these sporadic studies are limited to the analysis of its expression. Before the present invention was proposed, there was no RPA2 gene in related plants. reports on the biological functions of
In 1997, van der Knaap et al. (van der Knaap, et al. (1997) Expression of an ortholog of replication protein A1 (RPA1) is induced by gibberellin in deepwater rice. Proc Natl Acad Sci U S A, 94, 9979–9983.) used The method of differential display identified a gene whose expression was increased after flooding stress from the intermediary meristem of rice. Sequence analysis showed that the gene was rice replication protein A (OsRPA1b), but this study did not involve its function
In addition, Japanese laboratories have done some research on RPA genes of rice and Arabidopsis (Marwedel, et al. (2003) Plant-specific regulation of replication protein A2 (OsRPA2) from rice during the cell cycle and in response to ultraviolet light exposure.Planta,217,457–465.Ishibashi,et al.(2001)Two types of replication protein A70kDa subunit in rice,Oryza sativa:molecular cloning,characterization,and cellular& tissue distribution.Gene,272,335–343.Ishibashi,et al.(2006 )A Higher Plant Has Three Different Types of RPA Heterotrimeric Complex.J Biochem(Tokyo),139,99–104.), but these studies did not involve its function, let alone the RPA2 gene involved in fertility in rice and Arabidopsis controlled reporting

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Clone of gene OsRPA2c for controlling rice sterility and application of clone
  • Clone of gene OsRPA2c for controlling rice sterility and application of clone
  • Clone of gene OsRPA2c for controlling rice sterility and application of clone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: Screening and Character Identification of Mutants

[0059] We used the constructed mutant material 03Z11CI73 (ie osrpa2c) to conduct experiments, and transgenic the mutant library to the current generation (ie T 0 Generation) plants were harvested separately, and planted under normal cultivation conditions the next generation, namely T 1 Generation, each material (called a family) planted 2 rows, 10 plants in each row, 20 plants in total, and planted at a planting density of 5×8 inches. The phenotype of each family is recorded in the field throughout the rice growth period, and the phenotype of the mutant plants in the same family is consistent and conforms to the typical 3:1 segregation. The DNA of each individual plant is extracted in time (according to the method of conventional reporting) for PCR Positive detection, for all mutants in a family, the single PCR positive family is used as the material for follow-up research. The primers for positive detect...

Embodiment 2

[0063] Example 2: Observation of the meiosis process of pollen mother cells of wild type and osrpa2c mutants

[0064] In order to reveal the cause of the sterility of the osrpa2c mutant, we used the tablet method (refer to Wang et al., MER3 is required for normal meiotic crossover formation, but not for presynaptic alignment in rice. 2009, J Cell Sci, 122:2055-2063 Methods and Procedures) The meiotic progression of wild-type meiotic 11 and osrpa2c pollen mother cells was observed. In the wild type Zhonghua 11, at the leptotene stage (A in Figure 2), the chromatin condenses into thin and long thin threads; figure 2 Middle B), homologous chromosomes approach and begin synapsis; at pachytene ( figure 2 Middle C), the chromatin is further shortened and thickened, and the homologous chromosomes have completed synapsis, showing a thicker linear structure under the light microscope. Formation of bivalents, each chromosome of which contains two chromatids; then, in the ditene phas...

Embodiment 3

[0066] Example 3: Isolation of mutant T-DNA flanking rice genome sequence

[0067] The present invention utilizes TAIL-PCR technology (with reference to Zhang etc., Non-random distribution of T-DNA insertions at various levels of the genome hierarchy as revealed by analyzing13,804T-DNA flanking sequences from an enhancer-trap mutant library.2007, Plant J , 49:947-959 methods and steps) isolated the flanking sequence of the mutant T-DNA insertion site.

[0068] Through this example, the flanking sequence of the insertion site of the T-DNA of the mutant family was obtained, and the balast search tool (http: / / rice.plantbiology.msu.edu / blast.shtml) provided on the Rice Genome Annotation Project website was used to search The isolated flanking sequence was located in the fourth intron of the LOC_Os06g47830 gene on chromosome 6 of rice. Such as image 3 As shown in A, there is a full-length cDNA J033091F07 corresponding to it in the KOME website (http: / / cdna01.dna.affrc.go.jp / cDNA...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of plant gene engineering, and in particular relates to clone of gene OsRPA2c for controlling rice sterility obtained by separating, cloning and verifying functions, and application of the clone. The nucleotide sequence of the cloned OsRPA 2c is as shown in SEQ ID NO:1, and the protein sequence is as shown in SEQ ID NO:2. Researches indicate that rice male sterility can be caused by inhibiting expression of the gene; The cloned gene and protein coded by the gene can be applied to transgenic crop male sterility line and production of plant hybrid seeds.

Description

technical field [0001] The invention relates to the field of plant genetic engineering. Specifically, it involves the cloning, functional verification and utilization of a gene OsRPA2c involved in the control of homologous chromosome recombination in the process of rice meiosis. The invention utilizes a rice T-DNA insertion mutant library to isolate and clone a gene OsRPA2c that controls rice fertility by participating in the control of homologous chromosome recombination in the process of rice meiosis. The invention also relates to the application of the gene in the process of cultivating male sterile lines of crops and reverse breeding to obtain breeding parents and finally produce hybrid seeds. Background technique [0002] The sexual reproduction of plants is not only an important part of their life history, but also the seeds or fruits produced through the reproductive process of plants are often important food, which is of great significance to human beings. However,...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29A01H5/00
Inventor 吴昌银李兴旺常玉晓
Owner HUAZHONG AGRI UNIV
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com