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Rice OsGRDP1 protein related to disease resistance of plants and its encoding gene and application

A disease resistance, plant technology, applied in the field of plant genetic engineering, can solve problems such as not yet established

Active Publication Date: 2018-05-11
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it is known that the functions of the proteins encoded by the LMM gene are diverse, which to a certain extent reflects the complex mechanism of the formation of lesion-like plaques, and therefore a complete mechanism for the formation of allergic reactions has not yet been established.
However, most LMM mutants exhibit broad-spectrum disease resistance

Method used

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  • Rice OsGRDP1 protein related to disease resistance of plants and its encoding gene and application
  • Rice OsGRDP1 protein related to disease resistance of plants and its encoding gene and application
  • Rice OsGRDP1 protein related to disease resistance of plants and its encoding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Embodiment 1, the acquisition of rice OsGRDP1 protein and its coding gene

[0077] 1. Acquisition of mutant spl-D and detection of disease resistance traits

[0078] 1. The acquisition of mutant spl-D and its phenotype

[0079] Mutant spl-D was screened from the offspring of T-DNA transformed rice variety Aichi Asahi. Compared with the wild-type rice variety Aichi Asahi, the mutant spl-D spontaneously produces lesion-like spots, and the phenotype of the mutant is as follows: figure 1 Shown in A. Among them, WT is the wild-type rice variety Aichi Asahi, and spl-D is the mutant rice. Under long-day conditions (Beijing, May-October, about 12-14 hours of sunshine), there is almost no difference between the mutant and the wild type at the rice seedling stage. As the growth proceeds, after entering the tillering stage, lesion-like spots appear successively from the lower leaves to the upper leaves, and the lesion-like spots can extend to the leaf sheaths; under short-day...

Embodiment 2

[0102] Example 2, the subcellular localization of OsGRDP1 protein

[0103] 1. Using the PCR method, GFP-GRDP1-F (5'-ATA ggtacc ATGGACGGGGAGCAAGAG-3', the underlined part is the Kpn I site) as the forward primer, GFP-GRDP1-R (5'-TAT ggatcc GGTGCTCGCGTTG-3', the underlined part is the Bam HI site) was used as a reverse primer to amplify the open reading frame sequence of the OsGRDP1 gene from the cDNA of the rice variety Aichi Asahi. After the PCR product was recovered, it was connected into the pMD-18T vector (TaKaRa). After the sequencing was correct, it was digested with Kpn I and BamHI, and the digested product was connected into the plant subcellular localization expression vector pCG1301 (purchased from Protin (Beijing) Co., Ltd.) to obtain the recombinant expression vector pCG1301-OsGRDP1-GFP containing the OsGRDP1 gene. The structure of the recombinant expression vector pCG1301-OsGRDP1-GFP is described as: a recombinant plasmid obtained by inserting positions 1-2970 ...

Embodiment 3

[0105] Example 3, Obtaining and phenotypic identification of OsGRDP1 gene overexpression plants

[0106] The gene involved in this example is the OsGRDP1 gene of the rice variety Aichi Asahi obtained in Example 1, its nucleotide sequence is sequence 1 in the sequence listing, and the protein (OsGRDP1) shown in sequence 2 in the coding sequence listing . Sequence 1 consists of 2973 nucleotides, and Sequence 2 consists of 930 amino acids.

[0107] 1. Obtaining and identification of OsGRDP1 gene overexpression plants

[0108] 1. Construction of recombinant expression vector pCAMBIA1301-Ubi-OsGRDP1

[0109] (1) Utilize PCR method, adopt primer 5'-ata gagctc GTGCAGCGTGACCCGGT-3' (the underlined part is the Sac I site) and 5'-ata ggatcc AAGTAACACCAAACAACAGGGT-3' (the underlined part is the BamHI site), using the binary vector pUbiGUSPlus (Prutin Biotechnology (Beijing) Co., Ltd.) as a template to amplify the ubiquitin promoter region. After the amplified product was recovered...

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Abstract

The invention discloses a rice OsGRDP1 protein related to disease resistance of plants and its encoding gene and application. The OsGRDP1 protein is a protein with a sequence 2. The encoding gene of the OsGRDP1 protein is a DNA molecule with a sequence 1. Through TAIL-PCR, a T-DNA insertion site is cloned. Through insertion of T-DNA, the up-regulated expression of the OsGRDP1 gene is caused. Through overexpression of the OsGRDP1 gene in the wild-type rice, it is proved that the overexpression of the OsGRDP1 gene can cause spontaneous rice lesion mimic and can improve resistance to rice blast and bacterial blight. The OsGRDP1 gene has a good application prospect of improving crop immunity. Through the gene engineering, the expression level of the OsGRDP1 gene is adjusted and controlled to be used for molecular breeding and has a potential application value in improving crop disease resistance.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and relates to rice OsGRDP1 protein related to plant disease resistance, its coding gene and application. Background technique [0002] Hypersensitive response (Hypersensitive Response, HR) is one of the most common and effective defense responses in plants, which is characterized in that when plants are infected by pathogens, local cell necrosis occurs rapidly around the infection site, thereby limiting the pathogen to the outside world. Expansion of adjacent cells (Morel and Dangl, 1997). Although the role of anaphylaxis in plant disease resistance is widely recognized, its mechanisms are incompletely studied. In order to study the mechanism of allergic reaction, many researchers are isolating Lession Mimic Mutants (LMMs) of plants, hoping to first determine the gene controlling allergic reaction through genetic pathways, and then study the molecular function of the gene. Lesion-like ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8281C12N15/8282
Inventor 赵文生赵晓胜彭友良
Owner CHINA AGRI UNIV
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