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Application of OsAGO18 protein or encoding gene of OsAGO18 protein to regulation and control on resistance of plants on RDV (Rice Dwarf Virus) or virus in same family as RDV

A dwarf virus, a technology encoding a gene, applied in the application field of resistance

Active Publication Date: 2016-06-01
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this may not be a general phenomenon and the results may depend on the specific virus

Method used

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  • Application of OsAGO18 protein or encoding gene of OsAGO18 protein to regulation and control on resistance of plants on RDV (Rice Dwarf Virus) or virus in same family as RDV
  • Application of OsAGO18 protein or encoding gene of OsAGO18 protein to regulation and control on resistance of plants on RDV (Rice Dwarf Virus) or virus in same family as RDV
  • Application of OsAGO18 protein or encoding gene of OsAGO18 protein to regulation and control on resistance of plants on RDV (Rice Dwarf Virus) or virus in same family as RDV

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1. It was found that the OsAGO18 gene can be induced and expressed by various rice viruses

[0064] The OsAGO18 gene involved in this example is derived from rice (Oryza.sativaL.), the cDNA sequence of the OsAGO18 gene is shown in sequence 1 in the sequence table, and sequence 1 consists of 3267 nucleotides, wherein the 1-3267th is Coding sequence (ORF): Sequence 1 encodes the protein shown in Sequence 2 in the Sequence Listing, and Sequence 2 consists of 1088 amino acid residues.

[0065] 1. OsAGO18 protein accumulates after rice dwarf virus, rice black-streaked dwarf virus and rice stripe virus infection

[0066] The wild-type rice (Oryza.sativa L.) variety Nipponbare and the control rice (not infected by rice dwarf The leaves of the wild-type rice variety Nipponbare) infected by the virus were used as materials, the total protein was extracted with 2×SDS loading buffer, and the accumulation of OsAGO18 protein in the two rice plants was detected by Western bl...

Embodiment 2

[0102] Example 2, OsAGO18 gene participates in the defense response of rice against RDV

[0103] 1. Obtaining homozygous mutants of ago18 rice

[0104] ago18 mutant seeds (NF6013) purchased in Tos17 database

[0105] (https: / / tos.nias.affrc.go.jp / ~miyao / pub / tos17 / ).

[0106] The T-DNA insert sequence is:

[0107] tos17_tail6 (LB): 5'-AGGTTGCAAGTTAGTTAAGA-3'.

[0108] Insertion site such as image 3 Shown in B.

[0109] Identification of homozygotes for ago18 mutants using methods provided in the database

[0110] (https: / / pc7080.abr.affrc.go.jp / cgibin / tos17 / ricegenome.cgi?action=getTarget&chr=7&pos=16895280&primer=y&version=7).

[0111] Using the homozygous cDNA of the rice ago18 mutant (NF6013) identified above as a template, semi-quantitative RT was performed using the 6013_LP / 6013_RP primer pair, tos17_tail6(LB) / 6013_RP primer pair, and OsAGO18-F / OsAGO18-R primer pair. -PCR reaction. At the same time, the wild-type rice (Oryza. sativa L.) variety Nipponbare was set ...

Embodiment 3

[0150] Example 3, Enhanced disease resistance of transgenic rice overexpressing OsAGO18

[0151] 1. Construction of OsAGO18 gene plant expression vector

[0152] 1. Acquisition of rice cDNA template

[0153] According to the instructions, the total RNA of Nipponbare rice (OryzasativaL.japonica.cv.Nipponbare) was extracted with TRIzol Reagent of Invitrogen Company, and reverse-transcribed with the company's SuperScriptII reverse transcriptase. The primer used for the reverse transcription is an Oligod (T) primer with 16 nucleotides, and the rice cDNA template obtained by the reverse transcription is finally obtained.

[0154] 2. Acquisition of rice OsAGO18 gene

[0155] Using the rice cDNA obtained in step 1 as a template, PCR amplification was performed with the following primers.

[0156] AGO18cds-F: 5'-ATAATGGCGAGCCGAGGAGGAGGC-3' (positions 4-24 of this sequence are positions 1-21 of sequence 1, and ATA in the first three positions is a protective base);

[0157] AGO18cd...

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Abstract

The invention discloses application of OsAGO18 protein or an encoding gene of the OsAGO18 protein or a recombinant vector containing the coding gene to the improvement of the resistance of rice on virus diseases caused by an RDV (Rice Dwarf Virus) or a virus in the same family as the RDV. The amino acid sequence of the OsAGO18 protein is concretely a sequence 2 or a sequence 4 in a sequence table. Experiments prove that the plant disease resistance in the rice with overexpression of the OsAGO18 gene is enhanced; otherwise, the disease resistance of an ago18 mutant is reduced; and the result shows that the OsAGO18 protein coded by the gene achieves an important effect in the anti-RDV reaction of the rice.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of OsAGO18 protein or its encoding gene or a recombinant vector containing the encoding gene in regulating the resistance of plants to rice dwarf virus or similar viruses. Background technique [0002] The severe and intermittent outbreaks of these viral diseases in my country's rice and wheat production are mainly caused by rice stripe disease virus (RSV), rice dwarf virus (RDV), rice black-streaked dwarf virus (RBSDV), and southern rice black-streaked virus. Caused by dwarf dwarf virus (SRBSDV), rice sawtooth dwarf virus (RRSV), barley yellow dwarf virus (BYDV) and wheat yellow mosaic virus (WYMV), most viruses can infect corn, etc. in addition to rice and wheat Grass crops, maize is particularly sensitive to the virus, for example, maize rough dwarf disease caused by RBSDV infection has become the main virus disease of maize, resulting in large-scale reduc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00
CPCC12N15/82
Inventor 李毅戚益军吴建国杨志蕊魏春红
Owner PEKING UNIV
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