Application of OsAGO18 gene in improving rice stripe disease resistance of rice

A technology for leaf blight resistance, rice stripe, applied in the biological field

Active Publication Date: 2015-04-29
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 a

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  • Application of OsAGO18 gene in improving rice stripe disease resistance of rice
  • Application of OsAGO18 gene in improving rice stripe disease resistance of rice
  • Application of OsAGO18 gene in improving rice stripe disease resistance of rice

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0061] Example 1. It was found that OsAGO18 gene can be induced and expressed by rice stripe virus

[0062] The OsAGO18 gene involved in this example is derived from rice (Oryza.sativa L.). The cDNA sequence of the OsAGO18 gene is shown in sequence 1 in the sequence table. Sequence 1 consists of 3267 nucleotides, of which positions 1-3267 It is the coding sequence (ORF); sequence 1 encodes the protein shown in sequence 2 in the sequence table, and sequence 2 consists of 1088 amino acid residues.

[0063] 1. OsAGO18 gene mRNA accumulates after rice stripe virus infection

[0064] Take 0.5g each of wild-type rice (Oryza.sativa L.) Nipponbare and control rice (wild-type rice variety Nipponbare that is not infected by rice streak virus) infected by rice stripe virus (RSV). After medium grinding, the total RNA was extracted according to Invitrogen's TRIzol Reagent instructions (Invitrogen Trizol Reagent, cat No. 15596-018). After determining the total RNA concentration, take 10 μg of to...

Example Embodiment

[0100] Example 2. OsAGO18 gene participates in rice defense response against RSV

[0101] 1. Obtaining homozygote of ago18 rice mutant

[0102] ago18 mutant seeds (NF6013) are purchased in the Tos17 database

[0103] (Https: / / tos.nias.affrc.go.jp / ~miyao / pub / tos17 / ).

[0104] The T-DNA insert sequence is:

[0105] tos17_tail6(LB): 5'-AGGTTGCAAGTTAGTTAAGA-3'.

[0106] Insertion sites such as image 3 Shown in B.

[0107] Identify the homozygote of the ago18 mutant using the methods provided in the database

[0108] (Https: / / pc7080.abr.affrc.go.jp / cgibin / tos17 / ricegenome.cgi?action=getTarget&chr=7&pos=16895280&primer=y&version=7).

[0109] Using the homozygous cDNA of the rice ago18 mutant (NF6013) identified above as a template, semi-quantitative RT was performed with 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 as a control.

[0110] The ...

Example Embodiment

[0160] Example 3. Transgenic rice overexpressing OsAGO18 has enhanced disease resistance

[0161] 1. Construction of OsAGO18 Gene Plant Expression Vector

[0162] 1. Obtaining rice cDNA template

[0163] According to the instructions, the total RNA of Nipponbare rice (Oryza sativa L.japonica.cv.Nipponbare) was extracted with Invitrogen's TRIzol Reagent, and reverse transcription was performed with the company's SuperScript II reverse transcriptase. The primer used for reverse transcription is the 16-nucleotide Oligod (T) primer, and finally the rice cDNA template obtained by reverse transcription is obtained.

[0164] 2. Obtaining rice OsAGO18 gene

[0165] Using the rice cDNA obtained in step 1 as a template, PCR amplification was performed with the following primer pairs.

[0166] AGO18cds-F: 5’-ATAATGGCGAGCCGAGGAGGAGGC-3’ (positions 4-24 of the sequence are positions 1-21 of sequence 1, and the first three ATAs are protected bases);

[0167] AGO18cds-R: 5'-GACCTAGCAAAAGAACATGGACTTTTTC...

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Abstract

The invention discloses application of an OsAGO18 gene in improving the rice stripe disease resistance of rice, and provides application of an OsAGO18 gene, encoding protein of the OsAGO18 gene, or a recombinant carrier containing the encoding gene, in regulating and controlling rice stripe disease resistance of plants. The amino acid sequence of the gene OsAGO18 protein is specifically shown in the sequence 2 in a sequence table. The experiment shows that the plant disease resistance in rice with over-expression of the OsAGO18 gene is improved; on the contrary, the disease resistance of an ago18 mutant is degraded, which means that the OsAGO18 protein encoded by the gene plays an important role in rice stripe disease resistance of rice.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of OsAGO18 gene in improving the resistance of rice to rice stripe blight. Background technique [0002] Rice stripe leaf blight is a viral disease caused by rice stripe virus (Rice Stripe Virus, RSV), which is transmitted by SBPH as a vector, commonly known as cancer on rice. Diseased plants often withered and booted or the ears were small and deformed. At the beginning of the rice seedling disease, intermittent yellow-green or yellow-white short stripes appear at the base of the heart leaf, and later the lesion enlarges and merges, expanding into yellow-green stripes parallel to the veins, and the stripes remain green; after jointing, the disease occurs on the sword. Yellow-green stripes appear on the lower part of the leaves, and all types of rice are not dead, but the heading is deformed and the fruit is few. [0003] Rice Stripe Virus (RSV) is a representative sp...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00
Inventor 李毅戚益军吴建国杨志蕊郑立佳魏春红
Owner PEKING UNIV
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