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Phosphoinositide 3-kinase EST segment in common wild rice expressed by bacterial blight stress

A technology of bacterial blight bacteria and phosphoinositide, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, transferases, etc., can solve the problem of difficult acquisition of bacterial blight resistance genes and loss of bacterial blight resistance genes Resistance, no effective measures, etc.

Inactive Publication Date: 2012-08-15
云南省农业科学院生物技术与种质资源研究所
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0002] At present, the discovery of bacterial blight resistance genes is mainly based on the isolation and cloning of known disease resistance genes, and it is difficult to obtain new bacterial blight resistance genes
Due to the continuous variation of physiological races of the bacterial blight pathogen, the original bacterial blight resistance gene in cultivated rice gradually loses resistance, and currently there are no effective measures for the control of bacterial blight

Method used

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  • Phosphoinositide 3-kinase EST segment in common wild rice expressed by bacterial blight stress

Examples

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Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Obtaining the EST fragment of phosphoinositide 3-kinase expressed in common wild rice under the stress of bacterial blight

[0057] 1. Design primers to amplify the phosphoinositide 3-kinase EST fragment expressed in common wild rice under the stress of bacterial blight

[0058] According to the obtained phosphoinositide 3-kinase EST fragment information SEQ ID NO: 1, use the biological software Primer5.0 to design primers, which are composed of upstream primers and downstream primers. The base sequence of the upstream primers is as in the sequence listing As shown in SEQ ID NO: 2, the base sequence of the downstream primer is shown in SEQ ID NO: 3 in the sequence listing. The primers were synthesized by Huada Gene Company.

[0059] 2. Total RNA was extracted from common wild rice leaves inoculated with bacterial blight

[0060] (1) Processing of samples

[0061] After the common wild rice leaves were inoculated with the bacteria solution of bacterial bl...

Embodiment 2

[0093] Example 2 A method for detecting the differential expression of phosphoinositide 3-kinase EST fragments in common wild rice provided by the present invention comprises the following steps:

[0094] 1. Extraction of total RNA

[0095] 1) Processing of samples

[0096] A. Treatment of the samples to be tested: After inoculating common wild rice leaves with bacterial blight liquid, collect and inoculate the common wild rice leaves at 24 h, 48 h, 72 h, 96 h, and 120 h at intervals of 24 h. The leaves of wild rice were placed in a mortar and quickly ground with liquid nitrogen, and 1 ml of TRIzol extract was added to each 100 mg sample, and vortexed for 15 s. The sampling of each period was a treatment sample, a total of 5 treatment samples;

[0097] B. The treatment of the control group, using sterilized double-distilled water instead of the bacteria liquid of bacterial blight to inoculate the leaves of common wild rice as the control group, every 24 h, at 24 h, 48 h, 72 ...

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Abstract

The invention discloses a phosphoinositide 3-kinase EST segment in common wild rice expressed by bacterial blight stress. The nucleotide sequence of the EST segment is disclosed as SEQ ID NO:1. The invention provides the EST segment for the first time, and the EST directly participates in the anti-disease response process of common wild rice and is a gene directly related to bacterial blight resistance. The EST segment provides partial sequence information for separating and cloning the gene full-length cDNA sequence, and provides theoretical references for researching bacterial blight resistance mechanism of the phosphoinositide 3-kinase in common wild rice. The primer designed by using the EST segment can detection the expression condition of the EST segment by bacterial blight stress in common wild rice leaves, thereby obtaining the expression spectrum of the gene after expression by bacterial blight stress in common wild rice, and laying the foundation for recognizing the functions of the gene and deeply researching the bacterial blight resistance mechanism of the gene in common wild rice.

Description

technical field [0001] The invention belongs to the technical field of biomolecular cloning, and in particular relates to the common wild rice phosphoinositide 3-kinase expression sequence tag (Expressend Sequence Tag, EST) under the stress of bacterial blight, and primers are designed based on the EST fragment, and semi-quantitative RT -PCR detection method for differential expression of the phosphoinositide 3-kinase EST fragment in common wild rice. Background technique [0002] At present, the discovery of bacterial blight resistance genes is mainly based on the information provided by known disease resistance genes for isolation and cloning, and it is difficult to obtain new bacterial blight resistance genes. Due to the continuous variation of physiological races of bacterial blight pathogens, the original bacterial blight resistance genes in cultivated rice have gradually lost their resistance. However, there are no effective measures for the control of bacterial blight...

Claims

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Application Information

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IPC IPC(8): C12N9/12C12N15/54C12N15/11C12Q1/68
Inventor 程在全黄兴奇蒋春苗余腾琼殷富有张敦宇李定琴钟巧芳李维蛟王玲仙付坚蒋聪王波陈玲李娥贤罗红梅
Owner 云南省农业科学院生物技术与种质资源研究所
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