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Pathogen from rice, chemically inducible promoter and their use

A rice, nucleotide sequence technology, applied in the fields of organic chemistry, biochemical equipment and methods, applications, etc., can solve the problems of slow speed, signal transmission defense system, plant susceptibility, etc.

Inactive Publication Date: 2004-11-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In some cases, the plant's defense against the pathogen is not successful, causing the plant to be susceptible to the disease, but in this case, the plant also responds to the pathogen invasion, but the response is too slow or too mild, so that the plant It is too late to pass the signal to the defense system to effectively resist the pathogen

Method used

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  • Pathogen from rice, chemically inducible promoter and their use
  • Pathogen from rice, chemically inducible promoter and their use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Cloning of the POG promoter of the blast fungus infection-induced gene

[0031] 1. Purification of rice peroxyhydroxy fatty acid hydroxyl cyclooxygenase (POG) Prepare the rice variety Aichiasahi (Oryza sativa cv Aichiasahi), and when the 4-5 leaves are pulled out, the rice inoculated with non-compatible blast fungus Leaves were used as experimental materials, through selective extraction of membrane proteins, 20% saturation ammonium sulfate precipitation to remove impurities and 45% saturation ammonium sulfate salting out, DEAE-Toyopearl anion exchange, DEAE-Sepharose anion exchange, CM-Toyopearl (pH5 .1) Through the steps of cation exchange, CM-Toyopearl (pH4.5) cation exchange and ultrafiltration concentration, the infection-induced POG of Magnaporthe oryzae was purified, and the electrophoretic pure enzyme protein was obtained. The N-terminal amino acid sequence of the POG is determined by using a lysine-specific Lysyl endopeptidase enzymatic hydrolysis met...

Embodiment 2

[0037] Example 2 Inducible expression characteristics of POG gene

[0038]Aichiasahi (Oryza sativa cv Aichiasahi) was used as the tested rice variety. Use Magnaporthe grisea race 131# which is not compatible with this variety and race 007# which is compatible with the variety as inoculum. The preparation method of Magnaporthe oryzae spores is the same as that of Peng et al. (Can. J. Biol., 66: 730-735, 1988). When the 5th leaf of the rice seedlings is nearly fully unfolded, rice blast is used to spray and inoculate rice with spore suspension (106 / ml), and then cultivated in the dark at 100% at 28° C. for 36 hours and then cultured in the light, with 0.02% Tween Mock inoculation of -20 spray was used as control. Samples were taken at 0h, 8h, 16h, 24h, 36h, 48h, 60h, and 72h after inoculation, and the middle part of the fifth leaf was cut when sampling. Other physical and chemical treatments are as follows: JA treatment: Spray 100 μM JA on the rice leaf surface, and then seal...

Embodiment 3

[0040] Example 3 Obtaining of POG Promoter 5' End Gradual Deletion Derivative Variants

[0041] The upstream regulatory fragment of the 5-terminal of the POG gene was searched in the database and analyzed by software to infer its possible cis-element positions. Then, based on these positions, six POG promoter 5′-terminal gradual deletions were obtained by enzyme digestion and PCR. The fragments are respectively named as GPF (corresponding to nucleotides 163 to 2789 of SEQ ID.1), G200 (corresponding to nucleotides 807 to 2789 of SEQ ID.1), and G150 (corresponding to nucleotides 807 to 2789 of SEQ ID.1). 1286 to 2789 nucleotides), G100 (corresponding to SEQ ID.1 1635 to 2789 nucleotides), G75 (corresponding to SEQ ID.1 2038 to 2789 nucleotides), G42 (corresponding to SEQ ID.1 2038 to 2789 nucleotides), G42 (corresponding to SEQ ID.1 nucleotides ID.1 nucleotides 2363 to 2789). Among them, GPF is obtained by PCR amplification of plasmid POGH with artificially synthesized oligonuc...

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Abstract

The present invention provides one kind of DNA sequence segment, which is obtained from rice, can be infected by rice blast and induced by salicylic acid and jasmonic acid, and may be used as pathological and chemical inducing promoter. The DNA sequence segment is the transcriptional control area is rice hydroperoxl fatty acid hydroxylated epoxide hydrolase gene and features that it has the nucleotide sequence of the promoter from site-1 to site-2789 shown in SEQ ID No. 1. The functional analog obtained via inserting, replacing and / or deleting one or several nucleotide sequences into the nucleotide sequence may be used to reach the aim of the present invention. The present invention may be used as pathological and chemical inducing promoter, molecular mark probe, rice re-separating control sequence or relevant control sequence for other transgenic plant and in breeding disease resisting plant.

Description

field of invention [0001] The invention belongs to the field of plant genetic engineering, and the invention provides an inducible gene promoter that can be induced and activated by pathogens, salicylic acid (SA) and jasmonic acid (JA) to start its associated DNA transcription. Relating to vectors, hosts and cells comprising the inducible gene promoter or derivative variants thereof, the promoter provided by the present invention can be used as a pathogen-inducible promoter to drive the expression of a gene that confers pathogen resistance to a plant host, or It is used to induce the expression of the target gene by treatment with chemical substances such as salicylic acid and jasmonic acid. technical background [0002] Plants will be affected by hundreds of microorganisms during their growth. During the long-term interaction with these microorganisms, plants gradually evolve a variety of defense mechanisms against the attack of potential pathogens around them, so that they...

Claims

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Application Information

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IPC IPC(8): A01H1/00C07H21/04C12N15/11C12N15/29C12N15/63
Inventor 彭友良王扬秦庆明孙宗修
Owner CHINA AGRI UNIV
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