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Application of Rice Gene osdf1 and Its Regulatory Function of Disease Resistance

A rice and genetic technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of limited range of available disease-resistant resources and long breeding cycle, and achieve a wide range of available disease-resistant resources, short cultivation cycle, and easy operation. Simple and convenient effect

Active Publication Date: 2020-01-07
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to natural genetic isolation, traditional disease-resistant breeding has significantly limited the available range of disease-resistant resources. Only disease-resistant resources with close genetic relationships can be used, and multiple crosses and backcrosses are required. Therefore, the breeding cycle is long and requires A lot of manpower and material resources

Method used

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  • Application of Rice Gene osdf1 and Its Regulatory Function of Disease Resistance
  • Application of Rice Gene osdf1 and Its Regulatory Function of Disease Resistance

Examples

Experimental program
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Effect test

Embodiment 1

[0041] The present invention clones a rice gene, clarifies the broad-spectrum anti-bacterial regulation function of the gene by constructing overexpression transgenic rice for the first time, and reveals that the gene has a broad-spectrum negative regulation function on rice bacterial disease resistance. Therefore, the present invention named the gene Oryza sativa defense-related 1 (abbreviated as OsDF1). Because the overexpression of the OsDF1 gene leads to a significant decline in the resistance of rice to bacterial diseases, it is possible to create new rice materials with reduced resistance to rice bacterial diseases by constructing an overexpression of the OsDF1 gene in rice homozygous lines for gene function. analysis etc. The main steps for the cloning and functional analysis of the OsDF1 gene, and the creation and acquisition of new rice materials with reduced resistance to rice bacterial diseases include:

[0042] 1) Cloning and preservation of rice OsDF1 gene

[00...

Embodiment 2

[0075] According to the anti-bacterial negative regulatory function of the rice gene OsDF1 elucidated by the present invention as described above, a set of RNAi transgenic rice using the gene is established, and a new rice material with broad-spectrum resistance to rice bacterial diseases is created and obtained by using genetic engineering technology. Technology System. The main steps include:

[0076] (1) Construction and acquisition of OsDF1 gene RNAi structure

[0077] Design primer OsDF1-F2 (5'-ttggatcc gaa tct ttg gttgag aga agc-3' according to the OsDF1 sequence cloned in the present invention, the italic part is the Bam HI restriction site) (sequence shown in SEQ ID NO: 5), and OsDF1-R3 (5'-ttctcgag cat gct ctg aaa atg ctg ctg-3', the part in italics is the XhoI restriction site) (sequence shown in SEQ ID NO: 6). Using the pMD19-OsDF1 plasmid obtained in 1) of Example 1 as a template, and OsDF1-F2 / OsDF1-R3 as a primer pair, a 300bp OsDF1 fragment was obtained by PCR ...

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Abstract

The invention provides a rice disease control gene named as OsDF1. An open reading frame of the gene is 1512bp long, and the encoded protein is composed of 503 amino acids. The protein does not comprise any structural domain of known functions. By constructing overexpression transgenic rice of the gene and disease resistance analysis thereof, a broad spectrum control effect of the gene on the bacterial disease resistance of the rice is illustrated first. The overexpression transgenic rice plants widely reduce the resistance of bacterial leaf streak and bacterial leaf blight of rice, and the broad spectrum negative control function of the gene on the bacterial disease resistance of the rice is disclosed. The invention first discloses biological functions of the gene, and provides application of acquiring rice materials with general decrease of bacterial disease resistance by creating OsDF1 overexpression rice and application of acquiring broad spectrum bacterial disease resistant rice materials by creating OsDF1-RNAi rice. The gene is a novel gene resource applied to creating and breeding novel materials and novel varieties of rice with broad spectrum bacterial disease resistance.

Description

technical field [0001] The invention belongs to the field of biological technology and relates to a rice gene OsDF1 and the application of the disease resistance regulating function thereof. Background technique [0002] 1. Plant gene cloning technology [0003] There are many methods for plant gene cloning. With the increase of plant species whose genome sequence has been determined, the method of plant gene cloning based on database sequence has been more and more widely used. The operation steps of the method mainly include primer design based on conserved sequences, RNA extraction from target plant tissue, reverse transcriptase-polymerase chain reaction (RT-PCR), connection of PCR product and vector, bacterial transformation of the connection product, and plasmid extraction. , enzyme digestion test, sequencing analysis, etc. [0004] 2. Plant gene expression control and gene function analysis technology [0005] The function of plant genes is carried out through gene ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00A01H6/46
CPCC07K14/415C12N15/8205C12N15/8218C12N15/8281
Inventor 蔡新忠徐幼平郭伟毅
Owner ZHEJIANG UNIV
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