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Plant disease-resistant regulation and control gene UEP and application thereof

A technology for regulating genes and plants, applied in the fields of plant gene improvement, plant products, genetic engineering, etc., can solve the problems of long breeding cycle and limited range of disease-resistant resources, and achieve high resistance level, wide range of disease-resistant objects, Strong resistance to the effect

Active Publication Date: 2013-04-17
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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  • Plant disease-resistant regulation and control gene UEP and application thereof
  • Plant disease-resistant regulation and control gene UEP and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The present invention establishes a set of high-quality disease resistance regulatory genes cloned by the inventor UEP , using genetic engineering technology to create and obtain a technical system for disease-resistant plant materials with a high level of resistance, a wide range of disease-resistant objects, and at the same time promoting plant growth and development. The main steps include:

[0028] 1) benthamiana ( Nicotiana benthamiana ) UEP Cloning and preservation of genes

[0029] Nicotiana benthamiana provided by the invention ( Nicotiana benthamiana ) UEP Genes were cloned through the following steps. The primer NbUbi-F (5'- gc ggatcc atg cag atc ttc gtg aaa acc-3', the italic part is Bam HI restriction site) (sequence shown in SEQ ID: 3), and NbRlp-R (5'- gc gtcgac tca atc ​​ggc acc ggc ctt gtt g -3’, the italic part is Sal Ⅰ restriction site) (sequence shown in SEQ ID: 4). Total RNA was extracted from Nicotiana benthamiana leaves with TRIZ...

Embodiment 2

[0041] Example 2 High-level, durable, broad-spectrum disease-resistant transgenic UEP Acquisition of Genetic Tobacco Plants

[0042] The main operation steps include:

[0043] 1) UEP Construction and acquisition of gene expression constructs

[0044] Carrying Nicotiana benthamiana owned by the inventor's laboratory UEP vector of gene sequence Bam HI / Sal ⅠDouble enzyme digestion, recovery by electrophoresis and gel tapping UEP The ORF sequence of the gene was subcloned into the strong promoter of the plant expression vector pCHF3 - after the 35S promoter of cauliflower mosaic virus (CaMV), strong expression was obtained UEP Plant expression construct of the gene pCHF3:: UEP .

[0045] 2) Conversion UEP Agrobacterium acquisition of gene expression constructs

[0046] Aspirate 1~2 μl UEP Gene expression construct pCHF3:: UEP , add 40 μl EHA105 Agrobacterium competent cells, mix quickly, and use the Bio-Rad Gene Pulser II electric shock instrument at a vol...

Embodiment 3

[0057] Example 3 High-level, durable, broad-spectrum disease-resistant transgenic UEP Procurement of Genetic Tomato Plants

[0058] The main operation steps include:

[0059] 1) UEP Construction and acquisition of gene expression constructs

[0060] Carrying Nicotiana benthamiana owned by the inventor's laboratory UEP vector of gene sequence Bam HI / Sal ⅠDouble enzyme digestion, recovery by electrophoresis and gel tapping UEP The ORF sequence of the gene was subcloned into the strong promoter of the plant expression vector pCHF3 - after the 35S promoter of cauliflower mosaic virus (CaMV), strong expression was obtained UEP Plant expression construct of the gene pCHF3:: UEP .

[0061] 2) Conversion UEP Agrobacterium acquisition of gene expression constructs

[0062] Aspirate 1~2 μl UEPGene expression construct pCHF3:: UEP , add 40 μl EHA105 Agrobacterium competent cells, mix quickly, and use the Bio-Rad Gene Pulser II electric shock instrument at a volta...

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Abstract

The invention provides a plant disease-resistant regulation and control gene. An open reading frame of the gene is 471 bp in length; and the gene encodes a ubiquitin extended protein consisting of 156 amino acids. The N end of the gene coded product is a ubiquitin molecule comprising 76 amino acids and the C end of the gene coded product is 40S ribosomal protein s27a comprising 80 amino acids. The gene is a high-quality disease-resistant gene resource. A disease-resistant material obtained by the gene has high resistance and a wide disease-resistant spectrum and can promote growth of plants. The gene widely joints in the regulation and control on resistance of various diseases caused by various pathogens by translating various kinds of targeted proteins and regulating modification after translation and joints in the regulation and control on growth and development of the plants. Due to overexpression of a tobacco UEP gene, the resistance of tobacco to various diseases such as Leptosphaeria maculans, wildfire, viral disease and the like is obviously improved. The plant disease-resistant regulation and control gene is applicable to creation and breeding of disease-resistant plant materials and varieties which have high resistance and wide disease-resistant objects and can promote growth of the plants.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a plant disease resistance regulatory gene (UEP) and its application. Background technique [0002] 1. Plant gene cloning technology [0003] There are many methods for plant gene cloning. With the completion of the determination of various plant genome sequences, plant gene cloning methods based on database sequences have 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 tissues, reverse transcriptase-polymerase chain reaction (RT-PCR), ligation of PCR products and vectors, bacterial transformation of ligation products, and plasmid extraction. , enzyme digestion test, sequencing analysis, etc. [0004] 2. Plant transgenic technology [0005] It is used to introduce exogenous genes into target plants, so as to obtain transgenic plants carrying exogenous genes. Includi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00
Inventor 蔡新忠宋哓毅赖亿玉程维舜赵媛
Owner ZHEJIANG UNIV
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