Rice MeRING29 gene, coding protein, recombinant vector and application

A recombinant vector and gene technology, applied in the field of genetic engineering, can solve the problems of decreased resistance, complex disease resistance mechanism, frequent variation of physiological races of pathogenic bacteria, etc.

Active Publication Date: 2020-10-23
云南省农业科学院生物技术与种质资源研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many types of grass crops, there are many types of diseases, and the mechanism of disease resistance is complex. So far, many problems have not been studied clearly. Moreover, due to the frequent variation of physiological races of pathogenic bacteria, the resistance of new resistant varieties will decline after several years of use. or disappear, therefore, it is necessary to continuously mine and utilize disease-resistant gene resources

Method used

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  • Rice MeRING29 gene, coding protein, recombinant vector and application
  • Rice MeRING29 gene, coding protein, recombinant vector and application
  • Rice MeRING29 gene, coding protein, recombinant vector and application

Examples

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Effect test

Embodiment 1

[0045] Cloning of the MeRING29 gene

[0046] The transcriptome sequencing of wild rice verruca yunnanensis was carried out by Beijing Baimaike Biotechnology Co., Ltd., and a gene fragment annotated as Arabidopsis ubiquitin ligase was screened and published in NCBI (http: / / www.ncbi.nlm.gov) Blast comparison analysis was carried out on the Internet, and the gene sequence (shown in SEQ ID No.3) with a homology of 91.5% to the rice gene was obtained, and then according to the sequence screened by this section, through the RACE method (see Beijing Jumei Biotechnology Co., Ltd. disclosed kit operation manual) to obtain the gene for improving rice disease resistance according to the present invention, its nucleotide sequence is as shown in SEQ ID No.1 in the sequence listing, and the amino acid sequence of the protein encoded by it is as SEQ ID No .2 shown. Design specific primers according to the nucleotide sequence shown in SEQ ID No.3, RING-GSP1:5'-GCGAGGGAGTGGCCGGTGGAGTGC-3'(SEQ...

Embodiment 2

[0057] Functional Analysis of MeRING29 Gene

[0058] (1) Bacteria blight and exogenous hormone treatment of wild rice verruca in Yunnan

[0059] Use NA medium to activate bacterial blight pathogen Y8. Y8 is a highly pathogenic physiological race in Yunnan, provided by the Institute of Biotechnology and Germplasm Resources, Yunnan Academy of Agricultural Sciences, cultured in the dark at 28°C for 3 days, and washed with 5 mL of sterile water first. bacterial lawn, and then continue to add sterile water until OD 600 =0.6, the prepared bacterial solution was used to inoculate the leaves of O. yunnanensis at the booting stage. When the greenhouse temperature rises to 28°C, use scissors to pick up the prepared bacterial solution and cut off the top 1.0cm of the leaves of the wild rice verruca yunnanensis with normal growth and no obvious wounds as the infection treatment. The control group is inoculated with sterile water instead of the bacterial solution After the inoculation gr...

Embodiment 3

[0072] Application of MeRING29 gene in enhancing rice resistance to bacterial blight

[0073] (1) Construction of expression vector for overexpression of MeRING29 gene

[0074] According to the full-length cDNA sequence of MeRING29, primers containing a complete open reading frame (ORF) were designed, and a restriction endonuclease NcoI restriction endonuclease NcoI restriction site was added to the upstream primer MeRING29-OF of the ORF sequence of MeRING29 during design. The sequence is shown in the table as SEQ ID NO.8 shows: 5'-CCCATGGATGCCGACGCCTCGCCGCCAC-3', the ORF sequence of MeRING29. The downstream primer MeRING29-OR adds the restriction endonuclease BglII restriction endonuclease site sequence as shown in SEQ ID NO.9 in the table: 5 '-GAAGATCTTTCAAACTTGCCGAGCGATTG-3', using MeRING29-OF and MeRING29-OR as primers, the full-length cDNA sequence of MeRING29 was amplified by conventional PCR and gel recovery, and the PCR amplification product and the vector pCAMBIA2301 ...

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Abstract

The invention provides a rice MeRING29 gene, a coding protein, a recombinant vector and application, and relates to the technical field of genetic engineering. A full-length cDNA sequence of the YunnanOryza meyeriana Baill. MeRING29 gene is cloned for the first time, a plant disease resistance gene pool is enriched, and the rice MeRING29 gene has important value for further research on the rice disease resistance molecular mechanism and application thereof. The invention also provides a coding amino acid sequence of the MeRING29 gene and a recombinant vector containing the gene and application, and particularly relates to application in improving rice disease resistance. Rice plants transformed with the MeRING 29 gene have a significantly improved resistance to bacterial blight and rice blast.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a rice MeRING29 gene, encoded protein, recombinant vector and application. Background technique [0002] Rice is one of the most important food crops in the world, and more than half of the world's population depends on rice as the staple food. With the continuous growth of the world's population, the demand for food is also increasing. It is estimated that by 2030, my country's rice consumption will increase by more than 20% compared to now. In recent years, with the mass cultivation and promotion of super rice varieties, rice yield has reached a new high. However, problems also followed. In the process of breeding new rice varieties, a large number of potential good genes were filtered out due to directional selection, resulting in a narrow genetic basis of modern cultivated species, lack of resistance to diseases, insect pests and various abiotic stres...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N9/00C12N15/82A01H5/00A01H6/46
CPCC12N9/93C12N15/8281C12N15/8282C12Y603/02019
Inventor 付坚王波程在全陈越陈玲钟巧芳王玲仙柯学张敦宇肖素勤蒋聪殷富有
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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