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Rice disease resistance-related gene osdr11 and its application in rice disease resistance

A disease-resistance-related gene and rice technology, applied in the field of genetic engineering, can solve the problems of low plant disease resistance and environmental pollution

Inactive Publication Date: 2019-05-10
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to control the occurrence of diseases, people often use chemical preparations and crop rotation to improve the level of rice disease resistance, but chemical preparations often increase additional economic costs and cause environmental pollution; and crop rotation under the modern agricultural planting mode has no effect on plant resistance. The disease rate is not high (Helliwell and Yang, 2013)

Method used

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  • Rice disease resistance-related gene osdr11 and its application in rice disease resistance
  • Rice disease resistance-related gene osdr11 and its application in rice disease resistance
  • Rice disease resistance-related gene osdr11 and its application in rice disease resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Isolation and clone OsDR11 gene and gene structure analysis

[0027] 1. Determination of two different transcripts of OsDR11

[0028] In the previous research work of the present invention, the State Key Laboratory of Crop Genetic Improvement, where the inventor works, used cDNA chip technology to analyze and found that the cDNA clone EI39C8 derived from rice variety Minghui 63 was inoculated with Xanthobacterium bacillus in rice variety Minghui 63. The expression level was induced to increase 2.4-2.6 times (Zhou et al., 2002). The inventors of the present invention sequenced the insert fragment of the EI39C8 clone, and used the EI39C8 sequence as a template to analyze the insert sequence of EI39C08 by BLASTN method, and found that the encoded product was homologous to Arabidopsis LAMMER protein kinase. According to the prediction results of software such as GenScan, FGENESH (http: / / www.softberry.com) and ORF Finder (http: / / www.ncbi.nlm.nih.gov / gorf / gorf.htm...

Embodiment 2

[0034] Embodiment 2: Analysis of OsDR11 gene coding product

[0035] 1. Analysis of subcellular localization of OsDR11 gene-encoded product

[0036]The amino-terminal of OsDR11 has the same nuclear localization sequence as other family members, indicating that both OsDR11S and OsDR11L may be localized in the nucleus to play a role. Clone EI39C8 with cDNA, use 39C8-4F4 (5′-ATTCTAGAATGGAGTGCTTGGCCGAGAT-3′) and 39R-XBA1 (5′-CTTCTAGACCTTAATAGCACTGGACTTG-3′) to amplify the full-length cDNA of OsDR11S, and connect it to the pM999-YFP vector after single digestion with XbaI Above, pick positive monoclonal sequencing verification, construct OsDR11S subcellular localization vector. Using the cDNA clone EI114F3 as a template, use 39C8-4F4 (5′-ATTCTAGAATGGAGTGCTTGGCCGAGAT-3′) and F3R-XBA1 (5′-CTTCTAGACATACAAGCAACAAATGAGC-3′) to amplify the exogenous fragment by PCR, and then ligate it to pM999 after single digestion with XbaⅠ -On the YFP vector, pick the positive monoclonal sequencing ...

Embodiment 3

[0042] Example 3: Analysis of the expression pattern of the OsDR11 gene

[0043] 1. Analysis of the expression pattern of two different transcripts of OsDR11 in rice tissues

[0044] RNA samples from different tissues of indica rice variety Minghui 63 at different stages, including callus, roots and leaves at two tillering stages, leaves, leaf sheaths and young panicles at booting stage, flag leaves and panicles at heading stage, and at flowering stage Stamens and pistils were detected by real-time quantitative PCR after reverse transcription (see Figure 4 in A panel). The experimental results confirmed that the two transcripts were expressed in different tissues at different growth stages of rice, with higher expression in leaves, leaf sheaths at the booting stage and gametes at the flowering stage, and OsDR11S was expressed in mature vegetative organs such as leaves and leaf sheaths OsDR11L is more abundant than OsDR11S in young vegetative organs such as callus and reprod...

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Abstract

The invention relates to the technical field of plant gene engineering, in particular to identification and functional verification of two transcripts (OsDR11S and OsDR11L) produced by rice disease resistance-related gene OsDR11 due to different selective splicing patterns. The OsDR11 gene codes LAMMER protein kinase, OsDR11S has no autophosphorylation activity, and OsDR11L has autophosphorylation activity. The genes are used for controlling rice to resist diseases caused by xanthomonas oryzae, and the bacterial blight resistance abilities of transgenic rice plants for over expression of OsDR11S and transgenic rice plants for inhibiting OsDR11L expression are remarkably improved.

Description

technical field [0001] The invention relates to the technical field of genetic engineering. It specifically involves the isolation and cloning, functional verification and application research of a rice disease resistance-related gene OsDR11. OsDR11 is involved in rice disease resistance response. Two transcripts, OsDR11S and OsDR11L, can be produced due to different alternative splicing patterns. The ability of the transgenic plants overexpressing OsDR11S and the transgenic plants inhibiting the expression of OsDR11L to resist bacterial blight was significantly improved. Background technique [0002] Plants are attacked by a variety of pathogens during the growth process. Pathogens that make plants pathogenic include viruses, bacteria, fungi, and nematodes. When the pathogen invades the plant, it may lead to two results: (1) the host plant kills the pathogen or prevents its growth, and produces a disease resistance response; (2) the host plant cannot resist the invasion ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/12C12N15/54C12N15/82A01H5/00A01H6/46
CPCC12N9/12C12N15/8281
Inventor 王石平段柳
Owner HUAZHONG AGRI UNIV
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