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Application of rice gene OsTPI1-1 to rice disease resistance improvement

A gene and rice technology, applied in the functional verification and application field of rice disease resistance-related gene OsTPI1-1, can solve the problems of no pathogen specificity and low resistance

Active Publication Date: 2020-02-14
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to previous reports, most disease resistance-related genes may be less resistant than MR genes when acting alone, but due to a variety of reasons, disease resistance-related genes are also gene resources worthy of vigorous development: (1) most disease resistance-related genes The encoded products of genes do not need to interact directly with pathogenic bacteria, and they may have persistent resistance; (2) The disease resistance reactions involved in most disease resistance genes are not pathogen-specific, and their resistance may have a broad spectrum; (3) Relatively As far as MR genes are concerned, the resources of disease resistance-related genes are very rich

Method used

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  • Application of rice gene OsTPI1-1 to rice disease resistance improvement
  • Application of rice gene OsTPI1-1 to rice disease resistance improvement
  • Application of rice gene OsTPI1-1 to rice disease resistance improvement

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Screening for interacting proteins of XA3 / XA26

[0024] The previous research results of the present invention show that the main disease resistance gene Xa3 / Xa26 can mediate broad-spectrum and long-lasting resistance to bacterial blight (Sun et al., 2004), and this gene has been used in rice breeding in China for many years (Gao et al., 2010). However, the identified factors related to this gene or involved in the disease resistance response mediated by this gene are still very limited, and their mechanism of action is still unclear.

[0025] The Xa3 / Xa26 gene encodes a cell membrane-localized receptor protein kinase (Cao Yinglong, 2007), which includes an extracellular LRR domain, a transmembrane region, and an intracellular near-membrane and kinase domain (Sun et al., 2004). The intracellular juxtamembrane and kinase domains of kinases are thought to transmit signals received extracellularly to the intracellularly. In order to better utilize the disease ...

Embodiment 2

[0034] Example 2: OsTPI1-1 (i.e. OsTPI1.1) in the rice variety Mudanjiang 8 (namely OsTPI1.1, the name of the gene in this specification is the same below, all changed to (i.e. OsTPI1-1) is a gene, the purpose of the modification is due to the patent electronic application system Names and names with a decimal point in the name are not accepted) Determination of cDNA sequence and protein activity

[0035] 1. Determination of OsTPI1-1 cDNA sequence in rice variety Mudanjiang 8

[0036] In order to obtain the full-length cDNA of the OsTPI1-1 gene in Mudanjiang 8, referring to the previous method (Zhou et al., 2002), the researchers first extracted the total RNA in the leaves of Mudanjiang 8, and then took 5 μg of total RNA and added DNaseI (Invitrogen company) after removing genomic DNA as a template, using oligo(dT) 15 The total cDNA was obtained by reverse transcription with oligomeric primers and M-MLV reverse transcriptase (Promega, USA). The registration number of OsTPI1-...

Embodiment 3

[0040] Example 3: Functional verification of OsTPI1-1 gene

[0041]The present invention adopts the genetic transformation method mediated by Agrobacterium to further verify the function of the gene by overexpressing the OsTPI1-1 gene in the rice variety Mudanjiang 8 and inhibiting the expression of the OsTPI1-1 gene in the material Rb49 carrying Xa3 / Xa26.

[0042] 1. Construction of genetic transformation vector

[0043] The overexpression vector used in the present invention is pU1301 (Qiu et al., 2007), which is a maize ubiquitin promoter (P Ubi ) Agrobacterium-mediated genetic transformation vector. Using the total cDNA of rice variety Mudanjiang 8 as a template, the full-length cDNA of the OsTPI1-1 gene was amplified with the primers K16-OxF and K16-oxR described in Example 2-1. After connecting to the TA cloning vector pGEM-T easy (Promega, USA), the positive clones were sequenced, and the cloned plasmids without mutation were digested with KpnI and BamHI to cut out th...

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Abstract

The invention belongs to the technical field of plant gene engineering, and particularly relates to application of a rice gene OsTPI1-1 to rice disease resistance improvement. The gene OsTPI1-1 codesone cytoplasm TPI (triosephosphate isomerase); the nucleotide sequence of the gene is shown as SEQ ID NO:1 in a sequence table; and a protein sequence coded by the gene is shown as SEQ ID NO:2 in thesequence table. Through the obvious expression quantity increase of the OsTPI1-1, the resistance of a converted rice plant to bacterial blight is enhanced. Under the background of a major disease-resistant gene Xa3 / Xa26, the expression quantity of the OsTPI1-1 is obviously reduced, so that the converted rice plant loses the resistance to the bacterial blight. Functional verification proves that the gene OsTPI1-1 is a positive regulation factor in the rice bacterial-blight-resistant reaction; and the disease resistance of the rice can be improved through over-expression of the gene OsTPI1-1.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering. It specifically involves the functional verification and application of a rice disease resistance-related gene OsTPI1-1. The gene encodes a triose phosphate isomerase localized in the cytoplasm, which positively regulates rice resistance to bacterial blight and the main gene Xa3 / Xa26 for bacterial blight resistance. Background technique [0002] Rice is one of the most important food crops in the world, providing the staple food for more than half of the world's population (Zhang, 2007). Rice disease is an important factor limiting rice production, and the prevalence of disease often leads to the decline of rice yield and quality. Rice bacterial blight is a vascular disease caused by Xanthomonas oryzae pv. One of the bacterial diseases. [0003] According to the response speed and strength of plants to pathogenic bacteria infection, the resistance of rice to pathogenic bacte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/90C12N15/82A01H5/00A01H6/46
CPCC12N9/90C12N15/8281C12Y503/01001
Inventor 王石平刘艳艳
Owner HUAZHONG AGRI UNIV
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