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Application of gene Os11g0682000 and proteins encoded by Os11g0682000 in regulating resistance to rice bacterial leaf blight

A bacterial blight resistance and gene technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of narrow resistance spectrum, easy mutation of pathogenic varieties, and easy loss of variety resistance.

Active Publication Date: 2020-05-08
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the 45 rice bacterial blight resistance genes / loci reported so far (http: / / www.shigen.nig.ac.jp / rice / oryzabase / gene / list) show a narrow resistance spectrum Or the problem of poor utilization, only genes such as Xa3, Xa4, Xa21 and Xa23 are widely used in production
[0003] Because Xanthomonas pv.

Method used

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  • Application of gene Os11g0682000 and proteins encoded by Os11g0682000 in regulating resistance to rice bacterial leaf blight
  • Application of gene Os11g0682000 and proteins encoded by Os11g0682000 in regulating resistance to rice bacterial leaf blight
  • Application of gene Os11g0682000 and proteins encoded by Os11g0682000 in regulating resistance to rice bacterial leaf blight

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1. Method for knocking out rice gene Os11g0682000 based on pYLCRISPR / Cas9 system

[0092] 1. Sequence analysis and target sequence screening of rice gene Os11g0682000

[0093] The nucleotide sequence of the rice gene Os11g0682000 is shown in SEQ ID No.2, and the amino acid sequence of the encoded protein is SEQ ID No.1. Sequence analysis shows that the gene includes 2 exons in total, which are No. 636-1680 (first exon) and No. 4346-6198 (second exon) of SEQ ID No. 2, respectively.

[0094] In the present invention, the sequence on the first exon of the rice gene Os11g0682000 is used as the 0682000-T1 target sequence of the rice gene Os11g0682000 targeted knockout method based on the pYLCRISPR / Cas9 system, and the sequence on the second exon of the rice gene Os11g0682000 is used as the target sequence based on pYLCRISPR The 0682000-T2 target sequence of the rice gene Os11g0682000 site-directed knockout method of the / Cas9 system.

[0095] After a large number o...

Embodiment 2

[0120] Example 2. Application of pYLCRISPR / Cas9 technology-based targeted knockout method in rice varieties

[0121] 1. pYLCRISPR / Cas9 technology targeted knockout of Os11g0682000 gene

[0122] The recombinant Agrobacterium EH105-Cas9-0682000 was used to infect the callus induced by wild-type Nipponbare mature embryos, and the obtained rice transformed plants were respectively named NIP-Cas9-0682000; the specific methods of the experiment are as follows:

[0123] 1. The recombinant Agrobacterium EH105-Cas9-0682000 obtained in Example 1 was inoculated in YEB liquid medium (containing 50 μg / ml kanamycin and 20 μg / ml rifampicin), and cultured with shaking at 28°C and 200rpm until OD600 is 0.6-0.8; centrifuge at 5000rpm, 4°C for 5min, and resuspend the bacterial pellet in AAM liquid medium (acetosyringone concentration is 200μM / L, pH 5.2) until the OD600 is 0.6-0.8.

[0124] 2. Remove the glumes from the mature seeds of wild-type Nipponbare, soak them in 75% ethanol for 1 min, th...

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Abstract

The invention discloses an application of gene Os11g0682000 and proteins encoded by the Os11g0682000 in regulating resistance to rice bacterial leaf blight. The invention firstly discloses the application of the following proteins in regulating the resistance to the plant bacterial leaf blight: A1) a protein with the amino acid sequence shown in SEQ ID No.1; A2) a fusion protein obtained by linking a tag at the N-terminus or / and C-terminus of the amino acid sequence shown in the SEQ ID No.1; and A3) a protein which has 90% or above identity and the same function as the protein shown in the A1)and is obtained by replacement and / or deletion and / or addition of one or more amino acid residues on the amino acid sequence shown in the SEQ ID No.1. The invention further discloses a method for breeding a gene mutant plant with the enhanced resistance to the bacterial leaf blight. The method provides an efficient breeding manner for creating bacterial leaf blight resistant rice based on the gene Os11g0682000, and has important application value.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to the application of gene Os11g0682000 and its encoded protein in regulating rice bacterial blight resistance. Background technique [0002] Bacterial blight caused by Gram-negative bacteria Xanthomonas oryzae pv. oryzae is the most serious and widespread bacterial disease in rice production. It can occur in the whole growth and development stages of rice, and it is most serious in the seedling and tillering stages. When the disease occurs, it seriously threatens the production safety of rice. It will generally lead to a 20%-30% reduction in rice production, and it can reach 50% in severe cases. Sometimes even will fail. Practice has proved that using resistance genes to breed and plant disease-resistant varieties is the most economical and effective measure to control rice bacterial blight. However, most of the 45 rice bacterial blight resistance genes / loci rep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8281
Inventor 周永力李健敏卢家玲
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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