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Rice retrotransposon gene LOC_Os11g45295 and encoding protein and application thereof

A technology of transgenic plants and proteins, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of easy mutation of pathogenic varieties, narrow resistance spectrum, difficult to use, etc.

Active Publication Date: 2020-01-17
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 44 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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  • Rice retrotransposon gene LOC_Os11g45295 and encoding protein and application thereof
  • Rice retrotransposon gene LOC_Os11g45295 and encoding protein and application thereof
  • Rice retrotransposon gene LOC_Os11g45295 and encoding protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1. Method for knocking out rice retrotransposon gene LOC_Os11g45295 based on pYLCRISPR / Cas9 system

[0072] 1. Sequence analysis and target sequence screening of rice retrotransposon gene LOC_Os11g45295

[0073] The nucleotide sequence of the rice retrotransposon gene LOC_Os11g45295 is shown in sequence 2, and the amino acid sequence of the protein encoded by it is sequence 1 in the sequence listing. Sequence analysis shows that the gene includes 4 exons in total, which are sequence 2 1-26 (first exon), 1308-1642 (second exon), 2610-2709 ( Exon 3), positions 4359-4737 (exon 4).

[0074] The sequence on the second exon of the rice retrotransposon gene LOC_Os11g45295 was used as the target sequence of LOC_Os11g45295-T1 and LOC_Os11g45295-T2 in the targeted knockout method of the rice retrotransposon gene LOC_Os11g45295 based on the pYLCRISPR / Cas9 system.

[0075]After extensive screening, it was determined that the pYLCRISPR / Cas9 technology was used to target th...

Embodiment 2

[0106] Example 2. Application of pYLCRISPR / Cas9 technology-based targeted knockout method in rice varieties

[0107] 1. Targeted knockout of LOC_Os11g45295 gene by pYLCRISPR / Cas9 technology

[0108] The recombinant Agrobacterium EH105-Cas9-LOC_Os11g45295 was used to infect the callus induced by mature embryos of the rice variety Nipponbare (hereinafter referred to as wild-type rice), and the obtained rice transformed plants were respectively named NIP-Cas9-LOC_Os11g45295; the specific methods of the experiment are as follows:

[0109] 1. Inoculate the recombinant Agrobacterium obtained in Example 1 into YEB liquid medium (containing 50 μg / ml kanamycin and 20 μg / ml rifampicin), and culture it with shaking at 28° C. and 200 rpm until the OD600 is 0.6-0.8 ; Centrifuge at 5000rpm and 4°C for 5min, and resuspend the bacterial pellet with AAM liquid medium (acetosyringone concentration: 200μM / L, pH 5.2) until the OD600 is 0.6-0.8.

[0110] 2. Remove the glumes from the mature seeds...

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PUM

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Abstract

The invention discloses a rice retrotransposon gene LOC_Os11g45295 and an encoding protein and application thereof. The invention discovers application of the rice retrotransposon gene LOC_Os11g45295and the encoding protein thereof to regulation and control of plant disease resistance, and by destroying the biological functions of the encoding protein of the LOC_Os11g45295 gene, resistance of therice to xanthomonas oryzae can be significantly improved. A CRISPR / Cas9 technology is utilized to realize efficient fixed-point knockout of the LOC_Os11g45295 gene, and after rice subjected to fixed-point knockout is inoculated with xanthomonas oryzae IV, KS-6-6 and PXO61, the length of a speck is significantly decreased. The new functions of the LOC_Os11g45295 gene provide a new method for disease resistance breeding of plants, and the LOC_Os11g45295 gene has the very important application value in agricultural production.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to rice retrotransposon gene LOC_Os11g45295 and its encoded protein and application. Background technique [0002] Bacterial blight caused by Xanthomonas oryzae pIV.oryzae is an important bacterial disease that restricts rice production. It is a serious hazard to the rice planting industry, generally causing a 20%-30% reduction in rice production. % or so, seriously up to 50%. Breeding and planting disease-resistant varieties using resistance genes is the most economical and effective measure to control rice bacterial blight. However, most of the 44 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 inaccessibility, only genes such as Xa3, Xa4, Xa21 and Xa23 are widely used in production. [0003] Because the pathogenic variety of Xanthomonas oryza...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/84A01H5/00A01H6/46
CPCC07K14/415C12N15/8218C12N15/8281
Inventor 周永力李全林史晓荣卢家玲
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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