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Rice receptor kinase gene LOC_Os11g47290 as well as its encoded protein and application thereof

A protein and protein technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of easy mutation of pathogenic varieties, narrow resistance spectrum, and difficulty in utilization

Active Publication Date: 2020-02-14
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI +1
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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 receptor kinase gene LOC_Os11g47290 as well as its encoded protein and application thereof
  • Rice receptor kinase gene LOC_Os11g47290 as well as its encoded protein and application thereof
  • Rice receptor kinase gene LOC_Os11g47290 as well as its encoded protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1. Site-directed knockout method of rice receptor kinase gene LOC_Os11g47290 based on pYLCRISPR / Cas9 system

[0071] 1. Sequence analysis and target sequence screening of the rice receptor kinase gene LOC_Os11g47290

[0072] The nucleotide sequence of the rice receptor kinase gene LOC_Os11g47290 is shown in sequence 2, and the amino acid sequence of the encoded protein is sequence 1 in the sequence table. Sequence analysis showed that the gene includes a total of 7 exons, which are positions 1-1696 (the first exon), positions 2042-2941 (the second exon), and positions 3130-3278 of the sequence 2 sequence. position (third exon), position 3677-3890 (fourth exon), position 6492-6611 (fifth exon), position 6976-7019 (sixth exon), 7163- 7171 (the seventh exon).

[0073] The sequence on the first exon of the rice receptor kinase gene LOC_Os11g47290 was used as the 47290-T1 and 47290-T2 target sequences of the rice receptor kinase gene LOC_Os11g47290 site-directed kn...

Embodiment 2

[0101] Example 2. Application of site-directed knockout method based on pYLCRISPR / Cas9 technology in rice varieties

[0102] 1. Targeted knockout of LOC_Os11g47290 gene by pYLCRISPR / Cas9 technology

[0103] Recombinant Agrobacterium EH105-Cas9-47290 was used to infect callus induced by mature embryos of rice variety Nipponbare (hereinafter referred to as wild-type rice), and the obtained transformed rice plants were named as NIP-Cas9-47290; the specific experimental methods were as follows:

[0104] 1. The recombinant Agrobacterium obtained in Example 1 was inoculated into YEB liquid medium (containing 50 μg / ml kanamycin and 20 μg / ml rifampicin), and shaken at 28 ° C and 200 rpm until OD600 was 0.6-0.8 ; Centrifuge at 5000 rpm and 4 °C for 5 min, and resuspend the cell pellet with AAM liquid medium (acetosyringone concentration is 200 μM / L, pH 5.2) to an OD600 of 0.6-0.8.

[0105] 2. The mature seeds of the rice variety Nipponbare were removed from the glumes, soaked in 75% eth...

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Abstract

The invention discloses a rice receptor kinase gene LOC_Os11g47290 as well as an encoded protein and an application thereof. According to the application of the rice receptor kinase gene LOC_Os11g47290 and the encoded protein thereof in regulation and control of plant disease resistance, the resistance of rice to bacterial blight can be remarkably improved by destroying the biological function ofthe encoded protein of the LOC_Os11g47290 gene. The efficient LOC_Os11g47290 gene fixed-point knockout is realized by using a CRISPR / Cas9 technology, and after the rice is inoculated with bacterial blight bacteria GD1358 and V, the scab length is obviously shortened. The new function of the LOC_Os11g47290 gene provided by the invention provides a new method for disease-resistant breeding of plants, and the LOC_Os11g47290 gene has 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 a rice receptor kinase gene LOC_Os11g47290 and its encoded protein and applications. Background technique [0002] Bacterial blight caused by Xanthomonas oryzae pv.oryzae is an important bacterial disease restricting rice production. % or so, up to 50% in severe cases. Breeding and planting disease-resistant varieties with resistance genes is the most economical and effective measure to control rice bacterial blight. However, most of the 44 reported rice bacterial blight resistance genes / loci (http: / / www.shigen.nig.ac.jp / rice / oryzabase / gene / list) show narrow resistance spectrum Or the problem of difficulty in utilization, only genes such as Xa3, Xa4, Xa21 and Xa23 are widely used in production. [0003] Since Xanthomonas oryzae pathogenic varieties are easy to mutate, the co-evolution of rice and Xanthomonas oryzae (Xanthomonas oryzae pathogenic varieties) le...

Claims

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Application Information

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IPC IPC(8): C12N9/12C12N15/54C12N15/113C12N15/82A01H5/00A01H6/46
CPCC12N9/12C12N15/113C12N15/8218C12N15/8281C12N2310/20
Inventor 翟文学周永力李全林史晓荣卢家玲
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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