Rice Wd40-enriched repeat protein oswd40-141 and its encoding gene and application

A technology for rice bacterial blight and rice, which is applied in the fields of application, genetic engineering, plant genetic improvement, etc., and can solve the problems of easy loss of variety resistance, easy mutation, and difficulty in utilization.

Active Publication Date: 2021-04-27
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

Breeding disease-resistant varieties containing resistance genes is currently the most economical and effective measure to prevent and control rice bacterial blight. However, 42 rice bacterial blight resistance genes / loci have been reported so far (http: / / www.shigen .nig.ac.jp / rice / oryzabase / gene / list) most of them show a narrow resistance spectrum or are difficult to use, and only genes such as Xa3, Xa4, Xa21 and Xa23 are widely used in production
[0003] Because Xanthomonas oryzae pathogenic species (Bacterial blight) are easy to mutate, the co-evolution of rice and Bacterial blight leads to easy loss of variety resistance

Method used

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  • Rice Wd40-enriched repeat protein oswd40-141 and its encoding gene and application
  • Rice Wd40-enriched repeat protein oswd40-141 and its encoding gene and application
  • Rice Wd40-enriched repeat protein oswd40-141 and its encoding gene and application

Examples

Experimental program
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Effect test

Embodiment 1

[0079] Example 1. Method for targeted knockout of rice WD40-rich repeat protein gene OsWD40-141 based on Lb-CRISPR / Cpf1 system

[0080] 1. Sequence and analysis of rice WD40-rich repeat protein gene OsWD40-141

[0081] The sequence of the rice WD40-rich repeat protein gene OsWD40-141 is shown in SEQ ID No.2. Sequence analysis shows that the gene includes 14 exons, which are respectively the 144th-204th positions of the SEQ ID No.2 sequence ( First exon), 2093-2238 (second exon), 2313-2659 (third exon), 4910-5022 (fourth exon), 5092-5138 bit (fifth exon), 6715-6865 (sixth exon), 7015-7078 (seventh exon), 7255-7413 (eighth exon), 7527 -7648 (ninth exon), 8570-8644 (tenth exon), 14371-14424 (eleventh exon), 14505-14578 (twelfth exon) ), 15650-15724 (13th exon), 15885-16104 (14th exon).

[0082] In the present invention, the sequence on the promoter of the rice WD40-rich repeat protein gene OsWD40-141 is used as the OsWD40-141-T1 target sequence of the rice WD40-rich repeat pro...

Embodiment 2

[0094] Example 2. Application of targeted knockout method in rice varieties based on Lb-CRISPR / Cpf1 technology

[0095] The recombinant Agrobacterium EH105-Lb-OsWD40-141-1 was used to infect the callus induced by mature embryos of rice variety Nipponbare, and the obtained rice transformed plants were named NIP-Lb-OsWD40-141-1 respectively.

[0096] The specific method is as follows:

[0097] 1. Inoculate the recombinant Agrobacterium EH105-Lb-OsWD40-141-1 in YEB liquid medium (containing 50 μg / ml kanamycin and 20 μg / ml rifampicin), and culture it with shaking at 28°C and 200 rpm to OD600 0.6-0.8; centrifuge at 5000rpm, 4°C for 5min, resuspend the bacterial cell sediment concentration in AAM liquid medium (acetosyringone concentration is 200μM / L, pH 5.2) until the OD600 is 0.6-0.8, and obtain recombinant Agrobacterium resuspension liquid.

[0098] 2. Remove chaff from mature seeds of rice variety Nipponbare, soak in 75% ethanol for 1 min, then sterilize in NaClO solution (mix...

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Abstract

The invention discloses a rice WD40-rich repeat protein OsWD40‑141, its coding gene and application. The present invention first discloses the application of any of the following proteins in regulating plant bacterial blight resistance: A1) the protein whose amino acid sequence is SEQ ID No.1; A2) the amino acid shown in SEQ ID No.1 A fusion protein obtained by connecting the N-terminal or / and C-terminal tag of the sequence; A3) the amino acid sequence shown in SEQ ID No.1 through the substitution and / or deletion and / or addition of one or several amino acid residues and The proteins shown in A1) have more than 90% identity and have the same function. Further disclosed is a method for cultivating gene mutant plants with enhanced resistance to bacterial blight. The invention provides an efficient breeding method for creating bacterial blight-resistant materials based on the protein OsWD40‑141, and has important application value.

Description

technical field [0001] The invention belongs to the field of biotechnology. Specifically, it relates to rice rich in WD40 repeat protein OsWD40-141 and its coding gene and application. Background technique [0002] Rice bacterial blight is an important bacterial disease that restricts rice production. It is a serious hazard to the rice planting industry. Generally, it can cause a 20%-30% reduction in rice production, and it can reach 50% in severe cases. Breeding disease-resistant varieties containing resistance genes is currently the most economical and effective measure to prevent and control rice bacterial blight. However, 42 rice bacterial blight resistance genes / loci have been reported so far (http: / / www.shigen .nig.ac.jp / rice / oryzabase / gene / list) most of them showed a narrow resistance spectrum or were difficult to use, and only genes such as Xa3, Xa4, Xa21 and Xa23 were widely used in production. [0003] Because the pathogenic strain of Xanthomonas oryzae (Bacteria...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00A01H5/10A01H6/46
CPCC07K14/415C12N15/8218C12N15/8281
Inventor 周永力李全林卢家玲
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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