Rice WD40-rich repeat protein OsWD40-141 and coding gene and application thereof

A gene and protein technology, applied in the biological field, can solve the problems of easy mutation, narrow resistance spectrum, difficult to use and so on

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

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 th

Method used

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  • Rice WD40-rich repeat protein OsWD40-141 and coding gene and application thereof
  • Rice WD40-rich repeat protein OsWD40-141 and coding gene and application thereof
  • Rice WD40-rich repeat protein OsWD40-141 and coding gene and application thereof

Examples

Experimental program
Comparison scheme
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 and a coding gene and application thereof. The invention firstly discloses application of any one of proteins in regulation of plantbacterial leaf blight resistance; the proteins include A1) a protein with an amino acid sequence of SEQ ID No. 1, A2) a fusion protein obtained by connecting a tag to the N end or/and C end of the amino acid sequence shown in SEQ ID No. 1, and A3) a protein which has 90% or more identity with the protein shown in A1, has the same functionality, and is obtained by subjecting the amino acid sequenceshown in SEQ ID No. 1 to substitution and/or deletion and/or addition of one or several amino acid residues. Further disclosed is a method for growing a mutant plant having enhanced resistance to bacterial leaf blight. The invention provides an efficient breeding means to create a bacterial leaf blight resistant material based on protein OsWD40-141; the efficient breeding means 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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IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00A01H6/46
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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