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Method for improving broad-spectrum resistance of rice blast by using CRISPR/Cas9 mediated adenine base editing system

A technology of adenine base and rice blast disease, which is applied in the fields of agricultural production and production of rice germplasm resources

Pending Publication Date: 2019-04-19
YICHUN UNIVERSITY
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Problems solved by technology

The study did not identify any indels or other base transitions or transversions at the target site or potential off-target sites

Method used

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  • Method for improving broad-spectrum resistance of rice blast by using CRISPR/Cas9 mediated adenine base editing system
  • Method for improving broad-spectrum resistance of rice blast by using CRISPR/Cas9 mediated adenine base editing system
  • Method for improving broad-spectrum resistance of rice blast by using CRISPR/Cas9 mediated adenine base editing system

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Embodiment

[0043] The present invention is a method for improving rice blast broad-spectrum resistance by using CRISPR / Cas9-mediated adenine base editing system, using Nipponbare rice variety as the acceptor material, which uses CRISPR / Cas9-mediated adenine base Editing system for rice by Agrobacterium-mediated transformation Bsr-d1 A single base in the promoter region of the gene, -618 away from the start codon, site-specific substitution (A→G), identification and screening of biallelic mutant plants at this site after obtaining transgenic regenerated plants, screening out after self-fertilization The transgene-negative offspring are improved rice germplasm materials with broad-spectrum resistance to rice blast.

[0044] A method for improving rice blast broad-spectrum resistance by utilizing CRISPR / Cas9-mediated adenine base editing system according to the present invention, specifically comprises the following method steps:

[0045] (1) Determine the target site of the CRISPR / Cas9-me...

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Abstract

The invention discloses a method for improving the broad-spectrum resistance of rice blast by using a CRISPR / Cas9 mediated adenine base editing system, which uses Nipponbare rice as an acceptor material and is characterized in that a CRISPR / Cas9 mediated adenine base editing system is used for carrying out appointed replacement on a single base (A-G) which is -618 sites from the initiation codon of a promoter region of a rice Bsr-d1 gene through agrobacterium mediated transformation, after obtaining transgenic regenerated plants, identifying and screening are carried out to obtain double-allelic mutant plants of the site; and after selfing and fruiting, transgenic negative offspring, namely the improved rice planting material with broad-spectrum rice blast resistance, is carried out.

Description

Technical field: [0001] The technical field of agricultural production of the present invention specifically relates to the field of rice germplasm resource production in agricultural production, in particular to a method for improving the broad-spectrum resistance of rice blast disease by using the adenine base editing system mediated by CRISPR / Cas9. Background technique: [0002] Rice is one of the most important food crops in the world. About half of the world’s population uses rice as rations. Rice blast is one of the main diseases of rice. It is called "rice cancer" in the industry, which can cause a large reduction in rice production. The situation In severe cases, the reduction in production can reach 30% to 50%, or even no harvest. According to incomplete statistics, the loss of rice production due to rice blast in the world is more than 160 million tons. Therefore, improving the resistance of rice to blast fungus has always been an important issue in rice disease re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/46
CPCC12N15/8213C12N15/8282
Inventor 沈春修
Owner YICHUN UNIVERSITY
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