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Method for improving wheat scab resistance through genome editing

A scab and resistance technology, applied in genetic engineering, plant genetic improvement, botany equipment and methods, etc., can solve the problem of nutrient loss

Active Publication Date: 2022-08-09
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Editing wheat susceptibility genes can significantly improve the resistance of wheat to pathogens, however, the editing of susceptibility genes may lead to the occurrence of excessive immunity in wheat and loss of its own growth and development Nutrition in the process, so how to balance wheat disease resistance and maintain normal growth and development?

Method used

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  • Method for improving wheat scab resistance through genome editing
  • Method for improving wheat scab resistance through genome editing
  • Method for improving wheat scab resistance through genome editing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] Example 1. Preparation of recombinant plasmids

[0109] The genomic and coding sequences on chromosomes A, B and D of the TaCIPK14 gene were amplified from DNA and cDNA of wheat variety Fielder. The nucleotide sequence of TaCIPK14 in the A genome is represented by TaCIPK14-4A, the nucleotide sequence of TaCIPK14 in the B genome is represented by TaCIPK14-4B, and the nucleotide sequence of TaCIPK14 in the D genome is represented by TaCIPK14-4D. Amplification results showed that the full-length TaCIPK14 gene was 1335 bases without introns. In the wheat Fielder genome, the nucleotide sequence of the TaCIPK14 gene in the A genome (corresponding to the A chromosome group) is shown in SEQ ID No. http: / / plants.ensembl.org / index.html) in the wheat gene sequence similarity is 100%; the nucleotide sequence of the TaCIPK14 gene in the B genome (corresponding to the B chromosome group) is as shown in SEQ ID No. .2 (encoding the protein shown in SEQ ID No. 5), the similarity with ...

Embodiment 2

[0116] Example 2. Using gRNA1 and gRNA2 to obtain gene-edited wheat by Agrobacterium-mediated genetic transformation

[0117] 1. Agrobacterium-mediated genetic transformation of wheat

[0118] The gene editing recombinant plasmid VK005-06-g1g2 constructed in Example 1 was transformed into immature immature embryos of the wheat variety Fielder through Agrobacterium-mediated genetic transformation, followed by differentiation, screening, regeneration and rooting to finally obtain regenerated plants. The genetic transformation of wheat is completed by the genetic transformation platform of the State Key Laboratory of Crop Stress Biology in Arid Regions of Northwest A&F University, and the public can contact the platform for commercialization to complete the transformation operation.

[0119] 2. Detection of gene editing

[0120] 1. To T 0 Identification of regenerated plants

[0121] The test plants are: 22 T plants obtained in step 1 0 Generation of regenerated plants, Field...

Embodiment 3

[0175] Example 3. Analysis of resistance to wheat scab

[0176] The test plants were Fielder plants, homozygous mutant lines in which three copies of TaCIPK14 were simultaneously knocked out in wheat A, B, and D genomes, including KO-1, KO-4 and KO-5 prepared in Example 2.

[0177] Carry out the identification of resistance to scab on the test plant, and its steps are:

[0178] (1) The cultivation of the meristems selects the wild-type strain PH-1 of Fusarium graminearum, the pathogen of wheat scab, as the strain, and the wild-type strain PH-1 of F. Gifted by the teacher, recorded in "Li Yimin et al. Functional verification of histone deacetylase genes (HDACs) in Fusarium graminearum. Northwest A&F University, 2010, Master's thesis", the public can obtain from the applicant, only It can be used to repeat the experiment of the present invention, and it shall not be used for other purposes. The strain was cultured and activated on PDA solid medium, and a solid with Gibberella ...

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Abstract

The invention discloses a method for improving wheat scab resistance through genome editing. The invention provides application of TaCIPK14 protein or related biological materials thereof in regulating and controlling the resistance of plants to gibberellic disease. The TaCIPK14 protein is a TaCIPK14-4A protein (SEQ ID No. 4), a TaCIPK14-4B protein (SEQ ID No. 5) and / or a TaCIPK14-4D protein (SEQ ID No. 6), and the TaCIPK14 protein is a TaCIPK14-4A protein (SEQ ID No. 4), a TaCIPK14-4B protein (SEQ ID No. 5) and / or a According to the invention, three alleles of TaCIPK14 on wheat are simultaneously edited by using a CRISPR-Cas9 gene editing technology, so that a TaCIPK14 gene knockout mutant plant is successfully obtained. The TaCIPK14 gene knockout mutant shows resistance when inoculated with gibberellic disease bacteria, and the main agronomic trait evaluation result proves that the TaCIPK14 gene knockout mutant still maintains the main agronomic traits. The invention is of great significance to prevention and control of wheat scab.

Description

technical field [0001] The invention relates to the technical field of breeding of disease-resistant varieties, in particular to a method for improving the resistance of wheat scab through genome editing. Background technique [0002] Fusarium head blight (FHB) is a fungal disease that severely disrupts global wheat production, causing billions of dollars in damage to wheat production each year (McMullen et al., 2012). Since 2000, due to the influence of global warming, the prevalence of wheat scab has become more and more serious, and the epidemic years of wheat scab have increased significantly, and the yield loss of some fields has reached as high as 70% (Yerkovich et al., 2017). Since 2010, the occurrence area of ​​wheat scab has continued to increase. According to the data of the National Agricultural Technology Promotion Center, the average annual occurrence area of ​​wheat scab in my country from 2011 to 2015 reached 82 million mu, of which the incidence of wheat scab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H6/46A01H5/00C12N9/12C12N15/54
CPCC12N9/1205C12N15/8282C12N15/8218
Inventor 郭军何付新康振生孙慧琳柏星轩王策王炎峰
Owner NORTHWEST A & F UNIV