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The method of bnac2-gmyb28 gene editing to obtain seeds of low glucosinolate rape

A bnac2-gmyb28, 1. bnac2-gmyb28 technology, applied in the field of BnaC2-GMYB28 gene editing to obtain low glucosinolate rapeseed, can solve problems affecting the promotion and application of transgenic products

Inactive Publication Date: 2021-07-02
WUHAN ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the introduction of foreign genes, widespread public concerns about improved strains have been caused, which ultimately affects the promotion and application of genetically modified products

Method used

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  • The method of bnac2-gmyb28 gene editing to obtain seeds of low glucosinolate rape
  • The method of bnac2-gmyb28 gene editing to obtain seeds of low glucosinolate rape
  • The method of bnac2-gmyb28 gene editing to obtain seeds of low glucosinolate rape

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Experimental program
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Embodiment 1

[0050] 1. Experimental part:

[0051] 1.1. Determine the key genes for glucosinolate synthesis and determine the sgRNA target sequence for gene editing:

[0052] According to the sequence information in the reference genome of Brassica napus reference genome website (http: / / www.genoscope.cns.fr / brassicanapus / ), the key gene regulating glucosinolate synthesis in rapeseed was isolated from G120 material and named BnaC2-GMYB28 , the nucleotide sequence of BnaC2-GMYB28 is shown in SEQ ID No.1;

[0053] Gene expression analysis was carried out in tissues of rapeseed at different stages, and the results showed that BnaC2-GMYB28 was expressed in cotyledons of 7 days after sowing, young leaves of 30 days after sowing, mature leaves of 60 days and 90 days after sowing, and flower buds of 120 days of rapeseed , expressed in siliques pollinated 7 days and 14 days after flowering and ovules fertilized 18 days after flowering.

[0054] Submit the BnaC2-GMYB28 gene sequence to the CRISPR-...

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Abstract

The method for BnaC2-GMYB28 gene editing to obtain low glucosinolate rapeseed seeds, comprising the following steps: determining key genes for glucosinolate synthesis and determining the sgRNA sequence of the target gene; constructing an expression cassette; constructing a gene editing vector; transforming the gene editing vector into Agrobacterium; sowing rape seeds ; Agrobacterium activation preparation; explant preparation and dipping; explant transfer and culture. The present invention determines the endogenous gene that regulates the synthesis of glucosinolates in rapeseed seeds, uses the gene editing technology to directional mutation, and obtains individual plants of several different mutation types, whose seed glucosinolates are significantly reduced compared with the control, and after selfing The glucosinolate content of the obtained T1 family seeds was extremely significantly lower than that of the unedited recipient materials, indicating that this technology can quickly obtain non-transgenic exogenous fragments, low glucosinolate phenotype and stable genetic rapeseed.

Description

technical field [0001] The present invention designs the technical field of genetic engineering, particularly relates to the method for BnaC2-GMYB28 gene editing to obtain low glucosinolate rapeseed. Background technique [0002] Rapeseed is one of the three major oil crops in the world. The cake after oil extraction from rapeseed is rich in protein and is a good natural animal feed. However, the content of glucosinolate (glucosinolate for short) in rapeseed cake is high, and the rapeseed cake of its hydrolyzate is fed to poultry and livestock, which will cause toxicity to poultry and livestock, thus greatly limiting the use of rapeseed cake in feed use value. The content of glucosinolates in rapeseed determines the value of rapeseed meal as feed. Reducing the content of glucosinolates in rapeseed meal is an important goal in quality breeding of rapeseed. After decades of hard work by rapeseed breeders, the quality of rapeseed varieties has been greatly changed, among whi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/29
CPCC07K14/415C12N15/8205C12N15/8243
Inventor 万丽丽
Owner WUHAN ACADEMY OF AGRI SCI
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