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Rapid plant breeding method

A technology for plant and male sterility, applied in the field of breeding, can solve the problems of long-term cycle and limited large-scale application of haploid breeding

Active Publication Date: 2022-02-15
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the process of its reincarnation selection still needs a long period
On the other hand, although the gene editing method has been used to create a wheat induction line, its haploid induction efficiency can reach 10-20%, but because it requires complex operations such as glume clipping and detasseling of the induced female parent, it is limited. Large-scale application of haploid breeding

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Embodiment 1, rapidly create wheat DH line

[0082] 1. Using the CRISPR / Cas9 system to knock out wheat PLAs genes to obtain mutants and create wheat haploid induction lines

[0083] Using the CRISPR / Cas9 system to knock out the wheat A genome PLA-A gene (SEQ ID No.1), B genome PLA-B gene (SEQ ID No.2) and D genome PLA-D gene (SEQ ID No.3), In order to obtain a three-mutant wheat in which A genome PLA-A, B genome PLA-B and D genome PLA-D are simultaneously knocked out, hereinafter referred to as the wheat haploid induction line.

[0084] The specific preparation method of the wheat haploid induction line is as follows:

[0085] 1. Select the target

[0086] The knockout vector is designed as a single target, and the target sequence is as follows:

[0087] 5'-GACGGTGCTGACCATCGACG-3' (SEQ ID No. 6).

[0088] The sgRNA sequence and sgRNA coding DNA sequence designed for the target site sequence are as follows:

[0089] sgRNA sequence of target site: 5'-guuuuagagcuagaaa...

Embodiment 2

[0131] Embodiment 2, rapid induction of oily nauploid

[0132] 1. Using the CRISPR / Cas9 system to knock out the DMP gene of rapeseed to obtain mutants and create a rapeseed naploid induction line

[0133] Using the CRISPR / Cas9 system to knock out rapeseed DMP homologous genes BnDMP1A (SEQ ID No.7), BnDMP2A (SEQ ID No.8), BnDMP2C (SEQ ID No.9) to construct a three-mutant material, hereinafter referred to as oil menu Ploidy induction line.

[0134] The specific preparation method of the oily neuploid induction system is as follows:

[0135] 1. Select the target

[0136] Target sites were designed using the CRISPOR website (http: / / crispor.tefor.net / ). First, rapeseed was selected as the reference genome, and the sequence type of PAM was NGG. According to the "Doench 2016" score, the target sites with high editing efficiency and low off-target probability were selected. A total of 4 knockout targets were designed for the three genes, and the target sequences are as follows:

...

Embodiment 3

[0193] Embodiment 3, rapid induction Arabidopsis haploid

[0194] 1. Construct a vector expressing the sterility gene Ms7 combined with Cre / loxP recombinase and LhGR / pOp6 chemical induction system to create female dominant male sterile lines of Arabidopsis thaliana

[0195] 1. Construction of sterile vector

[0196] The specific construction method is as follows:

[0197] (1) Anther high expression promoter selection

[0198] According to the screening, the promoter p5126 highly expressed in maize anthers was selected to drive the Ms7 gene, the gene sequence is shown in SEQ ID No.14, and the p5126 promoter sequence is shown in SEQ ID No.15.

[0199] (2) Vector construction based on the Golden Gate method

[0200] A dominant sterile vector was constructed using the MoClo Toolkit kit (Kit#1000000044). Using the DNA of maize B73 as a template, the promoter p5126 sequence (SEQ ID No.15) was amplified using the primer pair p5126F / R, and cloned into the Level 0 vector pICH41295 ...

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Abstract

The invention discloses a rapid plant breeding method. The invention provides a method for obtaining a plant DH line, which comprises the following steps: hybridizing by taking a haploid induction line as a male parent and taking a heterozygous genotype dominant male sterile material or a photo-thermo-sensitive male sterile material or a sterile material subjected to chemical emasculation treatment as a female parent; and identifying and separating haploid from the F1 generation, doubling, and selecting a fertile doubled plant to obtain the target DH line. According to the rapid DH creation method provided by the invention, the period of age of pure line breeding of crops can be remarkably shortened, the workload is reduced, and the workload and cost of breeding are greatly reduced, so that the breeding efficiency of varieties is improved, and a technical support is provided for variety upgrading of new varieties.

Description

technical field [0001] The invention relates to the field of breeding, in particular to a rapid plant breeding method. Background technique [0002] Wheat, rice, etc. are self-pollinating plants, and they are also the most important food crops, which are widely grown all over the world. In recent years, my country's crop breeding level and variety performance have been continuously improved, which has continuously increased the unit yield and total output of my country's wheat, rice and other crops. With the continuous improvement of people's living conditions, the types of consumers' demands for food are constantly diversifying and the quality is further improved. High yield, disease resistance, and stress resistance must be continuously improved in processing quality, eating quality, and nutritional quality. In the final analysis, it is the demand for rapid replacement of new varieties. Traditional breeding methods for self-breeding crops such as wheat or rice generally ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/02A01H1/04A01H1/08
CPCA01H1/021A01H1/04A01H1/08
Inventor 陈绍江刘晨旭钟裕陈琛祁晓龙刘宗凯王雨文刘晋初
Owner CHINA AGRI UNIV
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