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A method for gene mapping of Brassica napus wax powder based on whole genome resequencing

A Brassica napus, the whole genome technology, applied in the field of bioinformatics, can solve the problems of low detection efficiency and heavy workload, and achieve the effect of qualified quality and sufficient sequencing data

Active Publication Date: 2022-03-04
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the method of mapping the wax powder gene of Brassica napus needs to detect all offspring individuals. This detection method has a large workload and low detection efficiency.

Method used

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  • A method for gene mapping of Brassica napus wax powder based on whole genome resequencing
  • A method for gene mapping of Brassica napus wax powder based on whole genome resequencing
  • A method for gene mapping of Brassica napus wax powder based on whole genome resequencing

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] The parental materials are the homozygous dimorphic line D0721, the temporary protection line D1577 and the restorer line D0936 grown in the Zhuanghang Experimental Base of the Institute of Crop Breeding and Cultivation, Shanghai Academy of Agricultural Sciences.

[0061] 1) The sterile plant of the homozygous two-type line D0721 of Brassica napus was used as the mother, and the temporary insurance line D1577 was used as the male parent, and crossed to obtain a complete sterile line, a complete sterile line and a restorer line D0936 with no wax powder traits. Hybridization to obtain the F1 generation;

[0062] 3) From the 15:1 segregation population of the F2-F5 generation, the plants with the wax powder trait were selected for selfing to obtain the F6 generation.

Embodiment 2

[0064] 1 source of material

[0065] The test materials are the homozygous two-type line D0721, the temporary protection line D1577 and the restorer line D0936 planted in the Zhuanghang Experimental Base of the Institute of Crop Breeding and Cultivation of the Shanghai Academy of Agricultural Sciences. Wax plants and all with the wax phenotype did not segregate populations.

[0066] 2 methods

[0067] 2.1 Library construction and sequencing

[0068] The genomic DNA in the sample is extracted, and the library is constructed after the electrophoresis test is qualified. Qualified DNA samples were first randomly fragmented into 350bp fragments by Covaris, and the TruSeq DNA LT Sample Prepkit kit was used to build a library. The DNA fragments underwent end repair, poly(A) tailing, sequencing adapters, purification, and PCR amplification. and other steps, and finally complete the library construction, and use the sequencer to perform double-end sequencing after the library is qua...

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Abstract

The invention provides a gene positioning method of brassica napus wax powder based on whole genome resequencing, which belongs to the technical field of biological information. The present invention establishes near-isogenic lines on the basis of segregating populations. Using mixed grouping analysis, non-wax plants in the 3:1 segregation population of near isogenic lines, all non-segregation populations with wax phenotype and 3 parents were selected, and 5 DNA pools were constructed for genome resequencing. The resequencing data was used for genetic association analysis, and the loci controlling the traits of wax powder were located in the 590663-1657546bp region of the A08 chromosome. The biggest feature of the method of the present invention is that it does not need to detect all progeny individuals, but conducts mixed pool analysis on the progeny of two extreme traits. Moreover, the sequencing data of the method of the present invention are sufficient and of acceptable quality, and can be used for the next step of analysis.

Description

technical field [0001] The invention relates to the technical field of biological information, in particular to a gene positioning method of brassica napus wax powder based on whole genome resequencing. Background technique [0002] Brassica napus L. is one of the three major types of rapeseed, which evolved from double diploidization of Chinese cabbage (AA, n=10) and Brassica napus (CC, n=9) through natural interspecific hybridization. A compound species. The surface of the plant is generally covered with a layer of wax. Wax is a mixture of long-chain fatty acids, which can prevent ultraviolet radiation damage, reduce excessive evaporation of water on the plant surface and protect plants from diseases and insect pests, and affect the photosynthetic efficiency and pollen fertility of plants. Generally speaking, the stems, leaves and siliques of Brassica napus are covered with wax powder, while the stems, leaves and siliques of the non-wax mutant are not covered with wax po...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6858A01H1/02A01H1/04G16B30/10G16B50/30
CPCC12Q1/6895C12Q1/6858A01H1/02A01H1/04G16B30/10G16B50/30C12Q2600/156C12Q2600/13C12Q2535/122C12Q2537/165
Inventor 张俊英朱吉风周熙荣杨立勇江建霞蒋美艳李延莉王伟荣孙超才
Owner SHANGHAI ACAD OF AGRI SCI
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