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Molecular marker brsf2604 primer for major qtl loci at flowering and maturation stages of Brassica napus and its application

A Brassica napus and molecular marker technology, which is applied in the fields of molecular biology and genetic breeding, can solve the problems of difficult rapeseed breeding application, high quality requirements, high technical difficulty, etc., and achieves convenient and rapid detection methods, clear locations, and improved selection. The effect of efficiency

Active Publication Date: 2022-03-15
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, AFLP markers also have some disadvantages: high cost, complex process, and technical difficulty; most of the markers are dominant markers; there are high requirements for DNA quality and restriction endonuclease quality
The genetic research results of flowering stage in Brassica napus all show that flowering stage is a quantitative trait controlled by multiple genes and has major gene effects (Zaman et al., 1995; Lagercrantz et al., 1996; Osborn et al., 1997; Cai Changchun et al., 2007) At present, there are some reports on the QTL mapping research on the flowering stage and maturity stage of rapeseed, but the QTL effect value usually detected is small and the repeatability is not good, so it is difficult to apply in rapeseed breeding

Method used

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  • Molecular marker brsf2604 primer for major qtl loci at flowering and maturation stages of Brassica napus and its application
  • Molecular marker brsf2604 primer for major qtl loci at flowering and maturation stages of Brassica napus and its application
  • Molecular marker brsf2604 primer for major qtl loci at flowering and maturation stages of Brassica napus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Construction and trait determination of segregating populations at flowering stage of rapeseed:

[0027] In the example, Shuang11, a rapeseed variety sequenced, was used to cross with another rapeseed variety 73290 with large genetic differences, and the F1 generation of the hybrid was selfed to generate the F2 population and its F2:3. Among them, the Yangluo Comprehensive Experimental Base of the Institute of Oil Plants, Chinese Academy of Agricultural Sciences planted F2 populations in 2009, and planted F2:3 family populations in 2010, 2011 and 2012. The survey data of traits showed that the average values ​​of flowering stage and maturity stage in the three environments were normally distributed, indicating that the quantitative genetic characteristics of flowering stage and maturity stage traits ( figure 1 , figure 2 )

Embodiment 2

[0029] Primer development and synthesis:

[0030] The SSR primers used by the applicant include two types: one is the published primer sequence in published articles and Brassica database (http: / / www.brassica.info / resource / markers / ssr-exchange.php); the other is It was developed by the applicant based on the sequences of cabbage and cabbage scaffolds, named BrSF and BoSF series respectively. The specific development method is to use SSR Hunter software to search for SSRs in each scaffold, and then use Primer3.0 software to design SSR primers. The self-developed InDe l primers are obtained by comparing the resequencing sequence of 73290 with the reference genome sequence of Zhongshuang11. First, the resequencing sequence of 73290 is positioned on the whole genome reference sequence of Zhongshuang11 by using BW A software , and then use samtools software to find InDel.

Embodiment 3

[0032] The screening process of primer polymorphism is as follows:

[0033] (1) Randomly select 10 strains of DNA from each parent and mix them in equal amounts as templates for screening primers.

[0034] (2) Utilize the primers after dissolving to carry out PCR amplification to the parental DNA,

[0035] reaction system:

[0036]

[0037] PCR reaction program:

[0038]

[0039] (3) Gel electrophoresis band pattern interpretation

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Abstract

The invention belongs to the technical field of molecular biology and genetic breeding, and discloses the molecular marker BrSF2604 primer and its application of the main QTL site of flowering stage and maturity stage of Brassica napus. The applicant passed F2 and F2 of Zhongshuang 11 and 73290 : 3 family segregation populations conducted field experiments and trait investigations to obtain phenotypic data of flowering period traits; combined with the genotype and genetic map of F2 segregation populations, QTL detection was carried out, and the C2 linkage group jointly controlled the flowering and maturity stages of rapeseed. The main effect gene locus has a contribution rate of 21.1% to the flowering period of rapeseed, an additive effect of ‑2.8, and a dominant effect of 0.3; a contribution rate of 10.4% to the maturity stage of rapeseed, an additive effect of ‑0.3, and a significant The sex effect was 0.4. By detecting the molecular markers related to the traits of flowering and maturity, the length of flowering and maturity can be predicted, so as to accurately and quickly screen early-flowering and early-maturing varieties.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and genetic breeding, and more specifically relates to a molecular marker BrSF2604 primer for main QTL loci in flowering stage and maturity stage of Brassica napus and its application. Background technique [0002] Rapeseed is the largest oilseed crop in my country, accounting for about 20% of the world's rapeseed production (Hu et al., 2016). Rapeseed oil is the largest source of domestic edible vegetable oil, accounting for 57.2% of the total domestic edible vegetable oil (Fan Chengming et al., 2018). my country's foreign dependence on domestic vegetable oil has exceeded 60%, and there is a tendency to continue to increase (Wang Hanzhong et al., 2014). In addition, rapeseed oil and diesel oil have relatively similar fatty acid composition, which is a kind of green and renewable energy. [0003] Although my country is the largest rapeseed oil producer in the world, the supply of veget...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 王汉中师家勤朱瑶瑶詹杰鹏王新发刘贵华
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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