Molecular marker BrSF0239 primer of main-effect QTL loci in florescence and maturity of Brassica napus and application thereof

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, small repeatability, high technical difficulty, etc., and achieves convenient, fast, accurate and rapid detection methods, and improved selection. The effect of efficiency

Active Publication Date: 2019-07-12
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 BrSF0239 primer of main-effect QTL loci in florescence and maturity of Brassica napus and application thereof
  • Molecular marker BrSF0239 primer of main-effect QTL loci in florescence and maturity of Brassica napus and application thereof
  • Molecular marker BrSF0239 primer of main-effect QTL loci in florescence and maturity of Brassica napus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Construction and trait determination of segregating populations at flowering stage of rapeseed:

[0028] In this 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 anthesis and maturity phenotypes of the two parents and segregating populations were identified after harvest at maturity. The test data showed that the mean 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

[0030] Primer development and synthesis:

[0031] 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

[0033] The screening process of primer polymorphism is as follows:

[0034] (1) Randomly select 10 strains of DNA from each parent and mix them in equal amounts as templates for screening primers.

[0035] (2) Utilize the primers after dissolving to carry out PCR amplification to the parental DNA,

[0036] reaction system:

[0037]

[0038] PCR reaction program:

[0039]

[0040]

[0041] (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 particularly discloses a molecular marker BrSF0239 primer for main-effect QTL loci in florescence and maturity of Brassica napus and an application thereof. Phenotypic data of florescence characters were obtained by field experiments and character investigation on F2 and F2: 3 pedigree segregated populations of Zhongshuang No. 11 (a kind of oilseed rape) and 73290 (a kind of oilseed rape); genotype and genetic map of the F2 segregated populations are combined for QTL detection, and the main-effect geneloci, on an A2 linkage group, for joint controlling the florescence and the maturity are obtained, specifically, the contribution rate to the florescence of oilseed rape is 17.2% with additive effect2.1 and dominant effect 0.9; and the contribution rate to the maturity of the oilseed rape is 9.4% with the additive effect 0.6 and the dominant effect 0.4. By detecting the molecular markers relatedto the characters in the maturity, the length of the maturity can be predicted, and then early-maturing varieties can be selected quickly and accurately.

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 BrSF0239 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, and occupies a very important position in the national edible oil supply (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 coun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 王汉中师家勤朱瑶瑶王新发刘贵华詹杰鹏
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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