Molecular marker primer of major gene locus of rape grain weight character and application of molecular marker primer
A technology of main effect genes and molecular markers, applied in the field of molecular biology and genetic breeding, can solve the problems of small QTL effect value, difficult rapeseed breeding application, high quality requirements, etc., to achieve accurate and rapid screening, convenient and fast detection methods, and location clear effect
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Embodiment 1
[0027] Construction and trait determination of rapeseed grain weight segregation population:
[0028] The segregation populations used in this example are the F2 derived from the parents of large and small rapeseed Zhongshuang 11 and 73290 (planted at the Yangluo Comprehensive Experimental Base of the Institute of Oil Plants, Chinese Academy of Agricultural Sciences in 2009) and its F2: 3 family line (2010, In 2011, it was planted at the Yangluo Comprehensive Test Base of the Institute of Oil Plants, Chinese Academy of Agricultural Sciences). The grain weight phenotypes of the two parents and the population were identified after harvesting at the maturity stage. The grain weight test data showed that the two parents had weak superparent segregation, indicating that the large-grain gene was mainly distributed in the genome of Zhongshuang 11; the population grain weight was normally distributed, which proved the quantitative genetic characteristics of the grain weight trait ( f...
Embodiment 2
[0031] Primer development and synthesis:
[0032] 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 InDel 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 using BWA software, and then Use samtools software to find InDel.
Embodiment 3
[0034] The screening process of primer polymorphism is as follows:
[0035] (1) Randomly select 10 strains of DNA from each parent and mix them in equal amounts as templates for screening primers.
[0036] (2) Utilize the primers after dissolving to carry out PCR amplification to the parental DNA,
[0037] reaction system:
[0038]
[0039] PCR reaction program:
[0040]
[0041] (3) Gel electrophoresis band pattern interpretation
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