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

Active Publication Date: 2020-05-05
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
At present, there are some reports on the QTL mapping research on rapeseed grain weight, but usually the detected QTL effect value is small and the reproducibility is not good, so it is difficult to apply in rapeseed breeding

Method used

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  • Molecular marker primer of major gene locus of rape grain weight character and application of molecular marker primer
  • Molecular marker primer of major gene locus of rape grain weight character and application of molecular marker primer
  • Molecular marker primer of major gene locus of rape grain weight character and application of molecular marker primer

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention belongs to the technical field of molecular biology and genetic breeding, and particularly discloses a molecular marker primer of a major gene locus of a rape grain weight character andan application of the molecular marker primer. According to the invention, field experiments and seed tests are carried out on F2:3 families of Zhongshuang 11 and 73290 to obtain phenotypic data of the grain weight character; and QTL detection is carried out by combining the genotype and genetic map of the F2 segregation population to obtain a major gene locus qSW.A7 and a molecular marker BoSF2344 for controlling the grain weight of rape on an A7 linkage population. According to the marker, genotype analysis is carried out on F3 generations derived from two parents, the average 1000-grain weight (4.28g) of a selected single plant carrying a favorable gene exceeds the average 1000-grain weight (3.95g) of a single plant carrying an unfavorable gene, and the seed test result shows that the proportion that the grain weight of single plants carrying a favorable marker is higher than the grain weight of single plants carrying the unfavorable gene is as high as 89.8%, so that selection efficiency of high-yield breeding can be greatly improved by using the marker for auxiliary selection.

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 primer for a main effect gene locus of rapeseed grain weight, and also relates to the application of the molecular marker in high-yield breeding of rapeseed. Background technique [0002] Rapeseed is the largest oilseed crop in my country, accounting for about 20% of the world's rapeseed output. 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 security strategy. In addition, rapeseed oil and diesel oil have relatively similar fatty acid composition, which is a kind of green and renewable energy. As my country's urbanization scale continues to expand and the area of ​​arable land further shrinks, increasing the oil yield per unit area of ​​rapeseed (= per unit...

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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