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Molecular marker primers of major gene loci of NSS (Number of Seeds per Silique) trait of rape and application of molecular marker primers

A molecular marker and molecular marker-assisted technology, which is applied in the fields of molecular biology and genetic breeding, can solve the problems of small QTL effect value, small repeatability, difficult rapeseed breeding application, high quality requirements, etc., and achieves convenient and rapid detection methods and low selection efficiency. , the effect of clear location

Active Publication Date: 2019-04-26
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 also some reports on the QTL mapping research on the number of kernels per corner 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 primers of major gene loci of NSS (Number of Seeds per Silique) trait of rape and application of molecular marker primers
  • Molecular marker primers of major gene loci of NSS (Number of Seeds per Silique) trait of rape and application of molecular marker primers
  • Molecular marker primers of major gene loci of NSS (Number of Seeds per Silique) trait of rape and application of molecular marker primers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Construction and trait determination of rapeseed segregated populations per corner:

[0026] The segregation populations used in this example are F2 (planted in 2009 at the Yangluo comprehensive test of the Institute of Oil Plants, Chinese Academy of Agricultural Sciences) derived from the parents of multi-grain and few-grain rapeseed Zhongshuang 11 (≈21 grains) and 73290 (≈11 grains). Base) and its F2:3 (planted at the Yangluo Comprehensive Test Base of the Institute of Oil Plants, Chinese Academy of Agricultural Sciences in 2010) and F2:4 family (planted at the Qinghai University Test Base in 2011). The phenotypes of the number of seeds per corner of the two parents and the two populations were identified after harvesting at the maturity stage. The test data of the number of seeds per corner shows that the two parents have weak superparental segregation, indicating that the multigrain gene is mainly distributed in the genome of Zhongshuang 11; Quantitative genetic tr...

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 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

[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 fields of molecular biology and genetic breeding and particularly discloses molecular marker primers of major gene loci of an NSS (Number of Seeds per Silique) trait of rape and application of the molecular marker primers. Phenotype data of the NSS trait are obtained through subjecting F2 and F2 :3 family separated colonies of Zhongshuang 11# and 73290 to field experiment and seed examination; QTL (Quantitative Trait Locus) detection is carried out with reference to genotypes and genetic maps of an F2 separated colony. The major gene loci, i.e., qSN.A7 and a molecular marker Ni201 for controlling NSS of the rape on an A7 linkage group are obtained. Through subjecting F3 derived from two parent plants to genotype analysis by the marker, a result indicates that an NSS mean of selected single plants carrying over favorable genes exceeds that of single plants carrying over unfavorable genes; proven by seed examination results, a mean of NSS of singleplants carrying over favorable markers is higher than that of single plant colonies carrying over the unfavorable genes by 88.0%; therefore, the efficiency of selection of high-yield seed breeding canbe increased greatly through carrying out auxiliary selection by using the marker.

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 the rape seed number per corner trait, 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 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 security strategy (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 compo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
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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