Brassica napus seed number per silique major gene locus close linkage molecular markers and applications thereof

A technology of molecular markers and rapeseed, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of ineffective discovery and utilization of favorable genes of germplasm resources, lack of independent intellectual property rights and breeding value genes and labeling, poor selection effect of complex quantitative traits, etc., to achieve the effect of convenient and fast detection method, accurate and fast screening, and saving production cost

Active Publication Date: 2018-03-13
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, compared with developed countries, there is still a big gap in the study of rapeseed molecular breeding in my country, which is mainly reflected in: the inability to effectively discover and utilize favorable genes in germplasm resources, the lack of genes and markers with independent intellectual property rights and breeding value

Method used

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  • Brassica napus seed number per silique major gene locus close linkage molecular markers and applications thereof
  • Brassica napus seed number per silique major gene locus close linkage molecular markers and applications thereof
  • Brassica napus seed number per silique major gene locus close linkage molecular markers and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: Construction of QTL-NIL

[0026] According to the strategy of fine positioning ( figure 1 ), NIL was constructed against qSN.A6. In this experiment, the hybrid F 1 During this period, the molecular markers BrSF47-389 and BnID108 on both sides of qSN.A6 were used to select heterozygous individual plants and double 11 consecutive backcrosses in the recurrent parents for 4 generations to obtain BC 4 f 1 generation. Then, 80 molecular markers (Yang et al., 2016) other than the target main effect QTL and evenly distributed on the 19 linkage groups of rapeseed were used for genetic background screening, from BC 4 f 1 In the population, select a single plant that is heterozygous for the target fragment and has a background recovery rate >95% to self-cross to obtain BC 4 f 2 seed. Field Planting BC 4 f 2 Generation of QTL-NIL segregation populations, using the above molecular markers to screen recombinant individual plants, and harvest test species at th...

Embodiment 2

[0027] Example 2: Development of Molecular Markers

[0028] First, use self-designed primers to compare with the genome sequence of Chinese cabbage or Brassica napus to determine the genomic region corresponding to the target QTL. This genomic region was searched for SSRs using MISA software. Using BWA software, the parental No.73290 resequencing sequence was mapped to the reference genome sequence of Shuang11 in the parent, and the InDel site of the target QTL interval was found by samtools software. Then, use Primer3.0 software to design SSR / InDel primers.

[0029] Using the magnetic bead method to extract the genomic DNA of the leaves of Shuang11 and No.73290 and QTL-NIL populations in the parents, using the self-developed SSR / InDel primers, and PCR amplification of the parental DNA, the products were in denaturing polyacrylamide gel After electrophoresis, staining and developing, the size of the bands was discriminated, and polymorphic primers were screened.

[0030] PC...

Embodiment 3

[0036] Example 3: Genotype analysis of QTL-NIL segregation population and screening of recombinant individual plants

[0037] (1) Using the magnetic bead method to extract the genomic DNA of the QTL-NIL isolated population;

[0038] (2) Using the molecular markers developed above to carry out molecular marker analysis on the genomic DNA of the QTL-NIL segregation population;

[0039] (3) Analyze the genotypes of these molecular markers in the QTL-NIL segregation population and screen recombinant individual plants.

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Abstract

The invention discloses brassica napus seed number per silique major gene locus close linkage molecular markers and applications thereof, wherein the close linkage markers can be used for the molecular marker assisted selection breeding of brassica napus and map-based cloning of the major QTL. According to the present invention, hybrid F1 and a parent strain are subjected to continuous backcross aplurality of generations, a near iso-genic line is constructed by combining a molecular marker assisted selection method, a heterozygous single plant with high background recovery rate is selected and is subjected to selfing to obtain a QTL-NIL separate population, the molecular marker of the target interval is gradually encrypted, the genotype is analyzed to find the exchange single plant, finally the target interval is reduced to achieve the physical distance of only 88 Kb, the molecular marker is gradually encrypted, the genotype is analyzed to find the exchange single plant so as to obtain the molecular markers such as BrSF46-28 and BrSF46-78, the genotyping is further performed on the expanded NIL population by using the two closely linked molecular markers, and the results show thatthe seed numbers per silique between different alleles have significant difference; and the molecular markers of the present invention can be used for the molecular marker assisted selection breedingof brassica napus, can improve the selection efficiency, and can accelerate the breeding.

Description

technical field [0001] The invention belongs to the technical field of molecular breeding of rapeseed, and in particular relates to a molecular marker closely linked with main effect QTL loci of seed number per corner of rapeseed and an application thereof. Background technique [0002] Rapeseed is one of the most important oil crops in the world, and it is also one of the oil crops with the highest oil production efficiency. my country is a big consumer of oil crops in the world, but its supply has been in short supply for a long time (Wang Hanzhong, 2010). In 2013, the proportion of my country's vegetable oil imports was as high as 64.7% (Wang Hanzhong and Yin Yan, 2014). Facing the serious shortage of domestic production and the increasingly high dependence on foreign countries, it is imperative to vigorously develop the rapeseed industry and increase the production of rapeseed to solve the consumption of edible vegetable oil in my country. Under the same planting densit...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/6895
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 王汉中师家勤杨玉花张江江刘贵华王新发
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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