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SNP markers closely linked with height trait of cabbage type rape, and application of SNP markers

A Brassica napus and marker technology, applied in the field of plant genetic engineering and biology, can solve problems such as unclear functional genes and their regulatory mechanisms

Active Publication Date: 2019-12-17
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In conclusion, apart from the cloning of a few dwarf genes, the functional genes and their regulatory mechanisms that play a decisive role in the establishment of rapeseed plant height are still unclear, and the relevant research needs to be further in-depth. dig further

Method used

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  • SNP markers closely linked with height trait of cabbage type rape, and application of SNP markers
  • SNP markers closely linked with height trait of cabbage type rape, and application of SNP markers
  • SNP markers closely linked with height trait of cabbage type rape, and application of SNP markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1 Obtaining of Rapeseed Dwarf Mutant DF09

[0086] The test material is NY18, which has the advantages of lodging resistance, disease resistance, cold resistance, crack angle resistance, large grain size, high yield and high combining ability. Screen NY18 large and plump seeds, soak the seeds in 1.0% EMS solution diluted in phosphate buffer (0.1M, pH=7.0) for 12 hours, rinse with tap water for 1 hour after the treatment, dry the seed surface moisture, and place the seeds (M 1 ) evenly sowed on the seedbed, and transplanted to the field when the seedlings were 35 days old. Self-bagging all individual plants at flowering stage, and harvest M at maturity stage 2 seed. M to be harvested 2 M 2 Family, 1 row for each family. Rapeseed at various stages of development, for M 2 The family was observed, and a single plant with a dwarf mutant phenotype was selected for self-bagging and harvested at maturity. 3 seed. Will M 3 Seed grows into M 3 Families, one line...

Embodiment 2

[0089] Example 2 Initial positioning of control loci for dwarf stem traits

[0090] NY18 was crossed with DF09, selfed and backcrossed to obtain P 1 ,P 2 , F 1 , F 2 , B 1 and B 2 Waiting for six basic generations, inspecting the plant height traits at the mature stage, using the plant major gene + polygene mixed genetic model (SEA-G6) for analysis, according to the principle of the minimum AIC value, 1MG-A is the most suitable inheritance for this trait Based on the model, it was preliminarily determined that the dwarf trait of DF09 was controlled by a pair of major genes.

[0091] F of NY18×DF09 2 There were 170 individual plants in the population, from which 24 high-stem extreme individual plants and 24 short-stem extreme individual plants were selected, DNA was extracted and mixed in equal amounts, two DNA mixed pools were constructed, and genome resequencing was performed together with the two parents. Based on the resequencing results of the two parents, the polym...

Embodiment 3

[0092] Example 3 Fine mapping of dwarf stem trait control loci

[0093] The Canadian material Holly was crossed with DF09, and a F containing 2536 individuals was obtained. 2 The population is planted according to the conventional cultivation method, and the young leaves are selected at the seedling stage, and the genomic DNA is extracted by the CTAB method. The plant height phenotype of all individual plants was counted at the final flowering stage.

[0094] Using the resequencing results of NY18 and DF09, SNPs were screened in the interval of the dwarf trait locus BnDwf.C9. The 200bp sequences of the upstream and downstream of the candidate SNP were extracted, and the PARMS PCR primers were designed. A typical PARMS PCR reaction system includes 5 kinds of primers: Allele 1 FAM fluorescent universal primer, Allele 2 HEX fluorescent universal primer, Allele 1 specific amplification primer (allele primer 1), Allele 2 specific amplification primer (allele Primer 2) and Locus-...

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Abstract

The invention discloses SNP markers closely linked with a height trait of cabbage type rape, and application of the SNP markers. By using an EMS induced mutation technology, a dwarf mutant DF09 with the plant height being 65 cm is obtained. The dwarf mutant DF09 is taken as a research material, a method of map-based cloning is adopted, and a dwarf trait control locus BnDwf.C9 is subjected to finemapping in the 132 Kb interval of a C09 chromosome. In the fine mapping interval, three SNP molecule markers of BnaC09-42, BnaC09-46 and BnaC09-54 which are closely linked with a dwarf trait are obtained. According to the SNP markers closely linked with the height trait of the cabbage type rape, and application of the SNP markers, through a five primer amplification blocked mutation system technology (the BnaC09-42, the BnaC09-46 and the BnaC09-54) or a conventional PCR amplification technology (BnaC09-46pcr), a dwarf material can be accurately screened according to gene types, thus the dwarfmaterial is applied to molecular identification of early target traits, the progress of dwarf breeding is accelerated, and a foundation is laid for clone of dwarf genes.

Description

technical field [0001] The invention belongs to the fields of plant genetic engineering and biotechnology. Specifically, the present invention relates to SNP markers closely linked to height traits of Brassica napus and applications thereof. Background technique [0002] Rapeseed is an important oil crop in my country, and rapeseed oil accounts for more than 55% of the total domestic vegetable oil, which plays an important role in ensuring the safety of edible oil supply in my country. Lodging can lead to a 10%-30% yield reduction in rapeseed and a significant reduction in oil content. Reducing the plant height can significantly enhance the lodging resistance of rapeseed, thereby improving the yield and quality, and is beneficial to the mechanized production of rapeseed. The "Green Revolution" of wheat and rice is one of the greatest achievements of crop breeding. The semi-dwarf varieties cultivated are widely planted around the world, and the yield has nearly doubled. The...

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 王晓东华玮张洁夫郑明陈锋张维胡茂龙彭琦陈松浦惠明
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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