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Cabbage type rape grain weight-associated molecular marker and preparation method and application thereof

Brassica napus and molecular marker technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve problems such as difficulty in finding tightly linked molecular markers, limited genetic differences and genetic recombination, and achieve Improve selection efficiency and accuracy, and speed up the effect of breeding process

Active Publication Date: 2015-07-29
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

With the development of molecular marker technology, some quantitative trait loci of rape seed weight have been located by linkage analysis (BASUNANDA et al.2010; FAN et al.2010; QUIJADA et al.2006; RADOEV et al.2008 ; S HI et al.2009; UDALL et al.2006; YANG et al.2012; ZHANG et al.2011), however, the traditional linkage analysis is based on the population constructed by crossing two parents, and the genetic differences and genetic recombination are very limited, so The QTL mapping interval is often large (10-20cM) and it is difficult to find closely linked molecular markers

Method used

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  • Cabbage type rape grain weight-associated molecular marker and preparation method and application thereof
  • Cabbage type rape grain weight-associated molecular marker and preparation method and application thereof
  • Cabbage type rape grain weight-associated molecular marker and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Extraction of total DNA from leaves by CTAB method

[0028] Specific steps are as follows:

[0029] A. Take 0.1 gram of fresh leaves and grind them, add 700 microliters of extract to grind them, then transfer them to a 1.5 milliliter centrifuge tube and place them in a constant temperature water bath at 65°C for 60 minutes, during which they mix 2-3 times;

[0030] B. Add an equal volume of phenol: chloroform: isoamyl alcohol (25:24:1, V / V / V), gently invert to mix well, centrifuge at 12000rpm for 10 minutes, gently suck the supernatant into Another 1.5 ml centrifuge tube; add an equal volume of chloroform: isoamyl alcohol (24:1, V / V) and re-extract once;

[0031] C. Add 1 ml of -20°C pre-cooled absolute ethanol, freeze at -20°C for no more than 30 minutes to precipitate DNA, centrifuge at 12,000 rpm for 10 minutes to precipitate DNA, pour off the ethanol solution in the centrifuge tube; wash with 75% ethanol for 2 -3 times, pour off the soaking solution, open the cap ...

Embodiment 2

[0034] Molecular Marker Analysis of Rapeseed Germplasm Population

[0035] Proceed as follows:

[0036] (1) adopt embodiment 1 to extract the DNA of 576 parts of rapeseed germplasm resources;

[0037] (2) The 576 materials contained the mapping parents of the high-density genetic linkage map constructed by our laboratory, Shuang11 (sequencing) and 73290 (resequencing), in order to determine the genetic position of the SSR markers used. A total of 91 SSR markers with a single locus inheritance pattern (CHEN et al. 2008) distributed uniformly on all chromosomes were selected for PCR amplification in 576 materials.

[0038] reaction system:

[0039]

[0040] PCR reaction program:

[0041]

[0042] (3) Perform polypropylene gel electrophoresis, development, staining and band pattern interpretation on PCR products.

[0043] Reagent preparation:

[0044] A.5×TBE

[0045] Tris-base 53.9g

[0046] EDTA 3.72 g

[0047] Boric acid 27.5g

[0048] Make up to 1 L with ultrapur...

Embodiment 3

[0086] Structural Analysis of Germplasm Resource Population

[0087] Use Structure2.3 software to analyze the group structure of the association analysis group. The results of Monte Carlo simulation (k from 2 to 20; the number of iterations is 4, and the number of repetitions is 10000) show that the group is most suitable for being divided into two groups, corresponding to winter and semiwinter / spring rapeseed varieties, while semiwinter and spring varieties were separated again when divided into 3 groups.

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Abstract

The invention discloses a cabbage type rape grain weight-associated molecular marker and a preparation method and an application thereof. The preparation of the molecular marker comprises the following steps: 1) by the utilization of 91 unit points of SSR markers uniformly distributed on an oilseed rape genetic map, 576 varieties of oilseed rape from all around the world undergo genotype and population structure analysis and association analysis is carried out with the combination of thousand grain weight data character data; 2) 14 high-quality SSR markers are developed near the marker BrSF7-181 by the utilization of cabbage and oilseed rape whole genome sequence information so as to carry out association analysis once again, wherein BrSF6-2025 has the highest significance level; and 3) by the utilization of the molecular marker, grain weight extreme varieties in genetic resource populations are verified. The invention provides a new genetic marker for molecular breeding of rape grain weight and also provides useful information for map-based cloning of cabbage type rape grain weight genetic locus.

Description

technical field [0001] The invention belongs to the field of rapeseed molecular breeding and biotechnology, and more specifically relates to a molecular marker associated with the grain weight of Brassica napus, a preparation method of a molecular marker associated with the grain weight of Brassica napus, and a method related to the grain weight of Brassica napus Application of molecular markers associated with grain weight in Brassica napus for high-yield breeding. Background technique [0002] Rapeseed is one of the most important rapeseed and energy crops. Its fatty acid carbon chain length and structure are close to those of fossil diesel, and it is a vegetable oil suitable for biodiesel production. At present and for a long time in the future, the whole world will face the severe situation of shortage of edible vegetable oil and fossil energy. Therefore, a substantial increase in the total supply of rapeseed oil is a major demand to ensure global food and energy securit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 王汉中师家勤李娜刘贵华王新发杨庆
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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