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Molecular marker in close linkage with rape crotch angle character QTL (Quantitative Trait Loci) and application

A branching angle and molecular marker technology, applied in the field of molecular biology and genetic breeding, can solve problems such as heavy workload, long process cycle, and constraints on the progress of QTL positioning, and achieve accurate and rapid screening, saving production costs, and convenient detection methods quick effect

Active Publication Date: 2016-09-28
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, the traditional QTL mapping method needs to rely on the construction of mapping populations and the development of molecular markers. The process cycle is long and the workload is huge, which seriously restricts the progress of QTL mapping.

Method used

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  • Molecular marker in close linkage with rape crotch angle character QTL (Quantitative Trait Loci) and application
  • Molecular marker in close linkage with rape crotch angle character QTL (Quantitative Trait Loci) and application
  • Molecular marker in close linkage with rape crotch angle character QTL (Quantitative Trait Loci) and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Construction and trait determination of rapeseed branching angle segregation population:

[0036] In this example, the F2 segregation population was constructed by crossing the large-branch angle material Huyou 19 and the small-branch angle material Purler. At the maturity stage of rapeseed, 277 lines and two parents were randomly selected from the F2 population to measure the branch angle. For the specific sampling and measurement methods, see the method of Wang Wenxiang et al. (2015).

Embodiment 2

[0038] Extraction of total DNA from leaves of parents and F2 populations:

[0039] Utilize CTAB method to extract total DNA from leaves, the specific steps are as follows:

[0040] (1) Take 0.5 grams of fresh leaves and put them into a 2 ml centrifuge tube, add 200 microliters of 2 times CTAB extract, then add steel balls and place them on an automatic grinder for grinding. After grinding, add 500 microliters to the centrifuge tube Put 2 times the CTAB extract in a constant temperature water bath at 65°C for 30 minutes, during which time mix 2-3 times;

[0041] (2) Add an equal volume of chloroform:isoamyl alcohol (24:1, V / V), invert gently to make it fully mixed, centrifuge at 10000rpm for 10 minutes, gently draw 400 microliters of supernatant and transfer to another 1.5ml centrifuge tube;

[0042] (3) Add 1 / 10 volume (40 microliters) of sodium acetate to the supernatant, then add 2 times the volume of -20°C pre-cooled absolute ethanol, freeze at -20°C for half an hour to a...

Embodiment 3

[0045] QTL identification of branching angle in Brassica napus:

[0046] (1) In the F2 population, 30 copies of DNA from individual plants with extremely large branching angles were selected and mixed to form a large-angle pool, and 30 copies of DNA from plants with extremely small branching angles were mixed to form a small-angle pool. The HiSeq 2500 performs whole genome sequencing on two mixed pools and two parents, each pool constructs a 350bp library, the sequencing depth of the two DNA pools is 40×, and the sequencing depth of the two parents is 16×;

[0047] (2) Align the sequences obtained from the small-angle pool and the large-angle pool with the Brassica napus Darmor genome using BWA software, and then use SAMtools software (Li&Durbin, 2009) for SNP analysis, referring to SNP- in Takagi et al. (2013) The index calculation method calculates the SNP-index of all SNPs detected in the small-angle pool and the large-angle pool, and the difference between the subtractions...

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Abstract

The invention discloses a molecular marker in close linkage with rape crotch angle character QTL (Quantitative Trait Loci) and application. The molecular marker is characterized by carrying out hybridization by taking cabbage type rape variety Shanghai oil 19 as a female parent and Purler as a male parent, establishing F2 segregation population through selfing of hybrid F1, and analyzing, thus obtaining a crotch angle locus qBA1.A06; designing a primer by utilizing an Indel marker at the boundary of the crotch angle locus qBA1.A06 so as to detect the parents and the F2 population, thus obtaining a molecular marker BAIndel76 and a molecular marker BAIdenl79 which are in close linkage with the crotch angle character QTL; identifying a rape gene type formed after hybridization of the two parents by utilizing a marker primer, and carrying out auxiliary selection by utilizing the marker, thus greatly increasing the selection efficiency. According to the molecular marker disclosed by the invention, a novel genetic marker is provided for molecular breeding of the rape plant type, and useful information is also provided for accurate positioning on the crotch angle character QTL of the cabbage type rape and map-based cloning on related genes.

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 closely linked to a QTL locus for the branch angle trait of rapeseed and its application. Background technique [0002] Rapeseed is the largest oil crop in my country, and its planting area and output rank among the top in the world. The oil production of rapeseed accounts for 55% of the entire domestic oil crops, and occupies a major position in the supply of domestic vegetable oil. Therefore, increasing the planting area of ​​rapeseed and increasing the yield of rapeseed are of great significance to ensure the security of edible vegetable oil supply in my country (Wang Hanzhong, et al. 2014). However, with the continuous rise of labor prices and production costs, the production efficiency of rapeseed has been significantly reduced, which has led to stagnant or even declining rapeseed acreage in recent years. Poo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 胡琼成洪涛王会汪文祥刘佳梅德圣郝梦宇付丽周日金
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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