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Molecular marker closely linked with oil content character of rapes and application

A molecular marker and oil content technology, applied in the fields of molecular biology and genetic breeding, can solve the problems of poor repeatability, small effect value, difficult rapeseed breeding application, etc., and achieve the effect of clear selection target, high difficulty and high cost.

Active Publication Date: 2012-11-07
武汉中油种业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are some reports on the QTL mapping of oil content in rapeseed. Usually, the number of QTLs detected is large, but the effect value 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 closely linked with oil content character of rapes and application
  • Molecular marker closely linked with oil content character of rapes and application
  • Molecular marker closely linked with oil content character of rapes and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Construction and character determination of rapeseed oil content segregated population

[0032] The population used in this example is the hybrid offspring of high and low oil content parents (zy036 with 50% oil content and 51070 with 36% oil content, respectively) - DH population. The DH isolates were harvested in Wuhan in 2010, Yangluo and Qinghai in 2011. The seed oil content of the parental and DH populations was determined by a near-infrared analyzer. The oil content data distribution results show that the oil content of the three experiments is a continuous normal distribution, and the variation range is very wide, which proves that the oil content is a quantitative trait ( figure 1 ).

Embodiment 2

[0033] Example 2. Extraction of total DNA from leaves of parental zy036, 51070 and DH isolated populations

[0034] The total DNA of leaves was extracted by CTAB method. The specific steps are as follows:

[0035] A. Get 0.1 gram of leaf fresh sample and put it into grinding, add 700 microliters of extract and grind, then put into 1.5 milliliter centrifuge tube and place in 65 ℃ of constant temperature water baths for 60 minutes, mix 2-3 times in between;

[0036] b. Add an equal volume of phenol:chloroform:isoamyl alcohol (25:24:1, volume ratio), invert gently to mix thoroughly; centrifuge at 12,000 rpm for 10 minutes to separate the layers, gently pipette the supernatant and transfer it to another. 1.5 ml centrifuge tube; add an equal volume of chloroform:isoamyl alcohol (24:1, volume ratio) to re-extract once;

[0037] c. Add 1 ml of -20°C pre-cooled absolute ethanol, freeze at -20°C for no more than 30 minutes to allow DNA precipitation; centrifuge at 12,000 rpm for 10...

Embodiment 3

[0039] Example 3. Development of primers and screening of polymorphisms

[0040] The primers used by the applicant include three categories: the first category is simple repeat sequence (SSR) primer sequences published in published articles and Brassica databases (http: / / www.brassica.info / resource / markers / ssr-exchange .php), including BN, BnEMS, BnGMS, BoGMS, BrGMS, BRAS, CB, CNU, EJU, ENA, FITO, IGF, MR, Na, Ni, Ol, Ra, niab, sN, sR and sS and many other series; The second category is based on the sequencing results of Chinese cabbage, cabbage and Brassica napus, the applicant uses MISA software to search for SSR sites on the whole genome sequences of Chinese cabbage, Brassica napus and Brassica napus, and develop simple repeat sequence (SSR) primers. Using the SNP function of SOAP software to predict SNP sites in the whole genome sequences of Chinese cabbage, cabbage and Brassica napus, and develop single nucleotide polymorphism (SNP) primers named BrSF, BrBAC, P, BoSF, SF, ...

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Abstract

The invention discloses a molecular marker closely linked with an oil content character of the rapes and an application of the molecular marker. The molecular marker BrSF34-123 closely linked with the oil content character of the rapes comprises the following screening steps: 1) hybridizing a cabbage type rape variety zy036 and 51070 so as to obtain a DH generation segregation population; 2) designing a primer to polymorphically screen a parent, and establishing a genetic linkage map; 3) carrying out an field experiment and harvesting for the DH generation segregation population to obtain a phenome database of the oil content; and 4) carrying out QTL (quantitative trait locus) detection by combining the genetic linkage map of the developed high-density molecular marker with a genotype of the segregation population and the phenome database of the oil content to obtain the molecular marker closely linked with the oil content character of rapes. Therefore, the marker is utilized to be conducted for marker-assisted selection, thereby being capable of quickly screening the strains with high oil content for breeding the oil content of the rapes, being definite in breeding selection target and saving cost.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and genetic breeding. More specifically, it relates to a molecular marker closely linked to the oil content trait of rapeseed, and also relates to the application of the molecular marker in rapeseed oil content breeding. Background technique [0002] Rapeseed is one of the most important oil crops and renewable 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 period of 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 security. As one of the four major oil crops in the world, the production of rapeseed has an important impact on ensuring the effective supply of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68A01H1/04
Inventor 王汉中孙美玉华玮刘静黄顺谋王新发刘贵华
Owner 武汉中油种业科技有限公司
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