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

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

Active Publication Date: 2013-06-05
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 Measurement of Rapeseed Oil Content Separation Population

[0032] The population used in this example is the offspring of hybrids of high and low oil content parents (zy036 with oil content of 50% and 51070 with 36% respectively)—DH population. DH isolates were harvested in Wuhan in 2010, Yangluo and Qinghai in 2011, respectively. Seed oil content of parents and DH populations was determined by near-infrared analyzer. The distribution results of oil content data show that the oil content of the three experiments are all in 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 segregation populations

[0034] The total DNA of leaves was extracted by the CTAB method, and the specific steps were as follows:

[0035] A. Take 0.1 gram of fresh leaves and grind them, add 700 microliters of extract to grind them, then put them in 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;

[0036] B. Add an equal volume of phenol: chloroform: isoamyl alcohol (25:24:1, volume ratio), gently invert to mix well; centrifuge at 12000rpm for 10 minutes to make layers, gently suck the supernatant into another 1.5 ml centrifuge tube; add an equal volume of chloroform: isoamyl alcohol (24:1, volume ratio) and 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 precipitate DNA; centrifuge at 12,000 rpm for 10 minutes, pour off...

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 the simple repeat sequence (SSR) primer sequence published in published articles and Brassica database (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 that the applicant uses MISA software to search for SSR sites in the whole genome sequence of Chinese cabbage, Brassica oleracea and Brassica napus based on the sequencing results of Chinese cabbage, Brassica oleracea and Brassica napus, and develop simple repeat sequence (SSR) primers. Use the SNP function of SOAP software to predict SNP sites in the whole genome sequences of Chinese cabbage, Brassica oleracea and Brassica napus, and develop single nucleotide polymorphism (SNP) primers, named...

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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 Patents(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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