Oil content character main gene locus of rape 6F313 and applications thereof

A technology of main effect genes and gene loci, which is applied in the field of molecular biology and genetic breeding, can solve the problems of small effect value and difficult application of rapeseed breeding, and achieve the effect of high difficulty, convenient and fast detection, and clear position

Active Publication Date: 2014-03-26
武汉中油种业科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there are some reports on the QTL mapping research on rapeseed oil content. Usually, the number of QTLs detected is large, but the effect value is small, and it is difficult to apply in rapeseed breeding.

Method used

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  • Oil content character main gene locus of rape 6F313 and applications thereof
  • Oil content character main gene locus of rape 6F313 and applications thereof
  • Oil content character main gene locus of rape 6F313 and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Construction of F2 Segregation Population of Combination of High and Low Oil Content Rapeseed Lines and Determination of Oil Content of F2 and F2:3 Families

[0039] The population used in this example is the offspring of the hybrids of high and low oil content parents (6F313 with oil content of 50% and Y817 with 35% respectively)—the F2 generation. The oil content of the parents, F1 and F2 seeds was determined by a near-infrared analyzer. The distribution of oil content data in the F2 generation segregation population shows that the distribution of oil content is continuous, and the variation distribution is normal, and the variation range is very wide, which proves that oil content is a quantitative trait ( figure 1 ).

Embodiment 2

[0041] Extraction of Total DNA from Leaves of Parents, F1 and F2 Segregation Populations

[0042] The total DNA of leaves was extracted by the CTAB method, and the specific steps were as follows:

[0043] A. An appropriate amount of leaf samples are taken from an ultra-low temperature refrigerator (-70°C), immediately put them into a frozen mortar, add liquid nitrogen and grind them into powder; quickly put them into a 50ml centrifuge tube, and add them to a water bath at 60°C for pre-treatment. Heat the extract solution (0.2M Tris-Cl, 0.25NaCl, 25mM EDTA, 0.5% (mass ratio) SDS, pH7.5), mix well, put it in a water bath at 60°C for 40min;

[0044] B. Take out the centrifuge tube, add an equal volume of chloroform: isoamyl alcohol (24:1, V / V), slowly invert the centrifuge tube 30-50 times to mix well, and centrifuge at 1300g for 10 minutes;

[0045] C. Take the supernatant in another centrifuge tube, add an equal volume of chloroform: isoamyl alcohol (24:1, V / V), and extract ag...

Embodiment 3

[0048] SNP microarray analysis of genotype and linkage analysis of 6F313, Y817 parents and F2 segregation population

[0049] The 60K Rapeseed SNP chip developed by Illumina was used to type the DNA samples of the two parents and the F2 segregation population of Rapeseed. After the polymorphic SNP sites were screened in the parents, their distribution in the F2 population was analyzed. Through the data analysis of the distribution in the F2 generation segregation population, according to the law of linkage exchange, the genetic map of rapeseed was constructed by using the population genotype data. The software used was Joinmap3.0, and the minimum LOD value was set to 2.5 to obtain the linkage map ( figure 2 It is the A10 chromosome linkage group map). Input the oil content data of 105 individual plants of the F2 population and the distribution of polymorphic primers into the computer, run the WinQTL cart4.0 software to perform correlation analysis on the data, and measure th...

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Abstract

The invention discloses an oil content character main gene locus of rape 6F313 and applications thereof. The method to find the locus comprises the following steps: (1) hybridizing a combination of rape 6F313 with a high oil content (50%) and rape Y817 with a low oil content (35%) so as to obtain F2 generation population with separated oil content character and a F2:3 family; (2) establishing a linkage map of F2 generation segregation population by utilizing the SNP chip of rape 60K, then obtaining the molecular label tightly linked with the oil content character and the main gene locus according to the oil content data of the F2 population and the F2:3 family. The invention discloses a main gene locus 6F313A10OC which locates on the A10 chromosome and controls the high oil content character of rape 6F313, and discloses two molecular labels 6F313A10OC-1 and 6F313A10OC-2 which are tightly linked with the main gene locus at the same time. The tightly linked molecular labels can detect whether rape 61616 or derivated species (line) contains a main gene QTL locus or not, thus can predict the oil content, so the breeding efficiency of high oil content rape is greatly improved.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and genetic breeding. More specifically, it relates to a main effect gene locus of rapeseed oil content and a molecular marker closely linked to the main effect gene locus, and also relates to the application of the molecular marker in rapeseed oil content breeding. Background technique [0002] 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 edible vegetable oil and feed protein in my country, improving food structure, and promoting the development of aquaculture and processing industries. Although my country's rapeseed planting area and output rank first in the world, it is far from meeting domestic consumption demand, and about 63% of edible vegetable oil depends on imports. With the growth of population and the improvement of people's living standards, it is estimated that the consumption of edi...

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 王汉中华玮刘静刘贵华王新发胡志勇杨红丽
Owner 武汉中油种业科技有限公司
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