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Rape oil content character major gene resistance bit and application

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

Inactive Publication Date: 2011-03-23
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

[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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  • Rape oil content character major gene resistance bit and application
  • Rape oil content character major gene resistance bit and application
  • Rape oil content character major gene resistance bit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Construction of high and low oil content rape strain combination F2 segregation population and determination of oil content

[0032] The population used in this example is the offspring of hybrids of high and low oil content parents (zy036 with oil content of 51% 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

[0033] Example 2. Extraction of total DNA from leaves of parent, F1 and F2 segregation populations

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

[0035] A. An appropriate amount of leaf samples are taken from an ultra-low temperature refrigerator (-70°C), put them into a frozen mortar immediately, 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 pretreatment. The heated extract (0.2M Tris-Cl, 0.25NaCl, 25mM EDTA, 0.5% (mass ratio) SDS, pH 7.5) was mixed evenly, and placed in a water bath at 60°C for 40 minutes;

[0036] 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 up and down to fully mix, and centrifuge at 1300g for 10 minutes;

[0037] C. Take the supernatant into another centrifuge tube, add an equal volume of chloroform: isoamyl a...

Embodiment 3

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

[0040] The screening of primers was carried out according to the reported SSR marker primers and the self-developed molecular markers of functional genes related to oil content. Screening results showed that there were 180 pairs of primers that were different between the parents. The polymorphism screening procedure is as follows:

[0041] (1) The DNA templates of 5 strains were randomly selected from each of the parents and mixed in equal amounts to form the DNA samples of the two parents for primer screening.

[0042] (2) PCR system and procedures:

[0043] 10×buffer(MgCl2 free) 1ul

[0044] dNTPs mixture (10mM) 0.2ul

[0045] Left primer (50ng / ul) 1ul

[0046] Right primer (50ng / ul) 1ul

[0047] Taq polymerase (5U / ul) 0.1u

[0048] MgCl2 (25mM) 0.8ul

[0049] Rapeseed total DNA 2ul

[0050] ddH2o 5.9ul

[0051]

[0052] total 10ul

[0053] PCR reaction p...

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Abstract

The invention discloses a rape oil content character major gene resistance bit and application. An exploring method of the bit comprise the following steps of: 1. using a combination of zy036 with high oil content (51 percent) and Y817 with low oil content (35 percent) for hybridization to obtain F2 generation groups with the separated oil content character; and 2. using parents for sieving polymorphic primers, building a genetic map through the distribution in the F2 generation separated group, obtaining molecular markers closely linked with the obtained oil content characters according to the oil content data of the F2 group, and obtaining the major gene resistance bit. The invention discloses two rape zy036 high-oil-content character control major gene resistance bits BnOCN22-1 and BnOCN22-2 on the same linking group, and simultaneously discloses two molecular markers closely linked with the major gene resistance bits. Whether the major gene resistance QTL bits are contained in zy036 and derivative varieties (systems) of zy036 or not is detected through the closely linked molecular markers, the oil content can be predicted, and the selective efficiency of oil content breeding 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 both rank first in the world, it is far from meeting the 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 consumpti...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11A01H1/04
Inventor 王汉中华玮刘静刘贵华王新发杨庆孙美玉黄顺谋
Owner 武汉中油种业科技有限公司