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Molecular marker of pot shattering resistance trait major gene locus of rapes and application

A technology of molecular markers and major genes, applied in the field of molecular biology and genetic breeding, can solve the problems of quantitative traits being easily affected by environmental conditions, long cycle, poor selection effect, etc., and achieve convenient, fast and high-cost detection methods , the effect of improving the selection efficiency

Active Publication Date: 2013-07-03
OIL CROPS RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Quantitative traits are susceptible to environmental conditions, so selection does not work well
The long cycle of traditional breeding methods is mainly caused by quantitative traits

Method used

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  • Molecular marker of pot shattering resistance trait major gene locus of rapes and application
  • Molecular marker of pot shattering resistance trait major gene locus of rapes and application
  • Molecular marker of pot shattering resistance trait major gene locus of rapes and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Construction of Rapeseed Anti-crack Angle Segregation Population and Determination of Characters

[0032] The populations used in this example are F2 and F2:3 populations of the offspring of rapeseed with high and no resistance to cleavage angle (Zhongshuang 11: cleavage angle resistance index 0.83; 73290: cleavage angle resistance index 0.48). The cleavage-horn resistance phenotypes of the parents, F2 and F2:3 populations were identified after harvesting at maturity. The results of anti-cracking angle index of F2 and F2:3 segregation populations showed that: the anti-cracking angle index of the two populations showed a continuous distribution, but the variation distribution did not show a typical normal distribution, which proved that the anti-cracking angle trait was a quantitative trait and there was a main effect gene locus ( figure 1 ).

Embodiment 2

[0033] Example 2. Parents, F2 and F2:3 Extraction of Total DNA of Segregation Population Leaves

[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 rapeseed leaf fresh sample and grind it, add 700 microliters of extract to grind it, then put it into a 1.5 milliliter centrifuge tube and place it in a constant temperature water bath at 65°C for 60 minutes, during which it mixes 2-3 times;

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

Embodiment 3

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

[0040] The SSR primers we used include two types: one is the primer sequences published in published articles and Brassica database (http: / / www.brassica.info / resource / markers / ssr-exchange.php), including CB, CNU, niab, MR, FITO, Ol, BRMS, BRAS, Ni, Na, BN, NGA, MB, BnGMS, BrGMS, BoGMS, BnEMS, and many other series; the other type is spliced ​​​​by our sequencing of cabbage and cabbage The scaffold sequences were developed and named BrSF and BoSF series respectively. The specific development method is to use SSRHunter software to search for SSRs in each scaffold, and then use Primer3.0 software to design SSR primers. We have synthesized a total of 3085 pairs of public and newly developed SSR primers through the biological company, and the screening results showed that 15% of the amplified products of the primers had length polymorphism differences between the parents. The polymorphism scree...

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Abstract

The invention discloses a molecular marker of a pot shattering resistance trait major gene locus of rapes and an application. The steps are as follows: 1) hybridizing double 11 variety and 73290 variety (which have obvious differences in the pot shattering resistance traits) in a cabbage type rape varieties, and carrying out selfing on progenies to obtain F2 and F2:3 segregation populations with the segregated pod shattering resistance traits; 2) carrying out polymorphic screening on parent DNA (deoxyribonucleic acid) by utilizing an SSR (simple sequence repeat) primer, and establishing a genetic linkage map by carrying out SSR molecular marker genotyping on the F2 progeny segregation populations; 3) carrying out field experiments and plant inquisition on the F2 and F2:3 segregation populations so as to obtain phenotype data of the pod shattering resistance traits; and 4) carrying out QTL (quantitative trait locus) detection by combining with the developed high-density molecular marker genetic linkage map and genotype and the phenotype data of the segregation populations, utilizing QTL Cart2.5 software to obtain the major gene locus Psr.A9 for controlling the pod shattering resistance of the rapes on a cabbage type rape A9 linkage group and also obtain the molecular marker BrSF0007-39 closely linked with the major gene locus. The selection efficiency of breeding with the pod shattering resistance can be greatly improved by utilizing the marker for assistant selection.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and genetic breeding, and in particular relates to the main gene locus of the brassica napus anti-cracking angle trait and its closely linked molecular markers, and also relates to the application of the molecular markers in the breeding of rape with high cracking-horn resistance . Background technique [0002] Rapeseed is the third largest oil crop in the world and the fifth largest crop in my country after rice, wheat, corn and soybean. About 13% of the world's vegetable oil comes from rapeseed. In addition to being the main raw material for edible oil, rapeseed is also an important industrial raw material and a major biological resource for renewable energy in the main member states of the European Union. The planting area and total output of my country's rapeseed account for about 30% of the world's total. It is not only related to the adjustment and optimization of my country's pla...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 王汉中华玮胡志勇刘贵华王新发师家勤黄顺谋杨庆
Owner OIL CROPS RES INST CHINESE ACAD OF AGRI SCI
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