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Seed number per pod character major gene site of rape and application thereof

A main effect gene, rapeseed technology, applied in the field of molecular biology and genetic breeding, can solve the problems of small repeatability of QTL effect value, difficult application of rapeseed breeding, etc., to achieve convenient and fast detection method, accurate and fast screening, saving The effect of production costs

Inactive Publication Date: 2012-11-21
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are also some reports on the QTL mapping research on the number of kernels per corner of rapeseed, but the QTL effect value usually detected is small and the repeatability is not good, so it is difficult to apply in rapeseed breeding

Method used

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  • Seed number per pod character major gene site of rape and application thereof
  • Seed number per pod character major gene site of rape and application thereof
  • Seed number per pod character major gene site of rape and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Construction and trait determination of rapeseed segregated populations per corner:

[0034] The isolated populations used in this example are the F2 and F2:3 populations derived from Shuang11 (≈21 grains) and 73290 (≈11 grains) of rapeseed with many and few grains. The phenotypes of the number of seeds per corner of the two parents and the two populations were identified after harvesting at the maturity stage. The number of seeds per corner species test data shows that the two parents have weak super-parental segregation, indicating that the multigrain gene is mainly distributed in the genome of Zhongshuang 11; the number of seeds per corner of the two populations is normally distributed, proving the trait of number of seeds per corner Quantitative genetic characteristics of ( figure 1 ).

Embodiment 2

[0036] Extraction of total DNA from leaves:

[0037] The total DNA of leaves was extracted by the CTAB method, and the specific steps were as follows:

[0038] (1) Take 0.1 g of fresh leaves and grind them, add 700 microliters of extract to grind them, then transfer them to a 1.5 ml centrifuge tube and place them in a constant temperature water bath at 65°C for 60 minutes, during which they mix 2-3 times;

[0039] (2) 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, 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;

[0040] (3) Add 1 ml of -20°C pre-cooled absolute ethanol, and freeze at -20°C for no more than 30 minutes to precipitate DNA;

[0041] Centrifuge at 12000rpm for 10 minutes to allow DNA to precipitate, discard the ethanol solution in the centrifuge tube; wash 2-3 times ...

Embodiment 3

[0044] Primer development and synthesis:

[0045] The SSR primers utilized by the applicant include two types: one is the primer sequences published in published articles and Brassica databases (http: / / www.brassica.info / resource / markers / ssr-exchange.php); the other The series were developed by the applicant based on the sequences of Chinese cabbage and cabbage scaffolds, 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. The self-developed SNP primers are obtained by comparing the resequencing sequence of 73290 with the reference genome sequence of Zhongshuang11. First, the resequencing sequence of 73290 is positioned on the whole genome reference sequence of Zhongshuang11 using BWA software, and then Use samtools software to find SNP. The SNP detection adopts the SNAP (single nucleotide amplified polymorphism) method, that is, a misma...

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Abstract

The invention discloses a seed number per pod character major gene site of rape and application thereof. The process comprises the following steps: (1) hybridizing by Brassica napus variety with obvious difference on each seed number per pod character; (2) carrying out polymorphism screening for parent DNA by public and developed SSR (Simple Sequence Repeat) and SNP (Single Nucleotide Polymorphism) primers, and building a genetic linkage map by molecular mark gene type analysis to an F2-generation segregation population; (3) obtaining the phenotype data of each seed number per pod character by field experiments and variety research to F2 and F2:3 segregation population; and (4) combining the gene type and the phenotype data of the segregation population to carry out QTL (Quantitative Trait Loci) detection. The major gene site qQN.A6 and the molecular marker BrSF50-18 for controlling seed number per pod of rape on an A6 linkage group are obtained. The F3 generation derived from two parents is analyzed by the marker, and an individual plant with the marker is kept. A variety research result shows that the ratio of the seed number per pod is 83.4% higher than the F3 individual plate of F2:F3 family mean value, and thus, the marker is used for assisted selection to greatly improve the selection efficiency of high-yield breeding.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and genetic breeding, and more specifically relates to a molecular marker of a main effect gene locus of the grain number per corner trait of Brassica napus, and also relates to the application of the molecular marker in high-yield breeding of rapeseed. Background technique [0002] Rapeseed is one of the most important oil and 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. In the form of continuous expansion of urbanization scale and further reduction of cultivated land area, continuous incr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/11C12Q1/68
Inventor 王汉中师家勤刘贵华王新发华玮黄顺谋
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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