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Molecular marker of major QTL (Quantitative Trait Locus) of soybean seed protein content and application thereof

A technology of protein content and molecular markers, which is applied in the field of genetic breeding and achieves good application prospects

Inactive Publication Date: 2013-04-17
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although a large number of major QTLs for soybean grain protein content have been identified at present, it is still a long-term goal and challenge for soybean geneticists and breeders to discover new major QTLs and markers closely linked to major QTLs

Method used

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  • Molecular marker of major QTL (Quantitative Trait Locus) of soybean seed protein content and application thereof
  • Molecular marker of major QTL (Quantitative Trait Locus) of soybean seed protein content and application thereof
  • Molecular marker of major QTL (Quantitative Trait Locus) of soybean seed protein content and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1. Acquisition of main effect QTL for soybean grain protein content

[0039] (1), 'NJRSXG' group construction

[0040] Hybrid F 1 , through the "single-seed transmission" method to construct the population and obtain 147 F 2:7 Generation recombinant inbred line ('NJRSXG').

[0041] The steps of the "single-seed transmission" method are as follows: a seed harvested from the female parent plant in the parental hybrid generation grows into a F1 generation single plant, and its self-crossing (that is, self-pollination) yields 1 F plant 2 generation seeds, the latter is planted to grow into a F2 generation line containing segregation traits, and each individual plant of it is self-fertile and harvests F 3 For generation seeds, the isolated plants of the same traits are harvested and threshed into bags, and each seed is planted separately in the next year. 3 F 4 Generation seeds, ... until the characters of different plants in each family are completely stable and ...

Embodiment 2

[0049] Example 2. Acquisition of major QTL tight marker Z56 for protein content in soybean grain

[0050] (1) Molecular marker development

[0051] Using the published soybean physical map information and the sequence information of the sequencing materials of the National Soybean Improvement Center, multiple molecular markers were designed in the Gm08:13000kb-17000kb region of chromosome 8.

[0052] (2) Phenotype determination of local materials

[0053] Soybean local materials for analysis consisted of 366 accessions from 6 ecological regions of the country, and dried soybean seeds were milled using a 1095 Knifetec sample mill (FOSS Tecator, Switzerland). Using VECTOR 22 / N Fourier transform infrared spectrometer to measure the spectrum of soybean seed powder, and use the established soybean protein content model to predict the protein content of soybean seeds.

[0054] (3) Molecular marker identification of local materials

[0055] Using CTAB method to extract genomic DNA...

Embodiment 3

[0060] Example 3. Application of Z56 Molecular Marker in Selection of Protein Content in Soybean Grain

[0061] (1) Genome amplification detection of both parents

[0062] The parents used to verify the QTL for soybean grain protein content were parent WT133 (Z56 allele band is 155bp, soybean seed protein content is 49.9%), parent XG30 (Z56 allele band is 185bp, soybean seed protein content is 46.9%), XG30 Although the seed protein content is smaller than that of WT133, it contains a potentiating site at the Z56 site.

[0063] (2), population amplification detection and marker analysis

[0064] Using the above two parents to cross, get F 1 After the seed is planted, it grows into a F 1 Generation single plant, its self-crossing is fruitful, and one F2 generation seed is harvested, and the latter grows into a F2 generation containing segregation traits. 2 Generation line, on F 2 Harvesting seeds from a single plant F 2：3 The adult plant row was planted, two replicates wer...

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Abstract

The invention discloses a molecular marker of the major QTL (Quantitative Trait Locus) of soybean seed protein content and application thereof. The major QTL locus of the soybean seed protein content is positioned on the No. eight chromosome Gm08:13000-17000 kb of a soybean. The forward primer sequence of the molecular marker is SEQ ID NO.1, the reverse primer sequence of the molecular marker is SEQ ID NO.2. In newly allocated hybridized combination, a Z56 primer pair is used for PCR (Polymerase Chain Reaction) amplification by taking the genomic DNA (Deoxyribose Nucleic Acid) of an F2 individual soybean seed to be measured as a template, amplification products are detected, and the soybean to be measured, which is included in the amplification products and has a specific DNA fragment with the size of 180-190bp, is a candidate material with high seed protein content. The molecular marker disclosed by the invention can be used for easily and fast screening the soybean seed high-protein materials and can accelerate the soybean seed high-protein content breeding.

Description

technical field [0001] The invention belongs to the field of genetic breeding, and relates to molecular markers and applications of soybean grain protein content main effect QTL. Background technique [0002] Soybean grain protein is the most important vegetable protein source in the world today, and its content is about 40%. Breeding soybean materials with high grain protein content is one of the important goals of soybean quality breeding. [0003] Traditional breeding for protein content in soybean grains is based on phenotypic selection. The determination of soybean grain protein content phenotype can be determined by chemical method and spectroscopic method. Determining the protein content of soybean seeds based on chemical methods requires destruction of soybean seeds and requires a lot of time and labor. Based on the spectroscopic method, the protein content of soybean seeds can be determined quickly and accurately. However, the spectroscopic method needs to establ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 盖钧镒张英虎赵团结
Owner NANJING AGRICULTURAL UNIVERSITY
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