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Molecular marker indel15-1 of soybean early-flowering-season major QTL (quantitative trait locus) and application thereof

A molecular marker, first flowering technology, applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of long breeding cycle and low selection efficiency

Inactive Publication Date: 2016-06-15
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Since molecular marker-assisted selection is not easily affected by environmental factors and the interference of traits showing recessiveness, the target trait gene can be selected from the molecular level, which overcomes the shortcomings of low selection efficiency and long breeding cycle in simple conventional breeding methods based on phenotype selection.

Method used

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  • Molecular marker indel15-1 of soybean early-flowering-season major QTL (quantitative trait locus) and application thereof
  • Molecular marker indel15-1 of soybean early-flowering-season major QTL (quantitative trait locus) and application thereof
  • Molecular marker indel15-1 of soybean early-flowering-season major QTL (quantitative trait locus) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Acquisition of the main QTL for the initial flowering stage of soybean

[0036] (1) 'NJZNRIL' group building

[0037] The hybrid F 1 , construct the population through the "single-seed transmission" method, and obtain 236 F 2:8 Generation of a recombinant inbred line population ('NJZNRIL').

[0038] The steps of the "single-seed transmission" method are as follows: a seed harvested from the female plant in the parental hybrid generation grows into a F 1 Generation single plant, its self-crossing (i.e. self-pollination) yields fruit and harvests 1 F 2 Generation seeds, which grow into a F containing segregating traits 2 Generation line, each individual plant of it self-fertilizes 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 an...

Embodiment 2

[0047] Example 2: Acquisition of the main QTL closely linked marker indel15-1 for soybean flowering stage

[0048] (1) Molecular marker development

[0049] Using the published soybean physical map information and the sequence information of the sequencing materials of the National Soybean Improvement Center, molecular markers were designed in the Gm15:48600kb-49850kb region of chromosome 15.

[0050] (2) Secondary population phenotype determination

[0051] (3) Secondary population molecular marker identification

[0052] Genomic DNA from leaves of NJZNRIL-derived secondary population soybean material was extracted by CTAB method, and the amplification experiment was carried out with indel15-1 primer pair. PCR reaction system (10ul), which contains 3ul DNA template (15ng), upstream and downstream primers (0.2mmol / L) each 1.5ul, 1.2ulMgCl 2 (2.5mmol / L), 0.24uldNTP (10mmol / L, N=A, C, G, T), 0.12ulTaq enzyme (5U / ul) and 1.4ulddH 2 O. PCR reaction program: denaturation at 95...

Embodiment 3

[0057] Embodiment 3: the application of indel15-1 primer pair in the selection of soybean initial flowering stage

[0058] (1) Genome amplification detection of both parents

[0059] Two groups of materials with longer and shorter flowering stages were randomly selected from the NJZNRIL population to verify the QTL for soybean flowering stage.

[0060] (2) Population amplification detection and marker analysis

[0061] The genomic DNA of leaves of each material was extracted by CTAB, and the amplification experiment was carried out with the indel15-1 primer pair. PCR reaction system (10ul), which contains 3ul DNA template (15ng), upstream and downstream primers (0.2mmol / L) each 1.5ul, 1.2ulMgCl 2 (2.5mmol / L), 0.24uldNTP (10mmol / L, N=A, C, G, T), 0.12ulTaq enzyme (5U / ul) and 1.4ulddH 2 O. PCR reaction program: denaturation at 95°C for 5 min; followed by 35 cycles of denaturation at 94°C for 30 s, annealing at 55°C for 40 s, extension at 72°C for 50 s; extension at 72°C for ...

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Abstract

The invention discloses a molecular marker indel15-1 of soybean early-flowering-season major QTL (quantitative trait locus) and application thereof. The soybean early-flowering-season major QTL is positioned on the 48600kb-49850kb interval of the soybean 15th chromosome Gm15, and the molecular marker is positioned on the 49588kb of the soybean 15th chromosome Gm15. The forward primer sequence of the molecular marker is Seq ID NO.1, and the reverse primer sequence is Seq ID NO.2. Among the two groups of materials with different early flowering seasons in the NJZNRIL population amplified by the primers and the newly prepared secondary population, the material containing 180bp specific DNA segment has a larger early-flowering-season phenotypic number. The molecular marker disclosed by the invention can be used for screening or identification of the length of the early flowering season for soybean varieties or lines, thereby enhancing the breeding efficiency.

Description

technical field [0001] The invention relates to soybean molecular marker assisted breeding, which belongs to the field of soybean genetic breeding, and in particular relates to the molecular marker indel15-1, which is the main effect QTL of the initial flowering stage of soybean, and its application. Background technique [0002] Soybean flowering stage is one of the important growth traits. It is a complex quantitative trait and is easily affected by environmental factors. So far, at least 70 soybean flowering stage QTLs have been published on Soybase, distributed in 20 linkage groups. The initial flowering stage of soybean is not only an important node controlling the transition from vegetative growth to reproductive growth in soybean, but previous research results also showed that the initial flowering stage of soybean has an impact on many yield-related traits. In different soybean producing areas, there are corresponding ecological types, so accurate prediction of soybe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 赵团结曹永策盖钧镒李曙光孔杰杰杨成凤
Owner NANJING AGRICULTURAL UNIVERSITY