Molecular marker InDeL_33 of main-effect QTL (quantitative trait locus) of soybean hundred-grain weight and application of molecular marker InDeL_33

A molecular marker, 100-grain weight technology, used in recombinant DNA technology, microbial assay/inspection, DNA/RNA fragments, etc.

Inactive Publication Date: 2016-05-04
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Domestically, the breeding of special soybean varieties is still relatively weak, but with the expansion of people's consumption demand for bean sprouts and natto, the corresponding breeding of small-grained soybean varieties has also been paid more and more attention.

Method used

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  • Molecular marker InDeL_33 of main-effect QTL (quantitative trait locus) of soybean hundred-grain weight and application of molecular marker InDeL_33
  • Molecular marker InDeL_33 of main-effect QTL (quantitative trait locus) of soybean hundred-grain weight and application of molecular marker InDeL_33
  • Molecular marker InDeL_33 of main-effect QTL (quantitative trait locus) of soybean hundred-grain weight and application of molecular marker InDeL_33

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Acquisition of main effect QTL for 100-grain weight in soybean

[0039] (1) 'NJRINP' group building

[0040]The hybrid F 1 , construct the population through the "single-seed transmission" method, and obtain 284 F 2:8 Generation of recombinant inbred lines ('NJRINP').

[0041] The steps of the "single-seed transmission" method are as follows: a seed harvested from the female plant in the present generation of parental parental hybridization grows into a F1 generation single plant, and its self-crossing (ie self-pollination) yields a F1 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 comple...

Embodiment 2

[0049] Example 2: Acquisition of the main QTL tight marker InDeL_33 for 100-grain weight in soybean

[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 Gm11:2261898bp-65p77664bp region of chromosome 11.

[0052] (2) Secondary population molecular marker identification

[0053] Use the kit to extract the genomic DNA of the leaves of NJRINP-derived secondary population soybean material, 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; exte...

Embodiment 3

[0058] Embodiment 3: the application of InDeL_33 molecular marker on the selection of soybean 100-grain weight

[0059] (1) Genome amplification detection of both parents

[0060] The parents used to verify the QTL of soybean 100-kernel weight were the parent Nannong 86-4 (the InDeL_33 allele band was 100bp, the 100-kernel weight was 18.46g), the parent NP-7 (the InDeL_33 allele band was 120bp, the 100-kernel weight is 8.16g).

[0061] (2) Population amplification detection and marker analysis

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Abstract

The invention discloses a molecular marker InDeL_33 of a main-effect QTL (quantitative trait locus) of soybean hundred-grain weight and an application of the molecular marker InDeL_33. The main-effect QTL of the soybean hundred-grain weight is located on a NO.11 chromosome Gm11:2261898bp-6577664bp of the soybean; an upstream primer sequence of the molecular marker InDeL_33 is represented as Seq ID NO.1, and a downstream primer sequence is represented as Seq ID NO.2. In a built secondary group, to-be-detected F2 single-plant genome DNA (deoxyribonucleic acid) is taken as a template, an InDeL_33 primer pair is adopted to perform PCR (polymerase chain reaction) amplification, an amplification product is detected, and a to-be-detected material with a specific DNA fragment in the size of 120bp in the amplification product is a candidate small-grain soybean material. By the aid of the molecular marker, large-grain and small-grain soybean materials can be screened simply and conveniently, and the small-grain soybean breeding process can be accelerated.

Description

technical field [0001] The invention relates to soybean molecular marker assisted breeding, belongs to the field of soybean genetic breeding, and in particular relates to the molecular marker InDeL_33 of the main effect QTL of soybean 100-grain weight and its application. Background technique [0002] Soybean 100-seed weight is one of the important indicators to measure the size of soybean kernels, and it is an important factor that constitutes yield. The 100-kernel weight of soybeans varies greatly. The 100-kernel weight of cultivated soybeans is usually between 6 and 55g, and the 100-kernel weight of wild soybeans is generally between 1 and 3g. The 100-grain weight below 12g is generally referred to as small-grain soybeans. Compared with larger soybeans, small soybeans have the advantages of high protein, high unsaturated fatty acids, crude fiber, high calcium and zinc content, low fat, sugar, and a suitable ratio of linolenic acid and linoleic acid, so they can be used a...

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 赵团结常芳国盖钧镒孔杰杰王吴彬
Owner NANJING AGRICULTURAL UNIVERSITY
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