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Indel/SNP molecular marker related to flavor character of fresh edible soybeans and application of Indel/SNP molecular marker

A molecular marker and fragrance technology, applied in the field of agricultural biology, can solve the problems of high cost, volatile and low accuracy of content identification

Pending Publication Date: 2021-12-17
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to its extremely low content, high volatility and instability, the identification of its content is costly, inaccurate and difficult

Method used

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  • Indel/SNP molecular marker related to flavor character of fresh edible soybeans and application of Indel/SNP molecular marker
  • Indel/SNP molecular marker related to flavor character of fresh edible soybeans and application of Indel/SNP molecular marker
  • Indel/SNP molecular marker related to flavor character of fresh edible soybeans and application of Indel/SNP molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 HRM molecular marker detection method of fresh soybean aroma major gene GmBADH2

[0025] 1. Experimental equipment

[0026] Ultra-low temperature refrigerator (Thermo, the United States); high-speed refrigerated centrifuge (Eppendorf, Germany); gene mutation detector ( 480Ⅱ, Switzerland); adjustable micropipette (Eppendorf, Germany); chemiluminescent gel imaging system (G:BOX, China); electrophoresis instrument (BIO-RAD, USA).

[0027] 2. Soybean Genomic DNA Extraction

[0028] The leaves of soybean seedlings were taken, and the whole genome DNA of soybean was extracted with a plant genome DNA extraction kit (Solarbio, China). After extraction, 1% agarose gel electrophoresis is used to detect the quality of the DNA; a micro-nucleic acid concentration analyzer is used to measure the DNA concentration.

[0029] 3. Primer design

[0030] Specific primers were designed for the GmB ADH2 gene segment of chromosome 5 of the Glycine max Wm82.a2.v1 genome in the P...

Embodiment 2

[0039] Example 2 Verification test of the molecular marker in hybrid parents (Danbohei and Quxian No. 1)

[0040] 1. Test materials and methods

[0041]Danbahei comes from the Institute of Agricultural Sciences, Zhejiang Academy of Agricultural Sciences, and has no fragrance. Quxian No. 1 comes from Quzhou Academy of Agriculture and Forestry, and has fragrance. The genomic DNA of Danbahei and Quxian No. 1 was extracted, tested according to the steps in Example 1, and the genotype of the samples was judged according to the melting curve peak.

[0042] The method of GC-IMS was used to measure the 2AP content of Danbahei and Quxian No. 1, specifically: picking fresh pods, boiling the water and putting the soybeans in, taking them out 5-10 minutes after the water boiled again, removing the shells and putting them in the GC-IMS. IMS incubator. The detection conditions of the gas chromatography ion mobility spectrometer are: analysis time 20min, chromatographic column type FS-SE-5...

Embodiment 3

[0048] Example 3 Verification Test of Soybean Aroma-Related Indel / SNP Molecular Markers in Hybrid Populations

[0049] 1. Test materials and methods

[0050] A hybrid combination was prepared with Danbohei and Quxian 1 as parents to obtain hybrid seeds, the pseudo hybrids were eliminated in the F1 generation, F1 generation seeds were obtained, and F2 generation plants were obtained by field sowing; Genomic DNA was extracted from a single plant, and detected according to the steps in Example 1, and the genotype of the sample was judged according to the peak of the melting curve. The determination of 2AP content is the same as implementation case 2.

[0051] 2. Results and analysis

[0052] Of the 117 F2 individuals, 26 individuals had a melting temperature of 80°C and were homozygous recessive genotypes. The average content of 2 AP was 6.19 μg / g, which was similar to that of the parent Quxian No. 1; 32 individuals The melting temperature of the 59 individual plants was 80℃ a...

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Abstract

The invention provides application of an Indel / SNP molecular marker in flavor character detection of fresh edible soybeans. The nucleotide sequence of the molecular marker is as shown in SEQ ID No.1, 11 basic groups are deleted at the 40th bp position, AGA at the 36bp position is mutated into GAG, GT at the 52bp position is mutated into TC, CCG at the 56bp position is mutated into AAT, and TC at the 62bp position is mutated into CT. The invention further provides a primer sequence for detecting the Indel / SNP site and an identification method based on a high-resolution dissolution curve (HRM), genotypes of the site can be distinguished according to different dissolution curve peaks of target DNA sequence fragments, and then homozygosis and heterozygosis of the fragrance-related functional gene GmBADH2 of a sample are judged. By utilizing the Indel / SNP marker and the detection method, the individual genotype can be judged in a fresh edible soybean seedling stage, the early selection and early elimination of the flavor character are realized, so that the Indel / SNP marker and the detection method have important practical values for improving the seed selection efficiency of the flavor quality of the fresh edible soybeans and saving the breeding cost.

Description

technical field [0001] The invention relates to an Indel / SNP molecular marker used for detecting homozygous and heterozygous GmBADH2 genes related to aroma traits of fresh soybeans, belonging to the field of agricultural biotechnology. Background technique [0002] Fresh soybeans (also known as vegetable soybeans, edamame), are a special type of edible soybeans with full grains during the bulging period and green pods when they are green. Consistent, easy to harvest greens at one time, can be fried, cold, canned or frozen for export. Fresh soybeans are rich in nutrients. According to research, fresh soybeans are rich in plant protein, unsaturated fatty acids, plant crude fiber and various minerals, vitamin A, vitamin C and vitamin E necessary for the human body, and the amino acids contained in them are complete. , rich in content, high in nutritional value, and good in taste, it is a safe and healthy food that is recognized as pollution-free or less polluted today. Among ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 袁凤杰钱琳琳金杭霞郁晓敏杨清华竹龙鸣傅旭军
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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