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SNP functional molecular marker and application of paddy rice fragrance gene

A technology of functional molecules and aroma genes, applied in the field of molecular biology, can solve problems such as inaccurate manual identification results, long phenotypic identification breeding cycle, and unsafe experimental operations, so as to speed up genetic improvement, eliminate aerosol pollution, reduce workload effect

Inactive Publication Date: 2017-09-29
YANGTZE NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a SNP molecular marker of rice aroma gene, which solves the problems of long breeding cycle, high breeding cost and low breeding efficiency in the existing fragrant rice rice aroma.
The invention also provides a primer set and KASP method for detecting SNP functional molecular markers, which solves the problems of unsafe experimental operation, complicated operation and inaccurate manual identification results, etc.

Method used

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  • SNP functional molecular marker and application of paddy rice fragrance gene
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  • SNP functional molecular marker and application of paddy rice fragrance gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Application of molecular marker K_08g badh2-1 in detecting the genotype of natural genetic material

[0039] 46 natural genetic materials were selected for verification, including 30 aromatic rice materials and 16 non-aromatic rice materials.

[0040] 1. Genomic DNA was extracted from rice leaves by simplified CTAB method.

[0041] (1) Freeze-dry or dry the rice leaves, put an appropriate amount of leaves into a 2.0mL centrifuge tube, add two steel balls, and grind on a tissue grinder;

[0042] (2) Add 750 μL of CTAB solution, oscillate to homogenate, and incubate at 65°C for 0.5-1 h;

[0043] (3) Cool to room temperature, add 750 μL of chloroform:isoamyl alcohol (24:1) in the fume hood and mix by inverting 3-4 times;

[0044] (4) Centrifuge at 12000rmp for 10min, transfer 500μL supernatant to a new 1.5mL centrifuge tube;

[0045] (5) Add an equal volume of isopropanol solution and shake gently to mix, precipitate at -20°C for more than 1 hour, centrifuge a...

Embodiment 2

[0055] Example 2 Application of Molecular Marker K_08g badh2-1 in Detecting the Genotypes of Recurrent Parents, Backbone Parents and Gene Donor Materials

[0056] The marker K_08g badh2-1 was used to detect 95 important recurrent parents, backbone parents and important gene donor materials. Method is with embodiment 1.

[0057] As a result, only 3 materials were marked K_08g badh2-1 genotype aroma materials ( figure 2 ), most recurrent parents, backbone parents and important gene donor materials do not have this genotype, which further shows that this genotype material has a great application prospect and scope. Select the genotype variation material from the natural genetic population as parents, so as to introduce the gene into the material that does not contain the target gene, and realize the genetic improvement of important recurrent parents, backbone parents and important gene donor materials. Cultivation provides important technical support.

Embodiment 3

[0058] Example 3 Phenotypic verification of isolated populations of molecular marker K_08g badh2-1

[0059] 1. Detect the individual plant genotype of the F2 segregation population of the donor parent Wuxiang japonica crossed with the recipient material Nipponbare; the method is the same as in Example 1.

[0060] 2. Collect homozygous genotype single plant seeds at the maturity stage of rice and dry them in the sun, beat rice with a rice huller to obtain brown rice samples for later use;

[0061] 3. Take 2g brown rice sample, put it in a 50mL centrifuge tube, add 10mL 1.7% KOH solution, close the lid tightly, soak for about 10min, and mix it upside down several times during the process;

[0062] 4. Open the cap of the centrifuge tube, and gently sniff the sample with the nose to see if there is any fragrance emitted. The experimental control includes the parent material, the standard fragrance variety Zixiangnuo and the non-scent variety Guangluai No. 4;

[0063] 5. At least ...

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Abstract

The invention discloses an SNP functional molecular marker and application of a paddy rice fragrance gene. The fragrance gene is badh2, a number-2 exon area inside the gene has an insert loss of 7bp basic group, the SNP functional molecular marker is K_08g badh2-1 and is positioned at 20380271 basic group of number-8 chromosome of the paddy rice, the polymorphism of the SNP functional molecular marker is G/C, and a sequence of the SNP functional molecular marker is SEQ ID No.1. An SNP site is detected by utilizing a KASP method, and whether the paddy rice sample has fragrance or not is determined by virtue of different fluorescent signals. The SNP functional molecular marker developed by the invention can be used for high-flux screening and identification of paddy rice seedling period or natural genetic materials, the operation is simple and convenient, the result is accurate and reliable, the genetic improvement of the character of the paddy rice fragrance quality is accelerated, the breeding workload is reduced, the breeding efficiency is high, and the cost is low.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a SNP functional molecular marker of rice aroma gene and its application. Background technique [0002] Traditional pedigree selection is a method commonly used by rice breeders. Through years of efforts by breeders, a large number of new rice varieties with high yield, high resistance and high quality have been bred and created. However, the traditional breeding model combined with phenotypic identification has shortcomings such as inaccurate control of phenotypes, large demand for genetic populations, high labor costs, and long breeding cycles, making it difficult to achieve large-scale commercial integrated breeding or genetic improvement of materials. With the development of molecular biology and bioinformatics, molecular marker-assisted selection breeding (Molecular Assisted Selectoin, MAS) has shown great technical advantages, realizing the combination of genetic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156C12Q2600/172
Inventor 李文博曾秀红汪建华陈发波高健江波
Owner YANGTZE NORMAL UNIVERSITY
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