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Molecular marker of rice gelatinization temperature gene ALK as well as primer and application of molecular marker

A molecular marker, gelatinization temperature technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial determination/inspection, etc., can solve the problems of difficult phenotype, rough, time-consuming and laborious, and achieve accurate detection Time saving and good primer specificity

Pending Publication Date: 2022-05-13
CROP INST SICHUAN PROVINCE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The gelatinization temperature of rice is an important cooking quality trait of rice. It is a complex quantitative trait controlled by multiple micro-effect genes, and is easily affected by other factors such as the environment. selection, and the gelatinization temperature of rice can only be identified after it is mature and harvested, and it needs to be roughened before identification, which is time-consuming and laborious

Method used

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  • Molecular marker of rice gelatinization temperature gene ALK as well as primer and application of molecular marker
  • Molecular marker of rice gelatinization temperature gene ALK as well as primer and application of molecular marker
  • Molecular marker of rice gelatinization temperature gene ALK as well as primer and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Primer Design

[0025] According to rice material 1892S (gene ALK G , alkali elimination value ASV=3.5, medium alkali elimination value) and Guang 8B (gene ALK T , Alkaline elimination value ASV=7.0, high alkali elimination value) of the ALK gene sequence difference, except for the differences of SNP4198A / G, SNP4329G / T and SNP4330C / T, Guang 8B material has 4 deletions in the 3'UTR of ALK gene compared with 1892S material base ATCG, so the primer ALK26 was designed ( figure 1 ), the amplified fragment of the primer ALK26 contains this difference, as a linkage marker in the ALK gene, the molecular marker primer is applied to the selective breeding of high alkali elimination value materials.

[0026] ALK26 forward primer sequence (SEQ ID NO.1) is as follows:

[0027] 5'-AGCCTCCCCTGAAGAAGCTTG-3';

[0028] ALK26 reverse primer sequence (SEQ ID NO.2) is as follows:

[0029] 5'-GAACACACAAACCGGAAGCT-3'.

Embodiment 2

[0030] Example 2 Using the Indel molecular marker ALK26 to identify the gelatinization temperature gene of rice materials

[0031] 1. Test material

[0032] Medium-alkali digestion value rice materials include: 1892S.

[0033] Rice materials with high alkali consumption value include: Guang 8B, Chuan 106B, Chuan Kang 606B, Chuan Lv 609B, Cheng Hui 3203, Cheng Hui 727, IR24, Jing 4155S, Chuan 658S, Bing 4114. Both Chuankang 606B and Chuanlu 609B were bred from the cross combination of Chuan 106B and Chuan 345B. Chuan 658S was bred from the cross combination of Chuanxiang 29B, Chenghui 177 and Shuhui 527, and was used as a two-line CMS material.

[0034] 2. DNA extraction

[0035] Take 3-5 g of rice seedling tissue, use a mortar to grind the sample, and use the CTAB method to extract plant genomic DNA.

[0036] 3. PCR amplification

[0037] The PCR amplification system was 25 μL, and the specific dosage of each reagent was as follows: 2 μL of 50–100 ng / μL DNA template, 1 μL...

Embodiment 3

[0043] Example 3 Comparison of Primer ALK26 Molecular Marker Identification and Alkali Elimination Value Phenotype Identification

[0044] 1. Test material

[0045] 1892S, Guang 8B, Chuan 106B, Chuankang 606B, Chuanlv 609B, Chenghui 3203, Chenghui 727, IR24, Jing 4155S, Chuan658S, Bing 4114.

[0046] 2. Phenotypic identification of alkali elimination value

[0047] Randomly take 6 grains of mature and full-bodied rice from the test materials and place them in a glass petri dish, and add about 10.0 mL of 1.7% KOH solution. Arrange the rice grains evenly in the petri dish so that the rice grains are completely soaked without shaking randomly, and then cover it. Move the culture dish smoothly to the incubator, and set the temperature at 30±2°C. Take it out smoothly after standing for 24 hours. Observe the decomposition of the endosperm of rice grains one by one, classify the alkali digestion value according to the standard, record the alkali digestion value of each grain of r...

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Abstract

The invention discloses a molecular marker of a rice gelatinization temperature gene ALK as well as a primer and application of the molecular marker. Nucleotide sequences of the molecular marker are shown as SEQ ID NO.3 and SEQ ID NO.4. The nucleotide sequence of the high-alkali elimination rice amplification product is as shown in SEQ ID No.3, the nucleotide sequence of the medium-alkali elimination rice amplification product is as shown in SEQ ID No.4, and the heterozygous genotype rice material is simultaneously amplified into bands as shown in SEQ ID No.3 and SEQ ID No.4. According to the invention, the identification of the rice molecular marker assisted breeding high / medium alkali elimination value material can be rapidly detected by using a simple and reliable method.

Description

technical field [0001] The invention relates to a molecular marker of a rice gelatinization temperature gene, in particular to a molecular marker of a rice gelatinization temperature gene ALK, a primer and application thereof. Background technique [0002] Rice quality mainly includes four aspects: milling quality, appearance quality, nutritional quality and cooking and eating quality. The evaluation indicators of these four quality traits cover a series of physical and biochemical characteristics. The genetic factors of rice itself are the main reason for determining these characters. Therefore, it is of great significance to study a series of genetic mechanisms of rice quality. So far, scientists have mapped QTLs for multiple genes related to rice quality, some of which have been cloned and functionally verified. Most of these cloned genes and QTLs have been developed into molecular markers. Molecular marker-assisted selection technology can improve the breeding efficien...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 刘利平高方远任光俊任鄄胜吕建群刘松
Owner CROP INST SICHUAN PROVINCE ACAD OF AGRI SCI
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