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Gene marker for distinguishing rice starch quality

A gene marker and gene technology, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of lower transcript content, lower intron splicing efficiency, lower amylose content, etc.

Active Publication Date: 2022-07-08
CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The transcript content and enzyme activity of Wx were decreased in japonica rice cultivars, which was due to the fact that in japonica rice Wx b The 5' splicing site of the first intron has a GT→TT mutation, which reduces the intron splicing efficiency and causes a decrease in amylose content (Isshiki et al., 1998.)

Method used

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  • Gene marker for distinguishing rice starch quality
  • Gene marker for distinguishing rice starch quality
  • Gene marker for distinguishing rice starch quality

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Screening of gene markers affecting rice starch gelatinization temperature

[0037] 1. The SNP site sequence screening of the ALK gene includes the following steps:

[0038] 1) extracting the genomic DNA of the rice plant to be tested;

[0039] 2) with primers selected from the following table, PCR amplification is carried out to rice genomic DNA;

[0040] ALK-SNP-F CGCAGCACAACAGCAAGGTGCG SEQ ID NO: 7 ALK-SNP-R GGTCTCTTCACCATTGGTACTTG SEQ ID NO: 8

[0041] PCR reaction system is calculated as 20μl: 10X PCR reaction buffer 2μ1, 25mM MgSO 4 0.8μl, 2mM dNTP 2μl, 5μM forward primer (-F) and reverse primer (-R) 1.2μl each, genomic DNA 20ng, KOD-Plus polymerase 0.4μl, plus ddH 2 O is made up to 20 μl.

[0042] The PCR reaction conditions were as follows: 94°C pre-denaturation for 2 minutes, 94°C denaturation for 15 seconds, 55°C annealing for 30 seconds, 68°C extension for 1 minute, a total of 35 cycles; 68°C incubation for 5 minutes. ...

Embodiment 2

[0062] Example 2: Screening of gene markers affecting amylose content of rice starch

[0063] 1. The SNP site sequence screening of the Wx gene includes the following steps:

[0064] 1) extracting the genomic DNA of the rice plant to be tested;

[0065] 2) with primers selected from the following table, PCR amplification is carried out to rice genomic DNA;

[0066] Wx-SNP-F GAGGTGGCCGGTGGTGTTGTCCTTC SEQ ID NO: 9 Wx-SNP-R GAAGATTCCATAATGTACCAGG SEQ ID NO: 10

[0067] PCR reaction system is calculated as 20μl: 10X PCR reaction buffer 2μ1, 25mM MgSO 4 0.8μl, 2mM dNTP 2μl, 5μM forward primer (-F) and reverse primer (-R) 1.2μl each, genomic DNA 20ng, KOD-Plus polymerase 0.4μl, plus ddH 2 O is made up to 20 μl.

[0068] The PCR reaction conditions were as follows: 94°C pre-denaturation for 2 minutes, 94°C denaturation for 15 seconds, 55°C annealing for 30 seconds, 68°C extension for 1 minute, a total of 35 cycles; 68°C incubation for 5 minutes.

[00...

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Abstract

The invention discloses a gene marker for distinguishing rice starch quality. The gene marker comprises an SNP (Single Nucleotide Polymorphism) site sequence SEQ ID NOs: 1-4 selected from an ALK gene and an SNP site sequence SEQ ID NOs: 5-6 selected from a Waxy gene. The gene markers can be used for distinguishing the rice starch quality and effectively guiding rapid breeding of high-starch-quality rice varieties.

Description

technical field [0001] The invention belongs to the field of gene detection, and relates to a gene marker for distinguishing rice starch quality, in particular a gene marker for distinguishing rice starch quality and its application. Background technique [0002] Rice (Oryza sativa L.) is one of the most important food crops in the world and is eaten by more than half of the world's population. With the improvement of people's living standards, people have higher and higher requirements for the quality of rice, not only the appearance of rice such as chalkiness and transparency, but also the taste of high-quality rice, which is directly related to the quality of rice starch. improvement. [0003] The quality of rice starch includes a number of indicators, such as gelatinization temperature (GT), gel consistency, amylose content, protein content and so on. Generally, when the gelatinization temperature of japonica rice is ≥6 and the gelatinization temperature of indica rice...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 韩斌司丽珍吕威赵强上官颖颖朱静洁
Owner CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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