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PCR primer for rapidly identifying weedy rice in early stage and application thereof

A weedy rice, fast technology, applied in the biological field, can solve the problems of cumbersome steps and inability to detect, and achieve the effect of simple and fast operation and accurate results.

Pending Publication Date: 2021-02-26
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the reported early weedy rice identification methods are all identified by molecular techniques such as PCR, and PCR identification primers are mainly designed through the deletion marker of the red peel gene Rc, but the weedy rice with white peel cannot be detected; A method for identifying weedy rice by detecting SNP sites in weedy rice by nested PCR and enzyme digestion, but the steps of this method are cumbersome

Method used

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  • PCR primer for rapidly identifying weedy rice in early stage and application thereof
  • PCR primer for rapidly identifying weedy rice in early stage and application thereof
  • PCR primer for rapidly identifying weedy rice in early stage and application thereof

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Effect test

Embodiment 1

[0048]The method for rapid and early identification of weedy rice specifically includes the following steps:

[0049]1) P-Bh4 primer design

[0050]The forward primer P-Bh4-F is: 5’-TGATATGCTTCCTGCTGTGCAC-3’,

[0051]The reverse primer P-Bh4-R is: 5'-GACGGCCACGTTGAGCATCGAT-3'.

[0052]2) DNA extraction

[0053]Select 10 samples from the rice field flooded with weedy rice, collect the leaves, ground them with liquid nitrogen, and extract the genomic DNA of the samples with a plant genomic DNA extraction kit;

[0054]3) PCR amplification of sample genomic DNA

[0055]The PCR amplification reaction system is: sterile double distilled water 10.7μL, 2×PCR Mix 12.5μL, primers P-Bh4-F and P-Bh4-R (10μmol / L) each 0.4μL, template DNA (10ng / μL) ) 1 μL; amplification conditions are: 95°C pre-denaturation for 5 minutes; 95°C for 30s, 64°C for 30s, 72°C for 30s, 30 cycles; 72°C extension for 7 minutes.

[0056]4) PCR product electrophoresis detection and result judgment

[0057]After the PCR, take 7μL of the product on a ...

Embodiment 2

[0059]The method for quickly identifying weedy rice and cultivated rice specifically includes the following steps:

[0060]1) Primer design

[0061]a. P-Bh4 primer design:

[0062]The forward primer P-Bh4-F is: 5’-TGATATGCTTCCTGCTGTGCAC-3’,

[0063]The reverse primer P-Bh4-R is: 5'-GACGGCCACGTTGAGCATCGAT-3'.

[0064]The size of the amplified product is 356bp.

[0065]b. P-Rc primer design:

[0066]The forward primer P-Rc-F is: 5’-CATTCTCCAGATGGACTACTCC-3’,

[0067]The reverse primer P-Rc-R is: 5'-GATGGCACCGACTTTTCGCGT-3'.

[0068]The size of the amplified product is 191bp;

[0069]2) DNA extraction

[0070]120 samples of weed rice and 20 samples of cultivated rice were selected. The young leaves were collected at the seedling stage and ground with liquid nitrogen. The genomic DNA of the samples was extracted with a plant genomic DNA extraction kit;

[0071]3) Double PCR amplification of sample genomic DNA

[0072]The reaction system of double PCR amplification is: sterile double distilled water 9.5μL, 2×PCR Mix 12.5μL, p...

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Abstract

The invention discloses a PCR primer for rapidly identifying weedy rice in an early stage and application thereof. A single PCR primer P-Bh4 and a double PCR primer P-Bh4 / P-Rc are designed in a deletion marker area of a black glume gene Bh4 and a red peel gene Rc of rice; PCR amplification is carried out on rice genome DNA by using the primers; and a product is subjected to agarose gel electrophoresis separation, and the weedy rice is identified according to an electrophoretic band. According to the PCR primer for rapidly identifying the weedy rice in the early stage and the application thereof, the weedy rice with black glume or red peel can be identified by detecting wild type genes of the black glume gene Bh4 and the red peel gene Rc, so that the weedy rice can be comprehensively and effectively identified; and the PCR primer can be used for rapidly identifying the weedy rice in a rice seedling stage, the operation is simple, the result is accurate, and batch detection is easy.

Description

Technical field[0001]The invention relates to the field of biotechnology, in particular to a PCR primer for rapid and early identification of weedy rice and its application.Background technique[0002]Weed rice is a kind of rice that continuously regenerates in rice fields, endangers rice production, and has weed characteristics. It is early maturing, easy to shatter, and has most red rice grains. It has strong biological competitiveness and is second only to barnyard grass in paddy fields. The third most vicious weed of Stephanotis. In recent years, weedy rice has become more and more serious in some rice areas in my country's Jiangsu, Hunan, Guangdong, Liaoning and other provinces, seriously affecting rice yield and quality, and threatening my country's rice production and food security. The main reason for the occurrence of weedy rice is the widespread promotion and application of light rice cultivation measures such as wheat intercropping, direct seeding rice, no-tillage or less-t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 夏启玉张雨良张丽丽贺萍萍赵辉郭安平
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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