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Method for synchronously detecting and identifying three major pathogen nematodes in tobacco root knot nematode diseases

A technology for tobacco root-knot nematodes and pathogenic nematodes, applied in the field of detection of plant disease pathogens, can solve the problems of cumbersome operation and long time-consuming identification process, and achieve the effects of convenient identification, wide application range and good stability

Active Publication Date: 2018-07-20
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the step-by-step PCR detection method, at least three PCR reactions are required to complete the identification of a sample, which makes the identification process cumbersome and time-consuming.

Method used

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  • Method for synchronously detecting and identifying three major pathogen nematodes in tobacco root knot nematode diseases
  • Method for synchronously detecting and identifying three major pathogen nematodes in tobacco root knot nematode diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] 1. Design of specific primers for the identification of three pathogenic nematodes of tobacco root-knot nematode: According to the genome sequence information of root-knot nematode incognita, root-knot nematode peanut and root-knot nematode javanica published by the National Center for Biotechnology Information (NCBI), Analyze with DNASTAR 7.0 software, find out the species-specific sequences of three kinds of root-knot nematodes, utilize primer Premier 5.0 software to design 6 PCR primers that are used for identification of three kinds of root-knot nematodes species:

[0064] TR1: 5'-AAGCAAAAGACGAAGCACCAAAA-3'

[0065] TR2: 5'-GAGGATTCAGCTCCCCAGCAC-3'

[0066] TR3: 5'-GAGTACGGATTTGAAATACAGGG-3'

[0067] TR4: 5'-TGAATGCTATGCCATCAGGAG-3'

[0068] TR5: 5'-CGATTGAACTGAGCCCAGACT-3'

[0069] TR6: 5'-TTTATTCGCAAGACAACACCC-3'

[0070] A DNA synthesizer synthesized the above six oligonucleotide primers.

[0071] 2. Preparation of DNA templates for PCR identification of the...

Embodiment 2

[0075] 1. Design of specific primers for the identification of three pathogenic nematodes of tobacco root-knot nematode: According to the genome sequence information of root-knot nematode incognita, root-knot nematode peanut and root-knot nematode javanica published by the National Center for Biotechnology Information (NCBI), Analyze with DNASTAR 7.0 software, find out the species-specific sequences of three kinds of root-knot nematodes, utilize primer Premier 5.0 software to design 6 PCR primers that are used for identification of three kinds of root-knot nematodes species:

[0076] TR1: 5'-AAGCAAAAGACGAAGCACCAAAA-3'

[0077] TR2: 5'-GAGGATTCAGCTCCCCAGCAC-3'

[0078] TR3: 5'-GAGTACGGATTTGAAATACAGGG-3'

[0079] TR4: 5'-TGAATGCTATGCCATCAGGAG-3'

[0080] TR5: 5'-CGATTGAACTGAGCCCAGACT-3'

[0081] TR6: 5'-TTTATTCGCAAGACAACACCC-3'

[0082] A DNA synthesizer synthesized the above six oligonucleotide primers.

[0083] 2. Preparation of DNA templates for PCR identification of tob...

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Abstract

The invention discloses a method for synchronously detecting and identifying three major pathogen nematodes in tobacco root knot nematode diseases. The method comprises the steps of PCR (polymerase chain reaction) primer design, DNA (deoxyribonucleic acid) template preparation, PCR amplification and product detection. The species identification which contains 6 PCR primers and is used for three kinds of meloidogyne is designed according to the species specific region sequence of meloidogyne incognita, meloidogyne arenaria and meloidogyne javanica; the identification stability is high; the identification accuracy is high; the designed 6 primers are combined to be used; the synchronous detection of three kinds of pathogen nematodes of tobacco root knot nematode diseases through once PCR canbe realized; the identification is convenient; the time is saved; an ordinary PCR instrument and an ordinary reagent are used; the detection cost is low; the popularization and the application are facilitated. A DNA preparation method aiming at disease root samples and soil samples is designed; the species identification aiming at meloidogyne at each development stage including ova, larvas and imagos can be realized; the use range is wide.

Description

technical field [0001] The invention belongs to the technical field of detection of plant disease pathogens, and in particular relates to a method for synchronous detection and identification of three main pathogenic nematodes in tobacco root-knot nematode disease. Background technique [0002] root-knot nematodes ( Meloidogyne spp. ) is the nematode with the largest variety, the widest distribution and the most serious damage among plant pathogenic nematodes. This kind of nematode mainly spreads in the form of soil, harms the root system of plants, and causes the roots of the affected plants to form tumor-like root knots, thereby causing root-knot nematodes that seriously affect the normal growth and development of plants. Plants mainly include more than 3,000 species of Solanaceae, Fabaceae, Cucurbitaceae, Gramineae, Umbelliferae, etc. Plants in temperate, subtropical and tropical regions are particularly severely damaged. Among them, tobacco is most seriously harmed b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686
CPCC12Q1/686C12Q1/6888C12Q2600/16C12Q2537/143C12Q2565/125
Inventor 李梅云吴文涛宋中邦张靖曾建敏王扬王丙武焦芳婵高玉龙吴兴富李永平李文正
Owner YUNNAN ACAD OF TOBACCO AGRI SCI