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Technology for rapidly detecting rice black-streaked dwarf viruses in plants by reverse transcription-loop-mediated isothermal amplification (RT-LAMP) method

A technology of RT-LAMP, rice black stripe dwarfing, applied in the field of agricultural science, can solve the problem that homology is difficult to distinguish

Inactive Publication Date: 2011-08-17
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since these two viruses have very similar characteristics in many aspects, traditional plant virus detection methods, such as electron microscope observation (similar size and shape of virus particles), traditional biological inoculation identification (similar symptoms, mediators, and host ranges) , serological detection (sequences still have high homology) and other methods are difficult to distinguish

Method used

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  • Technology for rapidly detecting rice black-streaked dwarf viruses in plants by reverse transcription-loop-mediated isothermal amplification (RT-LAMP) method
  • Technology for rapidly detecting rice black-streaked dwarf viruses in plants by reverse transcription-loop-mediated isothermal amplification (RT-LAMP) method
  • Technology for rapidly detecting rice black-streaked dwarf viruses in plants by reverse transcription-loop-mediated isothermal amplification (RT-LAMP) method

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Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1, optimization of rice black-streaked dwarf virus RT-LAMP reaction conditions

[0026] ①Design of rice black-streaked dwarf virus RT-LAMP primers.

[0027] According to the rice black-streaked dwarf virus reported by NCBI (AF227206.1, http: / / www.ncbi.nlm.nih.gov / nuccore / 183229399; EU784843.1, http: / / Www.ncbi.nlm.nih. gov / nuccore / 209867306) S10 nucleotide sequence, the relatively conservative region 950-1467bp of the virus nucleic acid was found after analysis by the software DNAstar, and the corresponding region 950-1467bp on EU523359.1 was used as a template reference sequence to design online LAMP primers Software primerExplorer V4 ( http: / / primerexplorer.jp / elamp4.0.0 / index.html ) to design primers. After uploading the template sequence, the system software calculates the preliminary LAMP primer combination (FIP, BIP, F3, B3), and then provides parameters such as the stability of the 3' end or 5' end of the primer and the primer dimer through the so...

Embodiment 2

[0040] Embodiment 2, the specificity test of rice black-streaked dwarf virus RT-LAMP reaction

[0041] In order to verify the specificity of the RT-LAMP method, the southern rice black-streaked dwarf virus (SRBSDV) of the same genus as RBSDV was selected as a control. The total RNA of RBSDV and SRBSDV was used as the template respectively, and the reaction was carried out with the RT-LAMP reaction system in ②. Result shows that when using RT-LAMP primers to amplify the RNA template of SRBSDV, there is no amplified band; and when the RNA of amplified RBSDV can amplify the band of interest, this shows that the RT-LAMP detection method established by the present invention has Very good specificity.

[0042] The application of a method for detecting rice black-streaked dwarf virus on plants includes: rapid diagnosis and identification of suspected diseased plants such as rice, corn and wheat.

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Abstract

The invention provides a rapid detection technology for rice black-streaked dwarf viruses in plants such as rice, wheat and corn and the like and an application method thereof in the diagnosis of diseases. By the method, the interference of southern rice black-streaked dwarf disease can be eliminated, and plant samples of the rice black-streaked dwarf viruses can be rapidly identified, so that targeted preventive strategies can be used, and the loss brought by the diseases is reduced.

Description

technical field [0001] The invention relates to a rapid detection technology of rice black-streaked dwarf virus on plants such as rice, wheat and corn and its application in disease diagnosis, belonging to the field of agricultural science and technology. Background technique [0002] Rice black-streaked dwarf virus (RBSDV) is a serious virus disease on rice, belonging to the plant Reoviridae Fijivirus genus (Fijivirus), the virion is spherical, mainly composed of SBPH spreads. In the early stage of field symptoms, the main symptoms are dwarf plants and dark green leaves. In the later stage, wax drop-like protrusions appear on the leaves, leaf sheaths and stems, and then black stripes form. This has a great impact on the yield of rice. Severely diseased fields even May result in no yield. In addition to infecting rice, the virus can also infect corn (maize rough dwarf disease), wheat (wheat green dwarf) and various grass weeds. It was popular in Jiangsu, Shanghai and Zheji...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 周彤周益军杜琳琳兰莹徐秋芳孙枫范永坚
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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