Reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for rapidly detecting southern rice black-streaked dwarf viruses in plant hopper

A technology of RT-LAMP and black-streaked dwarf virus, applied in the field of agricultural science, can solve the problems of high sequence homology and difficulty in distinguishing

Active Publication Date: 2011-08-31
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
  • Description
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  • 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 in size and shape of virus particles), traditional biological inoculation identification (similar in 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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  • Reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for rapidly detecting southern rice black-streaked dwarf viruses in plant hopper
  • Reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for rapidly detecting southern rice black-streaked dwarf viruses in plant hopper

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Experimental program
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Effect test

Embodiment 1

[0022] Embodiment 1, optimization of southern rice black-streaked dwarf virus RT-LAMP reaction conditions

[0023] ① Design of RT-LAMP primers for southern rice black-streaked dwarf virus.

[0024] According to the nucleotide sequence of Southern rice black-streaked dwarf virus S9 reported on NCBI (EU523359.1, http: / / www.ncbi.nlm.nih.gov / nuccore / 183229399; EU784843.1.http: / / www .ncbi.nlm.nih.gov / nuccore / 209867306) to design primers. Using a relatively conserved sequence (311-770 bp) of the virus as a template, primers were designed using the online LAMP primer design software primerExplorer V4 (http: / / primerexplorerjp / elamp4.0.0 / index.html). After uploading the template sequence, the system software calculates and obtains the preliminary LAMP primer combination (F3, B3, FIP, BIP), and then provides parameters such as the stability of the 3' end or 5' end of the primer and the primer dimer through the software. Compare and select multiple pairs of primers designed, and finall...

Embodiment 2

[0031] Embodiment 2, the specificity test of southern rice black-streaked dwarf virus RT-LAMP reaction

[0032] In order to verify the specificity of the RT-LAMP method, the rice black-streaked dwarf virus (RBSDV) of the same genus as SRBSDV was selected as a control. Using the RNA of the white-backed planthopper carrying SRBSDV and the RNA of the brown planthopper carrying RBSDV as templates, the RT-LAMP reaction system in Example 1 was used for the reaction. The results show that when using the RT-LAMP primers of this experiment to amplify the total RNA of SBPH carrying RBSDV, there is no amplified product; while carrying the total RNA of SBPH carrying SRBSDV, the target product can be amplified ( figure 2 ). This is consistent with the expected result, indicating that the RT-LAMP detection method established by the present invention has good specificity.

[0033] The invention discloses a method for rapidly detecting the southern rice black-streaked dwarf virus in planthop...

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Abstract

The invention provides a rapid detection technology for southern rice black-streaked dwarf viruses in plant hopper and a method for applying the rapid detection technology to disease diagnosis and prediction. By the method, the interference of rice black-streaked dwarf viruses can be eliminated, and whether the southern rice black-streaked dwarf viruses are carried in the plant hopper is rapidly identified, so that a pertinent control strategy is adopted to reduce the loss caused by diseases.

Description

technical field [0001] The invention relates to a rapid detection technology of southern rice black-streaked dwarf virus in planthoppers and its application in disease diagnosis and forecasting, and belongs 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 recent years, the disease has spread and occurred in Jiangsu and Zhejiang regions, and it is becoming more and more serious. In 2008, the diseased area in Jiangsu Province alone rea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 周彤周益军杜琳琳兰莹徐秋芳孙枫程兆榜
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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