Quick sensitive detection method of potato viruses

A potato virus, sensitive detection technology, applied in the field of plant virology detection, can solve the problems of affecting potato yield and quality, unable to formulate disease prevention measures, detection sensitivity can not better determine the infection of different viruses, etc.

Active Publication Date: 2014-04-30
云南省农业科学院生物技术与种质资源研究所
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AI Technical Summary

Problems solved by technology

[0008] During potato growth, the occurrence of potato virus diseases is common and seriously affects the yield and quality of potatoes. The main pathogens are potato virus Y, potato virus S, potato virus A and potato leafroll virus. Therefore, it is impossible to formulate effective disease prevention measures; in the study of disease infection cycle and early disease occurrence, when ...

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  • Quick sensitive detection method of potato viruses
  • Quick sensitive detection method of potato viruses
  • Quick sensitive detection method of potato viruses

Examples

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Embodiment 1

[0043] 1. Primer design:

[0044] According to the sequences of Potato virus Y, Potato S virus, Potato virus A and Potato leafroll virus in GenBank, select the more conservative regions in a single virus to design primers, optimize the internal pairing and aggregation effects of different primer pairs through experiments between different viruses, and screen Different virus detection primer pairs available for multiplex PCR (see Table 1).

[0045] 2. Coating virus-specific antibodies

[0046] Through testing and optimizing conditions, add polyclonal antibody dilutions of potato virus Y, potato virus S, potato virus A and potato leafroll virus at final concentrations of 1:5000, 1:5000, 1:5000 and 1:5000 respectively in the PCR tube After coating 100 μL at 4°C overnight or at 37°C for 1 h, suck out the liquid in the PCR tube, and treat the PBST solution with DEPC [phosphate buffer containing 0.01% Tween 20 (0.1 mol / L, pH 7. 0) was added to 0.2% DEPC (diethyl pyrocarbonate) and...

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Abstract

The invention discloses a method for potato Y virus, potato X virus, potato A virus and potato leaf roll virus in Yunnan potato main culture species Li potato No.6, Yun potato 401 and Xuan potato No.2 infected plants. The specific mixed antibody is utilized to entrap the potato Y virus, potato X virus, potato A virus and potato leaf roll virus in Yunnan potato main culture species Li potato No.6, Yun potato 401 and Xuan potato No.2 infected plants, so that the total RNA (ribonucleic acid) of the sample does not need to be extracted to perform random primer reverse transcription and specific virus primer compound PCR (polymerase chain reaction) reaction. The method combines quick specific immune precipitation and sensitive compound RT-PCR (reverse transcription-polymerase chain reaction), sets healthy potato random primer cDNA (complementary deoxyribonucleic acid) product and corresponding virus PCR specific segment positive plasmid as negative and positive controls, and thus, is quickly, accurately and sensitively used in detecting field potato plant samples, antiviral materials, field environment suspect infection sources and the like.

Description

technical field [0001] The invention belongs to the technical field of plant virology detection, and in particular relates to a rapid and sensitive detection method of potato virus. Background technique [0002] The field symptoms of potato virus disease are complex and diverse, and the common symptom types can be summarized as follows: (1) mosaic type. Light green, yellow-green and dark green mottled mottled leaves appear on the leaves (there are light mottled, heavy mosaic, shrunken mottled and macular mottled), and the leaves basically remain the same, or the leaves become smaller and wrinkled Shrinkage, dwarfing of plants. (2) Curly leaf type. Leaf margins curl upwards, even cylindrical, pale in color, hardened and leathery, and sometimes purplish red appears on the back of the leaves. (3) Necrotic type (or streak type). Leaf veins, petioles, stems and branches appear brown necrotic spots or conjoined streaks, and even leaves wither, die or fall off. (4) Arbuscular ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6804C12Q1/70C12Q2531/113C12Q2545/101
Inventor 丁铭卢训张仲凯
Owner 云南省农业科学院生物技术与种质资源研究所
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