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Method for synchronously detecting four whitefly transmitted gemini-viruses infecting tomatoes

A twin virus and simultaneous detection technology, applied in the field of agricultural biology, can solve the problems of difficult to distinguish viruses, loss of tomato production, and no distinction has been seen, and achieve the effects of simplifying operations, ensuring sensitivity and specificity, and reducing costs

Inactive Publication Date: 2015-05-27
INST OF PLANT PROTECTION GUANGXI ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2007, tomato yellow leaf curl disease caused by Chinese papaya leaf curl virus occurred in a large area in Henan Province, causing economic losses of more than 20 million yuan in Kaifeng County alone; the spread of the virus has brought great harm to the tomato production in these areas. loss
Traditional identification methods such as: biological inoculation method, electron microscope observation method, serological detection method, molecular biological method, etc. are difficult to distinguish due to the close characteristics of the viruses, and it has not been seen that the four viruses can be distinguished by using PCR once. Methods

Method used

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  • Method for synchronously detecting four whitefly transmitted gemini-viruses infecting tomatoes
  • Method for synchronously detecting four whitefly transmitted gemini-viruses infecting tomatoes
  • Method for synchronously detecting four whitefly transmitted gemini-viruses infecting tomatoes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Samples to be tested include:

[0028] 1 healthy tomato plants, 2 samples known to be infected with Chinese tomato leaf curl virus, 3 samples known to be infected with Chinese red thistle yellow vein virus, 4 samples known to be infected with Chinese papaya leaf curl virus, 5 samples known to be infected with Chinese tomato yellow koji Leaf virus samples, 6 samples known to be co-infected with Chinese tomato leaf curl virus, Chinese red thistle yellow vein virus, Chinese papaya leaf curl virus and Chinese tomato yellow leaf curl virus.

[0029]Perform PCR detection on each sample according to the following steps.

[0030] Extract the total DNA of the sample

[0031] Weigh 0.1g of the sample to be tested in a mortar, add 1ml of 0.5mol / L NaOH solution, grind it, transfer it to a 1.5mL centrifuge tube, centrifuge at 5000rpm for 5min, and dilute the supernatant with pH8.00.1mol / L Tris-HCl 50 times, as a template for PCR amplification;

[0032] PCR reaction

[0033] Re...

Embodiment 2

[0049] Samples to be tested include:

[0050] 1 healthy tomato plants, 2 samples known to be infected with Chinese tomato leaf curl virus, 3 samples known to be infected with Chinese red thistle yellow vein virus, 4 samples known to be infected with Chinese papaya leaf curl virus, 5 samples known to be infected with Chinese tomato yellow koji Leaf virus samples, 6 known to be co-infected with Chinese red thistle yellow vein virus and Chinese tomato yellow leaf curl virus samples, 7 known to be co-infected with Chinese tomato leaf curl virus and Chinese papaya leaf curl virus samples, 8 known to be co-infected Chinese tomato leaf curl virus, Chinese papaya leaf curl virus, and Chinese tomato yellow leaf curl virus samples, 9 samples known to be co-infected with Chinese red thistle yellow vein virus, Chinese papaya leaf curl virus, and Chinese tomato yellow leaf curl virus , 10 samples known to be co-infected with Chinese tomato leaf curl virus, Chinese red thistle yellow vein v...

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Abstract

The invention discloses a method for synchronously detecting four whitefly transmitted gemini-viruses infecting tomatoes. The method can be used for simultaneously identifying four viruses by utilizing four specific primers ToL-F / ToL-R, TY-F / TY-R, PaL-F / PaL-R, AY-F / AY-R especially for four viruses, namely, tomato leaf curl China virus, tomato yellow curlleaf China virus, papaya leaf curl China virus and ageratum yellow vein China virus by combining a PCR (polymerase chain reaction) detection technology. According to the method, after a total DNA (deoxyribonucleic acid) of a sample is extracted, the four whitefly transmitted gemini-viruses can be detected by only once PCR detection, and a simple method is provided for investigation and identification. Compared with the traditional method which can be used for detecting one virus by once PCR, the method has the advantages of reducing cost, saving time, guaranteeing sensitivity and specificity and simplifying operation, and is a specific, sensitive, convenient, economical and effective detection method.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and particularly relates to a method for synchronously detecting four kinds of Bemisia tabaci-transmitted geminiviruses infecting tomatoes, including Chinese tomato leaf curl virus, Chinese tomato yellow leaf curl virus, and Chinese papaya leaf curl virus and Chinese red thistle yellow vein virus. Background technique [0002] Whitefly-transmitted Gemini-viruses (WTGs) disease is one of the important diseases of tomato production in the world. It was first discovered in Israel and named Tomato yellow leaf curl virus. , TYLCV) name. Subsequent studies have found that the virus is not a virus but a type of virus. This type of virus has twin particle morphology and single-stranded circular DNA. It belongs to the Begomovirus genus of the family Geminiviridae. Composed of two-component DNA (DNA-A and DNA-B) or single-component DNA, the size of each DNA component is 2.5kb to 2.8kb, and some ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 李战彪秦碧霞徐鹏超蔡健和
Owner INST OF PLANT PROTECTION GUANGXI ACADEMY OF AGRI SCI
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