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Method for preventing and controlling tobacco mosaic virus disease

A tobacco mosaic virus, tobacco technology, applied in the directions of botanical equipment and methods, disinfectants, chemicals for biological control, etc., can solve the problems of high cost, difficult realization, difficult results, etc. Simple, convenient effect in field application

Inactive Publication Date: 2008-07-02
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, although many units are studying how to use siRNA to prevent and treat plant virus diseases, the cost of this RNA chemical synthesis method is very high, and it is difficult to inject RNA into a large number of plants, so the results are difficult to industrialize

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Embodiment 1: Utilize the phenol chloroform method to extract the RNA genome of tobacco mosaic virus; With the genome RNA of tobacco mosaic virus as template, reverse transcription obtains the cDNA of tobacco mosaic virus motor protein; Utilize PCR technology to isolate tobacco mosaic virus movement A fragment of 21-23bp in the range of 150-800bp on the protein cDNA; the obtained above-mentioned fragment is cloned into a plasmid vector capable of expressing RNA; the cloned fragment is expressed in the corresponding E. coli (E.coli) lacking RNase RNA; smash the cells expressing the purpose RNA, and make it into a suspension solution; the content of RNA in the solution is 2-20ng / ml, and the photoprotectant aromatic amino acid tryptophan accounts for 0.01-0.1% of the volume of the solution, dispersed Agent 1-2%, glycerin 2-5%, sterile water 96.99-92.9%. The dispersant is L602 or LH656. During preparation, glycerin, aromatic amino acid tryptophan, dispersant and sterilized...

Embodiment 2

[0011] Embodiment 2: In the present embodiment, the composition of suspension solution is: the content of RNA is 5ng / ml, photoprotectant aromatic amino acid tryptophan 0.05%, L602 dispersant 1.5%, glycerin 3.5%, sterile water 94.95% %. Others are the same as in Example 1.

Embodiment 3

[0012] Embodiment 3: In the present embodiment, the composition of suspension solution is: the content of RNA is 15ng / ml, photoprotectant aromatic amino acid tryptophan 0.08%, LH656 dispersant 1.8%, glycerol 4%, sterile water 94.72% %. Others are the same as in Example 1.

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Abstract

The invention discloses a method for preventing and curing tobacco mosaic virus disease which includes the following steps of extracting a RNA gene group of the tobacco mosaic virus; utilizing the RNA gene group of the tobacco mosaic virus to acquire a cDNA of movement protein of the tobacco mosaic virus by reverse transcription; utilizing a PCR technology to separate a section of a 21-23bp length in 150-800bp on the cDNA of the movement protein of the tobacco mosaic virus; cloning the section into a plasmid carrier that can express the RNA so as to lead the cloned section to be expressed as the RNA in a colon bacillus of an enzyme lacking the RNA; crushing a cell and making the cell into a suspension solution with the chroma of 2-20ng / ml; spraying the suspension solution on the leaf surface of the tobacco after suspension solution is diluted 1000 times. The method has the advantages of simple and convenient production, low cost and no harm to human, animals and higher animals. The invention is suitable for massive use on tobaccos. The prepared suspension solution overcomes the defects of wettable powder and emulsion; accessory ingredient is easy to disperse. The invention is more convenient to apply in fields and has no remnant and environment pollution.

Description

technical field [0001] The invention belongs to plant protection, in particular to a method for preventing and treating tobacco mosaic virus disease by using double-stranded RNA interference technology. Background technique [0002] The mosaic disease caused by tobacco mosaic virus is a very important disease on tobacco, tomato and other crops. It occurs commonly all over the world and causes serious losses. For example, it can occur continuously from the seedling stage to the field stage on tobacco. After the seedlings are infected, the veins of the new leaves become lighter in color, showing translucent "bright vein disease", and then form yellow and green mosaic disease; seedling stage Infected plants develop slowly and are of little economic value. In the field stage, the plants are affected. In addition to the symptoms of bright veins and mosaic leaves, blisters will form on the diseased leaves, and the thickness will be uneven; the leaf shape will also appear various ...

Claims

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Application Information

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IPC IPC(8): A01N63/00A01P1/00
Inventor 梁振普邱立友张小霞戚元成高玉千
Owner HENAN AGRICULTURAL UNIVERSITY
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