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Obtaining method of soybean mosaic virus infecting tobacco, and application of soybean mosaic virus infecting tobacco

A technology of soybean mosaic virus and tobacco, applied in the field of genetic engineering, can solve the problem of inability to verify the function of soybean anti-SMV candidate genes

Active Publication Date: 2014-04-30
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although tobacco is easy to transform, people have been unable to verify the function of soybean anti-SMV candidate genes by transforming tobacco because SMV cannot infect tobacco.

Method used

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  • Obtaining method of soybean mosaic virus infecting tobacco, and application of soybean mosaic virus infecting tobacco
  • Obtaining method of soybean mosaic virus infecting tobacco, and application of soybean mosaic virus infecting tobacco
  • Obtaining method of soybean mosaic virus infecting tobacco, and application of soybean mosaic virus infecting tobacco

Examples

Experimental program
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Effect test

Embodiment 1

[0048] 1. Obtaining SMV isolates capable of infecting Nicotiana benthamiana

[0049] (1) Thirty-seven SMV isolates were isolated and identified from soybean production areas across the country (Table 1). These isolates were artificially inoculated on the susceptible soybean variety Nannong 1138-2, and 0.1mol / L. Phosphate buffer solution with pH=7.3~7.4 (approximately 5ml of buffer solution per 1g of fresh leaves) plus a little corundum (approximately 600 mesh) is ground into a homogenate in a mortar.

[0050] (2) Cultivate Nicotiana benthamiana in an artificial climate chamber at 25°C, 16 hours of light, 8 hours of darkness, and 60% humidity to ensure that there are no aphids in the climate chamber to prevent cross-infection of strains caused by aphid transmission. At the stage of 5-6 leaves, artificially inoculate SMV.

[0051] The homogenate is applied to each tobacco leaf with a hairbrush, and the young leaves at the top are not inoculated. After the inoculation, the toba...

Embodiment 2

[0094] The SMV isolate submitted to CCTCC for deposit is S9, and the deposit number is CCTCC V201349. The isolate of SMV S9 came from the diseased leaves in the Hebei field. Due to the complex virus composition in the field, each diseased sample may be the product of one or more SMV strains co-infected, or even the result of multiple viruses co-infected. Therefore, each disease sample was isolated and purified by Topcrop single spot and identified by DAS‐ELISA serum to ensure that each isolate used for identification was a purified SMV. Because the strain identification results are easily affected by aphids, the environment, and especially temperature, all virus inoculation experiments in this experiment were carried out in insect-proof net rooms or greenhouses, and the temperature was controlled at 20‐30 °C. All SMV isolates were verified twice or more on 10 identification hosts to ensure the accuracy of strain identification results

[0095] 1 Collection and storage of dise...

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Abstract

The invention discloses an obtaining method and application of a soybean mosaic virus infecting tobacco, and an application of the soybean mosaic virus infecting tobacco. A soybean mosaic virus isolate having a preservation number of CCTCC NO:V201349 is used to invade Nicotina benthamiana to obtain the soybean mosaic virus infecting tobacco. The soybean mosaic virus infecting tobacco obtained through the method can be applied in the verification of anti-soybean mosaic virus candidate genes. The soybean mosaic virus capable of invading tobacco is obtained by screening from many isolates, and confirmation is carried out through the isolate invading tobacco, disease symptom investigation, Nannong 1138-2 backtie experiment, ELISA and an RT-PCR detection means. The soybean mosaic virus infecting tobacco is obtained in the invention, realizes the verification of the anti-SMV candidate gene function on the tobacco, and breaks the restriction that the genetic transformation in soybean is difficult.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and relates to a method for obtaining tobacco-infecting soybean mosaic virus and its application. Background technique [0002] Soybean mosaic virus (SMV) disease is one of the most important soybean diseases in the world. It is widely distributed and occurs in almost all soybean planting areas. SMV belongs to the genus Potyvirus (Potyvirus). After soybeans are infected with SMV, symptoms such as mosaic and necrosis will occur, resulting in reduced photosynthetic area and photosynthetic capacity of soybeans, dwarfing of plants, decreased growth, and brown spots on grains. 15% reduction in production, or even no harvest. [0003] Breeding disease-resistant varieties is the most effective way to control SMV. In addition to traditional disease-resistant breeding methods, soybean transgenic technology has become one of the methods to quickly cultivate disease-resistant varieties. Both transgenic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12Q1/70C12Q1/68G01N33/569C12R1/94
Inventor 智海剑高乐李凯任锐仲勇坤翟锐
Owner NANJING AGRICULTURAL UNIVERSITY
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