Tomato Mosaic Virus (ToMV) ELISA Kit (DEIAPV10)

Regulatory status: For research use only, not for use in diagnostic procedures.

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Size
1000T
Sample
host plans
Intended Use
The test can be used to detect ToMV in infected solanaceous crops.
Contents of Kit
500 wells
Coating antibody: 0.250 ml, 4°C
Detecting conjugate, alkaline phosphatase: 0.250 ml, 4°C
96-well ELISA plates: 5, Room temperature
Instruction: 1

1000 wells
Coating antibody: 0.50 ml, 4°C
Detecting conjugate, alkaline phosphatase: 0.50 ml, 4°C
96-well ELISA plates: 10, Room temperature
Instruction: 1

5000 wells
Coating antibody: 2*1.25 ml, 4°C
Detecting conjugate, alkaline phosphatase: 2*1.25 ml, 4°C
96-well ELISA plates: 50, Room temperature
Instruction: 1
Storage
All reagent components should be stored at the recommended temperature to assure their full shelf life. Do not store prepared working solution from day to day.
Sensitivity
Sensitivity of the ELISA is very high. The virus can be consistently detected in infected plant leaf tissues diluted at 1:2430 - 1:7290.
General Description
ToMV is found worldwide and economically damaging in glasshouse and outdoor tomatoes and many other plants. ToMV can cause yellowing and stunting of tomato plants, resulting in loss of stand and reduced yield. In addition, the virus may cause uneven ripening of fruit, further reducing yield. The virus is readily spread by handling and cultural operations. It also contaminates seeds and soil, but no natural vector is known. The best measure to control and reduce infection is to use certified disease-free seed and remove any infected plants, including the roots. Remove Also, discard any plants near those affected.
Standard Curve

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References


An inhibitor of viral RNA replication is encoded by a plant resistance gene

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA

Authors: Ishibashi, Kazuhiro; Masuda, Kiyoshi; Naito, Satoshi; Meshi, Tetsuo; Ishikawa, Masayuki

The tomato Tm-1 gene confers resistance to tomato mosaic virus (ToMV). Here, we report that the extracts of Tm-1 tomato cells (GCR237) have properties that inhibit the in vitro RNA replication of WT ToMV more strongly than that of the Tm-1-resistance-breaking mutant of ToMV, LT1. We purified this inhibitory activity and identified a polypeptide of approximate to 80 kDa (p80(GCR237)) using LC-tandem MS. The amino acid sequence of p80GCR237 had no similarity to any characterized proteins. The p80GCR237 gene cosegregated with Tm-1; transgenic expression of p80GCR237 conferred resistance to ToMV within tomato plants; and the knockdown of p80(GCR237) sensitized Tm-1 tomato plants to ToMV, indicating that Tm-1 encodes p80GCR237 itself. We further show that in vitro-synthesized Tm-1 (p80(GCR237)) protein binds to the replication proteins of WT ToMV and inhibits their function at a step before, but not after, the viral replication complex is formed on the membrane surfaces. Such binding was not observed for the replication proteins of LT1. These results suggest that Tm-1 (p80(GCR237)) inhibits the replication of WT ToMV RNA through binding to the replication proteins.

Inducible viral inoculation system with cultured plant cells facilitates a biochemical approach for virus-induced RNA silencing

ARCHIVES OF VIROLOGY

Authors: Tamai, Atsushi; Dohi, Koji; Mori, Masasi; Meshi, Tetsuo; Ishikawa, Masayuki

An inducible virus infection system was demonstrated to be an efficient protein expression system for inducing synchronous virus vector multiplication in suspension-cultured plant cells. A GFP-tagged tomato mosaic virus (ToMV-GFP) derivative that has a defect in its 130 K protein, a silencing suppressor of ToMV, was synchronously infected to tobacco BY2 cultured cells using this system. In the infection-induced cells, viral RNA was degraded rapidly, and a cytosol extract prepared from the infected cells showed RNA degradation activity specific for ToMV- or GFP-related sequences. In lysate prepared from cells infected by ToMV-GFP carrying the wild-type 130 K protein, sequence-specific RNA degradation activity was suppressed, although siRNA derived from the virus was generated. Furthermore, the 130 K protein interfered with 3'-end methylation of siRNA. The inducible virus infection system may provide a method for biochemical analysis of antiviral RNA silencing and silencing suppression by ToMV.

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