Inoculation of resistant and susceptible Nicotiana tabacum cultivars with TMV by Jeremiah Bell

In this lab we infected resistant and susceptible tobacco plants (Nicotania tabacum) with the Tobacco mosaic virus. The plants labeled as n/n were susceptible and the plants labeled N/N were resistant. We will be able to tell if a plant becomes sick by looking for small necrotic lesions. The necrotic lesions are caused by plant cells recognizing the replicase protein in TMV and initiating apoptosis in those cells preventing the virus from replicating. We inoculated leaves of n/n and N/N plants with a series of dilutions made from a high concentration sample of TMV. We used a high concentration (50 ng/μl), a medium concentration (10 ng/μl), a low concentration (2 ng/μl), and finally some leaves were just rinsed with water as a control. We used carborundum powder on the leaves to make a wound for the TMV to infect cells.

Preparing plants for infection with TMV by swabbing the leaves of N resistant and non-N resistant tobacco plants with carborundum powder.

Preparing plants for infection with TMV by swabbing the leaves of N resistant and N susceptible t tobacco plants with carborundum powder.

She is rubbing the carborundum powder into the leaves with her fingers before adding TMV to the leaves.

She is rubbing the carborundum powder into the leaves with her fingers before adding TMV to the leaves.

Finally we inoculated the plants with TMV via pipettes.

Finally we inoculated the plants with TMV via pipettes.

 

Rough

The plant leaf will wilt and die if rubbed to roughly with the powder.

TMV lesions

Here is a leaf with a few lesions created by the plants apoptosis response to the TMV. The lesions are circular and are white in the center with a decaying ring around it. This was seen in the N/N tobacco plants.

Virus test

Finally a virus test kit. You place a piece of leaf in the bag, grind it up into a juice, and place the TMV indicator into the fluid. One red line means a negative result and two red lines mean a positive result

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