Disease Development in Cultured Cells of Nicotiana tabacum L. var. Samsun NN Injected with Tobacco Mosaic Virus

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Cell chains were transferred from callus tissue shake cultures of N. tabacum var. Samsun NN, a local lesion host of TMV, to oil-chamber microscope slides. In each slide culture a single cell of one chain was directly injected with a bacteria-free aqueous suspension of TMV. A single cell of another chain was injected with heatinactivated TMV. A third cell chain was not inoculated. Injections were made with a hollow glass needle connected to a microinjection syringe and guided by a micromanipulator. Inoculated and uninoculated control cells showed no evidence of infection. Cells injected with infective TMV, and noninjected cells in the same chain, lived 4-10 days and slowly developed disease symptoms. Inclusions appeared at the nucleus, beginning 23-69 hours after inoculation, and gradually assumed the hexagonal disc form of TMV crystals. Transvacuolar cytoplasmic strands merged with the parietal cytoplasm and cytoplasmic streaming slowed to a stop. The cytoplasmic reticulum became less complex and eventually disappeared. Most plastids and other organelles gradually disappeared from the cytoplasm. Plastids remaining in the cytoplasm became much smaller. In some cells numerous spherical bodies of various size appeared in the vacuole. One cell contained an inclusion similar to an X-body.
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