A general method for the detection of potyviral gene products in plant protoplasts and tissue

A rapid immunostaining procedure for detecting potyviral antigens in individual protoplasts, isolated mesophyll cells, and epidermal strips of Nicotiana tabacum is described. Although all specific antibodies tested detected potyviral antigens in electroporated protoplasts, those against cytoplasmic inclusion (CI) protein provided the most useful results. The number of protoplasts reacting with anti-CI increased with time after inoculation, roughly in parallel with the accumulation of capsid protein, which was measured independently by enzyme-linked immunosorbent assay. Potyviral gene products were also detected in epidermal strips and mesophyll cells separated from systemically infected leaves, indicating that the immunostaining method is generally applicable and that it may prove useful for studying the movement of potyviruses from cell to cell in intact plants.