The susceptibility of tobacco protoplasts to infection with cowpea chlorotic mottle virus varied with the age both of the plants and leaves used as a source for the protoplasts. The efficiency of infection for freshly prepared and stored protoplasts could be increased by an improved direct inoculation procedure. Poly--ornithine, poly--arginine, poly--lysine and poly--lysine were all effective in inducing infection, the first stage of which may occur within 20 s of exposure to inoculum. The effects of temperature, injury and sodium azide on infection suggested that the limiting process during inoculation is not controlled by enzymes. The yield of virus per infected protoplast was decreased by some antibiotics but not affected by various other additives to the culture medium.


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