The Infectivity of Extracts made from Leaves at Intervals after Inoculation with Viruses

SUMMARY: When leaves are macerated at intervals after being inoculated with plant viruses, the infectivity of the extracts obtained decreases with increasing time until newly produced virus becomes detectable. Infectivity does not start to increase until approximately twice the time apparently needed for virus to multiply in the epidermis and spread from there to the mesophyll. Epidermal cells infected by inoculation seem to produce too few virus particles to be detected by infectivity tests or else the first-formed particles are unstable in vitro. No evidence was obtained, with Rothamsted tobacco necrosis virus (RTNV) in leaves of tobacco and French bean, that the initial decrease in infectivity occurs because of changes in virus particles that succeed in infecting and causing lesions. If such changes occur they are obscured by the inactivation of particles that do not multiply and cause lesions. Washing inoculated leaves removes 95 % of the inoculated virus, but only slightly decreases the numbers of infections, and adding ‘Celite’ to the inoculum greatly increases the numbers of lesions without increasing the amount of virus retained by washed leaves. Neither washing nor adding ‘Celite’ to the inoculum affects the rate at which the infectivity of successive extracts from inoculated leaves decreases. Infectivity continues to decrease after virus appears to have multiplied in and spread from the epidermis. Cells of Nicotiana glutinosa that are infected by tobacco mosaic virus spreading from inoculated epidermal cells die only a few hours after the infectivity of leaf extracts starts to increase: few cells seem to become infected from virus produced in these secondarily infected cells and, at 20°, infectivity reaches a maximum in 2 days. Mesophyll cells of French bean leaves at 22° seem to synthesize new RTNV particles within 5 hr. of becoming infected from the epidermis and to continue synthesizing for another 30 hr., when they probably contain about 106virus particles/cell. Although the cells then die, the virus spreads to further cells and the infectivity of leaf extracts increases for at least five days.