Infection of Turnip Protoplasts with Cauliflower Mosaic Virus

Infection of Protoplasts from Several Brassica Species with Cauliflower Mosaic Virus Following Inoculation Using Polyethylene Glycol

“…'Purple Top' (Table 1). Following the course of infection using fluorescent antibody staining, maximum virus accumulation occurred by 65 to 96 h, a time consistent with that previously reported (Furusawa et al, 1980). Protoplasts from all the other species became infected (Table 1) but in comparison with turnip protoplasts the percentage infections were less and varied more between experiments.…”

“…Unfortunately in the study of CaMV, so far the contributions made using protoplasts have been few. This paper describes an alternative technique to that published (Furusawa et al, 1980), with which protoplasts from a range of Cruciferous species can be readily infected with CaMV following PEG-mediated uptake of virus particles. The routine infection of a high proportion of the protoplasts inoculated permits the measurement of viral nucleic acid produced following viral multiplication.…”

“…Infections of turnip protoplasts with CaMV (Howell & Hull, 1978 ;Furusawa et al, 1980) and CaMV DNA (Yamaoka et al, 1982a) have been reported previously with, in one case, up to 80 ~ of protoplasts becoming infected (Furusawa et al, 1980). The inocula used in each of these studies contained the polycation, poly-L-ornithine.…”

Dissection of Cauliflower Mosaic Virus Transactivator/Viroplasmin Reveals Distinct Essential Functions in Basic Virus Replication

“…1F, right). Taking into account that 333 to 1,667 productive cells were analyzed in a single PCR (calculated from the number of transfected cells [10 6 ], the portion of transfected cells used for DNA extraction [one-third], the proportion of DNA analyzed [5 l/50 l], a protoplast survival rate of 20 to 50% as examined by observing the cell shape under the microscope, and an infection rate of 5 to 10% as determined by immunofluorescence [8]), transfection with pE4Pin and pAATav gave rise to between 1,200 and 6,000 copies of viral DNA accumulated in one productive cell. These levels are about 1 order of magnitude lower than the previously estimated CaMV DNA copy number per cell in CaMV-infected plants (26) or CaMV particle-infected protoplasts (25).…”

Reasons for the low accumulation level of aphid transmission factor protein in infected leaves with an aphid-non-transmissible cauliflower mosaic virus isolate, CM1841

“…Turnip protoplasts were prepared as described by Furusawa & Okuno (1978). Protoplasts were inoculated with CaMV and incubated as described by Furusawa et al (1980). Total proteins were extracted from CM1841-and CMBX-infected protoplasts (104 to 1.5 x 104 cells) at 24, 48 and 72hp.i., and accumulation of p18 was compared with that of IB protein using the Western blotting method.…”