Early events of tomato spotted wilt transcription and replication in protoplasts.
A protoplast transfection system for tomato spotted wilt tospovirus (TSWV) was established by PEG-mediated infection of tobacco protoplasts. Analysis of viral RNA synthesis revealed an asymmetric production of viral (v) and viral-complementary (vc) strands of all three genomic RNA segments and a separation in time of subgenomic transcription during the first 72 hr post infection (p.i.). Synthesis of vc-RNA was detectable 8-10 hrs prior to v-RNA production. During the first 72 h of infection the accumulation of S-RNAs exceeded M- and L-RNAs and the amount of vc-RNAs was larger than that of genomic v-RNAs. The subgenomic N-mRNA was first detected 5 hr p.i., followed by vc-S RNA and the mRNA for the nonstructural protein (NSs) 15 hr later. The two subgenomic mRNA species of the S RNA appeared to be regulated independently from each other and from other viral mRNA species. Defective interfering (DI) elements from L RNA associated with the virus inoculum, were not preferentially replicated at the expense of full-sized L-RNA.
Max-Planck-Institut für Züchtungsforschung, Abteilung Genetische Grundlagen der Pflanzenzüchtung, Köln, FRG.
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Steinecke
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Heinze
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Oehmen
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Adam
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Schreier
D007150:Immunohistochemistry; D018551:Solanum lycopersicum / Q000821:virology*; D011523:Protoplasts / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis / Q000096:biosynthesis; D017865:Tospovirus / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic; D014779:Virus Replication
D012367:RNA, Viral