Genomic regions of tomato leaf curl virus DNA satellite required for replication and for satellite-mediated delivery of heterologous DNAs.
Tomato leaf curl virus (TLCV) satellite DNA (sat-DNA) is a 682 nt, circular, single-stranded molecule that lacks an open reading frame (ORF) or an apparent promoter. It contains binding motifs for the TLCV replication-associated protein, but these are dispensable for replication. To identify the regions of the sat-DNA critical for replication, the entire sequence was scanned by deletion/replacement mutagenesis. Transient assays using Nicotiana benthamiana revealed that sequences within nt 296-35 (through nt 682) are essential for replication. Sequence deletions and replacements between nt 35 and 296 were tolerated but with a significant loss of infectivity, indicating that genome size strongly influences replication efficiency. Within the permissible region, inserts of 100-700 nt were retained in transient assays although with a slight reduction in replication. In addition, sat-DNA constructs containing short non-viral DNAs replicated and spread in tobacco plants, indicating their potential as gene-delivery vectors.
Li
Dongmei
D
School of Agriculture, Food and Wine, The University of Adelaide, Waite Campus, SA 5064, Australia.|CSIRO Plant Industry, PO Box 350, Glen Osmond, SA 5064, Australia.
Behjatnia
S A Akbar
SAA
Plant Protection Department, College of Agriculture, Shiraz University, Shiraz, Iran.|CSIRO Plant Industry, PO Box 350, Glen Osmond, SA 5064, Australia.
Dry
Ian B
IB
CSIRO Plant Industry, PO Box 350, Glen Osmond, SA 5064, Australia.
Randles
John W
JW
School of Agriculture, Food and Wine, The University of Adelaide, Waite Campus, SA 5064, Australia.
Eini
Omid
O
Department of Plant Protection, School of Agriculture, Zanjan University, Zanjan, Iran.|School of Agriculture, Food and Wine, The University of Adelaide, Waite Campus, SA 5064, Australia.|CSIRO Plant Industry, PO Box 350, Glen Osmond, SA 5064, Australia.
Rezaian
M Ali
MA
School of Agriculture, Food and Wine, The University of Adelaide, Waite Campus, SA 5064, Australia.
D001483:Base Sequence; D053839:Begomovirus / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology*; D004274:DNA, Recombinant / Q000235:genetics; D004276:DNA, Satellite / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D020868:Gene Silencing; D017930:Genes, Reporter; D005822:Genetic Vectors; D016679:Genome, Viral; D005966:Glucuronidase / Q000235:genetics; D018551:Solanum lycopersicum / Q000821:virology; D016296:Mutagenesis; D010935:Plant Diseases / Q000821:virology; D030821:Plants, Genetically Modified; D017384:Sequence Deletion; D014026:Nicotiana / Q000235:genetics / Q000821:virology; D014779:Virus Replication / Q000235:genetics
D004274:DNA, Recombinant; D004276:DNA, Satellite; D004279:DNA, Viral; D005966:Glucuronidase