In situ localization of the putative movement protein (pr17) from potato leafroll luteovirus (PLRV) in infected and transgenic potato plants.
The potato leafroll virus (PLRV) 17-kDa protein (pr17), the putative movement protein for this phloem-limited luteovirus, was localized on ultrathin sections of leaves from PLRV-infected and transgenic potato plants. The transgenic plants expressed the entire viral genome from a full-length cDNA copy (PLRVfl) or only the gene encoding pr17 (ORF4) under the control of the cauliflower mosaic virus 35S promoter. Virus-infected and PLRVfl-transgenic plants developed symptoms typical of virus infection, whereas pr17-transgenic plants did not display symptoms or ultrastructural alterations. Immunogold electron microscopy using an anti-pr17-serum detected pr17 in plasmodesmata, in virus-induced vesicles, in mitochondria, and in chloroplasts of phloem cells, in PLRV-infected as well as PLRVfl-transgenic plants. In addition, in transgenic plants, pr17 was expressed in mesophyll cells (which are not infected by PLRV under natural conditions) and localized to the same sites as in phloem cells, except in plasmodesmata. In contrast, in pr17-transgenic plants the protein was never observed on organelles, but was almost exclusively associated with plasmodesmata of all leaf cell types, indicating that the targeting of pr17 to plasmodesmata is an intrinsic property of the protein. These results support the role of pr17 in PLRV movement.
Schmitz
J
J
MPI für Züchtungsforschung, Carl-von-Linné Weg 10, Cologne, 50829, Germany.
Stussi-Garaud
C
C
Tacke
E
E
Prüfer
D
D
Rohde
W
W
Rohfritsch
O
O
D002352:Carrier Proteins / Q000032:analysis* / Q000235:genetics; D017796:Caulimovirus / Q000235:genetics; D002736:Chloroplasts / Q000737:chemistry; D004268:DNA-Binding Proteins; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007365:Intercellular Junctions / Q000502:physiology / Q000821:virology*; D017793:Luteovirus / Q000235:genetics; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D008928:Mitochondria / Q000737:chemistry; D010750:Phosphoproteins / Q000032:analysis / Q000235:genetics; D030821:Plants, Genetically Modified / Q000737:chemistry* / Q000648:ultrastructure; D011401:Promoter Regions, Genetic; D016601:RNA-Binding Proteins; D011198:Solanum tuberosum / Q000737:chemistry* / Q000648:ultrastructure; D014764:Viral Proteins / Q000032:analysis* / Q000235:genetics
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D014764:Viral Proteins; C083784:p17 nucleic acid-binding protein, potato leafroll luteovirus