| Autor: |
Vaira AM; Institute for Sustainable Plant Protection, IPSP-CNR, Strada delle Cacce 73, 10135, Torino, Italy.; USDA-ARS, USNA, Floral and Nursery Plant Research Unit, 10300 Baltimore Ave, Beltsville, MD, USA., Lim HS; USDA-ARS, USNA, Floral and Nursery Plant Research Unit, 10300 Baltimore Ave, Beltsville, MD, USA.; Department of Applied Biology, Chungnam National University, Daejeon, 305-764, Republic of Korea., Bauchan G; USDA-ARS, BARC, Electron and Confocal Microscopy Unit, 10300 Baltimore Ave, Beltsville, MD, USA., Gulbronson CJ; USDA-ARS, USNA, Floral and Nursery Plant Research Unit, 10300 Baltimore Ave, Beltsville, MD, USA.; Oak Ridge Institute for Science and Education (ORISE) Postdoctoral Fellow, USA., Miozzi L; Institute for Sustainable Plant Protection, IPSP-CNR, Strada delle Cacce 73, 10135, Torino, Italy., Vinals N; Institute for Sustainable Plant Protection, IPSP-CNR, Strada delle Cacce 73, 10135, Torino, Italy., Natilla A; USDA-ARS, BARC, Molecular Plant Pathology Laboratory, 10300 Baltimore Ave, Beltsville, MD, USA.; Present address: Arc Horizon, LLC, Innovation Park, 1736 West Paul Dirac Dr., Tallahassee, FL, USA., Hammond J; USDA-ARS, USNA, Floral and Nursery Plant Research Unit, 10300 Baltimore Ave, Beltsville, MD, USA. |
| Abstrakt: |
The Lolium latent virus (LoLV) major coat protein sequence contains a typical chloroplast transit peptide (cTP) domain. In infected Nicotiana benthamiana leaf tissue, LoLV coat proteins can be detected at the chloroplast. In transient expression, several N-terminal deletions of the CP sequence, increasing in length, result in disruption of the domain functionality, markedly affecting intracellular localization. A yeast two-hybrid-based study using LoLV CP as bait identified several potentially interacting Arabidopsis host proteins, most of them with chloroplast-linked pathways. One of them, an ankyrin repeat protein, was studied in detail. The N. benthamiana homologue (NbANKr) targets chloroplasts, is able to co-localize with LoLV CP at chloroplast membranes in transient expression and shows a robust interaction with LoLV CP in vivo by BiFC, which has been confirmed by yeast two-hybrid data. Silencing NbANKr genes in N. benthamiana plants, prior to challenging with LoLV by mechanical inoculation, affects LoLV infection, significantly reducing the level of viral RNA in young leaves, compared to levels in control plants, and suggesting an inhibition of virus movement. Silencing of NbANKr has no obvious effect on plant phenotype, but is able to interfere with LoLV infection, opening the way for a new strategy for virus infection control. |
