PMID- 26484673
OWN - NLM
STAT- MEDLINE
DCOM- 20160429
LR  - 20190202
IS  - 1553-7374 (Electronic)
IS  - 1553-7366 (Print)
IS  - 1553-7366 (Linking)
VI  - 11
IP  - 10
DP  - 2015 Oct
TI  - Rescue of a Plant Negative-Strand RNA Virus from Cloned cDNA: Insights into 
      Enveloped Plant Virus Movement and Morphogenesis.
PG  - e1005223
LID - 10.1371/journal.ppat.1005223 [doi]
LID - e1005223
AB  - Reverse genetics systems have been established for all major groups of plant DNA 
      and positive-strand RNA viruses, and our understanding of their infection cycles 
      and pathogenesis has benefitted enormously from use of these approaches. However, 
      technical difficulties have heretofore hampered applications of reverse genetics 
      to plant negative-strand RNA (NSR) viruses. Here, we report recovery of 
      infectious virus from cloned cDNAs of a model plant NSR, Sonchus yellow net 
      rhabdovirus (SYNV). The procedure involves Agrobacterium-mediated transcription 
      of full-length SYNV antigenomic RNA and co-expression of the nucleoprotein (N), 
      phosphoprotein (P), large polymerase core proteins and viral suppressors of RNA 
      silencing in Nicotiana benthamiana plants. Optimization of core protein 
      expression resulted in up to 26% recombinant SYNV (rSYNV) infections of 
      agroinfiltrated plants. A reporter virus, rSYNV-GFP, engineered by inserting a 
      green fluorescence protein (GFP) gene between the N and P genes was able to 
      express GFP during systemic infections and after repeated plant-to-plant 
      mechanical passages. Deletion analyses with rSYNV-GFP demonstrated that SYNV 
      cell-to-cell movement requires the sc4 protein and suggested that uncoiled 
      nucleocapsids are infectious movement entities. Deletion analyses also showed 
      that the glycoprotein is not required for systemic infection, although the 
      glycoprotein mutant was defective in virion morphogenesis. Taken together, we 
      have developed a robust reverse genetics system for SYNV that provides key 
      insights into morphogenesis and movement of an enveloped plant virus. Our study 
      also provides a template for developing analogous systems for reverse genetic 
      analysis of other plant NSR viruses.
FAU - Wang, Qiang
AU  - Wang Q
AD  - State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang 
      University, Hangzhou, China.
FAU - Ma, Xiaonan
AU  - Ma X
AD  - State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang 
      University, Hangzhou, China.
FAU - Qian, ShaSha
AU  - Qian S
AD  - State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang 
      University, Hangzhou, China.
FAU - Zhou, Xin
AU  - Zhou X
AD  - State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang 
      University, Hangzhou, China.
FAU - Sun, Kai
AU  - Sun K
AD  - State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang 
      University, Hangzhou, China.
FAU - Chen, Xiaolan
AU  - Chen X
AD  - State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang 
      University, Hangzhou, China.
FAU - Zhou, Xueping
AU  - Zhou X
AD  - State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang 
      University, Hangzhou, China; State Key Laboratory for Biology of Plant Diseases 
      and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural 
      Sciences, Beijing, China.
FAU - Jackson, Andrew O
AU  - Jackson AO
AD  - Department of Plant and Microbial Biology, University of California, Berkeley, 
      California, United States of America.
FAU - Li, Zhenghe
AU  - Li Z
AD  - State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang 
      University, Hangzhou, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20151020
PL  - United States
TA  - PLoS Pathog
JT  - PLoS pathogens
JID - 101238921
RN  - 0 (DNA, Complementary)
RN  - 0 (RNA, Plant)
SB  - IM
MH  - DNA, Complementary/genetics
MH  - Immunoblotting
MH  - Microscopy, Fluorescence
MH  - Plant Diseases/virology
MH  - Plant Viruses/*genetics
MH  - RNA, Plant/genetics/*isolation & purification
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
MH  - Rhabdoviridae/*genetics
MH  - Rhabdoviridae Infections/*genetics
MH  - Sonchus/*virology
PMC - PMC4616665
COIS- The authors have declared that no competing interests exist.
EDAT- 2015/10/21 06:00
MHDA- 2016/04/30 06:00
PMCR- 2015/10/20
CRDT- 2015/10/21 06:00
PHST- 2015/07/26 00:00 [received]
PHST- 2015/09/22 00:00 [accepted]
PHST- 2015/10/21 06:00 [entrez]
PHST- 2015/10/21 06:00 [pubmed]
PHST- 2016/04/30 06:00 [medline]
PHST- 2015/10/20 00:00 [pmc-release]
AID - PPATHOGENS-D-15-01733 [pii]
AID - 10.1371/journal.ppat.1005223 [doi]
PST - epublish
SO  - PLoS Pathog. 2015 Oct 20;11(10):e1005223. doi: 10.1371/journal.ppat.1005223. 
      eCollection 2015 Oct.