Helicase Domain Encoded by Cucumber mosaic virus RNA1 DeterminesSystemic Infection of Cmr1 in Pepper

Collection with item attached
2012
Item details URL
http://open-repository.kisti.re.kr/cube/handle/open_repository/479778.do
DOI
10.1371/journal.pone.0043136
Title
Helicase Domain Encoded by Cucumber mosaic virus RNA1 DeterminesSystemic Infection of Cmr1 in Pepper
Description
This work was supported by a grant (code: 0636-20110005) from theVegetable Breeding Research Center through Agriculture Research Center(ARC) program from Ministry for Food, Agriculture, Forestry andFisheries, Republic of Korea and by Basic Science Research Programthough the National Research Foundation of Korea (NRF) funded by theMinistry of Education, Science and Technology (2011-0005310). Thefunders had no role in study design, data collection and analysis,decision to publish, or preparation of the manuscript.
abstract
The Cmr1 gene in peppers confers resistance to Cucumber mosaic virus isolate-P0 (CMV-P0). Cmr1 restricts the systemic spread of CMV strain-Fny (CMV-Fny), whereas this gene cannot block the spread of CMV isolate-P1 (CMV-P1) to the upper leaves, resulting in systemic infection. To identify the virulence determinant of CMV-P1, six reassortant viruses and six chimeric viruses derived from CMV-Fny and CMV-P1 cDNA clones were used. Our results demonstrate that the C-terminus of the helicase domain encoded by CMV-P1 RNA1 determines susceptibility to systemic infection, and that the helicase domain contains six different amino acid substitutions between CMV-Fny and CMV-P1. To identify the key amino acids of the helicase domain determining systemic infection with CMV-P1, we then constructed amino acid substitution mutants. Of the mutants tested, amino acid residues at positions 865, 896, 957, and 980 in the 1a protein sequence of CMV-P1 affected the systemic infection. Virus localization studies with GFP-tagged CMV clones and in situ localization of virus RNA revealed that these four amino acid residues together form the movement determinant for CMV-P1 movement from the epidermal cell layer to mesophyll cell layers. Quantitative real-time PCR revealed that CMV-P1 and a chimeric virus with four amino acid residues of CMV-P1 accumulated more genomic RNA in inoculated leaves than did CMV-Fny, indicating that those four amino acids are also involved in virus replication. These results demonstrate that the C-terminal region of the helicase domain is responsible for systemic infection by controlling virus replication and cell-to-cell movement. Whereas four amino acids are responsible for acquiring virulence in CMV-Fny, six amino acid (positions at 865, 896, 901, 957, 980 and 993) substitutions in CMV-P1 were required for complete loss of virulence in 'Bukang'.
provenance
Made available in Cube on 2018-09-28T13:08:30Z (GMT). No. of bitstreams: 0
language
English
author
Kang, Won-Hee
Seo, Jang-Kyun
Chung, Bong Nam
Kim, Kook-Hyung
Kang, Byoung-Cheorl
accessioned
2018-09-28T13:08:30Z
available
2018-09-28T13:08:30Z
issued
2012
citation
PLOS ONE(7): 8
issn
1932-6203
uri
http://open-repository.kisti.re.kr/cube/handle/open_repository/479778.do
Funder
농림수산식품부
Funding Program
융복합연구센터지원
Project ID
1545002723
Jurisdiction
Rep.of Korea
Project Name
Vegetable Breeding Education & Research
rights
openAccess
type
article


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