Molecular and Biological Characterization of Ralstonia Phage RsoM1USA, a New Species of P2virus , Isolated in the United States

The first -infecting bacteriophage from soil of the United States, designated RsoM1USA, was isolated from a tomato field in Florida. Electron microscopy revealed that phage RsoM1USA is member of the genus in the family with an icosahedral head of about 66 nm in diameter, a contractile tail of about...

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Published inFrontiers in microbiology Vol. 10; p. 267
Main Authors Addy, Hardian Susilo, Ahmad, Abdelmonim Ali, Huang, Qi
Format Journal Article
LanguageEnglish
Published Switzerland Frontiers Media S.A 19.02.2019
Subjects
characterization
isolation
Microbiology
Myoviridae
P2virus
Ralstonia phage
Ralstonia solanacearum
Ralstonia phage
isolation
Ralstonia solanacearum
characterization
Myoviridae
P2virus
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Abstract The first -infecting bacteriophage from soil of the United States, designated RsoM1USA, was isolated from a tomato field in Florida. Electron microscopy revealed that phage RsoM1USA is member of the genus in the family with an icosahedral head of about 66 nm in diameter, a contractile tail of about 152 nm in length, and a long "neck." Phage RsoM1USA infected 12 of the 30 tested species complex strains collected worldwide in each of the three species: , , and . The phage completed its infection cycle 180 min post infection with a burst size of about 56 particles per cell. Phage RsoM1USA has a genome of 39,309 nucleotides containing 58 open reading frames (ORFs) and is closely related to Ralstonia phage RSA1 of the species . The genomic organization of phage RsoM1USA is also similar to that of phage RSA1, but their integrases share no sequence homology. In addition, we determined that the integration of phage RsoM1USA into its susceptible strain K60 is mediated by the 3' 45-base portion of the threonine tRNA (TGT), not arginine tRNA (CCG) as reported for phage RSA1, confirming that the two phages use different mechanism for integration. Our proteomic analysis of the purified virions supported the annotation of the main structural proteins. Infection of a susceptible strain RUN302 by phage RsoM1USA resulted in significantly reduced growth of the infected bacterium , but not virulence in tomato plants, as compared to its uninfected RUN302 strain. Due to its differences from phage RSA1, phage RsoM1USA should be considered the type member of a new species with a proposed species name of
AbstractList The first -infecting bacteriophage from soil of the United States, designated RsoM1USA, was isolated from a tomato field in Florida. Electron microscopy revealed that phage RsoM1USA is member of the genus in the family with an icosahedral head of about 66 nm in diameter, a contractile tail of about 152 nm in length, and a long "neck." Phage RsoM1USA infected 12 of the 30 tested species complex strains collected worldwide in each of the three species: , , and . The phage completed its infection cycle 180 min post infection with a burst size of about 56 particles per cell. Phage RsoM1USA has a genome of 39,309 nucleotides containing 58 open reading frames (ORFs) and is closely related to Ralstonia phage RSA1 of the species . The genomic organization of phage RsoM1USA is also similar to that of phage RSA1, but their integrases share no sequence homology. In addition, we determined that the integration of phage RsoM1USA into its susceptible strain K60 is mediated by the 3' 45-base portion of the threonine tRNA (TGT), not arginine tRNA (CCG) as reported for phage RSA1, confirming that the two phages use different mechanism for integration. Our proteomic analysis of the purified virions supported the annotation of the main structural proteins. Infection of a susceptible strain RUN302 by phage RsoM1USA resulted in significantly reduced growth of the infected bacterium , but not virulence in tomato plants, as compared to its uninfected RUN302 strain. Due to its differences from phage RSA1, phage RsoM1USA should be considered the type member of a new species with a proposed species name of
The first Ralstonia-infecting bacteriophage from soil of the United States, designated RsoM1USA, was isolated from a tomato field in Florida. Electron microscopy revealed that phage RsoM1USA is member of the genus P2virus in the family Myoviridae with an icosahedral head of about 66 nm in diameter, a contractile tail of about 152 nm in length, and a long “neck.” Phage RsoM1USA infected 12 of the 30 tested R. solanacearum species complex strains collected worldwide in each of the three Ralstonia species: R. solanacearum, R. pseudosolanacearum, and R. syzygii. The phage completed its infection cycle 180 min post infection with a burst size of about 56 particles per cell. Phage RsoM1USA has a genome of 39,309 nucleotides containing 58 open reading frames (ORFs) and is closely related to Ralstonia phage RSA1 of the species Ralstonia virus RSA1. The genomic organization of phage RsoM1USA is also similar to that of phage RSA1, but their integrases share no sequence homology. In addition, we determined that the integration of phage RsoM1USA into its susceptible R. solanacearum strain K60 is mediated by the 3′ 45-base portion of the threonine tRNA (TGT), not arginine tRNA (CCG) as reported for phage RSA1, confirming that the two phages use different mechanism for integration. Our proteomic analysis of the purified virions supported the annotation of the main structural proteins. Infection of a susceptible R. solanacearum strain RUN302 by phage RsoM1USA resulted in significantly reduced growth of the infected bacterium in vitro, but not virulence in tomato plants, as compared to its uninfected RUN302 strain. Due to its differences from phage RSA1, phage RsoM1USA should be considered the type member of a new species with a proposed species name of Ralstonia virus RsoM1USA.
The first Ralstonia -infecting bacteriophage from soil of the United States, designated RsoM1USA, was isolated from a tomato field in Florida. Electron microscopy revealed that phage RsoM1USA is member of the genus P2virus in the family Myoviridae with an icosahedral head of about 66 nm in diameter, a contractile tail of about 152 nm in length, and a long “neck.” Phage RsoM1USA infected 12 of the 30 tested R. solanacearum species complex strains collected worldwide in each of the three Ralstonia species: R. solanacearum , R. pseudosolanacearum , and R. syzygii . The phage completed its infection cycle 180 min post infection with a burst size of about 56 particles per cell. Phage RsoM1USA has a genome of 39,309 nucleotides containing 58 open reading frames (ORFs) and is closely related to Ralstonia phage RSA1 of the species Ralstonia virus RSA1 . The genomic organization of phage RsoM1USA is also similar to that of phage RSA1, but their integrases share no sequence homology. In addition, we determined that the integration of phage RsoM1USA into its susceptible R. solanacearum strain K60 is mediated by the 3′ 45-base portion of the threonine tRNA (TGT), not arginine tRNA (CCG) as reported for phage RSA1, confirming that the two phages use different mechanism for integration. Our proteomic analysis of the purified virions supported the annotation of the main structural proteins. Infection of a susceptible R. solanacearum strain RUN302 by phage RsoM1USA resulted in significantly reduced growth of the infected bacterium in vitro , but not virulence in tomato plants, as compared to its uninfected RUN302 strain. Due to its differences from phage RSA1, phage RsoM1USA should be considered the type member of a new species with a proposed species name of Ralstonia virus RsoM1USA.
Author Ahmad, Abdelmonim Ali
Addy, Hardian Susilo
Huang, Qi
AuthorAffiliation 2 Department of Plant Protection, Faculty of Agriculture, University of Jember , Jember , Indonesia
1 Floral and Nursery Plants Research Unit, United States National Arboretum, United States Department of Agriculture–Agricultural Research Service , Beltsville, MD , United States
3 Department of Plant Pathology, Faculty of Agriculture, Minia University , El-minia , Egypt
AuthorAffiliation_xml – name: 1 Floral and Nursery Plants Research Unit, United States National Arboretum, United States Department of Agriculture–Agricultural Research Service , Beltsville, MD , United States
– name: 3 Department of Plant Pathology, Faculty of Agriculture, Minia University , El-minia , Egypt
– name: 2 Department of Plant Protection, Faculty of Agriculture, University of Jember , Jember , Indonesia
Author_xml – sequence: 1
  givenname: Hardian Susilo
  surname: Addy
  fullname: Addy, Hardian Susilo
  organization: Department of Plant Protection, Faculty of Agriculture, University of Jember, Jember, Indonesia
– sequence: 2
  givenname: Abdelmonim Ali
  surname: Ahmad
  fullname: Ahmad, Abdelmonim Ali
  organization: Department of Plant Pathology, Faculty of Agriculture, Minia University, El-minia, Egypt
– sequence: 3
  givenname: Qi
  surname: Huang
  fullname: Huang, Qi
  organization: Floral and Nursery Plants Research Unit, United States National Arboretum, United States Department of Agriculture-Agricultural Research Service, Beltsville, MD, United States
BackLink https://www.ncbi.nlm.nih.gov/pubmed/30837978$$D View this record in MEDLINE/PubMed
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Keywords Ralstonia phage
isolation
Ralstonia solanacearum
characterization
Myoviridae
P2virus
Language English
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Edited by: Ramy Karam Aziz, Cairo University, Egypt
This article was submitted to Virology, a section of the journal Frontiers in Microbiology
Reviewed by: Jessica Sacher, University of Alberta, Canada; Hany Anany, Agriculture and Agri-Food Canada (AAFC), Canada
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Snippet The first -infecting bacteriophage from soil of the United States, designated RsoM1USA, was isolated from a tomato field in Florida. Electron microscopy...
The first Ralstonia -infecting bacteriophage from soil of the United States, designated RsoM1USA, was isolated from a tomato field in Florida. Electron...
The first Ralstonia-infecting bacteriophage from soil of the United States, designated RsoM1USA, was isolated from a tomato field in Florida. Electron...
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StartPage 267
SubjectTerms characterization
isolation
Microbiology
Myoviridae
P2virus
Ralstonia phage
Ralstonia solanacearum
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Title Molecular and Biological Characterization of Ralstonia Phage RsoM1USA, a New Species of P2virus , Isolated in the United States
URI https://www.ncbi.nlm.nih.gov/pubmed/30837978
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