Isolation and Characterization of a Novel Jumbo Phage from Leaf Litter Compost and Its Suppressive Effect on Rice Seedling Rot Diseases

Jumbo phages have DNA genomes larger than 200 kbp in large virions composed of an icosahedral head, tail, and other adsorption structures, and they are known to be abundant biological substances in nature. In this study, phages in leaf litter compost were screened for their potential to suppress rice seedling rot disease caused by the bacterium Burkholderia glumae, and a novel phage was identified in a filtrate-enriched suspension of leaf litter compost. The phage particles consisted of a rigid tailed icosahedral head and contained a DNA genome of 227,105 bp. The phage could lyse five strains of B. glumae and six strains of Burkholderia plantarii. The phage was named jumbo Burkholderia phage FLC6. Proteomic tree analysis revealed that phage FLC6 belongs to the same clade as two jumbo Ralstonia phages, namely RSF1 and RSL2, which are members of the genus Chiangmaivirus (family: Myoviridae; order: Caudovirales). Interestingly, FLC6 could also lyse two strains of Ralstonia pseudosolanacearum, the causal agent of bacterial wilt, suggesting that FLC6 has a broad host range that may make it especially advantageous as a bio-control agent for several bacterial diseases in economically important crops. The novel jumbo phage FLC6 may enable leaf litter compost to suppress several bacterial diseases and may itself be useful for controlling plant diseases in crop cultivation.

Complete Genome Sequence of the Cluster F1 Mycobacteriophage KingMidas

Subcluster F1 bacteriophage KingMidas was isolated from soil collected in Providence, Rhode Island, using Mycobacterium smegmatis mc2155 as the host. The genome is 57,386 bp and contains 105 predicted protein-coding genes but no transfer-messenger RNAs or tRNAs. This siphovirus has an icosahedral head, with a genome 99.1% identical to that of F1 mycobacteriophage Scottish.

Characteristics of two myoviruses induced from the coastal photoheterotrophic bacterium Porphyrobacter sp. YT40

ABSTRACT In this study, we characterized two induced myoviruses from one marine photoheterotrophic bacterium Porphyrobacter sp. YT40 belonging to the Sphingomonadales family in Alphaproteobacteria. The genome sequence of prophage A is ∼36.9 kb with an average GC content of 67.1%, and its core or functional genes are homologous to Mu or Mu-like phages. Furthermore, induced viral particles from prophage A show a knob-like neck structure, which is only found in bacteriophage Mu. The genome size of prophage B is ∼36.8 kb with an average GC content of 65.3%. Prophage B contains a conserved gene cluster Q-P-O-N-M-L, which is unique in P2 phages. Induced viral particles from prophage B display an icosahedral head with a diameter of ∼55 nm and a 130 ± 5 nm long contractile tail. To our knowledge, this is the first report that characterizes the induced P2-like phage in marine Alphaproteobacteria. Phylogeny analyses suggest that these two types of prophages are commonly found in sequenced bacteria of the Sphingomonadales family. This study sheds light on the ongoing interaction between marine bacteria and phages, and improves our understanding of bacterial genomic plasticity and evolution.

Characterization of the Salmonella bacteriophage vB_SenS-Ent1

The bacteriophage vB_SenS-Ent1 (Ent1) is a member of the family Siphoviridae of tailed bacteriophages and infects a broad range of serovars of the enteric pathogen Salmonella enterica. The virion particle is composed of an icosahedral head 64 nm in diameter and a flexible, non-contractile tail of 116 × 8.5 nm possessing terminal fibres. The adsorption rate constant at 37 °C is 6.73 × 10−9 ml min−1. Latent and eclipse periods are 25 and 20 min, respectively, and the burst size is 35 progeny particles per cell after 35 min at 37 °C. Sequencing revealed a circularly permuted, 42 391 bp dsDNA genome containing 58 ORFs organized into four major transcriptional units. Comparisons with the genome sequences of other bacteriophages revealed a high level of nucleotide sequence identity and shared orthologous proteins with the Salmonella phages SETP3, SE2 and KS7 (SS3e) and the Escherichia phages K1G, K1H, K1ind1 and K1ind3.

Classification of 17 newly isolated virulent bacteriophages of Pseudomonas aeruginosa

Seventeen virulent bacteriophages specific to Pseudomonas aeruginosa strains were isolated by screening various environmental samples. These isolated bacteriophages were grouped based on results obtained from restriction fragment analysis of phage genomes, random amplification of polymorphic DNA (RAPD) typing, morphology observations under transmission electron microscope, and host range analysis. All 17 bacteriophages are double-stranded DNA viruses and can be divided into 5 groups based on DNA restriction profiles. A set of 10-mer primers was used in RAPD typing of phages, and similar conclusions were obtained as for restriction fragment analysis. One phage was randomly selected from each of the 5 groups for morphology observations. Four of them had an icosahedral head with a long contractile tail, belonging to the Myoviridae family, and one phage had an icosahedral head with a short tail, thereby belonging to the Podoviridae family. Host range experiments were conducted on 7 laboratory strains and 12 clinical strains of P. aeruginosa. The results showed that 13 phages had the same infection profile, killing 8 out of 19 tested P. aeruginosa strains, and the remaining 4 phages had different and unique infection profiles. This study highlights the diversity of bacteriophages specific to P. aeruginosa in the environment.

Fighting foam with phages?

Seventeen (17) phages infective for the mycolata were isolated from six samples of activated sludge using 21 prospective hosts from the genera Dietzia, Gordonia, Nocardia, Rhodococcus, Tsukamurella and Mycobacterium. Their morphology indicated that they were all members of the viral family Siphoviridae, but they varied in the size of the icosahedral head and length of non-contractile tail, suggesting they were different. This was confirmed by host-range studies with 47 strains of mycolata, which showed that each phage had a unique host-range, and this was polyvalent in the majority (15/17) of cases, with 12 phages infective for hosts representing two or three of the genera Gordonia, Nocardia and Rhodococcus. The potential for use of these phages in the control of foaming and other applications is discussed.

Phylogeny of the Major Head and Tail Genes of the Wide-Ranging T4-Type Bacteriophages

ABSTRACT We examined a number of bacteriophages with T4-type morphology that propagate in different genera of enterobacteria,Aeromonas, Burkholderia, andVibrio. Most of these phages had a prolate icosahedral head, a contractile tail, and a genome size that was similar to that of T4. A few of them had more elongated heads and larger genomes. All these phages are phylogenetically related, since they each had sequences homologous to the capsid gene (gene23), tail sheath gene (gene 18), and tail tube gene (gene 19) of T4. On the basis of the sequence comparison of their virion genes, the T4-type phages can be classified into three subgroups with increasing divergence from T4: the T-evens, pseudoT-evens, and schizoT-evens. In general, the phages that infect closely related host species have virion genes that are phylogenetically closer to each other than those of phages that infect distantly related hosts. However, some of the phages appear to be chimeras, indicating that, at least occasionally, some genetic shuffling has occurred between the different T4-type subgroups. The compilation of a number of gene 23 sequences reveals a pattern of conserved motifs separated by sequences that differ in the T4-type subgroups. Such variable patches in the gene 23sequences may determine the size of the virion head and consequently the viral genome length. This sequence analysis provides molecular evidence that phages related to T4 are widespread in the biosphere and diverged from a common ancestor in acquiring the ability to infect different host bacteria and to occupy new ecological niches.

Structural Analysis of gp6 and gp6/gpl5/gpl6 Complex by Cryo-Electron Microscopy At 10 Å Resolution

The mechanism of encapsidation of DNA into a bacteriophage head is a most intriguing problem. The portal protein is essential for the assembly of tailed double-stranded DNA (dsDNA) bacteriophages. These turbine-like homo-oligomers consist of 12 or 13 subunits surrounding a central channel. Portal oligomers are located at the vertex of the icosahedral head that binds to the phage tail. It was shown that gp6 portal protein from bacteriophage SPP1 has 13 subunits, prior to their incorporation into the viral procapsid structure. After packaging of the viral DNA inside the phage capsid, additional proteins have been found attached to the portal oligomer: gpl5 and gp 16. A complex of gp6/gpl5/gpl7 forms the connector structure that provides the interface for attachment of the phage tail. We here present the three-dimensional (3D) reconstructions of the gp6 wild-type protein alone and of the gp6/gpl5/gpl6 complex - both at 10Å resolution - based on electron cryo-microscopy using the angular reconstitution single particle methodology.

Isolation of a virulent bacteriophage from a Propionibacterium species in the sheep rumen

Propionibacterium is a facultative anaerobe associated with the rumen epithelium, the presence of which may influence the anaerobic environment through oxygen scavenging, as well as providing a source of propionate. Factors such as bacteriophages that influence Propionibacterium populations may therefore be important regulators of rumen function. This study describes the isolation and identification of a ruminal Propionibacterium bacteriophage. Sheep rumen fluid was screened for Propionibacterium species and 3 isolates were identified and characterised. One isolate, PA1, was used as an indicator strain to screen for the presence of Propionibacterium-specific virulent bacteriophages. A virulent bacteriophage, PB2, was isolated from clear plaques on a lawn of PA1 cells and was shown by transmission electron microscopy to be a siphovirus-like particle comprising an icosahedral head 50 nm in diameter and a tail 140 nm in length. The bacteriophage was visibly attached to and within PA1 cells, and was shown to infect all 3 ruminal isolates of Propionibacterium and 4 of 6 clinical isolates of P. acnes. Restriction mapping of bacteriophage PB2 demonstrated a 30.8 kb genome.

Spontaneous induction of bacteriophage during growth of Azospirillum brasilense in complex media

Spontaneous induction of bacteriophage occurred during growth of Azospirillum brasilense ATCC 29145 in complex media incubated at 27 or 30 °C under agitated and nonagitated conditions. Phage replication on strain 29145 was stimulated by divalent cations and inhibited by NaCl or temperatures greater than 37 °C. The phage morphology as revealed by electron microscopy showed an icosahedral head of 60 nm and a tail of 208 nm. The host range was limited to A. brasilense strains 29145 and 29710. Other Azospirillum strains tested did not produce phage under any of the conditions tested.