Characterization and genome analysis of phage AL infecting Pseudoalteromonas marina
Xinran Zhang1, Fang Zhang2, Ye Mi3
1College of Marine Life Sciences and Institute of Evolution and Marine Biodiversity, Ocean University of China, Qingdao, China.
Abstract:
Although Pseudoalteromonas is an abundant, ubiquitous, marine algae-associated bacterial genus, there is still little information on their phages. In the present study, a marine phage AL, infecting Pseudoalteromonas marina, was isolated from the coastal waters off Qingdao. The AL phage is a siphovirus with an icosahedral head of 53 ± 1 nm and a non-contractile tail, length of 99 ± 1 nm. A one-step growth curve showed that the latent period was approximately 70 min, the rise period was 50 min, and the burst size was 227 pfu/cell. The genome sequence of this phage is a 33,582 bp double-stranded DNA molecule with a GC content of 40.1 %, encoding 52 open reading frames (ORFs). The order of the functional genes, especially those related to the structure module, is highly conserved and basically follows the common pattern used by siphovirus. The stable order has been formed during the long-term evolution of phages in the siphovirus group, which has helped the phages to maintain their normal morphology and function. Phylogenetic trees based on the major capsid protein (mcp) and genome-wide sequence have shown that the AL phage is closely related to four Pseudoalteromonas phages, including PHS21, PHS3, SL25 and Pq0. Further analysis using all-to-all BLASTP also confirmed that this phage shared high sequence homology with the same four Pseudoalteromonas phages, with amino acid sequence identities ranging from 44 % to 71 %. In particular, their similarity in virion structure module may imply that these phages share common assembly mechanism characteristics and infection pathways. Pseudoalteromonas phage AL not only provides basic information for the further study of the evolution of Pseudoalteromonas phages and interactions between marine phage and host but also helps to explain the unknown viral sequences in the metagenomic databases.
Insights
Researchers isolated marine phage AL, a siphovirus infecting Pseudoalteromonas marina. This discovery enhances understanding of marine phage-host interactions and viral evolution in Pseudoalteromonas bacteria.
Area of Science:
- Marine microbiology
- Virology
- Bacteriophage research
Background:
- Pseudoalteromonas bacteria are abundant marine microbes associated with algae.
- Limited knowledge exists regarding the phages that infect Pseudoalteromonas species.
- Understanding these phages is crucial for marine microbial ecology.
Purpose of the Study:
- To isolate and characterize a novel marine phage infecting Pseudoalteromonas marina.
- To analyze the genomic and morphological features of the isolated phage.
- To investigate the evolutionary relationships of the new phage with other Pseudoalteromonas phages.
Main Methods:
- Isolation of phage AL from coastal waters.
- Morphological characterization using electron microscopy.
- Genome sequencing and analysis of open reading frames (ORFs).
- Phylogenetic analysis using major capsid protein (mcp) and whole-genome sequences.
- BLASTP analysis for sequence homology comparison.
Main Results:
- Isolation and identification of marine phage AL, a siphovirus.
- Characterization of phage AL's morphology (icosahedral head, non-contractile tail) and growth curve parameters (latent period, rise period, burst size).
- Determination of a 33,582 bp double-stranded DNA genome with 52 ORFs and conserved gene order.
- Phylogenetic and homology analyses revealed close relationships with four other Pseudoalteromonas phages (PHS21, PHS3, SL25, Pq0).
- High sequence homology suggests shared assembly mechanisms and infection pathways.
Conclusions:
- Phage AL provides essential data for studying Pseudoalteromonas phage evolution and marine phage-host dynamics.
- The conserved gene order in siphoviruses aids in maintaining morphology and function.
- The findings contribute to understanding unknown viral sequences in metagenomic datasets.
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