G20 Gene Research Articles

T4-Like Cyanophages of Lake Baikal: Genetic Diversity and Biogeography

The work deals with investigation of genetic diversity and biogeography of T4-like cyanophages from the shallow bay of the Posolsk Sor (Lake Baikal), based on analysis of the g20 marker gene. High diversity of g20 gene fragments and their uniqueness were revealed. The greatest similarity was noted with the previously obtained sequences from Lake Baikal and with those from freshwater ecosystems: oligotrophic Lake Green, Lake Round, oligomesotrophic Lake Ancy, and mesotrophic Lake Bourget. From the point of view of biogeography, it was determined that the phage sequences were similar to the previously obtained ones from different Lake Baikal ecotopes than to those from other ecosystems.

Survey of cyanomyovirus abundance in Shantou coastal waters by g20

To understand the genetic diversity and population changes in cyanophages in the coastal waters of Shantou, northeast South China Sea, we used the capsid assembly protein gene g20 as a marker of the abundance and phylogeny of natural cyanomyovirus communities. The abundance of total viruses, heterotrophic bacteria, and picophytoplankton in the coastal waters was monitored with flow cytometry. Hydrological parameters (NO3−, NO2−, NH3, soluble reactive phosphorus, total dissolved nitrogen, total dissolved phosphorus, dissolved oxygen, chemical oxygen demand, temperature, salinity, and chlorophyll a concentration) and microbial abundance (total viruses, total bacteria, Prochlorococcus, Synechococcus, and eukaryotes) were measured in the upper and lower layers at four sampling sites in the research area. In the direct viral counts, cyanomyoviruses accounted for 1.92% to >10% of the total viral community. A phylogenetic analysis showed that the g20 sequences in the Shantou coastal waters were very diverse, distributed in eight distinct operational taxonomic units, including the newly formed Cluster W. The g20 gene copies inferred from real time PCR assay indicated that cyanomyoviruses were correlated significantly with the heterotrophic bacteria numbers and the nitrate and chlorophyll a concentrations. These results suggest that cyanomyoviruses are ubiquitous and are an abundant component of the virioplankton in Shantou coastal waters.

Genetic diversity of cyanophages of the myoviridae family as a constituent of the associated community of the Baikal sponge Lubomirskia baicalensis

Molecular-genetic analysis of cyanophages of the family Myoviridae from the associated community of the endemic Baikal sponge Lubomirskia baicalensis was carried out based on the g20 gene fragment. A large cyanophage diversity according to the g20 marker gene was found in the sponge. The Baikal sponge cyanophages were shown to be similar to those inhabiting plankton. Moreover, specific cyanophage groups that are significantly different from all of the known groups inhabiting the Lake Baikal were revealed.

Molecular genetic diversity of the Myoviridae family cyanophages in Lake Khövsgöl (Mongolia)

The diversity of the Myoviridae family cyanophages that inhabit Lake Khovsgol was studied by analyzing g20 gene fragments. It was shown that the gene sequences of cyanomyoviruses are diverse and similar to those of the viruses from Lake Baikal. The presence of closely related virus strains in these lakes is associated with the geographical proximity, direct water connection, and similar hydrochemical parameters of the lakes.

High abundances of cyanomyoviruses in marine ecosystems demonstrate ecological relevance

The distribution of cyanomyoviruses was estimated using a quantitative PCR (qPCR) approach that targeted the g20 gene as a proxy for phage. Samples were collected spatially during a > 3000 km transect through the Sargasso Sea and temporally during a gyre-constrained phytoplankton bloom within the southern Pacific Ocean. Cyanomyovirus abundances were lower in the Sargasso Sea than in the southern Pacific Ocean, ranging from 2.75 × 10(3) to 5.15 × 10(4) mL(-1) and correlating with the abundance of their potential hosts (Prochlorococcus and Synechococcus). Cyanomyovirus abundance in the southern Pacific Ocean (east of New Zealand) followed Synechococcus host populations in the system: this included a decrease in g20 gene copies (from 4.3 × 10(5) to 9.6 × 10(3) mL(-1) ) following the demise of a Synechococcus bloom. When compared with direct counts of viruses, observations suggest that the cyanomyoviruses comprised 0.5 to >25% of the total virus community. We estimated daily lysis rates of 0.2-46% of the standing stock of Synechococcus in the Pacific Ocean compared with c. < 1.0% in the Sargasso Sea. In total, our observations confirm this family of viruses is abundant in marine systems and that they are an important source of cyanobacterial mortality.

Portal protein diversity and phage ecology

Recent whole-genome cyanophage sequencing (Sullivan et al., 2010) has revealed that the sequences of some genes do not match those published previously as single-gene sequences derived from DNA fragments PCR-amplified from phage isolates. Specifically, the g20 gene sequences for S-SSM7 (gene GI:189397276, protein GI:189397277) and Syn33 (gene GI:189397306, protein GI:189397307) in Sullivan and colleagues (2008) were incorrect. The revised sequences do not alter g20 tree topology or the conclusions of Sullivan and colleagues (2008). The source of error for these gene sequences is not certain, but could have its origin in either a clerical error, PCR contamination and/or mixed phage stocks. On this latter point, it has recently come to light that triplicate plaque purifications do not always result in pure stocks for these cyanophages, possibly due to the fluid consistency of the agarose plates (0.28% agarose) required for growth of the host. More reliable purification procedures have subsequently been developed. However, as mixed phage stock preparations could influence host range, we recommend that published host range data (Sullivan et al., 2003) be verified before their use in further research.

New phylogenetically distinct cyanophages found in the coastal Yellow Sea by Qingdao

Genetic composition of natural cyanophage population in the coastal Yellow Sea area near Qingdao was investigated using the g20 gene as a genetic marker. Amplification of this gene by PCR yielded sequences, which were cloned and subjected to the restriction fragment length polymorphism (RFLP) analysis. Sixteen clones exhibiting different RFLP patterns were sequenced and analyzed. The sequence analysis indicated a diversity of cyanophages covering sixteen g20 genotypes. Their phylogenetic analysis revealed two distinct phylogenetic groups. One group (7clones) localized with the cyanophage P77 in the cluster III described before. Second group (9 clones) did not localize with any of the cyanophage described so far and was designated as a novel W cluster. The study revealed a genetic diversity of cyanophages in the coastal Yellow Sea and confirmed the genetic differences between cyanophage populations in Western and Eastern Pacific.

Seasonal Variations in Virus-Host Populations in Norwegian Coastal Waters: Focusing on the Cyanophage Community Infecting Marine Synechococcus spp

Viruses are ubiquitous components of the marine ecosystem. In the current study we investigated seasonal variations in the viral community in Norwegian coastal waters by pulsed-field gel electrophoresis (PFGE). The results demonstrated that the viral community was diverse, displaying dynamic seasonal variation, and that viral populations of 29 different sizes in the range from 26 to 500 kb were present. Virus populations from 260 to 500 kb and dominating autotrophic pico- and nanoeukaryotes showed similar dynamic variations. Using flow cytometry and real-time PCR, we focused in particular on one host-virus system: Synechococcus spp. and cyanophages. The two groups covaried throughout the year and were found in the highest amounts in fall with concentrations of 7.3 x 10(4) Synechococcus cells ml(-1) and 7.2 x 10(3) cyanophage ml(-1). By using primers targeting the g20 gene in PCRs on DNA extracted from PFGE bands, we demonstrated that cyanophages were found in a genomic size range of 26 to 380 kb. The genetic richness of the cyanophage community, determined by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified g20 gene fragments, revealed seasonal shifts in the populations, with one community dominating in spring and summer and a different one dominating in fall. Phylogenetic analysis of the sequences originating from PFGE and DGGE bands grouped the sequences into three groups, all with homology to cyanomyoviruses present in cultures. Our results show that the cyanophage community in Norwegian coastal waters is dynamic and genetically diverse and has a surprisingly wide genomic size range.

Genetic diversity of marine Synechococcus and co‐occurring cyanophage communities: evidence for viral control of phytoplankton

Unicellular cyanobacteria of the genus Synechococcus are a major component of the picophytoplankton and make a substantial contribution to primary productivity in the oceans. Here we provide evidence that supports the hypothesis that virus infection can play an important role in determining the success of different Synechococcus genotypes and hence of seasonal succession. In a study of the oligotrophic Gulf of Aqaba, Red Sea, we show a succession of Synechococcus genotypes over an annual cycle. There were large changes in the genetic diversity of Synechococcus, as determined by restriction fragment length polymorphism analysis of a 403- bp rpoC1 gene fragment, which was reduced to one dominant genotype in July. The abundance of co-occurring cyanophage capable of infecting marine Synechococcus was determined by plaque assays and their genetic diversity was determined by denaturing gradient gel electrophoresis analysis of a 118-bp g20 gene fragment. The results indicate that both abundance and genetic diversity of cyanophage covaried with that of Synechococcus. Multivariate statistical analyses show a significant relationship between cyanophage assemblage structure and that of Synechococcus. These observations are consistent with cyanophage infection being a major controlling factor in picophytoplankton succession.

Genetic diversity and population dynamics of cyanophage communities in the Chesapeake Bay

AME Aquatic Microbial Ecology Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials AME 34:105-116 (2004) - doi:10.3354/ame034105 Genetic diversity and population dynamics of cyanophage communities in the Chesapeake Bay Kui Wang, Feng Chen* Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 East Pratt Street, Suite 266, Baltimore, Maryland 21202, USA *Corresponding author. Email: chenf@umbi.umd.edu ABSTRACT: In order to understand the genetic diversity and population dynamics of cyanophages in estuarine waters, the viral capsid assembly (g20) gene was used as a gene marker to monitor genetic variations of natural cyanomyovirus communities in the Chesapeake Bay, USA. Unique and diverse g20 sequences were found. Only 1 of 15 g20 genotypes was closely related to the known cyanomyovirus isolates. Most of the g20 genotypes in the bay were not related to the g20 clonal sequences recovered from open-ocean waters. Terminal-restriction fragment length polymorphism (T-RFLP) based on the g20 gene was developed to investigate spatial and temporal distribution of cyanomyovirus communities in the bay. The T-RFLP profiles of the g20 gene demonstrated that the cyanomyovirus population structures in the bay were more dynamic seasonally than spatially. Seasonal variation in the cyanophage community appeared to correspond to changes in host-cell density, which in turn was mainly affected by water temperature. This study represents the first effort to monitor both cyanophage titer and genetic diversity over time and space. The results of our study suggest that cyanophages could play a significant role in regulating Synechococcus biomass and population structure in the Chesapeake Bay. KEY WORDS: Cyanophage · Synechococcus · Natural virus community · Phylogenetic diversity · T-RFLP Full article in pdf format NextExport citation RSS - Facebook - Tweet - linkedIn Cited by Published in AME Vol. 34, No. 2. Online publication date: February 04, 2004 Print ISSN: 0948-3055; Online ISSN: 1616-1564 Copyright © 2004 Inter-Research.

The physical environment affects cyanophage communities in British Columbia inlets.

Little is known about the natural distribution of viruses that infect the photosynthetically important group of marine prokaryotes, the cyanobacteria. The current investigation reveals that the structure of cyanophage communities is dependent on water column structure. PCR was used to amplify a fragment of the cyanomyovirus gene (g) 20, which codes for the portal vertex protein. Denaturing gradient gel electrophoresis (DGGE) of PCR amplified g20 gene fragments was used to examine variations in cyanophage community structure in three inlets in British Columbia, Canada. Qualitative examination of denaturing gradient gels revealed cyanophage community patterns that reflected the physical structure of the water column as indicated by temperature and salinity. Based on mobility of PCR fragments in the DGGE gels, some cyanophages appeared to be widespread, while others were observed only at specific depths. Cyanophage communities within Salmon Inlet were more related to one another than to communities from either Malaspina Inlet or Pendrell Sound. As well, surface communities in Malaspina Inlet and Pendrell Sound were different when compared to communities at depth. In the same two locations, distinct differences in community composition were observed in communities that coincided with depths of high chlorophyll fluorescence. The observed community shifts over small distances (only a few meters in depth or inlets separated by less than 100 km) support the idea that cyanophage communities separated by small spatial scales develop independently of each other as a result isolation by water column stratification or land mass separation, which may ultimately lead to changes in the distribution or composition of the host community.

Genetic diversity and temporal variation in the cyanophage community infecting marine Synechococcus species in Rhode Island's coastal waters.

The cyanophage community in Rhode Island's coastal waters is genetically diverse and dynamic. Cyanophage abundance ranged from over 10(4) phage ml(-1) in the summer months to less then 10(2) phage ml(-1) during the winter months. Thirty-six distinct cyanomyovirus g20 genotypes were identified over a 3-year sampling period; however, only one to nine g20 genotypes were detected at any one sampling date. Phylogenetic analyses of g20 sequences revealed that the Rhode Island cyanomyoviral isolates fall into three main clades and are closely related to other known viral isolates of Synechococcus spp. Extinction dilution enrichment followed by host range tests and PCR restriction fragment length polymorphism analysis was used to detect changes in the relative abundance of cyanophage types in June, July, and August 2002. Temporal changes in both the overall composition of the cyanophage community and the relative abundance of specific cyanophage g20 genotypes were observed. In some seawater samples, the g20 gene from over 50% of isolated cyanophages could not be amplified by using the PCR primer pairs specific for cyanomyoviruses, which suggested that cyanophages in other viral families (e.g., Podoviridae or Siphoviridae) may be important components of the Rhode Island cyanophage community.