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Kelman, D., Kashman, Y., Hill, R. T., Rosenberg, E., & Loya, Y. (2009). Chemical warfare in the sea: The search for antibiotics from Red Sea corals and sponges. Pure Appl. Chem., 81(6), 1113–1121.
Abstract: Marine sponges and corals are widely recognized as rich sources of novel bioactive natural products. These organisms are frequently colonized by bacteria. Some of these bacteria can be pathogenic or serve as beneficial symbionts. Therefore, these organisms need to regulate the bacteria they encounter and resist microbial pathogens. One method is by chemical defense. Antimicrobial assays performed with extracts of 23 Red Sea corals and sponges against bacteria isolated from their natural environment revealed considerable variability in antimicrobial activity. Soft corals exhibited appreciable activity, sponges showed variability, and stony corals had little or no activity. Among the soft corals, Xenia macrospiculata exhibited the highest activity. Bioassay-directed fractionation of the extract indicated that the activity was due to a range of compounds, one of which was isolated and identified as the diterpene desoxyhavannahine. Among the sponges, Amphimedon chloros exhibited strong activity. Bioassay-directed fractionation resulted in the isolation of the pyridinium alkaloid antibiotics, the halitoxins and amphitoxins. These compounds showed selective activity against specific bacteria, rather than being broad-spectrum. They were highly active against seawater bacteria, whereas bacteria associated with the sponge were resistant. This selective toxicity may be important in enabling certain bacteria to live in close association with their sponge host while it maintains a chemical defense against microbial pathogenesis. The halitoxin-resistant bacteria were identified by 16S rRNA gene analysis as Alphaproteobacteria, closely related to other Alphaproteobacteria isolated from various marine sponges. The study of microbial communities associated with sponges and corals has important implications for the production of symbiont-dcrived bioactive compounds and for the use of corals and sponges as source material for microbial diversity in screening programs for natural products.
Keywords: marine natural products; antimicrobial activity; sponges; corals; Red Sea; terpenes; alkaloids; halitoxins
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Zhao, Y. L., Wang, K., Jiao, N. Z., & Chen, F. (2009). Genome sequences of two novel phages infecting marine roseobacters. Environ. Microbiol., 11(8), 2055–2064.
Abstract: P>Two bacteriophages, DSS3 Theta 2 and EE36 Theta 1, which infect marine roseobacters Silicibacter pomeroyi DSS-3 and Sulfitobacter sp. EE-36, respectively, were isolated from Baltimore Inner Harbor water. These two roseophages resemble bacteriophage N4, a large, short-tailed phage infecting Escherichia coli K12, in terms of their morphology and genomic structure. The full genome sequences of DSS3 Theta 2 and EE36 Theta 1 reveal that their genome sizes are 74.6 and 73.3 kb, respectively, and they both contain a highly conserved N4-like DNA replication and transcription system. Both roseophages contain a large virion-encapsidated RNA polymerase gene (> 10 kb), which was first discovered in N4. DSS3 Theta 2 and EE36 Theta 1 also possess several genes (i.e. ribonucleotide reductase and thioredoxin) that are most similar to the genes in roseobacters. Overall, the two roseophages are highly closely related, and share 80-94% nucleotide sequence identity over 85% of their ORFs. This is the first report of N4-like phages infecting marine bacteria and the second report of N4-like phage since the discovery of phage N4 40 years ago. The finding of these two N4-like roseophages will allow us to further explore the specific phage-host interaction and evolution for this unique group of bacteriophages.
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Eissler, Y., Wang, K., Chen, F., Wommack, K. E., & Coats, D. W. (2009). ULTRASTRUCTURAL CHARACTERIZATION OF THE LYTIC CYCLE OF AN INTRANUCLEAR VIRUS INFECTING THE DIATOM CHAETOCEROS CF. WIGHAMII(BACILLARIOPHYCEAE) FROM CHESAPEAKE BAY, USA. J. Phycol., 45(4), 787–797.
Abstract: Numerous microalgal species are infected by viruses that have the potential to control phytoplankton dynamics by reducing host populations, preventing bloom formation, or causing the collapse of blooms. Here we describe a virus infecting the diatom Chaetoceros cf. wighamii Brightw. from the Chesapeake Bay. To characterize the morphology and lytic cycle of this virus, we conducted a time-course experiment, sampling every 4 h over 72 h following viral inoculation. In vivo fluorescence began to decline 16 h after inoculation and was reduced to < 19% of control cultures by the end of experiment. TEM confirmed infection within the first 8 h of inoculation, as indicated by the presence of virus-like particles (VLP) in the nuclei. VLP were present in two different arrangements: rod-like structures that appeared in cross-section as paracrystalline arrays of hexagonal-shaped profiles measuring 12 +/- 2 nm in diameter and uniformly electron-dense hexagonal-shaped particles measuring similar to 22-28 nm in diameter. Nuclei containing paracrystalline arrays were most prevalent early in the infection cycle, while cells containing VLP increased and then declined toward the end of the cycle. The proportion of nuclei containing both paracrystalline arrays and VLP remained relatively constant. This pattern suggests that rod-like paracrystalline arrays fragmented to produce icosahedral VLP. C. cf. wighamii nuclear inclusion virus (CwNIV) is characterized by a high burst size (averaged 26,400 viruses per infected cell) and fast generation time that could have ecological implications on C. cf. wighamii population control.
Keywords: algal virus; Chaetoceros; diatom; viral ecology; virus
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Bachvaroff, T. R., Place, A. R., & Coats, D. W. (2009). Expressed Sequence Tags from Amoebophrya sp Infecting Karlodinium veneficum: Comparing Host and Parasite Sequences. J. Eukaryot. Microbiol., 56(6), 531–541.
Abstract: Parasitic dinoflagellates of the genus Amoebophrya play important roles in the ecology of estuaries and open ocean environments. Little is known of the cell and molecular biology of Amoebophrya, but the genus is intermediate on phylogenetic trees between apicomplexans and typical dinophycean dinoflagellates. Here, we constructed four cDNA libraries, from different stages after infecting the host, Karlodinium veneficum, with Amoebophrya sp. These libraries were used to generate 898 expressed sequence tags (ESTs), with sequences attributed to either the host or parasite, based on AT bias, codon usage, and occurrence during infection. Overall, 209 sequences were attributable to the parasite and 685 to the host. The 50 putative parasite sequences with good protein matches in GenBank were used to find the same protein from host ESTs. For 26 genes, both host and parasite sequences were identified, of which 20 encoded ribosomal proteins. PCR for seven predicted parasite and two host genes were used to confirm attributions. The most common host and parasite ESTs were compared to see if multiple gene copies were present. The host plastocyanin gene had multiple sequence variants, but parasite rps27a contained only one polymorphism, likely due to an amplification error. Amplification, cloning, and sequencing of five parasite protein-coding genes suggested that the parasite has a single sequence for each gene, but three host genes were found to have multiple variants. The genome of Amoebophrya sp. infecting K. veneficum appears to have an organization more similar to other eukaryotes than to the tandem gene arrangements found in dinoflagellates.
Keywords: Expressed genes; parasitism; Syndiniales; transcriptome
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Chen, F., Wang, K., Huang, S. J., Cai, H. Y., Zhao, M. R., Jiao, N. Z., et al. (2009). Diverse and dynamic populations of cyanobacterial podoviruses in the Chesapeake Bay unveiled through DNA polymerase gene sequences. Environ. Microbiol., 11(11), 2884–2892.
Abstract: Many podoviruses have been isolated which infect marine picocyanobacteria, and they may play a potentially important role in regulating the biomass and population composition of picocyanobacteria. However, little is known about the diversity and population dynamics of autochthonous cyanopodoviruses in marine environments. Using a set of newly designed PCR primers which specifically amplify the DNA pol from cyanopodoviruses, a total of 221 DNA pol sequences were retrieved from eight Chesapeake Bay virioplankton communities collected at different times and locations. All DNA pol sequences clustered with the eight known podoviruses that infect different marine picocyanobacteria, and could be divided into at least 10 different subclusters (I-X). The presence of these cyanopodovirus genotypes based on PCR-amplification of DNA pol gene sequences was supported by the existence of similar DNA pol genotypes with metagenome libraries of Chesapeake Bay virioplankton assemblages. The composition of cyanopodoviruses in the Bay also exhibited distinct winter and summer patterns which were likely related to corresponding seasonal changes in the composition of cyanobacterial populations. Our study suggests that diverse and dynamic populations of cyanopodoviruses are present in the estuarine environment. The PCR method developed in this study provides a specific and sensitive tool to explore the abundance, distribution and phylogenetic diversity of cyanopodoviruses in aquatic environments. Linking the dynamics of host and viral populations in the natural environment is critical to broader characterization of the ecological role of virioplankton within microbial communities.
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