| Contributors | Affiliation | Role |
|---|---|---|
| Jiang, Sunny | University of California-Irvine (UC Irvine) | Principal Investigator |
| Kinkade, Danie | Woods Hole Oceanographic Institution (WHOI BCO-DMO) | BCO-DMO Data Manager |
Bacterial total genome DNA was extracted from the single species Pseudo-nitzschia culture. 16S rDNA pyrosequencing were carried out using the bTEFAP method based on the Titanium reagents and protocol for Genome Sequencer FLX System.
| File |
|---|
diatom_microbiome.csv (Comma Separated Values (.csv), 176 bytes) MD5:262860733979b675a15baeb087ba1f47 Primary data file for dataset ID 564378 |
| Parameter | Description | Units |
| project_type | dimensionless | |
| attributes | dimensionless | |
| NCBI_bioproject_id | dimensionless |
| Dataset-specific Instrument Name | Seqman II |
| Generic Instrument Name | Automated DNA Sequencer |
| Dataset-specific Description | A Seqman II (version 9.0 DNASTAR Inc, Madison, WI, USA) was used for sequencing of bacterial 16s rDNA. |
| Generic Instrument Description | General term for a laboratory instrument used for deciphering the order of bases in a strand of DNA. Sanger sequencers detect fluorescence from different dyes that are used to identify the A, C, G, and T extension reactions. Contemporary or Pyrosequencer methods are based on detecting the activity of DNA polymerase (a DNA synthesizing enzyme) with another chemoluminescent enzyme. Essentially, the method allows sequencing of a single strand of DNA by synthesizing the complementary strand along it, one base pair at a time, and detecting which base was actually added at each step. |
| Dataset-specific Instrument Name | Genome Sequencer FLX System |
| Generic Instrument Name | Automated DNA Sequencer |
| Dataset-specific Description | Genome Sequencer FLX System was used for pyroseqencing of pseudo-nitzschia microbiome. |
| Generic Instrument Description | General term for a laboratory instrument used for deciphering the order of bases in a strand of DNA. Sanger sequencers detect fluorescence from different dyes that are used to identify the A, C, G, and T extension reactions. Contemporary or Pyrosequencer methods are based on detecting the activity of DNA polymerase (a DNA synthesizing enzyme) with another chemoluminescent enzyme. Essentially, the method allows sequencing of a single strand of DNA by synthesizing the complementary strand along it, one base pair at a time, and detecting which base was actually added at each step. |
| Dataset-specific Instrument Name | Light Microscope |
| Generic Instrument Name | Microscope - Optical |
| Generic Instrument Description | Instruments that generate enlarged images of samples using the phenomena of reflection and absorption of visible light. Includes conventional and inverted instruments. Also called a "light microscope". |
| Dataset-specific Instrument Name | Plankton Net |
| Generic Instrument Name | Plankton Net |
| Dataset-specific Description | 20um mesh size plankton net was used for sampling. |
| Generic Instrument Description | A Plankton Net is a generic term for a sampling net that is used to collect plankton. It is used only when detailed instrument documentation is not available. |
| Dataset-specific Instrument Name | GeneAmp 2700 PCR system |
| Generic Instrument Name | Thermal Cycler |
| Dataset-specific Description | GeneAmp 2700 PCR system (Applied Biosystems, Foster City, CA) was used for PCR amplification. |
| Generic Instrument Description | A thermal cycler or "thermocycler" is a general term for a type of laboratory apparatus, commonly used for performing polymerase chain reaction (PCR), that is capable of repeatedly altering and maintaining specific temperatures for defined periods of time. The device has a thermal block with holes where tubes with the PCR reaction mixtures can be inserted. The cycler then raises and lowers the temperature of the block in discrete, pre-programmed steps. They can also be used to facilitate other temperature-sensitive reactions, including restriction enzyme digestion or rapid diagnostics.
(adapted from http://serc.carleton.edu/microbelife/research_methods/genomics/pcr.html) |
| Website | |
| Platform | Santa Cruz Warf |
| Start Date | 2011-06-15 |
| End Date | 2011-06-15 |
| Description | Sampling location for the project 'Diatom Microbiome' (PI: Sunny Jiang). A 20um mesh plankton net was used to collect sample. |
This project seeks to elucidate the interactions between viruses, bacteria and toxic bloom-forming marine diatom genus Pseudo-nitzschia in the coastal marine environment. The cosmopolitan Pseudo-nitzschia sp. (Bacillariophyceae) causes harmful algal blooms in various parts of the world. Recent genome sequences of two marine diatoms revealed that bacterial genes contribute up to 5% of their genome make-up, highlighting the close association of marine diatoms with bacteria in evolutionary history. So far, few have looked at the interactions of diatoms with their epibiotic bacteria and none have studied the interactions of virus, bacteria and phytoplankton using direct experimental approach.
This project investigates the physiological, biochemical, genetic and ecological interactions of algae-microbe associations in order to decipher the influences of microbes on algal bloom dynamics and toxigenesis. By incorporating methods in phylogeny, physiology, biochemistry, comparative genomics and metagenomics, the research seeks answers to the following questions: 1) What are the epiphytic bacteria associating with different species of Pseudo-nitzschia and how do they change at different stages of the bloom? 2) How do these bacteria affect the physiology of the Pseudo- nitzschia hosts and how does the algal host regulate these microbial interactions? 3) Can viruses promote a bloom by selectively lysing algicidal bacteria, while creating opportunities for growth- promoting bacteria or vice versa? This project is guided by the preliminary research on microbe- algae interaction, where significantly different epibiotic bacterial communities were found to associate with two Pseudo-nitzschia species. Bacterial associates of one Pseudo-nitzschia sp. stimulated the growth of the algal-host but acted as a pathogen on another Pseudo-nitzschia sp. This project will expand the work on epibiotic bacterial community diversity and dynamics by sampling additional species of Pseudo-nitzschia and investigating the changes in epibiotic and planktonic bacterial community structure following the initiation, peak and decline phases of Pseudo-nitzschia blooms in coastal oceans. Algal host responses to epibionts association will be investigated using physiological experiments of binary and multi-culture experimental approaches. The chemical communication between algal host and epibiont bacteria will be addressed by looking at the algal exudate excretions and bacterial enzyme secretions using established LC-MS and enzyme assays. To illustrate the influences of viruses on bacterial-algal interaction, bacteriophages specific to epibiotic bacteria will be isolated and included in the physiological experiments.
Related Publications:
Sison-Mangus, M., Jiang, S., Tran, K. N., Kudela, R. M. Host-specific adaptation governs the interaction of the marine diatom, Pseudo-nitzschia and their microbiota. ISME Journal 8, 63-76 doi:10.1038/ismej.2013.138
Rowe, J.; Jiang, S. Interactions of bacterial microbiome with bloom forming marine diatom pseudo-nitzchia. Ocean Science Meeting. Honolulu, Hawaii. 2014. (http://www.sgmeet.com/osm2014/viewabstract.asp?AbstractID=17958)
Sison-Mangus, M.; Jiang, S.; Tran, K.; Kudela, R. Factors that influence the association between marine diatom pseudo-nitzschia and their bacterial associates. Ocean Science Meeting. Honolulu, Hawaii. 2014. (http://www.sgmeet.com/osm2014/viewabstract.asp?AbstractID=17910)
Sison-Mangus, M., S. C. Jiang. The microbiota of marine diatoms: is it influenced by algal host phylogeny? American Society for Liminology and Oceanography, Ocean Science Meeting, Salt Lake City, Utah, Feb. 20-24, 2012 (http://www.sgmeet.com/osm2012/viewabstract2.asp?AbstractID=12577)
Sison-Mangus, M. P., Tran, K., Jiang, S. Growth stimulation and killing of pseudo-nitzschia by non-native epibiotic bacteria. American Society for Liminology and Oceanography, Aquatic Science Meeting, San Juan, Pureto Rico, Feb. 13-18, 2011 (https://www.sgmeet.com/aslo/sanjuan2011/viewabstract2.asp?AbstractID=8790)
Tran, K., M., Sison-Mangus, S. C. Jiang. Bacterial diversity associated with toxic and non-toxic pseudo-nitzschia species. American Society for Liminology and Oceanography, Aquatic Science Meeting, San Juan, Pureto Rico, Feb. 13-18, 2011 (http://www.sgmeet.com/aslo/sanjuan2011/viewabstract2.asp?AbstractID=9063)
| Funding Source | Award |
|---|---|
| NSF Division of Ocean Sciences (NSF OCE) |