Biogeophysical interactions control the formation of iron oxide mat microbial communities in acidic geothermal springs
Ecological succession of microbial communities in their natural environment is a complex process that is not well understood. Microbial ecosystems are difficult to examine in part due to the small size of microbial cells and small scale of key biogeochemical processes, where microscopy and molecular techniques must be employed to track the activities microorganisms in their environment. However, modern molecular (e.g., community genomics) and microscopy techniques (e.g., fluorescence in situ hybridization) have allowed for microbial communities to be studied in greater detail. This study focused on the successional development of iron oxide mat microbial ecosystems that occur within high temperature (65-75° C), acidic (pH ~ 3), geothermal spring outflow channels in Yellowstone National Park. These iron oxide microbial communities contain both microorganisms from the domains Archaea and Bacteria that either use inorganic sources of energy (e.g., iron) and fix carbon dioxide (lithoautotrophs) or utilize organic sources of energy and do not fix carbon dioxide (organoheterotrophs). Glass microscope slides were placed in the outflow channel to provide a substrate for microbial growth and total iron deposited was measured as well as community development by using 16S rRNA genes. Results suggest inter-spring variation of iron deposition likely caused by differences in flow and seasonal variations due to increases UV radiation causing an increase in viral pressure in summer months. Lithoautotrophs were often found to be more dominant in early stages of community development, whereas organoheterotrophs were more abundant when the community reached a pseudo-steady state. Thus, iron oxide microbial community development is a dynamic process.
Joseph Yavitt
Faculty: Project Co-PI
Does a glass slide act as a suitable experimental substrate for the geochemical processes involved? It seems that the rocks in the pools are a much larger source of iron than a glass slide?
Jake Beam
Dear Dr. Yavitt,
Thank you for your inquiry and interest in my research. The glass slide (as SiO2) mimics the rhyolitic substratum (which is predominantly SiO2) that these iron oxide microbial mats form on initially when a new geothermal spring source appears. The ferrous iron that is utilized by iron-oxidizing microbes in this geothermal environment originates from the overlying thermal waters. The images on the poster represent mature iron oxide microbial mats that contain significant amounts of accreted iron oxide. You are correct in that iron oxidation and subsequent accretion of Fe oxides is faster on iron oxide surface than on a SiO2 glass slide surface. I hope this answers your question.
Thanks,
Jake
Debashish Bhattacharya
Faculty
What role do stochastic events and seasonal changes play in the recruitment of taxa on your slides, or are these environments very stable and recruitment well understood?
Jake Beam
Dear Dr. Bhattacharya,
Thank you for interest and question. These acidic geothermal springs are relatively stable over time, which has been confirmed by > 10 years of thorough geochemical sampling (e.g., dissolved ions and gases) and analysis by our laboratory. However, it is important to note that we do not have data loggers in the field and stochastic events (e.g., high winds) could produce small temporal changes in aqueous gas concentrations or temperature for instance, which might result in the preferential recruitment of specific microbial taxa. In essence, stochastic events probably do effect the colonization of microbes to the slides but on the time scales we are interested in, I don’t believe that they play a major role or significantly effect the overall pattern of microbial recruitment to the slides.
I do believe, however, that seasonal changes do effect the recruitment of taxa to the slides. We have preliminary evidence that high UV pressure in the summer months effects the community as well as the function of the community (e.g., iron accretion) most likely due to viral attack (see Fig. 3).
We are only beginning to understand the formation and assembly of these iron oxide microbial ecosystems. The temporal community dynamics (Fig. 4) is initial data, but I have more time points in sequencing now using iTags (Illumina sequencing) so I believe these will ultimately shed light on the assembly of these ecosystems.
It is also worth noting that we have metagenomes of these “mature” iron oxide ecosystems from 3 generations of sequencing technology (i.e., Sanger, 454, and Illumina). These metagenomes encompass approximately 7 years of data and have revealed that the mature communities are relatively stable over these time periods (i.e., from year to year or season to season taxon abundances do not significantly change). I hope this addresses your question.
Thanks,
Jake
Daniel McGarvey
IGERT Alum
The observed succession of (1) autotrophic colonizers and (2) subsequent heterotroph colonization does substantiate your conceptual model, in a qualitative sense. I wonder: how will you quantitatively test step (3) niche partitioning? This is a difficult mechanism to “prove” in more well-known systems. How does one approach this in a thermal spring?
Jake Beam
Dear Dr. McGarvey,
Thank you for your comments and question. Indeed, you are correct that niche partitioning is difficult to quantitatively determine, even in macroecology and probably even more so in microecology.
As for your first comment, I completely agree, to an extent, about the qualitative observation of stages 1 and 2 of mat development in the conceptual model. The 16S rRNA gene sequences observed in Fig. 4, 28 days are almost all exclusively related to an autotrophic archaeon, which has been determined by our lab. Currently, I am collaborating with scientists at Lawrence Livermore National Laboratory to better understand autotroph/heterotroph colonization by utilizing natural carbon isotopic fractionation by individual community members in situ (nanoSIMS), which will provide better quantitative support for the conceptual model.
We hypothesize that depth-wise gradients in dissolved oxygen exist in these microbial mats and that aerobes will be more abundant near the mat/water interface whereas anaerobes will be more abundant further away from the mat/water interface. We approached this question by utilizing Clark-type oxygen microelectrodes specifically designed for these high temperature ecosystems and also careful dissection of the iron oxide mats coupled to taxon specific PCR and reverse transcriptase PCR of a gene that encodes a heme copper oxidase for oxygen respiration (see Bernstein et al., 2013, Environmental Microbiology). We found that 1) oxygen gradients do exist in these microbial mats and oxygen penetrates about 0.5 mm into the mat (10 % of the total mat depth) and 2) aerobic microbes are generally more abundant and active (mRNA expression of an oxygen respiration gene) at the mat/water interface. In the future, I hope to carefully dissect these mats to determine depth-wise distribution of specific taxa in these microbial mats utilizing multiple molecular techniques (e.g., 16S rRNA gene PCR and fluorescence in situ hybridization on mat thin sections) as we have already determined the extent to which oxygen penetrates these iron oxide microbial mats. I hope this brief explanation helped explain how I will determine this conceptual model.
Thanks,
Jake
Daniel McGarvey
IGERT Alum
Interesting, Jake. Thanks for the thoughtful reply!
Jake Beam
Thanks again for your interest and comments Dr. McGarvey!
Volker Radeloff
Faculty: Project Co-PI
Hi Jake,
Thanks for your video and poster. You did a really nice job explaining what you do, and I was very impressed by that, and learned a lot. What I wasn’t quite as clear about is “why should I care?” I’m not asking that question in a confrontational manner, I just would like to hear (or read) from you how this would be relevant beyond the system that you study?
Thanks,
Volker
Jake Beam
Dear Volker,
Thank you for your comments and question. Your question is completely valid and I often encounter this question in other situations as well and it may be difficult to envisage other applications other than the geothermal system presented here or in similar geothermal habitats.
These geothermal ecosystems contain a few dominant community members or “ecotypes” as compared to a microbial assemblage in a mesophilic stream channel that could contain upwards of hundreds to thousands of “ecotypes”. These differences in the number of taxa present is due to the phyiscochemical constraints that exist in geothermal environments. This allows us to begin to understand community assembly in more detail because we are only dealing with a few taxa as compared to hundreds or perhaps thousands of taxa. Thus, it makes studying these communities “easier” due to the low amount of taxa present. I guess basically what I am getting at is that these geothermal ecosystems provide excellent case studies and techniques and hypotheses built around the assembly of these communities can be utilized and is directly applicable to all environments (e.g., a mesophilic stream channel). Furthermore, the field of microbial ecology is in its infancy and determining how microbial communities assemble is fundamental to our understanding of ecosystem processes as it is for macroecology. So it stands to reason that studying the ecological succession of any microbial community (in this case in a geothermal environment) will begin to shed light on the factors controlling community assembly in any given environment. I could go on forever, but it basically boils down to laying down a foundation for future studies on the ecological succession of microbial communities. I hope I addressed your question (and I didn’t find it confrontational at all).
Thanks,
Jake
Virginia Anderson
Partner: Other
Biofilms are so interesting and those in hot springs are fascinating to look at in Yellowstone Park. Your poster objectives were very clear and made the great graphics easy to follow.Can you tell me if members of the new candidate phylum “Geoarchaeota” have been found to exhibit quorum sensing?
Jake Beam
Dear Dr. Anderson,
Thank you for your comments and question. I’m glad that you found my poster easy to follow and understood the objectives clearly.
Your question is very interesting, and as much as I know, quorum sensing in the domain Archaea hasn’t been studied in much detail. As for the “Geoarchaeota”, I am not aware of any mechanisms that they may use to signal to themselves or other thermophiles. Keep in mind too that the “discovery” of the “Geoarchaeota” was only made very recently by our laboratory (see Kozubal et al., 2013, ISME J) and the genomic analysis performed didn’t include an exhaustive search on all potentially functional attributes. However, I would hypothesize that not only do the “Geoarchaeota” have mechanisms of signalling one another, but other thermophiles in these communities would have similar mechanisms of signalling reminiscent of bacterial quorum sensing. Would be very interesting to study in these communities and is potentially an important mechanism of microbial recruitment to the glass slides. Thanks you for the idea, it is not something I had thought of previously.
Thanks,
Jake
Virginia Anderson
Partner: Other
Thank you for your candid answer and it was a premature question. I will read the article you suggested. The MSU Center for Bioengineering has strong ties and excellent resources for those of us who teach introductory microbiology. I use BIOFILMS:THE HYPERTEXTBOOK for research background with undergrads.
Jake Beam
Hi Dr. Anderson,
I don’t think it was a premature question at all. Yes, the Center for Biofilm Engineering at MSU has great resources and wonderful people to collaborate with, some of whom are working on this project with me. They also have great user facilities at the CBE, which I often take advantage of. Thanks again for your interest and comments, they are much appreciated!
Jake