Interview with the author: Broadening the taxonomic scope of coral reef palaeoecological studies using ancient DNA

Imagine being able to take a small handful of sediment from the bottom of the ocean, and from that seemingly lifeless material, be able to reconstruct the dynamic history of life in that area over the past decades and millenia. No, that’s not quite science fiction– advances in DNA sequencing technology have now made this possible. Hidden inside that sediment is what’s known as palaeoenvironmental DNA (aeDNA), or ancient fragments of DNA that come from the organisms that used to occupy the area, which when sequenced and combined with fossil records, allows us this amazing insight into the past.

In this blog post, we go behind the scenes with Dr. Maria del Carmen (K-le) Gomez Cabrera to talk about their recent publication in Molecular Ecology using aeDNA in a coral reef off the coast of Australia to paint a picture of the historic and complex communities that have inhabited these diverse ecosystems over the past hundreds of years.

Coring marine sediments using an aluminium pipe. Acknowledgement Dr. Brian Beck

What led to your interest in this topic / what was the motivation for this study? 
I have previously study the coral-zooxanthellae symbiosis and for this I have used molecular techniques, since it is impossible to identify the symbiont otherwise, I spent endless hours in the lab extracting tiny amounts of DNA. Then I changed fields and started working with Prof. Pandolfi and the Marine Palaeoecology Lab at The University of Queensland. The tools of the trade are rock hammers and chisels. I ended up surrounded by an inordinate amount of large rocks. But been dependant on only fossils to reconstruct the past leaves a lot of the story out since soft bodied organisms are very unlikely to leave a fossil record, this was very frustrating for me. When I attended a talk by Prof. Alan Cooper (a co-authors of this study) on ancient DNA from plaque in ancient human teeth, and considering my background, I decided we needed to try this on coral reefs. Ancient environmental DNA opened a new world for us to study ancient marine ecosystems of which we only know the story that fossilised organisms tell.

What difficulties did you run into along the way? 
Been the first study of its kind that we have undertaken, we ran into many difficulties. Extracting DNA from our samples was the first hurdle. Although we had a well-resourced molecular lab, we could not use it for this study since any trace of modern DNA was a potential contaminant. We had to repurpose a room in a remote campus where no molecular work was carried out, to use it as a clean space for subsampling the sediment cores, we then sent these samples to the Australian Centre for Ancient DNA to be processed. Making sense of the data was also difficult; there is little genetic information about most marine organisms. Although a steep learning curve, now we are better prepared and it is incredible the amount of DNA sequences been generated around the world on marine organisms at present that will greatly benefit this type of studies in the future.

CT scan of a core with coral fragments marked in colour. Each colour represents a different coral genera. Acknowledgement Dr. George Roff

What is the biggest or most surprising finding from this study? 
That we managed to actually extract workable ancient environmental DNA from reef sediment cores that were not even collected with this purpose in mind. This really blew our minds and opened so many new possibilities to answer important ecological questions that otherwise would have remained unanswered. Moreover considering that these sediment cores were collected from tropical environments, we were really pushing the limits of this technique.

Moving forward, what are the next steps for this research?
This study was a proof of concept, it allowed us understand the capabilities of this technique in the context of coral reefs. We are now establishing a new line of research, incorporating a team of amazing PhD students to explore ecological interactions in the past between key marine organisms such as coral and seaweed, we are also working on a more accurate picture of ecological baselines of natural resources such as fish before European colonization of the Australian continent. These studies will give managers better tools to assist with the management of the Great Barrier Reefs and its resources.

Dr. K-le Gomez Cabrera in the field. Acknowledgement Dr. George Roff

What would your message be for students about to start their first research projects in this topic? 
Read profusely and cover many subjects, do not stick just to your particular area of research because your eureka moment may come from something you read on a different field (think about the connection between ancient DNA in plaque from human teeth and coral reefs’ biodiversity). Take the time to understand the capabilities of the techniques you are planning to use. Researching ancient DNA is very expensive, so you need to really know what you want to achieve and how before you start extracting DNA, preparing DNA libraries or even collecting samples.

What have you learned about science over the course of this project? 
I’ve learn that lateral thinking is very important for the scientific endeavour. That it is crucial not to be boxed in your small bubble in your field of research but to keep thinking big, reading broadly, and scouting for opportunities to apply new approaches from other fields.

Describe the significance of this research for the general scientific community in one sentence.
This study opened new avenues of research that can be used to help us understand meaningful ecological interactions between tropical marine organisms hundreds of years in the past that would not have been possible with traditional methods.

Describe the significance of this research for your scientific community in one sentence.
By incorporating ancient environmental DNA into palaeoecological studies of coral reefs, we can better understand ecological interactions involving soft bodied organisms, a feat not possible with traditional palaeoecological tools.

Citation
Maria del Carmen Gomez Cabrera, Jennifer M. Young, George Roff, Timothy Staples, Juan Carlos Ortiz, John M. Pandolfi, & Alan Cooper. (2019). Broadening the taxonomic scope of coral reef palaeoecological studies using ancient DNA. Molecular Ecology, 28(10), 2636-2652. https://onlinelibrary.wiley.com/doi/10.1111/mec.15038

Summary from the authors: Extra‐pair mating in a socially monogamous and paternal mouth‐brooding cardinalfish by Theresa Rueger

Rueger, Theresa, Hugo B. Harrison, Naomi M. Gardiner, Michael L. Berumen, and Geoffrey P. Jones. “Extra‐pair mating in a socially monogamous and paternal mouth‐brooding cardinalfish.” Molecular Ecology. 2019. 28-10: 2625-2635

Link to paper: https://onlinelibrary.wiley.com/doi/full/10.1111/mec.15103

In this study, we followed more than 500 Pajama cardinalfish on reefs in Papua New Guinea. Cardinalfish stay close to each other in pairs for long periods of time, often for several years. Looking at the babies they produce with genetic markers, we found that most of them do exclusively breed with that partner. However, contrary to expectations, we also saw some sneaking behaviour. When presented the chance, both males and females take the opportunity to mate with other individuals.

Photo by Chris Hamilton

The findings are remarkable because the males brood the eggs in their mouths. They can’t feed during that time and their swimming ability is compromised, so brooding is very costly for them. That puts females in an advantageous position, because they can produce eggs quicker than the male can brood them and they can go and give eggs to another male. The males can offset that advantage by eating some or all of the eggs and accept eggs from another female. Also, in some rare cases males can fertilize eggs that another male is brooding, saving all the energy they would need to use to brood them themselves.

Photo by Chris Hamilton

It’s a complicated mating system, which is something we can only find out by spending lots of time observing the fish and using genetic analysis to identify parentage. Our tests reveal the complex nature of social groups in fishes and how promiscuity could upturn theories for how monogamy arose. – Theresa Rueger

Check out other media cover of the paper at ScienceDaily and follow on Theresa on twitter @TheresaRueger

Interview with the author: Heritable variation in bleaching responses and its functional genomic basis in reef-building corals

Given how quickly the environment is changing, a major question in evolutionary biology and conservation is the degree to which natural populations will be able to adapt to rapidly changing conditions. Corals, which form the foundation of biodiverse marine systems, are particularly threatened by rising ocean temperatures, with many species susceptible to coral bleaching. Understanding how genetic variation in natural populations is linked to coral bleaching responses may help us better understand and predict how increases in ocean temperatures will affect coral populations, and by extension, the health of our oceans.

In this blog post, we go behind the scenes with Dr. Katherine E. Dziedzic to talk about their recent publication in Molecular Ecology on heritable variation in bleaching response in a reef-building coral. Find out more about Dr. Dziedzic’s research by checking out their website or following them on twitter: @cnidiariangal

Photo from Katherine Dziedzic

What led to your interest in this topic / what was the motivation for this study? 
One of the most pressing questions for coral reef ecosystems is whether they are going to be able to adapt to changing ocean conditions, specifically rising ocean temperatures. Previous studies have shown variation in bleaching susceptibility across coral species, populations and geographic regions, but estimates of heritability of thermal tolerance only existed for coral larvae using controlled genetic crosses. When I started graduate school, I was interested in the same questions, but wanted to focus on adult corals that were already experiencing the effects of climate change. I wanted to determine if there was heritable variation in thermal tolerance in natural populations of corals, and therefore set out to use quantitative genetics and genomics to explore mechanisms of adaptation in a population of corals in the Caribbean.

What difficulties did you run into along the way? 
Field studies can be tricky, especially when trying to recreate environmental conditions and conduct heat stress experiments in lab settings. I chose Orbicella faveolata as the focal species for this experiment because there were genomic resources available (e.g. linkage map and genome), and it is a major reef building coral in the Caribbean. However, this species wasn’t as abundant as I had initially expected and therefore wasn’t able to collect the number of colonies I had initially intended (100 vs. 43 used in the experiment). Additionally, when I sequenced my RNAseq libraries I had low coverage and had to re-sequence my libraries multiple times. This took a lot of time and resources, but it made the results better.

What is the biggest or most surprising finding from this study? 
My co-authors and myself were surprised to see the estimate of heritability of thermal tolerance so high (h2 = 0.58). Heritability of thermal tolerance has been shown to be high in coral larvae, but no estimates existed for adults prior to this study. This was surprising, but we were excited to find hope for possible adaptation in this population. Also, when we explored gene expression across contrasting phenotypes, we found an interesting pattern where heat-tolerant corals expressed genes at higher levels constitutively and down-regulated these genes during heat stress. This pattern has been documented before, but it was surprising to see it as the dominant pattern of expression for all heat-tolerant corals in this study.

Moving forward, what are the next steps for this research?
I would love to expand this analysis to other coral species as well as other populations of corals to see if there is heritable variation in other reefs. Through genomic studies and population genetic surveys, we can use methods, like those used in this study, to determine which coral species have the capacity to adapt, and which populations globally and regionally have enough genetic variation for selection to act on. Using this type of information, we can determine which conservation actions (i.e. assisted evolution, restoration of certain reefs, resilience-based management, etc.)  can be put in place to protect and preserve these populations and thus the genetic variation in thermal tolerance.

What would your message be for students about to start their first research projects in this topic? 
If possible, I would suggest doing preliminary trials with temperature treatments and the species of interest to ensure that your stress temperature shows enough phenotypic variation and is biologically relevant for the area in which they were collected. I would also practice library preps and sequencing a small number of samples to ensure no issues with sequencing – troubleshooting ahead of time can really save a lot of time later when you’re in full production mode. And of course, enjoy your time in the field! If you have the opportunity to do scientific diving, don’t forget to look up and enjoy the view! It’s hard not to become over focused on the research project, so make sure you take a moment to appreciate the beauty.

Photo from Katherine Dziedzic

What have you learned about science over the course of this project? 
I’ve learned that science is not a straight, smooth path – there are twists and turns, speed bumps, and dead-ends. And that things fail and it is totally okay! Most of my research, particularly this project, ended up on Plan C or D, and I was really discouraged when I had to re-start or re-do aspects of it. But having to continually troubleshoot the experimental design and sequencing issues have been valuable teaching moments and have helped me become a better scientist. We only see and evaluate the final product, but it’s important to recognize and appreciate the messy part in the middle – the part that made the final product what it is. Science is messy, but that’s what makes is so fun and rewarding!

Describe the significance of this research for the general scientific community in one sentence.
This study contributes to the growing body of evidence that natural populations of coral reefs possess genetic variation in thermal stress responses that may potentially support adaptive responses to rising ocean temperatures.

Describe the significance of this research for your scientific community in one sentence.
This study provides crucial data for models aiming to predict corals’ adaptation to ocean warming, and identifies genetic markers for thermal tolerance that may be useful for restoration efforts as conservation biologists work to reverse the global degradation of coral populations resulting from changing ocean conditions.

Citation
Dziedzic, K. E., Elder, H., Tavalire, H., & Meyer, E. (2019). Heritable variation in bleaching responses and its functional genomic basis in reef-building corals (Oribicella faveolata). Molecular Ecology, 28(9), 2238-2253. https://onlinelibrary.wiley.com/doi/full/10.1111/mec.15081