BioPOD
BioPOD
The forgotten Kingdom: Inside the World of Fungi
In this episode, Nitara interviews Dr Ester Gaya, a researcher working at Royal Botanic Gardens, Kew, in the Fungi genome project.
Hello and welcome to BioPol, the official podcast for the school of biological scientists here at the University of India. I'm Vagilia, and today Manushw and Tara are sitting down with Hester Gagia to talk about fungi. In this episode, we explore why fungi are such an important and often overlooked group of organisms, how scientists study fungal diversity using modern sequencing approaches, and why large-scale projects like those at Q are transforming our understanding of the fungal tree of life. So now over to you, Manusri Mittara.
SPEAKER_02:Hello and welcome to BioPod, the official podcast for the School of Biological Sciences here at the University of Edinburgh. I am Manushri. And I'm Mitarra. We're from the BioPar team. Today we're chanting with Dr. Esther Gaya, senior mycologist based at Q Gardens, currently working on sequencing fungal genomes. Fungal genomes from Q's fungarium and collections at the National History Museum and Royal Botanic Gardens, Edinburgh, are being sequenced to understand fungi and lichen diversity and evolution. Welcome to the podcast, Esther. It's great to have you with us. With Astanan, thank you for inviting me. Now onto the interview. Our listeners would love to know more about your journey with science before and during this project. How did you develop an interest in science?
SPEAKER_03:Well, in science and in particular in fungi, I developed an interest at an early age. I'm Spanish. I grew up in Barcelona, in Spain. And I think Spanish culture society is very mycophilous. This means it's a society that likes fungi. There are some societies that are a bit scared of fungi. And Spanish culture is the opposite. It's something that is integrated in our culture. And since early age, I would go out and foray and collect fungi with my family. And I started as a budding taxonomist, a person that classifies organisms since a very early age. Obviously, we were focused on edible fungi. And as I grew up older, I started getting more interested in other types of fungi, not just the ones that we put in the pot or in the paella, but in other types of fungi. And as I reached university, I went for biology. I focused mostly on biodiversity studies and mycology, the study of fungi.
SPEAKER_01:Do you think that people in the UK like fungi in general?
SPEAKER_03:Yeah, I think uh the British Mycological Society has a long, long story, uh history. And the British culture is less prone to eat fungi, that's a fact. But that doesn't mean that they are not interested. And actually, there's a lot, a lot of amateurs and people right now foraging for fungi and just interested in fungal diversity. Actually, nowadays, and it's not just in the UK, I feel there is a bit of a new wave of fungal and interest at different levels from the roots. These things like, for example, the um all things fungi festival that happened a few weeks ago in Sussex with more than a thousand attendants. And it's basically all things fungi from medicinal fungi, edible fungi that talks about fungi in terms of research, there's a bit of everything. So there's a lot of interest from the wider public now on fungi. And it's something new. Something new when I started in uh my undergrad studies in biology, and I was telling people I was looking at fungi people were like, What? Mycology, what is this? This has changed, but also at different levels, there's lots, lots of new companies, new startups that are trying to develop um new materials, biomaterials, new sources of food, medicines all around fungi. So there's, I think there's a big peak interest in the UK, but also globally. Things have changed. Everyone wants to do fungi now. Honestly, it's been a bit uh sedendipitous. My my group of expertise is a bit of a special one because I did my PhD on lichens, lichen evolution. It's a lichenized fungi. That means it's not just a single organism. In lichens, we have fungi, but also they establish a symbiosis with other organisms, mainly an algae or um cyanobacteria, and other microbes. It's a it's a bit of a mix of different things, right? But we call them lichenized fungi because the main pion is the this main body of the lichen, is the fungus. So I studied for many years since my PhD, uh lichen evolution and diversity, and I did some fungal genomics with lichens. But working at Kew and being so close to the largest collection of fungi in the world, we have a really, really massive dry collection of fungi, more than 1.25 million specimens. That's a lot of fungi coming from all over the place. We have a fantastic global collection, but also a UK collection. Even if my group of expertise were like it, it was almost uh impossible to ignore such a fantastic collection. And for several years, we attempted to sequence that collection. Actually, we have a small publication from 10 years ago where we sequence a bunch of genomes from some of our fungi. And since then, that was back in 2016. I've been trying to raise funds to do the sequencing up scale to attack that collection as a whole. And almost two years now, the DEFRA approached us and they were very interested in the idea because, as you may know, with climate change, global change, globalization, there's lots of uh invasive species that are arriving to the UK. There's lots of new pests and diseases that are not well understood, and some of them are fungi. And honestly, there's lots of work on bacteria and other groups and other pests, but uh insects as well, but not so much on fungi. One thing I should clarify for those that are not so familiar with fungi and how we study them fungi are very, very diverse. They have different shapes and colors and flavors, and uh, they produce lots of different compounds, pigments, and some of those compounds, and what we call secondary metabolites, are the ones that have turned into antibiotics, for example. Um, they have a huge um chemical diversity. That's how they communicate with the environment and with other organisms chemically, they are like chemical factories, but they have a huge, huge plasticity. So their phenotypes, their looks, their morphology, are very misleading. So for many, many years we we studied the morphology, the looks, and we try to categorize and classify them based on that. But when we started sequencing, when the DNA sequencing revolution happened more than 20 years ago now, and we started sequencing fungi, we realized that what we were using to classify them, that the characteristics, the characters that we were using were very misleading, and things that would look alike and that we could cluster together, when we sequence the DNA, we realized that they were completely different things, that they came from different ancestors, and sometimes things that look very, very radically different when you sequence the DNA, they come together. They are very closely related. If you add up this complicated story that you realize that you need to do the sequencing, the DNA sequencing of fungi to really understand how they diversify and how they evolve. If you add up that to the revolution that has happened the last few years with e-DNA, that means sequencing samples of the environment globally. You get, for example, a sample of soil or you get a sample of water, and you sequence everything that is in there. Bits and pieces of the genomes and the DNA sequences that you can get in that body, that could call meta-barcoding or metagenomics. You sequence things that are invisible, right? And you see you may sequence fungi that are invisible. They are hidden to the naked eye. You only see them under the microscope, and you just have pieces of DNA from them. So if you add that to the complication that fungi may have by different plasticity or phenotypes, we realize that we needed reference sequences, things that have a physical voucher, a physical individual. And this is what, for example, are the type specimens that we hold in our collection. These are the specimens that were used for the very first time when a new species of fungus was described. Those specimens were used as a reference. The person that said, Wow, Eureka, I found a new fungal species, they use that specimen as a reference. We realized that we sequence those type specimens for which we have a physical collection where people can come and see how it looks like, and then we sequence their genome, their genetic code, the whole sequence. We can use that as a backbone to place and identify all that cryptic diversity that our colleagues are sequencing out there in the wild, in the environment, or people that are trading, as I said earlier, invasive species, new pests, that we don't know what they are. It could be that some of those unknowns were already collected once 100 years ago, 200 years ago, and that we have them in our collection. So, in a nutshell, this project aims at putting a tiny bit of order in the chaos of thousands of thousands of sequences of fungi, of unknown fungi out there.
SPEAKER_02:Since you mentioned like it took you quite a few years to end up getting funding for this project, why do you think fungal sequencing has been ignored so far?
SPEAKER_03:I would say it's it hasn't been ignored. We we've been sequencing, because as I said, maybe 20 years ago we realized that without DNA, it was very difficult to understand fungi and to tidy them up and relate to them one to each other. But we were sequencing bits and pieces of the genome. We were not doing whole genome sequencing. That took a while. They are complicated organisms because, as I mentioned earlier, they produce lots of compounds, lots of chemicals. Some of those chemicals may inhibit the sequencing, and we have seen that, for example, with uh long-rid biosequencing. It's quite challenging. It has proved more challenging with uh fungi than with other organisms. And we guess it is mostly due to these compounds that may act as inhibitors. But it's possible, and we're doing, we're still doing it with sequencing. It's just how we'd say that sometimes we call fungi the forgotten kingdom. People tackle first animals and plants and things that you can see, but evolving an interest on things that most of the time you don't see, it's difficult. There's a bit of both difficulty, technical difficulties in sequencing them, but also as I said earlier, fungi have been kind of rediscovered recent in recent times, but for a long time they were a bit neglected. It was just a handful of us looking at them. And obviously, we have some fungal species that have been model systems for a long time. Yeasts are fungi, right? And and we use yeasts in many purposes, and uh and there's some fungal species that are very, very well known and very well studied, but they were just a handful. And obviously, we have penicillium, right? Or antibiotics uh come from, but the majority of fungi were not well studied and a bit neglected. That's that's the reality. And as I said, it's for technical reasons, but also the fact that people don't see them. Because I would like to highlight that mushrooms, um, touch tools, what you see when you go in the woods, that's the tip of the iceberg, right? If you dig up down underground, you will see the actual organism. We know that some of these fungi that produce mushrooms, not all of them uh produce mushrooms, are some of the largest, biggest organisms on earth. This one recorded in in the North America in in Colorado that goes uh several hectares and is all underground. It's the mycelium of the fungus that connects the different different trees, and that can be huge. But you don't see that, you just see the little mushrooms. And mushrooms basically are the reproducing structures, is they carry the spores, like flowers carry pollen, right? So they produce the spores that will disperse and they are used in reproduction. Yeah, many many fungi don't even have that. You have lots of fungi that are hidden inside plants, not just uh associating with the roots, but in their leaves, in their stems. You have fungi everywhere. These fungi are endophytes. We call them endophytes, and they are what we call um asymptomatic fungi. They seem to live in symbiosis with the plants, but they don't cause any symptoms. You don't know they are there until you sequence a bit of your plant and then find that it contains fungi. So they can be anywhere, and you can also have pathogens, right? Pathogens that grow on the leaves of plants, they uh cause massive damages in our crop production, but also you have animal pathogens. We are full of fungi ourselves, and we have, I think the uh that series, the The Last of Us, uh uh generated a bit of uh open imagination of people on the fungal pathogens. I think that was a bit of an exaggeration, but some some fungi can cause very, very weird, interesting uh infections, and the plastic ones that people find yeah kind of uh funny are the ones that attack some some arthropods, uh the entomopathogens, that, for example, the zombie fungi that control ants and make them climb on top of branches of trees, and then they let the spores of the fungi that may grow on the antenna of the ants, let them fall on the rest of the ant colony, and then infect the rest of the ants to continue the cycle and disperse the spores of the fungi. So this is an example of the zombie, zombie fungi, right? So they do yeah, all types of funny things, and those ones obviously don't make they don't grow mushrooms, they have other shapes. But some of the um campans that are used to control those animals are very, very interesting. And some of them have already been used in for medicinal, for medical purposes. Some of the classic examples that we all always mention, cyclosporines and some of the medication, for example, that is used to control multiple sclerosis was originally discovered in fungi. The drugs that are used for organ transplants to allow your body to accept a new organ come from fungi as well, from some of these groups that colonize other animals, the same group. They are they produce very, very uh interesting, potentially useful compounds. Um, you asked me if um why they haven't been sequenced before, but yeah, I think I gave a very, very long answer.
SPEAKER_02:This is very informative, actually. Q is partnering with uh the National History Museum and the Royal Botanic Gardens, Edinburgh, on this project. Why was it important for these three institutions to work together on this?
SPEAKER_03:Basically, they are the largest collections in the country. We at Kiyu we have mostly non-lichen fungi. NHM has the largest collection of lichens in the world, actually. So they create most of them lichenized fungi. They have fantastic type reference collections, and the same for RBG. RBG does a lot of research on lichens, mostly lichen ecology. They have very interesting work on like the effect of climate change and lichens as bioindicators. So historically, they have evolved into having a very nice lichen collection and also other types of fungi as well. And also these three institutions, because we have those collections, we have a lot of expertise. The main fungal experts and taxonomic experts work in those institutions. So that's one of the things that we also raised. If you want to sequence as much diversity as possible of fungi, just go for these three institutions. We have the best collections, and we have also experts in them that can help you and guide you on what we can select for sequencing and revise the specimens.
SPEAKER_01:So experts have estimated million species of fungi to exist in the world, but maybe only 150,000 or so have been discovered. Do you have any guesses why so few have been discovered? Do you think it's related back to that sense that it's been morphological classification for so long?
SPEAKER_03:Yeah, it's exactly we have found recalculating and re-estimating that number of potentially unknown fungi. A couple of years ago, we recalculated that figure. Now we got a review paper and we set it to 2.5 to 3 million species. Some people say that it could be more, but it seems we we what we did was revise all the methods that have been used in the past to assess the number of species. And we did some proxies as well with plants and animals, and we came up with a revised number. Obviously, that can change in the future, but we like to play with that. And as you said, we have about 155,000, a bit more now. We may be close to 200,000 described fungi that's named fungi, recorded and published, accepted. This isn't that's it's what you mentioned. They most of them you don't see. So it's very difficult to describe diversity of things that you don't see, right? And that's why we we are trying to do the Pungarium sequencing project, as I mentioned, to put some references out there that may help us to describe uh new diversity, hidden species that we don't see, and it's difficult to find them, but also cryptic diversity. So two things. First of all, things that we haven't never ever collected that are out there and nobody has discovered it yet. And the second thing is things that we have already collected, things that we may have already described, may contain hidden diversity. And this is something that has been underestimated. I always like to put an example of colleagues of mine found that one single lichen species that had been collected for many years actually was not just a single species, one single species actually uh represented more than 400 different species. And they found that this just went, they sequenced many, many specimens that looked alike and that they they were named equally, and they realized that actually there was what we call cryptic diversity. Diversity that you only detect. When you do sequencing, DNA sequencing, and then sometimes you compare those that back that DNA sequencing, that's what they did. They went back to the original collections and started digging, and then they finally found other features and traits that maybe they had missed when they had described the single species, so they could tell apart also physically those species. It's quite striking, like from one thing to 400, that's what happens. That's why the number of unknowns is so big. I think only the insect group beats us. I think they also have lots of unknowns there. But I think fungi is very difficult. And with the estimates are maybe wrong at the same time. There was a publication a few years ago that were claiming six million of unknowns. What we know is that there's lots of things that we haven't discovered yet. And also things that we have collected, but we haven't discovered. Because, as I mentioned earlier, we have a largest collection of fungi with lots of hidden envelopes and hidden samples away that maybe nobody has touched since the person that collected them 100 years ago brought them back to Kew or to RBGE or to NHM to the Natural History Museum. And that has happened, has already happened, that someone takes that specimen, that fungus, and realizes that it's a new species that nobody else has described before, and maybe nobody else has collected before. Because maybe that specimen comes from a place that unfortunately doesn't exist anymore, and this is gonna happen more and more. Habitats and ecosystems are being destroyed, and some of the fungal species that come from very, very specific habitats that can only live in those places may not exist anymore in the wild.
SPEAKER_02:How do experts estimate like the number of undiscovered species since we only know how much we know?
SPEAKER_03:It's basically on previous studies, it's a combination of different things. For example, we have lots of data on soil fungi. So one of the things they do is assessing if we have so many soil samples and so many sites, and we use that as a proxy to estimate the global diversity of fungi, not just in soil. There's also sites that have been studied for many, many years, and we have very good estimates of the number of fungi in there, and then we can extrapolate that globally to other sites. The problem is that some of those long-term studies have been done mostly in temperate habitats, and then when you extrapolate to other habitats, you may be biasing a little bit because if you assume that the tropical site might have the same diversities, it's a bit tricky, we don't know. But we add up all the evidence. Soil diversity, sites that have been studied long term, and also the proportion with diversity of plants. We have a ritual, so we assume that six to one and nine to one, and then you estimate if you have so many plants and so many other things in other places, how many potential fungi you may find in there. Also, based on it's this is quite complicated. We can also use higher taxonomic level diversity in different groups of fungi, and based on that higher taxonomic diversity, potentially estimate number of species in each group. So combining all that evidence, we can end up with an average, but it's all a bit esoteric. It's not hardcore evidence.
SPEAKER_01:So fungi have such an incredible amount of diversity, and as you've mentioned, so much hidden and unknown diversity. How will the sequencing project help with applications and uses of fungi? You've mentioned how they're used in medicine, like antibiotics and possibly identification of disease-causing fungi. How will the sequencing project help with those use cases?
SPEAKER_03:So we don't know yet. We hope they will help. The discovery of compounds is a bit funny because, from one side, many of the drugs that we use today that can be synthesized, mirroring plants or fungal compounds were a bit ad hoc, just by chance. It's a bit like the antibiotic, the penicillium, right? It was a mistake and then they discovered it. So there's a lot of that at hoc. Ah, let's see here. Oh, this group of fungus seem to do lots of things. Maybe we look into that. So it's a bit like that. And then on the other hand, big pharma companies they can't waste time looking for different things, right? So what they've done for many, many years is use monosystems, use things that can grow in culture, like things that can grow easily in vitro and work on them and then maybe modify them and change the conditions, and see what else can they produce. So basically, we have a handful of fungi that have been very, very, very well studied because they can be manipulated in in vitro, and at oxen the discoveries that here and there, oh maybe, yeah, I don't know. Yeah, soil, there was the historical cladium found in soil, and then it was very useful later on. But it's been a bit like that. So, with the this project, I hope that we can provide a data set that can be more systematically data mined. The main issue is that we these are historical collections. In some cases, what we consider ancient DNA, DNA that is very, very degraded. They've been sitting there for a long time, or maybe they were collected in not the best conditions and they didn't dry properly, and the DNA degraded, and it's basically a molecule that you could call ancient DNA, very damaged. So that means that we have technical challenges, and our genomes won't be perfect. These are not classic long-read genomes, very well assembled to chromosome to chromosome, and annotated. No, no, no, no. It's almost the opposite. But there will be lots of them, lots and lots from different groups that maybe were never ever sequenced because they cannot be grown in the lab in a Petri dish, and then you don't have so much biomass, and it's difficult. But we are using Illumina short rate sequencing, which allows to sequence those fragmented samples of DNA. Um, so that's what we get. Now, with our pilot data, what we have demonstrated is that those genomes may be patchy. Then putting all the bits and pieces together may be a bit more tricky to know what's the order, one thing after the other. But we are getting high gene recovery. So the number of single-copy genes you would expect at the novel genome assembly, that number is pretty high, even if the genome is not perfectly sequenced. So we're getting between 86 to 90 percent Busco gene recovery. So this is very high. So my hope is that even if our sequence of all our fungi is not perfect, we will recover lots of genes that then can be investigated. So if you look through all the different species of fungi that will be sequencing, that they will be well curated, that we know that they are the original specimen that was used to describe that species. So it's for sure that species on top of that, you'll get lots of genes, and you can do what we call comparative genomics. You can compare all those genes across different groups of fungi, and then you can design bioassays, and then go to the lab and use that gene for knock-out experiments, knockout experiments, and see what function might be encoding for, etc. etc. And that's how we hope we can look for new molecules and new functions across many different groups of fungi that may not be culturable, may not be able to manipulate. What aim to do is a library, a reference library, genetic reference library, where you can go and see this gene, what is this gene might be doing here? And you can look through. And if you think you see patterns across many, many different things, and you see things that are there all over, may inform us of the relevance and importance of that gene involved in different functions across different groups of fungi as well.
SPEAKER_02:Some of our listeners may not be familiar with the Illuminati sequencing that you mentioned. Could you offer us an explanation for that, like how it works?
SPEAKER_03:So very, very basic, very basic. You have your sample, right? You go into the lab, you grind it, like like a like a mini Pima tube, you pulverize everything, and then you use a protocol that allows you to extract the DNA of your sample, you isolate your DNA, and then you take that and send it for sequencing. What happens with Illumina short read sequencing, they they kind of cut down the whole sequence. And instead of sequencing the whole thing in one piece, they sequence lots of bits and pieces. And sometimes what they do is they sequence many, many, many times the same piece, the same fragment over and over and over. So you have what we call depth of coverage. And then afterwards, it's like a puzzle. They give you all the bits and pieces that have been cut first and sequenced, and then you have to uh scaffold them, put them all on top of each other, and try to find the bits that you can connect, the ends of each of those reads that may be connecting with each other, and then you put together back your puzzle and your sequence, and then you can look at the sequence at the end as it was original in your sample. But that allows, as you said, for what we call high throughput sequencing, allows for a lot of sequencing, getting lots, lots, many, many, many copies of the same bits, and also allows for sequence for samples that are not that good that may have the DNA damage to be sequenced.
SPEAKER_02:Since there's such a large amount of data and information in this project, what's the strategy to collect and manage it?
SPEAKER_03:This is something that if there are any students and physicists listening, any experiment, any piece of work you want to do, first think data management. Because it's the most, most, most challenging bit. And I have a fantastic team. We are working very well, we're doing lots of fantastic work, but still now I find data management the most challenging thing. So I would like to highlight you can do the fanciest experiment in the world. If you don't data manage properly, your data will be useless. You won't be able to do anything with that. And this is that's essential. Those Excel spreadsheets, they have to be perfect. Your database has to be perfect. So what we've done with this project is where we are still it's not finalized, but we're developing a customized database that will allow to process all the information. But right now we use individual spreadsheets for each of the steps of the process. And I'm I'm very, very annoying. I chase everyone to make sure that the data is clean and is tidy and is up to date so we can merge it and put it together in a database that we won't be lost. But I recommend for anyone that starts a new project. I learned a bit a lot about data management. There's lots of different tools, you can do it in different ways, but get a bit literate about how you want to manage that data because it's very complicated. Most important thing, you have to have a sample ID, an identifier that doesn't let you lose the sample. That anyway goes that sample, you can trace it, you can find anything in your workflows and your pipelines. And that's challenging, is what we call data correction. And if you can get a bit of training on database development, that's super useful.
SPEAKER_02:What kind of troubleshooting have you had to do over the course of this project?
SPEAKER_03:Troubleshooting, well, that's why we convinced our funders that we could do it. Before this project, we had a small um project part of our PAFTOL program at Kiw. PAFTOL means the plants and fungal trees of life. Is a that's a long-term program and part of our science strategy. So we aim to reconstruct a plant and fungal trees. Part of the program allowed us to do a pilot study, and we tested different types of fungi from a collection, and we did develop some did some troubleshooting with the DNA extractions and with the sequencing. So we created a data set that demonstrated that we could upscale this type of sample processing, and that's how we got additional funding to do it a big scale.
SPEAKER_02:Has climate change impacted the quest to collect sequences in any way?
SPEAKER_03:That's an interesting question. I wanted to add something to that I haven't mentioned before. In addition to fungal collections, plant collections as well, any any collections you may have, the collections have an added benefit. They represent an untapped source of metadata, of history that we may not be aware of. These collections contain specimens that have been collected for many, many different years, and they can tell you how fungi have changed the behavior through time. And there's lots of publications, for example, that have used collection data determined shifts in seasonality, the time when fungi, for example, produce mushrooms, shifts on the seasonality of fruit in body production linked to climate change, for example. You see fungi producing bodies earlier or later due to global warming, for example. We can see changes in distribution. We can see species moving around, species that were not recorded in the past in some places now, they are present, and that your collection may tell you that. So it's these specimens have rich, very rich data that is kind of not, again, is a bit neglected, that can tell us a lot of the change they are undergoing in relation to climate change and global change in general. And there's lots already of interesting data, especially on protein body seasonality. It shows how that has mostly expanded because it gets warmer and fungi-like, warm and wet. So if those conditions go together, you will see more mushrooms in at those times. That's just again the tip of the iceberg because I think there's lots of other interesting information in there that we are missing that nobody's looking into. And I hope with this project again we can grind and wrangle that information and see if we see any trends, any patterns, any changes on, for example, plant pathogens. Have they changed through time? Are they becoming more virulent? If we can sequence the genomes of the same species again and again, we can also see that. We can also see if uh the genetic code has changed through time with historical collections.
SPEAKER_01:Since you have so many samples, are there any particular ones that you'll prioritize for sampling first, maybe from particular regions or ecosystems, or will you target the ones that are possibly cryptic species that you guess might be more than just one?
SPEAKER_03:Well, my answer is a bit prosaic, uh, because we started with the samples that we thought we would be more successful first during the this first year. So we started with big chunky mushrooms, bracket fungi, things that were big that we could resample if we had to, if we would fail in the lab, that we could go back to the collection and get more samples. So we've started with big things, but now we're slowly progressing and moving more into smaller fungi and symbiont, things that associate with plants and with animals that are trickier, but they might be more interesting. We don't guide ourselves taxonomically, we don't pay so much attention to what species we're looking at them. We look into the fungi more from the lifestyle perspective, the nutritional mode. So if they are mycorrhizal, sabrovs, if they they cause disease, if they are symbiotic, beneficial, or harmful, we try to cover all the different types of fungi that might be out there. And that includes different species, etc.
SPEAKER_02:Okay, as we wrap up today's episode, what are your hopes for the future of the fungi genome sequencing project? Like could this project and the information and knowledge we gain from it change how we classify or group fungi in the future?
SPEAKER_03:I hope so. I think it's going to happen because as I explained earlier, understanding lichen diversity so um fungal diversity is so challenging. My guess is that there will be some species that we have been naming for many, many years, and when we sequence the reference collection, we will realize that the actual species is something completely different. So, yes, my hope is that we will put some order, at least help understanding better classification of fungi and tidy up some of these, the least known groups. But also, I hope that the massive effort we've done with this project can be used as an example by other collections globally, that some of our colleagues can don't need to reinvent the wheel and use our project as an example and use our protocols to replicate that and use. Imagine all the collections all over the world of fungi, and if they can be sequenced, the um wealth of data would be amazing and incredibly informative. So I hope that what we put out there and we'll make everything publicly available can be used by others and replicated so we can get more and more information about fungi. Thank you, Esther, for joining us today. Thank you for inviting me. I hope you know a bit more about fungi now.
SPEAKER_02:And thank you, listeners, for tuning in to this episode.
SPEAKER_00:Thank you for listening to today's episode, and we hope you enjoyed the podcast and learned a bit more about the fascinating world of fungi and why they matter. Don't forget to subscribe to our podcast wherever you're listening to it, and to follow us on Twitter at Balapotadimbra as well as on Instagram and Facebook. Otherwise, enjoy your day and see you next time.