 Good afternoon or good morning, good evening, good night, depending where you are Under globe we have we have people from all over the world and logging in today And you're very welcome to this webinar posted by Femmes UC search and by Oxford University Press or publisher My name is John Morrissey Based in University College Cork in Ireland and I'm editor-in-chief of Femmes UC search And as you know, I'm going to be joined in a webinar today by Professor Daniela Donneri from University of Manchester in the UK And by Professor Tony Gibaldum from the Institute for Research and Biomedicine and the Supercomputing Centre in Barcelona in Spain So you're all very welcome Just by way of introduction and to give people time to be joining the webinar I'm just going to give a little bit of information about about the Femmes in the journal and the context of these webinars Let's see if I can advance my slides The webinar is hosted by the Federation of European Microbiology Societies Which is very well known of course to all All my colleagues in Europe or might be less well known in other parts of the world So Femmes is a federation of more than 50 microbiology societies from all countries in Europe and Femmes is really a it's a community of microbiologists It's a scientific society of course not for profit and Femmes invests resources in science and in scientists and in people and it's got a mission to try and help build the scientific Community of microbiologists and to help microbiologists have the maximum impact from our research I just want to mention a couple of upcoming events before we get on to the webinar itself Femmes holds a Congress every two years, but of course this year. We've had to change a lot of things because of COVID-19 and in fact the Femmes Congress that was due to be held in Maastricht in the summer Has now been merged with the ASM Congress and there will be a new Congress the first-ever World Microbe Forum Which is going to be held at 20 to 24th of June and this is an online Congress and You can find out more on the Femmes website worldmicrobeforum.org or indeed on the ASM website as well Now Femmes used to search who are The hosts for today's webinar and I'll say a bit about the journal in a moment We're hosting a symposium within the World Microbe Forum beyond the frontier with synthetic yeast I'm chairing that session and we've got some great speakers that you can see there I think Saki is still holding our hope that there will be an Olympics this year We all hope there'll be an Olympics, but in any case he's going to talk about yeast Olympians So that should be quite interesting. So that will be in the World Microbe Forum I'll mention The major yeast Congress has taken place this year. It's a joint meeting between ICY 15 and ICY GMB and This is due to be held in Vienna in August It will be held in Vienna in August. I guess it's still a chance It's not clear yet whether it can really be physical or online the plan is still for a physical meeting I just want to mention For people who are thinking of attending that meeting that there is a call for papers for Femmes yeast research I'm called a spirit of yeast and you can see more about that on either the conference website or on the journal website That kind of brings me to journals because one of the major activities of Femmes carries out to help provide opportunities for scientist research and to generate income that we invest back into our research community is we publish seven journals and You can see them listed across there. The latest two journals are microbes and micro life These are fully open access journals the first five you see listed are Hybrid journals, so you have a choice of free to publish or open access and as you wish and Of all of those journals clearly the most important of these is Femmes yeast research And Femmes used to search Of which I'm editor-in-chief Carl Monroe is deputy editor-in-chief We publish articles right across the spectrum of yeast biology and we have them special series of articles and issues I'll just draw your attention to The retrospective series that's edited by Terry Cooper Two articles I I list here my root intimacy of genomes Which is a retrospective by Bernard Dujan is actually one of the most highly read articles ever in the journal incredibly popular and Our most recent retrospective is one that would be of interest to a lot of people and it's got a certain relevance today to today's webinar Because it's a retrospective of Carl Singer and Carl Singer himself unfortunately passed away a few years back, but it was written by his son Harry and Terry Cooper based on interviews and writings and Memories and and so forth certainly for those of us of a certain vintage we have very good memories of Micro manipulators and dissecting yeast head trads and carrying out classical genetics in the era before genome sequences and a lot of a lot of us will have worked with micro manipulators developed by Carl Singer and That retrospective is a really interesting look back as well at some of the characters and some of the yeast conferences that took place and In especially I would say in the in the last 20 or 30 years of what is known as the last century So I can recommend that article to you the journal also publishes regular thematic issues on Topical issues We list three of them there and the most recent of these is the one on yeast genomes That was only published in this gen this January We had one synthetic biology published in a latter part of last year and an anniversary issue last year I have both of which are available on the website to see The thematic issue on yeast genomes was edited by Jean-Marc de Ramme and technically versus a Delft many of you in the yeast community will know John mark very well and He assembled an impressive list of authors to Write these review articles because he and some of the authors listed on the right on the slide here and and that thematic issue was launched in January and As with all our thematic issues all papers in that thematic issue are free to read for three months from the launch Regardless of whether they're on an open access license or not. You can see these articles But by all these authors now if you log on and so you can see the top two names on the list are Tony gavaldon and Daniela del nere and I'm very pleased to have Daniela and Tony With me today for this webinar It's our second Femmes used to search webinar. We had one on synthetic biology before Christmas that that was very well received It's part of a whole series of webinars that Femmes organizes and you can view all the past webinars if you go to the Femmes YouTube channel and This is a recording of this particular webinar will be up there as well in due course so Daniela del nere is Native of Italy, but she's been working in the United Kingdom for quite a few years now and She's an expert on Many aspects of sacramuses genomics genetics evolution population biology and biotechnology and And What you can see in front of you is a particular article that she published in that thematic issue And I know her talk today Will draw on some of that article, but I know it's going to pick up on some other themes as well And her second speaker that you can see is Tony gavaldon Tony is Well, I'll describe Tony as a computational biologist You might disagree with me. I'm not sure but Tony has done tremendous work on yeast evolution and on reconstructing the evolution of Sacramuses and other yeasts and has given us some really interesting insights and how we how we got to where we are with species and maybe on where we're going and Tony's a review article in the And yeast genomes Ti was based on hybridization and the origin of news new yeast lineages And I'm sure that's going to be part of what he talks about today as well and maybe some other things in addition So at this point now, I'm hoping there's somebody in the background Sarah has got control of my screen and Is we're going to pass over to Daniela Who's going to give the first presentation and maybe just to let people know what what's going to be Happening and Daniela is going to give her presentation for 20 to 25 minutes Tony will then give his presentation for similar length of time and then we'll have a discussion where I'll moderate the questions Under right hand side of your screen You've got a panel and you'll see somewhere in there at the opportunity to ask questions So if you have questions feel free to write them at any stage in that panel on the right and When we get to the end of the two presentations I'll take a look at the questions then and I'll then I'll pull out once that maybe well, I'll ask questions to the relevant speakers Based on what you submit. So please as questions occur to you throw them down The chat function is closed as far as I know So it's not possible to chat to each other And so the speakers aren't going to be answering questions in real time or anything like that In fact, the speakers don't see the questions. I'm the only person who sees the questions I see that there's about 240 people in attendance with which is great So in case I forget at the end, I'll just say thank you all for taking the time To attend a webinar. I'm sure it's going to live up to your expectations and as they say Without any further ado, Daniela, the floor is yours. Thank you very much Thank you very much John and board meeting members for inviting me to present the FEMS-YES webinar Today I'm going to sort of describe in a sort of light touch way The taxonomy of the saccharomyces genus and I will show you some experimental data on protein-protein interaction and transcriptome in hybrids and discuss some strategy to exploit the extent biodiversity for biotechnological uses So this now summarizes the evolution of the taxonomic rearrangement in the Cervizia genome and the saccharomyces Cervizia sensor-strict group was determined in the 1970 and and compasses pieces which are related to fermentation, industrial fermentation and it was separated from the saccharomyces Cervizia sensor-latu group which encompasses pieces more recently or later So back then the taxonomy was relying on a few morphological traits and few physiological aspects to call these pieces but with the event of more molecular tools some of the imitation of the original taxonomic classification were overcome in 1998 from the 21 species originally assigned to the saccharomyces Cervizia sensor-strict group which went down to 14 and in 2003 several other species were reclassified and mainly due to the work of Kutzen and Robnett they established that the sensor-strict group was a monophyllating group distinct from the sensor-latu and most of these sensor-latu species were now reclassified in new genals So for example, saccharomyces Castelli is now now Movia Castelli and saccharomyces Exeggius or Ulcerivaxii are now at Kazakhstan So and other things which happened in around about 2000 was the discovery of a couple of new species saccharomyces Micati and saccharomyces Curicevi They were discovered in Japan and then another species saccharomyces Carbicanus which was discovered in South America now yeast It does reproduce both sexually and sexually So given it has sexual reproduction The scientists found very useful to use the biological species concept to discriminate between species So all the species listed here in 2003 solve Timeline are species which are reproductively isolated from each other The hybrids between these pieces are are sterile However, later on when more genomic studies came out Leity and a Lewis group found that at the Carbicanus sequence genome sequence was Very very similar to those of paradoxes. There was really Not many many differences. So they they decided that saccharomyces Carbicanus is actually indeed a Subpopulation of paradoxes South American paradoxes sometimes they call it. So Well, I'd like to point out here is that a Carbicanus is reproductively isolated due to Translocations originally that were found for translocations between Carbicanus and paradoxes my group has Sequence that using Value the Carbicanus genome. We found actually a translocation of five and there are eleven inversion as well in this genome So Carbicanus is still a different biological species because it's still reproductively isolated But it has pretty much the same sequence as paradoxes and the four on the base is also a few genetic sequence Species concept. Carbicanus is now not considered species anymore in this group in 2011 we also had the discovery of Arborecola from China Bay and the discovery of Iberianus from Diego Lipkin and Joseph Hayo from Patagonia Argentina and the Iberianus that was quite excited. But finally we found the other parent of the Liberia East Pastorianus, which we knew one pair of Cervizia and the other one was still Debatable and now we know that the other planet is Iberianus and more recently my group Discovered a new species again a saccharomyces urea and We stumbled across the species in a very fortuitous sort of way and I can tell you quickly the story. I have been Enrolled in Coordinated a field course in biodiversity and ecology for undergrad students at the University of Manchester So we took this bunch of students about 30 of them in southern France in a region called Santobar about 1000 meter altitude and On the left the left today of these two weeks residential course So the student were pretty much free to do what they wanted and most of them did dangerous things like canyoning and You know and stuff Decided to do research. So myself with my research associate at the time Samina Aziz we went around the area and we discovered patches of oak trees which were quite high up was one thousand meter above sea level and we Got some samples and when Samina came back from the trip He she managed to isolate it 200 colony at that point We we didn't really know what much to do with these colonies So we sent them to your Robertson and Steve Jones In the what it was the issue for the research and we're asking could you please have a look at the ideas and check them out There's anything interesting. So we only sent three samples out of the 200 collected I'm sure enough but Steve came back and he said well actually something is really all we want I mean it kind of made the novel However, I left you got two samples, you know, we can't really Describe a new species because you need two independent samples or strain so Samina send them another five frames and That was from another bar and another oak one from soil up on bark and again, it came up That's an isolate which it could be no and And that's when we got really excited and so he asked me I just go on send the entire 192 samples that we got see what we got and You know with our this may everything else came out as saccharomyces paradoxes So there was nothing else that and so we were extremely lucky that within the first day sample we picked We found these two isolates. So first of all Samina looked whether it's a crime Is Yuri's a new biochip a species which is because they Spore viability is almost zero when we cross it with so busy a Mikari or three different population of paradoxes So the highest for what we saw was 3.1 percent And at that point I used to get teased quite a lot I mean Johnny Lee tea would come to me said oh, I mean it's gonna be another kind of kind of story in reality You know, you probably just discovered the European But I mean, thanks for that was not the cat is because when we did some sequencing Illuminous sequence and we did a phylogeny based on 101 Concarinated genes evolutionary concern that we saw that the saccharomyces you it does indeed Form a monophilated group close to me Gary We also did some pack bio sequencing to look a structural variation between the two strains and we saw that there is Collinearity between the two isolates of Yuri When we looked at them compared to some visa we saw that there were One and two translocations between these two species Interestingly one translocation of Yuri was in common with both strains of Mikari and these second part shows the That Mikari Chromosome versus the Yuri and the red is New translocation presence in the Yuri and the black is the other translocation that make happy 1815 has so The fact that saccharomyces Yuri did share one translocation with both Mikari isolates 15 and 16 It does point out to some sort of share evolutionary history between this this this species so We also looked at the biochemical and phenotypic characteristics of Yuri and Really we saw they can utilize maltose, but more importantly he can also utilize many times to to a good level As far as concern growth of different temperature the isolates were growing pretty well at 16 Called the temperature Somewhere in the middle here at 30 are compared to some easy which is the green the green line 37 they were not able to grow at all And only said we have a very good growth. So it's it's conclusion We show that saccharomyces Yuri is a new species brought both from the biological and phylogenetics The point of view we show that both trains have to the secret translocation compared to Cerveza and one is in common with with Mikari 18 and 16 Both trains of these new species show same mitochondria profiling and actually cox two and cox three Extrusion from polyphonic analysis shows that they are similar to the sort of paradoxes mitochondria And they can't ferment it's tires. They're more cryopolar and Cerveza and they do not grow at 37 And most of the community probably already know that the u-ray Saccharomyces Yuri name was chosen in memory of professor. You're a piscar and he's a sound contribution to these genetics and molecular biology So going back to the taxonomic rearrangement I just want to point out another Story which I call it that by anus saga. So so by anus went through a different rearrangement again In the 2011 there were two sort of variants of this by an uncle you've all come back or by anus With a you've all been more homogeneous generalized and then then the other it was only Thanks to the discovery of you by anus that to this taxonomy was resolved as well And it turns out that to that saccharomyces By anus variant you are me is actually true species and It's now called saccharomyces round while by anus by anus is a real hybrid And we each contain about two third of the genome of you by one third of the genome of you by anus And a little bit of intergression from so easier. So now of course He needs a picture. I'll just show you a couple of my braids May the main idea is known for that sense of stricter group, but The reality is that we've got tons of different hybrids between all these Species both my interest pieces level and interest pieces level and they're all sort of been isolated from From different subspecies so why Like the species I did eyes to this piece I guess so So readily well civilization, you know, it's a sweet mechanism to reshuffle the genome introduce novelty barrier It can combine advantageous trade from both parents And on and that it's sorry enough. We all know that but I mean also what we We Looked in my lab is is that hybrids has actually two diverged problems and the diverged protein itself could be a source of variation a source where the evolution could actually Act upon to lead to new adaptation and If you think, you know in hybrids to genomes and the four to problem coexist with this little Sort of the cartoon representing different products also, what we know is that the majority of the function is that are carried out by protein complexes and the correct assembly of a product in complex and the correct interaction is is key for Carrying to carry out the proper biological function. So you can imagine when you hybridized two different species you will have Assembly of product complexes in the hybrid. So one question is was what's happening to these assembly in the hybrid Will he just majority be Unispecific so these different prominent members co evolved the co evolved in our parent genome And therefore they might be able to preferentially stick with each other. So having to sort of Unispecific product complexes present in the hybrid. Oh, the other possibility though is Can't all it was different part of a orthologous member Create camera it product complexes and If you can create camera it probably comes to you immediately can see that there could be a substrate for variation And also the ability or not to form a functional camera in complex. It will have an effect on evolutionary trajectory in terms of gene retention and gene loss in the hybrid genome. So the study that we carried out we we we studied six different product complexes with precipitation and mass spectrometer and we show that actually Protein primary program can form naturally in the hybrid at least in four cases up to six that we looked The next question was what effect Suspended complex may have on a phenotype of the hybrid. So you can mention all these different combinations Could lead to a same fitness a loss of an increase of this fitness and also You could imagine that this could be environmental dependence. So one of the complex that we studied the trick to trick tree We are we look at what happened when we forced the cell to use a different complex So for example, we we deleted This this this trip two from some reason the trip three from Uvarum in this lab hybrid and we created two different chimeric combination And we created some unispecific combination and then what we look is that what happens to these hybrid In absent a trip to fan in terms of growth and as you can see here All the hybrids are very different phenotypic sort of display of growth and Interestingly the one which was growing Sort of back and the other was the one which was harboring the chimeric combination So this is done in monoculture where we also did One-to-one competition using facts and again the The hybrid which was out completed the other was the one which was having the chimeric complex So so these data points out that we have a phenotypic variation of different protein assembly And this could be important to adapt to to adaptation and to Genome evolution in itself going forward The other variation that you have in the hybrids is given from the mitochondria. So mitochondrial dna is usually Inherited be from both parents when you create a hybrid. However, within very Few generation the hybrid will kick out one or the other mitochondria So in recent years several studies has been that have been carried out to try to understand How wow why and how the hybrids size which mitochondria to keep And is this some sort of an environmental They are environmental cues that they That they help the hybrid to make this decision and and Is there any impact on the fitness of the hybrids when they host different mitochondria? And we now know from different groups from different studies That there are an impact on fitness and different nutritionally and also different temperature So my group specifically was interested to understand how the retention of the different mitochondria Can actually affect the expression of the nuclear genome in itself and What we did we we created some love hybrids between servisium and varum One harbour in the servisium one harmony you want mitochondria And we did some RNA seed data both in fermentative and non-fermentative condition and up to different temperature one Well higher one more hour so interactively we we saw Difference to nuclear expression due to the different mitochondria. These are circular plot and they can you can see for example in in in these I will look at We compare the hybrid which has the servisium mitochondria with a hybrid. We just got the varum mitochondria And for the servisium you can see the why there is no difference at all in any condition in the mitochondria with the servisium and with the hybrid servisium mitochondria We see a bunch of gene here up regulated when in the condition of non-fermented the condition are at warmer temperature You know, you can then look in mind these genes and they were mainly involved in their spiratory chain and mitochondria translation So there are and there are other examples like that both from the servisium and the varus in genome. So We also then decide to look at the co-expression profile throughout this this condition and it was interesting to see that There were a little specific cluster which were mapping a different cellular by chemical cellular pathway, for example In the respiratory chain complex the majority of the allele which had a shared co-expression profile where from servisium while the The in the lipid bias things is for example the majority of allele which did have a concerted expression where where there you are. So again this this data You know brings about the notion that the cell prefer to use either one parent or set or the other parent or set of the alleles According to the different biochemical pathway used So in conclusion, you know, we we show that chymaric product action are possible new source of natural variation that generate now the phenotypes Different mitochondria are also those also do also have any important fitness and nuclear gene expression and So the next question would be how we can use all these natural phenotype variation to improve Perhaps biotechnological traits in ivory. We know the hybrids are vastly used in fermentation and brewery, for example So I mean myself and Ed Lewis set up with collaboration with Catherine Smart and back then what used to be sub Miller Now is is a the in both We set up to see whether we can actually exploit, you know, breathing um over selection to improve Traits a strain for specific traits of industrial importance So the problem which We encounter when we deal with the hybrids that actually start out and if they're sterile, they're not genetically tractable as they most of you probably already know are the hybrids are And through meiosis in it to when goes through meiosis produce mainly majority dead spars because there is sequence divergence Which brings lack to the combination in the two set of homologous chromosome However, if instead of starting with a hybride, which is a diploid We start with a hybride which is a tetra ploid For which we have two set of chromosomes of one species and two set of chromosomes of a different species Then this time can happily go through meiosis and it can produce healthy Deployed hybrids spores, which contains one set of chromosomes from one species and one set of chromosomes from the other species This process has already been Showed by Duncan Gregg and Ed Lewis back in 2002 that tetrapodization Is actually breaking speciation barrier and you can actually have By an old sport So what we planned to do then was To incorporate as much possible biodiversity in tetra ploid hybrids are Then look at their diploid progeny for scramble traits So, uh, just a quick interruption. Are you keeping an eye on the time there? We just have a couple of minutes Okay, right. So i'm going to go very quickly here. Um, so we have two inter pieces Deployed that we can create in the lab Species n species b what we can do we can delete the major type locals on one and the other and they will They will Feel that they are actually deployed later And if they feel that they are made that they will actually create the third of hybrids So this first hub you can go through meiosis and the spores which will be deployed Some of them will be mat A and mat alpha. We'll have a sort of Scramble trait between all these species and again These if you do a random meeting and through meiosis, you can come to for example An f12 generation which you have a lot of scramble trait and at this point you can see you can actually have big phenotypic screening to understand that what which one of these will do better than the other in specific condition So my lab did the servise kudratzeri in urea hybrids at labs Focus on ubaianus and uvarum hybrids. We also introduced different mitochondria on each hybrids either servise or kudrat saving urea or servisia and so we have We have two different level of variation We have actually three variation of different strains variation of different species and variation of different mitochondria now That is just to show you different sort of a different Condition how big is that phenotypic space of this progeny and what you can do now you can take the 20 best performer and the 20 bottom performance and you can do sort of qtl type of studies, which we did Using a multiple approach So we we studied in my lab low temperature growth and maltes and acidic acid. So now I got a couple of slides with Results of these Which i'm gonna go sort of as quick as possible first things we look is that i might have come to have a profound effect on the qtl landscape There were very little qtl share between different mitotypes only this this yellow show a sharing between two A mitotype and that was pretty much happening in all conditions and for all hybrids that we looked um, also we were able to overlap qtl regions um and From different hybrids and that allows identification a lot of cold causal genes And you can see for example these on a low temperature We got all uh hybrids at least one mitotype which overlap with this core q6 gene which is Monoxidase mitogonal monoxidase involving the respiration and fatty acid and That so far so on. I don't have time now about similar the story of maltose and acidic acid. We had uh pretty much identified a specific qtl on qtl were Validated by the classic recycle amount and it goes is and all the leaves showed a significant difference in growth in a tetraploid Background but the key thing is that we now wanted to see whether these were specific to the hybrids or they were actually qtl which would come out also in a normal diploid Cervizia pieces and when we look at the same Sort of qtl in the parent species moving this time diploid Cervizia strain This qtl a significant is appeared Apart from these one which actually showed an inverse trend. So in this case the one of leel which was Defective or less good in in the tetraploid is actually better in the in parent So that still You know support the notion that there are Some lots of these qtl are actually hybrids specific So these are some conclusion The important thing is to to look at the for the idea of using these are Breathe in genetics and what do they apply to to hybrids and these are lines We managed to to look at a qtl Which are independent and dependent mitochondria origin we identify qtl hybrids specific And that we have a show a possibility of developing hybrids for for You know mitochondria purposes So the this is the last slide is another example of A nice but biotechnological project that we have in our lab. We exploit the saccharomyces urea And the difference of flavor profile that this train had to hybridize it with An age train to produce some craft there. So we didn't do it alone cloud water brew was was on board with this project and We we created different urines visa hybrid and These are the different sort of flavor profile that we found for for the parents and the hybrids. So The the urea had lower production of esters but a little bit more of spice and clove and more phenolic Taste also the primary urea doesn't remember you well. So the hybridization with the L strains Created a strain which had a fission fermentation. He had a much better sweet sugar attenuation and a different spectrum of flavor and now cloud water is is is Making up more of these awesome hybrid After hopefully the lockdown This is the knowledge man's life. I got to sense a lot of people but in particularly samina nz who is being Instrumental in the discovery of saccharomyces urea and the characterization of urea genome and also in the curial study Samina now has moved on from my lab. He's got her own position in the university staff for sure as a senior lecturer um Proping work is done by elizabeth. Hi, uh, uh, blow the review with me and she is an expert on Is biodiversity Sarah work on the mitochondria costantina walks on the beer and finerico work on the curial and on the beer with costantina Thank you so much. And these of course are my collaborator without them. Nothing would ever be done Thank you so much for listening. I'm sorry for a few minutes late That that that's all right daniel. I thank you very much that that was um, that's really interesting and I can see questions Coming in on my screen there that uh, that'll be holding to to ask you later If uh, if more people have questions for daniel, you know, you can be writing them now if you want in the questions panel there on your right Thank you. Um Enjoy that and I I think um, I think the hybridization or the discussion of hybrids is going to lead very nicely into Mr. Tony gavaldon's talk Uh, as I mentioned at beginning, Tony is based at the barcelona super computing center at the institute for research and biomedicine in in barcelona um, and uh, I won't use up any of your time now with any further introductions. Tony The floor is yours. I look forward to your presentation. Thank you So, thank you john. I just want to confirm that you see my slides and you hear me Yes, uh, yes, Tony. We do Okay, great. So as john said, I'm going to talk about uh hybridization and the origin of images And he's someone is more interested. Yeah, you can read the the review that was mentioned before on this special issue Of fans. Uh, yes research So let's start with this picture which I took In the old times when we were allowed to travel to the countries this picture I took in the in the airport of and In student in estocole and it depicts car from lignet who is very well known as the father of taxonomy He's also well known for going in the famous sentence Natura non facet salto Which means that nature does not progress in jobs. It was the idea that you can find gradients between different species and so Nature, you know, there was a natural gradient between Most differences you can find in nature this idea was embraced very heavily by child star winning his theory for the origin of species and in his mind also Species were evolving step by step and small steps and selection You know selecting those small changes that were a bit more affordable affordable than others However, already in those times the good friends of child star wind were You know telling him that maybe he was a bit You know taking a risk Raising that idea so heavily because you know nature at least in the area in the eyes of haxley Nature was indeed making small jumps So in modern genetics also this idea of you know step by step Evolution was was taken forward and most of the models They envision population Populations evolving by you know accumulating stations and then selection entry Making these populations moving slowly Through the genotype space or at least not slowly, but at least in small steps through the genotype space and then Regarding on the fitness, you know, choosing which direction they go genotype space However, we now know that there are actually jumps in evolution are sometimes called quantum leaves in which you know Species can jump in this genotype space by meaning of these jumps. They don't accumulate just tiny mutations or mutations, but these can be huge radical changes and here are some examples like gene or genome duplications processes of symbiosis could be considered as these Hybridization if what is one of them and lateral gene transfer all these are examples in which, you know, the jumps in the genotype space can be radically belong and therefore also the changes in fitness can be also Very large as a result So today I'm going to talk about hybridization and hybridization is it's a concept that was explored much earlier than the advent of genomics and it's a concept that is shall explore in the framework of the concept of the species. You all know the example of the horse and the donkey that they can breed and have a mule which is sterile and this is proofs in the geological species concept that horses and donkeys are different species However, I want to to emphasize that hybrids can also considered you know By mating within the same species the populations are, you know, divergent of these can also considered hybrids and just to refer to the famous book of Gregor Mendel He was referring as hybrids to the crosses of different varieties So this is another cartoon showing, you know, the idea, you know, a genetics insight into hybridization we can envision a species as a, you know, a set of different populations that are interconnected. They can exchange genes and genetic material and therefore there's kind of gradient in the genetic diversity within the species. So there's a continuous space of genetic diversity joining all these different populations that are somehow connected. We could consider the concept of a species like that but those populations can get you know isolated from one another by whatever barrier you can imagine And these populations may evolve differently for some time as depicted here But after some time these barriers can be removed by other islands that That separate but then there is a there is a lance joining them or something You can imagine whatever scenario and then the two populations can start, you know, exchanging genes or genetic material if they are still compatible And this is what is called, you know Hybrid zone usually is the area where they mix and the hybrids are the the offspring between making So what happens afterwards? So it can be, you know, different things can happen Imagine one of these hybrids cannot back cross with one of the parenthals but they can with another parent can imagine the hybrid between the red and the yellow has been an orange as a mixture of the of the genetic characteristics of the two species and it Can't back cross to one of the species after some time What this will lead to is genetic integration have Mostly red organizing mostly red genome Maybe of the yellow genome retain and particularly they will be retained if they confer a selective advantage That given you have heard of many examples of this happening in saccharomyces So but what can also happen is that the hybrid cannot back cross with either of the parents and therefore this hybrid is doomed to get extinct It must undergo You know sexual reproduction such as in the case of you But if it's in an organism that can have a sexual reproduction then Can survive as long as it fits the environment So it's a particular Couple of particular considerations for hybrids in fungi. I mean Because historically hybrids have been studied in animals and plants and not so much Fungi and this is because they are very difficult to detect. First of all, the species concept in microbes You know, that is perfectly clear and everybody agrees And so therefore it's also difficult to study hybrids because it's A difficult concept for microbes and also hybrids are difficult to recognize because Basically share the same morphology as the parents and if they may have of course Physiological differences, but it's true also that within a species different strains of the same species can have huge Physiological differences as well. So it's very difficult sometimes to recognize only from the outside characters But it's nevertheless suspected because you know in microbes We find less presciotic barriers as compared to animals and plants And also they had the ability to reproduce them. So they were sort of suspected And they were You know started to be discovered as as it was suspected with the advent of of genomics So in recent years with more and more genomes coming out we have been discovering A higher number of hybrids and Danielle already mentioned The huge number of hybrids that we find within the saccharomyces species Complex and just to mention that many of these hybrids they thrive fermentative environments and industrial or human-made environments Also some some work from from my group in the past we Show evidence that actually the whole so-called post-colonial duplication plate was actually Duried from an ancient hybridization between two different images that hybridize and because of these A sexuality and these incompatibilities between the genomes a tetraplonization favored recovery of Of the ability to reproduce sex This is what we think it happens in the origin of the whole genomic So nowadays this saccharotune from the review and is It's showing you know Where hybrids have been found within the saccharomycotina? And this is of course not complete. We have a lot of hybrids within saccharomyces, but also in psychosaccharomyces and In the different plates a lot of hybrids within the candida plate in pica plate And this picture is already out of date and i'm trying to kind of keep track of new hybrid species Or or you know species with hybrids that are described And one way i'm doing it is by asking in twitter and and i was really pointed out to too many new examples that were Our that arose during the last Last year So i want to remark that that we have in this these hybrid species Which means i call hybrid species You know species for which all the identified members are hybrids and sometimes even the Specimen that was used to describe the species is hybrid, but also we have a species with hybrids Species in which we have Homocybous parental species that can form a hybrids and we find you know a fraction of the strains from that species are actually hybrids most of these hybrids Have been identified from industrial or clinical environments But this is slightly reflecting assembling bias these are the environments that we are more interested in but I bet they likely have a similar important role natural environments Remember we are very much interested in in the role of hybrids Barolence and pathogenesis because we want to understand we are studying fungal pathogens And there we realize that within a single place you have a bunch of highly related species And that can be you know Can be commensals of humans or can be exposed to humans, but only from those Bed of species only Few of them are able to colonize or infect humans. I want to understand What are the genomic determinants of this ability to infect humans? pathogenesis And in several cases we have found that hybrids are enriched This is an example of the candida parapsylosis clades Clay that was you know divided into three different species parapsylosis orthopsylosis and metapsylosis Some years ago and we have found that both Candida metapsylosis and candida orthopsylosis are mostly hybrids in the case of candida metapsylosis all And clinical isolates we have Investigated so far are hybrids and we never found And the homozygous parenters among clinical isolates and the case of orthopsylosis This is similar, but we do find only one of the parenters some clinical isolates, but There's a bias towards only one of the parenters and these they constitute A minimal part of the clinical isolate. So the majority of the clinical isolates are actually hybrids These hybrids are You know present everywhere around the globe and we think there's some connection The fact that they are hybrids and the fact that they are able To colonize better humans and causticis as compared to their host Have proposed that hybridization may be a root in the emergence of clinical Or pathogenic species So since we proposed that we have been finding more And more hybrids among candida pathogens Most of these rare candida pathogens that are not so common in the clinics But are sometimes are very recalcitrant and very resistant To antifungal drugs and when we have gone and sequenced those genomes we have found that they have been They originated through hybridization. This is case for instance of candida inconstipation And also in the case of candida albicans, we can detect in the genome signatures that can be explained by an ancient hybridization In the common ancestor of all candida albicans So also candida albicans we can derive from an ancient hybridization And we don't know whether this was related to colonization with humans or not But he's he's very intrigued So conclusions from this first part Is that you know saccharomycotina saccharomycotina hybrids can be found everywhere Of course, if you look for them, if you look in some environments, you don't find them, but I I think they would find Every you know everywhere in many different environments Uh hybrids can be found recursively So for some of the species as I will talk about like candida alfopsilosis, the same hybrid has been formed Convertingly several times between the same two parental species So if we think as This has a lot of important implications in terms of species concepts Hybrids can form a new lineage and we consider this hybrid as a new species because it evolves independently from the others We could say that that species can be can be appearing again and again through evolution as long as the two parental Enegies are there and can hybridize and form a new hybrid So as I said some defined species are hybrid species. So the type specimen is Hybrid and all the members that have been analyzed are hybrids And they we also have hybrids between non-hybrid species so they They are shown to to successfully colonize new niches and as I explained this is a property a typical property Of hybrids that they sometimes show transversive phenotypes that are different from those existing Or interesting combinations of phenotypes present in the parental the parental lineages and that That's why they are, you know, sometimes colonizing new niches such as those created by by human also we can see And as humans as a new interesting niche for these And finally probably human related activities such as globalization trade environmental tradition climate change industry drags all these things probably is promoting the formation of novel hybrids by, you know Moving species around in the world and changing the environment and under stress new niches for hybrids to survive So in the second part of my talk if I have Time I will talk about, you know, how what we are doing in our lab to to study how these hybrids We are very interested in how they evolve once they are formed How they evolve and of course how how they can survive in the first place because most theory says That these hybrids should have a lot of negative epistatic interactions because you're putting together two genomes that have been able Separately for a long time and accumulate the mutations and be optimizing these complexes and all these things For some time and then you Mix up the two things together. So how can they Survive or or what are the constraints they have to evolve further? And in the review I talk about a concept that I think is important and I defined for the first time in that review Uh, which I call hybrid genetics This is opposed to hybrid zones, which you may have heard of which is Referring to the areas in which to At the this the geographical distribution of two species overlap and therefore hybrids between these two Close to the species can be so this is an geographical. Let's say Hybrid zone, but I'm referring to the the hybrid genetic zones, which is, you know maps in different Um in different axes or in different Dimensions so one dimension is the genetic divergence We can define the hybrids according to the genetic divergence between the genomes that that form the hybrid like The hybrid or the two lineages that form the hybrid and also the degree of gene flow between these two populations that form the hybrids So whether these two genomes are exchanging material very often or very rare And according to these parameters you can define Different hybrid zones you will define a species zone the typical species zone With populations that are highly related to each other and along with a lot of genetic change And you can find different hybrids in different zones And depending on what's the genetic divergence and what's the likelihood that this species exchange material And so on and according to this you can have different expectations regarding, you know, how frequently You're expected to find these hybrids How viable you expect them to be, you know, how much Seek or healthy you expect a typical hybrid to be This is related to how many incompatibilities you may have in their genomes and also the transgressiveness is, you know, how different Is the phenotype expected to be in respect to that of the So it's just a theoretical framework to interpret the different types of hybrids So I will talk about two different stories very briefly Not to make my talk very long and one relates to hybrids which are clearly in this zone between species that are, you know, highly divergent Genetically and also that they do not exchange much And in that in that area because there is a lot of genetic divergence The theory predicts that they have a lot of negative study interactions And also you can think of it's not just the proteins that interact but also The regulation of the genomes Genome is regulated with transcription factors and all these other factors that ensure that the gene is expressed in the right moment At the right level and then you put these two systems together And and mix them out. So our question here was to to understand whether these hybrids were suffering From what is called a transcriptional shock in the hybrids This idea put forward by Barbara Mclintock For plans in which, you know, when you create hybrids Because of interactions between the regulatory systems And hybrids may experience the shock in which genes are dysregulated And with respect to them So what we did was to set up this experiment in which we took The saccharomyces freshly made saccharomyces cerevisiae saccharomyces barum Hybrid which is highly divergent species as Daniela Explained already and then we put them, you know, we grow them independently in normal temperature and also low temperatures And we wanted to compare, you know, expression of the genes When they are in the in the pure species background genetic background and when they are put together In the hybrids hybrid context or in the pure Strain context and also would compare expression between orthologs and expression between Omnologs within the hybrid and also Because we had the different temperature. We wanted to compare, you know, how much The hybrid impact gene expression as compared to another type of shock in this case Or a thermal shock. We wanted to know whether hybrids were inducing a higher shock In transcription as compared And this is basically the results we got so that The top Here on the top you have one minus correlation, which means the lower is the more similar is the patterns of expressions of genes And in the given comparison and the color code Indicated comparison. So as you can see when we compared genes with The pattern of expressions of genes in the in the pure background To that in the hybrid we saw the highly correlated And the correlation was much higher. I mean, we're altering their expression much less that when you induce thermal shock So in this case the hybrid shock was smaller was smaller than And and also what another thing to see is that when we compared expression between the orthologs You have a given amount of divergence But when you compare this expression of the same genes in the hybrid context And they have much lower differences. So kind of hybridization Instead of increasing the differences between the expression of the genes of different species It is somehow buffering differences So this is a high contrast with what has been observed in animal hybrids or hybrids Or several crop species in which Even for more highly related species people has seen a lot of misregulation Of the trial of the transcript. We still don't understand why Why is that we don't know what's the fundamental differences in the absence of transposons? Maybe The different layers of regulations. We still don't understand that these shows that at least at the transcriptional level Hybridization is not a big issue. This hybrid may explain why they can be formed In in another project in the group. We are investigating another type of hybrids, which I will classify here So the divergence between the parenthas is much lower. It's like three five percent Sequence divergence between the parenthal genomes and there's no genetic exchange between Because they well actually the species are thought to be asexual and we don't see this And these are a candidate hybrids and we investigated several of them in this in this range And one of them is candidate orthopsilosis, which I mentioned to you that Have evidence from gelding butler's group that it has formed several times independently by the crossing of the same two parenthal lineages We know that because sometimes you have the mitochondrion of one parent sometimes a mitochondrion of your parents Sometimes you have a recombinant mitochondrion and we can define at least four independent hybrid clays we We and others have sequenced several of these clinical strains and you can see that the different clays have different patterns of loss of heterosegocity And by investigating these patterns we expect to learn You know how these hybrids evolve and what are the constraints that determine this Of loss of heterosegosity after a hybrid is born because we think this gives us an opportunity like the same You know experiment has been repeated by evolution several times and have access to natural hybrids And this work isn't going so I hope I would be able to talk About this in a future seminar And with this I will finish and I will of course thank the people in my group who participated Grace I thought mostly Veronica Michelle and and ran Havana. Sun are two busy students And well Both of them just finished their busy very recently and also collaborators. Don't look out at that No sec and this picture is the last picture We took when we were able to socially or fear interact closely and this picture will be more up to date And with this I think I'm happy to answer your questions. Thank you Tony thank you very much Again, that was that was a great presentation And raises many intriguing questions Okay, so I'm I'm going to look at these questions and do do my best now to answer that to ask them to you um I'll direct them to The person they've been directed to but maybe the other person might like to answer as well because some of the questions that came to Daniela I think will also apply to Tony um But Daniela, this was a question that came for you And it is which factors let the hybrid strains being protagonists in some industrial processes like beer And non-hybrid strains prevailing in others. I mean, this is wine baking and fuel ethanol Do you want and I think I'll come to Tony after you answer because I think Tony might like to comment on that as well Yeah, so I'm not sure I understand that is which hybrid I'm more relevant for Uh, brewery processes No, no, no. So I think the question is in some um, let's say some industrial processes Hybrids are very common like brewing whereas in other industrial processes It's a the question is the hybrids are rare and and why is that? Well, I mean in uh In beer we we find both bolder because we've got hybrids from the lager and The ales strains are usually syrups. So they're not hybrid in wine. We have a syrups, but we also have hybrid syrups so You know At least this is for this to so to two processes um I guess that's in uh, uh, it all comes down to Uh, whatever environmental situation the the the yeast is is finding itself in, you know, we know that from the fermentation the hybridization brings together the um ability of ferment a low temperature Which comes from the solubianus parent and the ability to ferment which is come from the syrup is apparent and That is ideal for the fermentation in in in the lager And to some degree we see that in the urea and The Saccharomides is a lestrain hybrid in which we were able to improve fermentation but adding the cryo A bit of tolerance from urea and a bit of different aroma profile Oh, okay. Thanks, uh, Tony do you want to comment on that as well and maybe a related thing you could could answer because it's come up is They take stent. I know you addressed it a little bit, but the extent to which human activity Either kind of creates hybrids if I phrase it like that or or selects for them Okay, so so I first regarding the first question I I agree with what daniella said and I also want to note that many of these non hybrid species And or strains that are present in these in these years or wine They do have regions of interagression So I think they are in a way. They are they are highly evolved. They were hybrids in the past and they in the end they retain So I think the process of hybridization actually favor colonizational Of this but I would say the same also is it's a question of contingency and opportunity You have a niche Whatever species that can thrive and how to compete the others will be there not necessarily a hybrid hybrid simply because they Shuffle genes and properties They may create You know things that are suited for new environments for which species have not been adapted for a lot And regarding the other question. Yeah, it's difficult to quantify Yeah, I would like, you know, one idea happens to go back in collections But yeah, collections are not that old because human activity, you know started Trading and so on a long time ago, but it would be good You know this but actually it's by common sense that you know if you are disturbing the environment I mean, we know this is happening, you know, the polar bears and the grizzly You know because of climate change and and impact they are now encountering each other hybridizing So we see this in in another species and for sure in microbes is happening Even more because of the ability to be sparse and to be, you know associated with crops and products and Okay, thanks, Tony and that is a question here that I suspect that might not be an answer to but Uh, it's to do I guess with a species concept And the question is what percentage of variability between species suggests a novel species? Um, it says in genomic and phenotypic level Yeah, okay, so To be honest, I don't remember on the top of my mind, but I think a few percentage, you know I mean, it's probably already enough to establish a new a new species. Uh, um the The um The problem of biological species concept versus what was called maybe phylogenetic species concept is Of our choice I mean, we we should decide which concept to choose and we should stick to it Um, so for the saccharomyces genus I can see that It makes sense right now to use the phylogenetic species concept because we can gain all the information Genomic information very quickly with this new generation sequencing and we can draw trees and understand Understand the sort of relationship DNA relations very quickly. Also, we know that the primarily barrier to to To reproduction is given by sequence divergence. Um, so that also would make sense To stick with that concept and thirdly saccharomyces The saccharomyces genus does not outcross very often. So does not do meiosis very often so I mean using the Phylogenetic species concept might make sense to me, but we need to stick but On what we choose having said that You know going back on on the case of saccharomyces cariocannus that is definitely And by a very different species because if you cross a bit paradoxus you would not get that Viables four and that majority and that is because it's got several uh structure variation 11 inversion five translocations that is enough to take down the entire small viability. So but Sequence divergence wise is is is a South American paradoxus and if we decided that with what is counts We just need to stick to that for our classification Well, uh, I guess I guess there's not a great history of a taxonomous agreeing to stick to to the same thing But but uh, thanks daniella and there's a question here for tony. Um It says that in your review that that that you talked about there You you mentioned that the mechanism and rates determining patterns of loss of heterosugosity In the hybrids are rather poorly characterized And the question is What is a good measure of LOH? I mean in hybrids, uh, because they are highly heterosidous And then I mean depending on the divergence, no if the if the two parental lineages are 10% Diversions, so there is an expectation, you know having, you know 10 snips every 100 And that's you can plot these along the genome and when you have an event of loss of heterosugosity You see clearly a drop in this in these levels of heterosugosity So this is the this is what I what I call loss of heterosugosity in the hybrids, which is no it's loss of these heterosugous Is it's true that they are they are hard To define no because yeah, you have regions where you don't have snips No, you have 90 Expectation of 90 positions without any heterosugous positions So you don't know really when this LOH boundary is But within even if we take rough Approximations to these, you know, LOH we can see different patterns and different hybrids and still we don't know I look at this candidate Orthopsylosis hybrids and I see a lot of LOH blocks But I don't know whether they represent, you know, how much time of evolution does this represent And for snips, you know, we have some idea of how many snips may accumulate for whatever number of generations And we don't know these basically for the hybrids Yeah, actually I had a question that just thinking about the LOH that struck me in the two Two kind of case studies you showed at the very end In a way, I thought that they were maybe leading to slightly contradictory conclusions not question the data, of course, but But from the from looking at transcriptomes, it looked like, you know, there was no negative impact of hybridization Whereas I guess loss of heterozygosity Would especially if it's the same regions which are lost multiple times Would suggest that there is a negative impact on that part on that part of the genome Yes, well, actually this is because I didn't Talk about what our results were in the second part But I can I can tell you that what we see is actually that there is not a very strong pattern Of selection in the in the in the patterns of LOHs So we see there is a higher overlap than expected by chance But we don't see, for instance, functional enrichment or enrichment, you know, for proteins that interact with mitochondria Things like that. We don't see we think there is a lot of drift Going on in this in this in these hybrids Which also in this case of these these candidate hybrids We don't see a bias towards one of the two parenternas We see that they have more roughly 50 percent of each of the two parenternas So it seems that they have been coexisting And they don't have much of a problem coexisting for a long time and they accumulate LOHs But maybe not because they have to survive because there is few regions that are You know always LOHs or always heterocycles Okay, thanks, Tony I've been taking the tax task but by a well-known taxonomist for bashing taxonomists, so I won't do that anymore, Andre I'm going to go to another question here for Daniella This is a question for Daniella about creating the allot tetraploids It is doesn't the deletion of the mat locus Make the yeast behave as mat A And how would a yeast make it once the mat locus was deleted? Okay, so so what you do you got two deepwood strain That we made the we hybridized two species We made a diploid and then we hybridized two different strains Belonging to these two different species and we make another diploid So we deleted the mat A On on the first diploid and we deleted the mat alpha On on the other diploid now these two deepwood perceive themselves as being halfway And then we we then can cross them. So we have a math alpha deleted in one diploid and a mat A LOH was deleted in the other Cross them and we have a tetraploid in that way. I hope that's clear Okay, great. Thank you You know some of the questions that people have asked are very long and detailed but but we will actually we will actually record all of these questions and I can pass them to the speakers afterwards and They might be able to answer them separately Um, so so there's another question for tony here. That's a short question. So I can I can ask it tony. It says The fact that candida species of asexual reproduction Uh, sorry does the fact that candida species have asexual reproduction And their transcriptional mechanisms are so conserved explain Why hybrids keep the transcriptional architecture? Yeah, well the the the transcription experiment was not done on a candida was That I showed was for scarmises. We saw we did the same for candida hybrids and and actually the results are very similar But I think it's nothing is not the specific for the candida to have the transcription And also despite the fact that they are described as asexual We don't really know and from the genomes. There is there is evidence that That they can exchange genomic material. So somehow They find the way to make sometimes probably we don't know where where or when They do these and probably they do this so readily that we consider them as fully Asexual but from the genomes you see that they can they can exchange material um Thanks, tony um There's a question here it says for both so either of you can answer this maybe but It says if hybrids are unable to reproduce and propagate Why do they occur naturally? Well, the hybrid they do propagate asexually So, uh, I mean we can keep a line of hybrid Propagating by mitosis That's how pastoralans This is kind of pastoralans, which is the language is does not correlate. So there's not under 10 miles at all So this got almost the best of the two worlds here But if If we go I'm thinking on a more sort of sort of evolutionary sort of You know in a world where You know an organism is only able to reproduce sexually A hybrid is a dead end. So a mule which is a cross between a horse and a donkey Is a hugely wasteful from an evolutionary point of view. I mean if you think you have All these energy put into Mate the mule the donkey and the horse and then you have to make sure that the the zygote Uh, uh is actually formed and then the actually the zygote develops And and then when develops is is actually Alive, which is the mule. So and then the mule is dead. It can go on over. So for species which are only sexual reproductive hybridization is Very wasteful and that's why for a lot of species of these species. There are some reinforcement of the of the What we call, you know pre mating barrier. So you're trying not even to move the mate because it's going to be completely wasteful um, but in the case of organism left like he's that can reproduce also A sexually then if the hybrids can reproduce a sexually can go on. I mean it will be problematic if There are adverse condition and he has to Make his tetra there because then the spores won't be viable. So if in that case is is is certainly Unadvantaged Okay, and actually tony I was struck as well. I mean you you mentioned that uh Example in wine yeasts. I mean there's there's a ton of examples of integrations And and so and you said that is probably all originated as hybrids So so that would um, I guess that would I don't know. I've speculated now Maybe you can correct me, but that would indicate a maybe a biological Function for hybrids as well. Wouldn't it because I mean it's it's a way of genetic exchange Yes, I actually I mean it's difficult to say but I do think it's a It's a shortcut for evolution in a way No, because you you you bring one trait or one gene or one allele that is you know was Developed by adaptation in another species drinking Into another species and you mix two this is way faster than expecting, you know that trait to evolve independently again And I actually do think that many of the huge adaptations we see in nature And I'm not only now talking about micro organisms I think this also happens for for many other organisms they relate to hybridization now We see this with the genome sequencing, you know, even in humans, you know With denisovans with neardentals, there is you know, there's an ancient hybridization and you know all europeans or asians You know, we derived from these hybrid in origin that we had You know genes from from the two populations and then some remain so And now people is seeing that these you know alleles that have come from neardentals or denisovans They are related to you know Resistance to some disease or adapt adaptations and so on. So I think it's a it's a shortcut For adaptation and I think nature uses it often at least more than we At least I was expecting in the beginning. Now we are seeing this Yeah Yeah, that's a very interesting way of thinking about it Um, I had a question and someone's asked something similar here daniella for you. It's a it's on a different topic No, it's it's coming back to your isolation of Sacrifice is a urii Is it I mean you you you described particularly fortuitous circumstances when you were afraid to go canyoning and and hence you As you identified a new yeast But you know, you only had 200 isolates and it makes one wonder I mean are are there many other species out there or Maybe conversely is a surprising that no one else has identified saccharomyces urii in in other habitats Okay, so uh, yes, uh, I mean I was Thinking exactly the same when we were thinking how it's possible that you know, I mean relatively inexperienced in this field God that bits take bits of soil and finally new species. I mean we were quite Um gobsmacked. I think there is more stuff out there to be found Uh, I think that our isolation method is is is skewing somehow towards the isolation of always the very same species like the paradoxes from the wild And uh, I mean now, I mean when it's high or pioneer the the protocol at low temperature We managed to get the kudriethedia. They much more in varum the kind of tolerance pieces and It has to say, I mean one of the idea that I also developed in the review together with Haia was Try to understand how much is in that just simply looking at the EDNA signature in sort of metagenomic type studies and People who've been trying before they found that really little Of of these saccharomyces genus. They can actually be seen if seen at all I mean, I remember a study of Duncan Gray if you look at the metagenomics and You know, lots of basidomyceres Very little paradoxes inspired by the father when he goes on isolation Paradoxes that are quite a lot So Haia developed a technique in which we actually Enriched for the ITS one of these genus, uh, which is specific Band size 850 base per and all the other fungal are are actually much lower So by enriching that we were able to see a little bit more of the community That there is that around and we did see a metagenomic signature on the kudri in europe We had a few reads about on new bayanos in two different samples. We had several new rate And and we we saw pretty much everything except for alborecola And also, I mean, I think I'm okay to say that I mean the work has been by archive Now the group of brian gibson has found another strain of saccharomyces urea in in germany And I believe he's also Made some beautiful beer with it. So, I mean clearly there is There is now, you know, more stuff coming out from from this biodiversity studies Okay, great. Thanks We've come to the 90 minutes that that I said that we would stick to so I'm sorry that I didn't get to everybody's question There was actually quite a few questions that I didn't get to but but as I said, uh, we will collate the questions And we can pass them on to daniella and tony You'll also find daniella and tony's contact details Um on the internet easily enough if you if you want to contact them yourself And to ask any further questions. I'm sure they'll answer you as best they can Um, I'd like to take the opportunity to To thank all the people who attended first of all, we had I don't know. We had 260 or 270 people We still have over 200. I think Um in attendance. So thank you very much for logging on and listening Uh, I didn't say it but there were quite a few very nice comments for daniella and tony In the questions to thank you for the talks Both of you and the people I think really appreciated it And I really appreciated the talks. I thought it was really interesting um, I've really enjoyed the discussion and um, I'd love to chat longer because I think it's really interesting and and It's very clearly an exciting topic with a lot of really new things happening That by your groups and I know by other people and in the community as well So I think we can kind of watch this space for more exciting developments So so with that, um, I will just say again Thank you to everybody. I'll say thank you to um to fems and op people behind the scenes for for doing the logistics and organizing it sarah and uh joseph and and other people and um Remember to send fems yeast research your best yeast papers. We're very happy to take them And um, keep an eye on the journal for forthcoming special issues and interesting articles and based on other new things in the pipeline for 2021 You can see us on the website. Follow us on twitter. Uh, et cetera. Okay. So with that, um We will finish. I don't know. I don't know how we actually end this or I keep talking till everyone is gone But so sarah I'd say you you can uh, you can terminate us At any point now. Uh, thank you very much