What Happened to the Transgenic American Chestnut?

<v Speaker 1>Okay, welcome to the crime pace, but botany doesn't podcast.
<v Speaker 1>I'm here on a cold West Texas day with Eric Carlson, PhD,
<v Speaker 1>student at sunny ESF A College of Environmental Sciences and
<v Speaker 1>Forestry and lovely, smooth, loving, caring nurturing Syracuse, New York.
<v Speaker 1>Does that sound like a Do you think that's an
<v Speaker 1>adequate way to sum up Syracuse? Would you call it nurturing?
<v Speaker 1>Is it tender?
<v Speaker 2>Yeah? I would call it tender. Yeah, that's a beautiful description.
<v Speaker 1>It is a tender. Okay, that's great. Do you guys
<v Speaker 1>have any massage parlors there?
<v Speaker 2>I'm sure there's a few.
<v Speaker 1>There's probably quite a few anyway. Okay, So Eric, I
<v Speaker 1>just want to ask before we get started, what happened?
<v Speaker 1>What happened? You work with American chestnuts. Everybody was so
<v Speaker 1>excited about this transgenic chestnut that was able to resist
<v Speaker 1>Chryphinectara parasitica, the blight by I believe it was digesting
<v Speaker 1>secreting enzymes that digest the oxidalic acid that the fungus produces.
<v Speaker 1>But then everything there was I saw this in the news,
<v Speaker 1>and of course they didn't explain what happened. But they
<v Speaker 1>said that there was some discrepancy and you know, data
<v Speaker 1>was wrong and the whole thing had to be stopped.
<v Speaker 1>What happened.
<v Speaker 2>Yeah, So about a year ago we made a discovery
<v Speaker 2>that one of our lines that we were using for
<v Speaker 2>government approval referred to as Darling fifty eight, it was
<v Speaker 2>actually a closely related line called Darling fifty four. And
<v Speaker 2>so basically it was a big naming issue where we
<v Speaker 2>needed to go back into our documents and change the
<v Speaker 2>names from Darling fifty eight to Darling fifty four, which
<v Speaker 2>is a little embarrassing but definitely not the end of
<v Speaker 2>the world.
<v Speaker 1>And so what with Darling fifty eight and Darling fifty four,
<v Speaker 1>I mean, does anybody know they? First off, they both
<v Speaker 1>have this allele right that codes for the digestion of
<v Speaker 1>oxalic acid that renders the fungus just the minor pest
<v Speaker 1>instead of this death now right, they both have that
<v Speaker 1>copy of that allele. Isn't an allele? Can you tell us?
<v Speaker 1>Tell us again, give us a refresher on and what's
<v Speaker 1>going on here with this transgenic chestnut tree.
<v Speaker 2>Sure, American chestnut is highly susceptible to a necrotrophic fungus
<v Speaker 2>called Cryphenectria parasitica. The fungus attacks the tree by subcreting
<v Speaker 2>oxalic acid, which is toxic to tree tissues. And so
<v Speaker 2>what we did is we inserted a gene from wheat
<v Speaker 2>that codes for an enzyme called oxalate oxidase. And what
<v Speaker 2>that enzyme does is it oxidizes that oxalic acid and
<v Speaker 2>converts it into hydrogen peroxide and CO two, which are
<v Speaker 2>not harmful to the plant. And both Darling fifty eight
<v Speaker 2>and Darling fifty four have the exact same copy of
<v Speaker 2>that gene and it's actually in the same exact clonal background,
<v Speaker 2>so they were that gene was inserted into a bunch
<v Speaker 2>of cells and there are independent events within those cells.
<v Speaker 2>And so both of these fifty four and fifty eight
<v Speaker 2>came from that same experiment, So they're basically genetically identical
<v Speaker 2>except for the place where that gene landed in the genome.
<v Speaker 1>Okay, so this wow, Okay, So it's not it's at
<v Speaker 1>different locations, different losi in the genome, and that was
<v Speaker 1>that's the only difference, the main difference between fifty eight
<v Speaker 1>and fifty four. Yep, that's right, all right, And this
<v Speaker 1>is a dominant Allele, right, and so to refresh everybody's
<v Speaker 1>memory to get dominant recessive, you know, a whole mental thing.
<v Speaker 1>He's fucking around with the pea flowers, right, the quote
<v Speaker 1>godfather of modern genetics. And so this is a dominant,
<v Speaker 1>it's not recessive. So you only need one copy. You know,
<v Speaker 1>you can be heterozygous for this trait if you're a
<v Speaker 1>chestnut tree, and it'll come true, but you know you've
<v Speaker 1>got to you need that one copy. And if you
<v Speaker 1>only stand a fifty to fifty chance of inheriting that copy,
<v Speaker 1>if say, a transgenic chestnut breeds with a non transgenic
<v Speaker 1>chestnut that's still susceptible to the blight, is that correct?
<v Speaker 2>Yeah, that's right. Chestnuts are diploid, so they've got two
<v Speaker 2>sets of chromosomes, and this oxogene is only inserted into
<v Speaker 2>one of those sets of chromosomes. And so when the
<v Speaker 2>tree with that single gene goes through myosis to produce
<v Speaker 2>gametes either pollen or egg cells, only fifty percent of
<v Speaker 2>those halfloid cells will have that gene insertion. So therefore
<v Speaker 2>only fifty percent of the offspring from those trees will
<v Speaker 2>inherit the gte.
<v Speaker 1>Okay, cool and then when you guys are in I
<v Speaker 1>got a question though, too, when you're inserting these genes,
<v Speaker 1>I mean, are Darling fifty eight? Are they all the
<v Speaker 1>same phenotype? They're all the same clone.
<v Speaker 2>Yes, they're They're all the same clone. That's right, because
<v Speaker 2>the cells are called somatic embryos, and they're all clonally
<v Speaker 2>identical to each other.
<v Speaker 1>Okay, so there's a lack of genetic diversity, but I'm
<v Speaker 1>sure you guys, I mean, I'm sure that was one
<v Speaker 1>of the first thing you guys thought of, and you were,
<v Speaker 1>you know, planning how to basically get more genetic diversity
<v Speaker 1>using this single clone and then create you know, a
<v Speaker 1>good gene pool that all has this single copy of Eele.
<v Speaker 2>Right, Yeah, that's right. We're not making a bunch of
<v Speaker 2>copies of that original transgenic event. What we do from
<v Speaker 2>that original transformant is we get pollen from it, and
<v Speaker 2>then we can take that pollen and cross it onto
<v Speaker 2>wild type mother trees in designated field plots, and then
<v Speaker 2>they'll produce chestnuts, fifty percent of them being transgendic and
<v Speaker 2>fifty percent of them not.
<v Speaker 1>Okay, but that and and that's the part. It's basically
<v Speaker 1>having to then find which ones got the gene and
<v Speaker 1>which ones didn't. That's like the what's what takes time?
<v Speaker 2>Right, and each generation of out crossing, you're incorporating more
<v Speaker 2>and more genetic diversity from those mother trees.
<v Speaker 1>Would it have been possible? I mean, fuck, I guess
<v Speaker 1>I should have listened to our podcast that we did
<v Speaker 1>two or three years ago on this, But would it
<v Speaker 1>have been possible to just take a bunch of different
<v Speaker 1>individual phenotypes and of wild American chestnuts, pure American chestnuts,
<v Speaker 1>not crossbread with Chinese which is resistant to Cryphonecta parasitica.
<v Speaker 1>Chinese chestnuts are resistant to Cryphonectria parasitica because they've had
<v Speaker 1>ten million years of coevolution in the same ecosystem. Would
<v Speaker 1>it have been possible to take that single copy you
<v Speaker 1>know that that what is it the oxogene? You said, yes,
<v Speaker 1>would it? Would it have been possible to take that
<v Speaker 1>in just in certain to like a number of different
<v Speaker 1>phenotypes of wild American chestnut and so you had more
<v Speaker 1>to work with, not just this one fifty eight clone
<v Speaker 1>or what.
<v Speaker 2>Yeah, that is possible, And that's actually something we've been
<v Speaker 2>working on for a few years now, is getting more
<v Speaker 2>of those sematic embryos into culture, so we can do
<v Speaker 2>new transformations into new clonal backgrounds. The Darling line though,
<v Speaker 2>comes from one sematic embryo line called Ellis, And that's
<v Speaker 2>a really important line because we got the genome sequence
<v Speaker 2>and annotated for that for that clone. So that makes
<v Speaker 2>genome analysis of our transgenic events much simpler. Having that
<v Speaker 2>annotated gene, you.
<v Speaker 1>Have the whole genome, every single coding and non coding
<v Speaker 1>region of that genome basically map.
<v Speaker 2>Yep, and it's publicly available on phytozone.
<v Speaker 1>Okay. And what was the name of that you said.
<v Speaker 2>It's yeah, it's called phytosomee.
<v Speaker 1>The specific line was began with an E.
<v Speaker 2>Yeah, it's called Ellis.
<v Speaker 1>Ellis Okay, cool, all right, So what are the differences
<v Speaker 1>between fifty eight and fifty four? Are these basically just
<v Speaker 1>these were different experiments where you were shooting that oxogene
<v Speaker 1>in there and seeing where it would end up or
<v Speaker 1>were you was there like control for you were able
<v Speaker 1>to control where it ended up or what.
<v Speaker 2>Yeah. So the method we use for inserting the gene
<v Speaker 2>is called agrobacterium mediated transformation. And is that No, it's not.
<v Speaker 2>It's kind of more of the classical genetic transformation method.
<v Speaker 2>You use this soil bacteria called agrobacterium. In nature, it's
<v Speaker 2>the causal agent of crown gall disease, and so it
<v Speaker 2>inserts in nature, it will insert tumor inducings which express
<v Speaker 2>all these vital hormones that cause these cancers like gross
<v Speaker 2>called gulls. And so what genetic engineers is they took
<v Speaker 2>out those tumor inducing genes and then you can insert
<v Speaker 2>your genes of interest into that agrobacterium and then it
<v Speaker 2>will insert those genes into plants for you. And that's
<v Speaker 2>a semi random process. Those insertions go in each independent line,
<v Speaker 2>the insertion will be in a different place. So you
<v Speaker 2>have to kind of go through and determine where those
<v Speaker 2>insertions are after you do them, you can't target them.
<v Speaker 1>And where does it? Does it matter where on the
<v Speaker 1>genome that this gene is inserted from what you can tell?
<v Speaker 1>And how does it affect other things? You know? So,
<v Speaker 1>and first off, how many base pairs are in the chestnut?
<v Speaker 1>How many base pairs long as the chestnut genome?
<v Speaker 2>It's sixteen megabases.
<v Speaker 1>Sixteen yes, yep, sixteen million letters. Okay, and so anyway.
<v Speaker 2>I'm sorry, I think it might be billion, sixteen billions
<v Speaker 2>more more cracky because each chromosome is several million holy shit.
<v Speaker 1>Okay, all right, well I could check that. We could
<v Speaker 1>check that later. But but yeah, so depending I mean,
<v Speaker 1>does how much does location uh of insertion on the
<v Speaker 1>genome matter?
<v Speaker 2>It matters a lot. And they're actually there's a name
<v Speaker 2>for it, they called position effects, and they can have
<v Speaker 2>a few different differences between them depending on where they're
<v Speaker 2>in asserted. It can have effects on a gene expression level.
<v Speaker 2>So if the insertion goes somewhere that's like a tightly
<v Speaker 2>bound like U chromatin, you're not going to get good
<v Speaker 2>expression because it's like tightly bound ding a. But if
<v Speaker 2>it's somewhere maybe further towards like the ends of the chromosomes,
<v Speaker 2>places where there's more active transcription, you're going to get
<v Speaker 2>higher levels of expression. And you can also have a
<v Speaker 2>diletorious effects if those insertions land in like the middle
<v Speaker 2>of a gene or the cause like deletions of like
<v Speaker 2>parts of the chrome zone. That's something that can happen too,
<v Speaker 2>but that's that's generally not what happens.
<v Speaker 1>So location matters a bunch, it does, Okay, all right,
<v Speaker 1>by the way, a megabase. Yeah, its million base pairs
<v Speaker 1>so that was was it sixteen megabases long?
<v Speaker 2>Maybe maybe it's gigabases.
<v Speaker 1>Gigabaite, Okay, either way, we can I'm looking online. It
<v Speaker 1>says the American chestnut genome is approximately seven hundred and
<v Speaker 1>three million base pairs long based on the assembled sequence
<v Speaker 1>available on fytozone.
<v Speaker 2>Okay, so that that is the correct number.
<v Speaker 1>Seven hundred and three megabase. Okay, all right, Well either way,
<v Speaker 1>so so location matters a bunch. So there was obviously
<v Speaker 1>a reason why Darling fifty eight was chosen as opposed
<v Speaker 1>to Darling fifty four. Why was that?
<v Speaker 2>So with Darling fifty eight, the tDNA insertion dot OXO
<v Speaker 2>insertion was in a space between genes where it wasn't interrupting,
<v Speaker 2>and you have like the native gene sequences in the chromosome.
<v Speaker 1>So that was the.
<v Speaker 2>Reason why Darling fifty eight was originally chosen.
<v Speaker 1>Right, Because you're just you're just slapping this stuff in anywhere.
<v Speaker 1>That's kind of where the lack of control comes in,
<v Speaker 1>I guess, which is a little worrying. And I'm sure
<v Speaker 1>the technology will be improved, but this gene, you know,
<v Speaker 1>you can't really control too much where this this gene
<v Speaker 1>ends up, and if it ends up in the middle
<v Speaker 1>of the instruction manual for another you know of another
<v Speaker 1>gene that's coding for something really important, it renders that
<v Speaker 1>gene useless basically, or it'll just fuck things up or what.
<v Speaker 2>Yeah, I mean potentially that's something that could happen, and
<v Speaker 2>I can understand why it sounds a little bit scary,
<v Speaker 2>but in the grand scheme of genetic changes, it's actually
<v Speaker 2>really small, especially when you can compare it to traditional breeding,
<v Speaker 2>or especially hybrid breeding, where you're crossing two different species together.
<v Speaker 2>You can have massive genetic changes where there's entire restructurings
<v Speaker 2>of chromosomes and things of that nature that interrupt thousands
<v Speaker 2>of genes at a time, as opposed to maybe just
<v Speaker 2>like one or two.
<v Speaker 1>Yeah, there's a lot more control than because you're only
<v Speaker 1>inserting one gene. Versus if you're hybridizing American chestnuts with
<v Speaker 1>Chinese chestnut, you don't know what the fuck you're going
<v Speaker 1>to get. You're inserting a lot of different genes with
<v Speaker 1>absolutely no control of where they end up.
<v Speaker 2>Yeah, the scale of changes is completely different between the two.
<v Speaker 2>Is much smaller. With the insertion with Agrobacterium.
<v Speaker 1>Yeah, so, but any anyway, I mean, you guys picked
<v Speaker 1>fifty eight for a reason, and then there was a
<v Speaker 1>mistake where it was actually fifty four, which of course
<v Speaker 1>there's no way to tell until you look at the
<v Speaker 1>you analyze it, right, you look at the DNA and see, oh,
<v Speaker 1>this is actually how do you think that happened?
<v Speaker 2>Yeah, So, Darling, fifty eight and fifty four were developed
<v Speaker 2>at the same time. They were the transformation as occurred
<v Speaker 2>in the same experiment, and they were regenerated at the
<v Speaker 2>same time, and they were both promising events. We were
<v Speaker 2>developing both of them at the same time because they
<v Speaker 2>both had a single copy of that oxogene. They had
<v Speaker 2>high expression. We measured high levels of mRNA expression, and
<v Speaker 2>they had high levels of blight tolerance when we screened
<v Speaker 2>them with anoculation essay. So we were developing both at
<v Speaker 2>the same time, and at a certain step in the
<v Speaker 2>process you put them into highlight growth chambers to produce
<v Speaker 2>pollen to go on the field and make those crosses
<v Speaker 2>like we were talking about. And so we were doing
<v Speaker 2>crosses with fifty eight and fifty four at the same time,
<v Speaker 2>and so there's kind of two main possibilities of where
<v Speaker 2>this switch happened. It either happened during the propagation stage
<v Speaker 2>like coming out of tissue culture and prop getting in pots,
<v Speaker 2>or it happened during the pollen collection stage where a
<v Speaker 2>tube of fifty four Pallen was mislabeled as fifty eight.
<v Speaker 1>But the dishes got swapped. It's all happened. Maybe the
<v Speaker 1>petriet dishes got swapped. Maybe the pots. You know, how
<v Speaker 1>are the weed laws in New York. Nobody was blunted
<v Speaker 1>when they were doing this, probably because right, it wasn't
<v Speaker 1>the the you know, the pot. How long has pot
<v Speaker 1>been legal?
<v Speaker 2>And yeah, I'm not sure the last few years. But
<v Speaker 2>this this switch that happened that this was eight years ago.
<v Speaker 1>Eight year see, so it couldn't have been. Nobody was
<v Speaker 1>blunted when they were doing it. Was a simple mistake.
<v Speaker 1>I can see myself making a fucking mistake like that.
<v Speaker 2>It's okay, These types of mistakes happened all the time.
<v Speaker 1>Happened.
<v Speaker 2>It's it's not great that it happens, but they definitely
<v Speaker 2>do happen, and everyone's out a story of something like
<v Speaker 2>this happening.
<v Speaker 1>Yeah, you know, you know, sometimes the shit hits the
<v Speaker 1>fan and what I don't think this, you know, I
<v Speaker 1>don't think it deserved this because they the way they
<v Speaker 1>made it sound in the press. And of course, when
<v Speaker 1>you're doing science, you should always be one hundred percent
<v Speaker 1>on top of your shit. I'm joking around for any
<v Speaker 1>negative nancies out there in the artists. But but you know,
<v Speaker 1>they made it sound like, oh, it doesn't work, the
<v Speaker 1>gene doesn't work, this transgendic. It's all been a mess.
<v Speaker 1>It just it's it was. It's back to just you know, slamming,
<v Speaker 1>you know, poling from Chinese chestnuts into into egg cells
<v Speaker 1>of American chestnuts and just hoping for the best. Back
<v Speaker 1>to that because we just have to abandon which is
<v Speaker 1>that's shit for brain, that's not you know. So it
<v Speaker 1>turns out it's a simple mistake to condemn those I mean,
<v Speaker 1>that's my opinion, to condemn the whole project and grant
<v Speaker 1>that I'm only hearing your set of the story, but
<v Speaker 1>this just doesn't sound. It's my hopes are still with us,
<v Speaker 1>because I want I want to see chestnuts at some point,
<v Speaker 1>even if they're transgenic. I want to see them, you know,
<v Speaker 1>these towering monolithic beasts that have been able to get
<v Speaker 1>around that ten years, the ten million years that they
<v Speaker 1>missed out on of co evolving with this fungus, you know.
<v Speaker 2>Yeah, And and this this name switch, it has no
<v Speaker 2>going on all the data that we've collected over the
<v Speaker 2>years on these trees, because even though there was this switch,
<v Speaker 2>it was it's all consistent data. It wasn't like some
<v Speaker 2>was fifty eight and some was fifty four. Luckily everything
<v Speaker 2>is just fifty four, so the data is still completely valid.
<v Speaker 2>And that data shows that these trees have much higher
<v Speaker 2>levels of blight tolerants than wild type American chestnuts, and
<v Speaker 2>also that they're very safe. We did lots of safety
<v Speaker 2>testing showing that they're not harmful to things like insects
<v Speaker 2>like caterpillars or bumblebees or wood frog tadpoles, and so
<v Speaker 2>that's all the stuff that is important to know for
<v Speaker 2>things like government regulators and of course the public too.
<v Speaker 2>The public wants to biologies.
<v Speaker 1>So you're wrecking So you're not wrecking a fucking ecosystem
<v Speaker 1>any more than it's already been wrecked. I mean, the
<v Speaker 1>whole ecosystem on the East Coast has been fucking wrecked.
<v Speaker 1>I mean, you know, the HDC as I refer to
<v Speaker 1>it has been out there for how long? With five
<v Speaker 1>hundred years? About a little five hundred and twenty, So
<v Speaker 1>you know they've had they got a head start on
<v Speaker 1>the East coast before they could really start ruining the
<v Speaker 1>ecosystems of the West coast. But but you know, yeah,
<v Speaker 1>you want to you want to test, You want to
<v Speaker 1>make sure there's because everything in an ecosystem is connected.
<v Speaker 1>You want to make sure you're not causing a cascade
<v Speaker 1>of shitty effects. How long is the oxogene? How many
<v Speaker 1>base pairs are we talking?
<v Speaker 2>So the coding region is six hundred and seventy five
<v Speaker 2>base person.
<v Speaker 1>Oh, it's nothing so small in the non coding region.
<v Speaker 2>Well, are you talking about the promoter and the terminator? Yes, yeah,
<v Speaker 2>you know, I'm not sure for for Darling, I think
<v Speaker 2>it's probably in the realm of like eight hundred something
<v Speaker 2>like that. So the promoter about the same for the terminator.
<v Speaker 1>And those would be of course as this as this
<v Speaker 1>tree becomes part of the ecosystem, and you know pollen
<v Speaker 1>has exchanged over generations, those non coding regions will be
<v Speaker 1>subject to change. But the oxogene will be selected for.
<v Speaker 2>Well, I hope not, because those non coding regions, they're
<v Speaker 2>kind of what regulate the expression of the gene. So
<v Speaker 2>the selection would probably be to keep those intact so
<v Speaker 2>they can keep that gene functioning properly. So if those
<v Speaker 2>ended up getting mutated and the oxygen stopped working, then
<v Speaker 2>those offspring would probably be selected against.
<v Speaker 1>Okay, yeah, okay, I see that. So but either way,
<v Speaker 1>I mean a lot of this just sounds like you
<v Speaker 1>create the pollen, you create the phenotype that's resistant, you
<v Speaker 1>create a couple of them, you put them back into
<v Speaker 1>the ecosystem, and then you know where there's the highest
<v Speaker 1>diversity of remaining chestnut trees, and then you just let
<v Speaker 1>you let natural selection do its work, and anything that
<v Speaker 1>doesn't inherit that oxygene will eventually get blight and you know,
<v Speaker 1>be terminated and just reduced to you know, continuously you know,
<v Speaker 1>sprouting stumps and anything that inherits it will then become
<v Speaker 1>full trees and keep you know, it's going to take
<v Speaker 1>a while, but maybe in five hundred years you could
<v Speaker 1>have this back. But you know, you've got to start somewhere, right.
<v Speaker 2>Yeah, And that's and that's a good point too, that
<v Speaker 2>this is it's a starting point. So there will need
<v Speaker 2>to be selection on the front end as well. So
<v Speaker 2>we're doing these crosses with darling and producing lots of offspring,
<v Speaker 2>and we do see variability in the offspring in a
<v Speaker 2>bunch of different things. And that's what you expect at
<v Speaker 2>the beginning of a breeding program, especially when you're using
<v Speaker 2>diverse genotypes for crosses. You see lots of variation. You
<v Speaker 2>see variation in height.
<v Speaker 1>What kind of work? Yeah, so variation in height. And
<v Speaker 1>then I mean, are you seeing any trees that you know,
<v Speaker 1>when they become a dults that look like because Chinese
<v Speaker 1>chestnuts are just these they're cool, they're great, they're you know,
<v Speaker 1>produce edible fruits, but edible seeds, but they're not that
<v Speaker 1>impressive of a tree. I mean, is it You're not
<v Speaker 1>getting anything that looks like that? Are you?
<v Speaker 2>No? Well, I, like I said, there is actually a
<v Speaker 2>bit of variability. So some of the offspring are quite
<v Speaker 2>a bit shorter than like the non transient offspring, and
<v Speaker 2>then some are the same. And what you need to
<v Speaker 2>do is select for the offspring that have the good
<v Speaker 2>growth characteristics to carry forward into the breeding program. It's actually,
<v Speaker 2>it's probably actually a pretty common thing in any genetically
<v Speaker 2>engineered plant. There's actually this example of this GMO clover
<v Speaker 2>in New Zealand where they were modifying it to have
<v Speaker 2>increased tannins in the leaves to reduce bloating in the
<v Speaker 2>livestock that eat the clover and in these original trains because.
<v Speaker 1>That increased tannons would reduce bloating.
<v Speaker 2>Yes, they increased tannons in the leaf tissue reduces the
<v Speaker 2>bloating and actually the methane production within livestock that eat it.
<v Speaker 2>But their GMO version had a reduction of vigor in
<v Speaker 2>like the early stages it was it didn't grow as
<v Speaker 2>well as a non transgendic but through several generations of
<v Speaker 2>selective crossing and selecting for vigor, they were able to
<v Speaker 2>regain all of that performance of the original line. And
<v Speaker 2>that's typical for any type of project using these molecular
<v Speaker 2>tools to create new events, you need to do hard
<v Speaker 2>selection at the beginning, and that's kind of the step
<v Speaker 2>that we're at now that we're starting to produce lots
<v Speaker 2>of offspring. We're going through a selection stage and getting
<v Speaker 2>picking out the best growers that have the best play
<v Speaker 2>resistance and then using their pollen for the future crosses.
<v Speaker 1>It's going to be interesting to see because it's kind
<v Speaker 1>of one of these things where you just put it
<v Speaker 1>out there, you do the first steps, and then the
<v Speaker 1>ecosystem kind of does the rest. Like the ecosystem is
<v Speaker 1>gonna select what what's going to thrive best and what won't.
<v Speaker 1>But again, the ecosystem on the East Coast is so wrecked,
<v Speaker 1>it's like, you know, what will be the selection pressures.
<v Speaker 1>They're not going to be the same ones that they
<v Speaker 1>were seven hundred years ago.
<v Speaker 2>Right, There's definitely gonna be some strong selection going on
<v Speaker 2>in order to get trees that are fit for the environment.
<v Speaker 1>Okay, so where so when? So tell me let's get
<v Speaker 1>into the melodrama. So what happened? This was discovered you guys,
<v Speaker 1>you know, honed up to it, brought it up, you know, Oops,
<v Speaker 1>we made a mistake, and then what was the blowback?
<v Speaker 1>What happened?
<v Speaker 2>Yeah, So as soon as we found out this labeling error,
<v Speaker 2>we went and told and we notified the regulatory agencies
<v Speaker 2>right away and let them know that we intended to
<v Speaker 2>correct our paperwork and uh and get that sorted out
<v Speaker 2>prior to deregulation. And there really weren't any issues with
<v Speaker 2>the regulatory agencies. Uh, and we have now submitted that
<v Speaker 2>paperwork revising our original petition. But there were some groups
<v Speaker 2>who decided to use this as a way to attack us,
<v Speaker 2>this labeling issue. And yeah, so there are a few
<v Speaker 2>publications published some negative stories about our stuff that didn't
<v Speaker 2>show us in the best light. But there's always people
<v Speaker 2>looking to jump on you like that when you're working
<v Speaker 2>with this genetically modified technology.
<v Speaker 1>Right right? That man? So what and regarding that, I mean,
<v Speaker 1>what were what was some of the press that was generated,
<v Speaker 1>because again, the way that I read it was not
<v Speaker 1>it didn't sound like, oh we just you know, both
<v Speaker 1>we make step two things that were both resistant to
<v Speaker 1>chestnut blight. Just a simple error. They made it sound
<v Speaker 1>like the whole project was doomed and had to be abandoned.
<v Speaker 2>Yeah, yeah, that's definitely what was being said for a
<v Speaker 2>while there, especially, you know, you'd see these activists organizations
<v Speaker 2>going out and people from that organization giving interviews to
<v Speaker 2>lots of different publications looking to to write about this.
<v Speaker 1>And.
<v Speaker 2>You know, there's there's always this these misunderstandings and fear
<v Speaker 2>about the technology, and when something complex like this happens
<v Speaker 2>in error, people don't really understand what that actually means,
<v Speaker 2>and so it could it was easily spun into something
<v Speaker 2>that it wasn't that this was like a complete failure
<v Speaker 2>and we had to start all over when really it
<v Speaker 2>was just an issue of a name change.
<v Speaker 1>Mm hmm. Yeah, Yeah, we got the the tags got swapped,
<v Speaker 1>but both are still resistant. What was do you think
<v Speaker 1>it was like anti GMO blowback or do you think
<v Speaker 1>it was just other organizations that were working on similar
<v Speaker 1>things with chestnuts? Like what? What? What did what could
<v Speaker 1>you tell? I mean, where where who were all the
<v Speaker 1>who were all the negative Nazis? Not who? But but
<v Speaker 1>what were they saying?
<v Speaker 2>Yeah, there's other people that work with chestnuts that don't
<v Speaker 2>haven't historically used the molecular breeding tools that we used,
<v Speaker 2>that kind of use that as a way of saying
<v Speaker 2>that their way was still the better way and that
<v Speaker 2>we you know, yeah.
<v Speaker 1>That seems ridiculous. That seems you're never I mean, it's
<v Speaker 1>going to take forever and you're not going to get
<v Speaker 1>pure American chestnuts by just back cross breeding. But what
<v Speaker 1>was their anti gm O blow too? Though? I mean,
<v Speaker 1>was it some of that?
<v Speaker 2>Yeah, some of the there were publications like the Washington
<v Speaker 2>Posts actually had an article and you saw members of
<v Speaker 2>the anti GMO organization giving interviews to those people.
<v Speaker 1>And oh God Jesus Christ, Yeah, that's always a bummer.
<v Speaker 1>I'd love to get my science from from those people.
<v Speaker 1>So what's the what is the what's the future of
<v Speaker 1>this project?
<v Speaker 2>Then we're continuing on. UH, We're gonna do what we've
<v Speaker 2>been doing, which is use this technology and uh improve
<v Speaker 2>the American chestnut. This darling tree was never supposed to
<v Speaker 2>be a silver bullet. We never claimed it was. We're
<v Speaker 2>still working on new lines that use this same gene,
<v Speaker 2>this oxygene, using new promoters and also in new cell
<v Speaker 2>lines like we discussed before. And we're working on developing
<v Speaker 2>new genes to genes other than oxo to combine with
<v Speaker 2>OXO potentially and UH to create more and more resistant trees.
<v Speaker 2>We're continuously developing biotechnology and American chestnuts so we can
<v Speaker 2>have resistance a chestnut blatant. We're also looking at other
<v Speaker 2>diseases too, like phytothra root rot is a major disease American.
<v Speaker 2>We're looking at genes for that as well.
<v Speaker 1>The Phytophtha that which is a relative of brown algae
<v Speaker 1>that became parasitic. It's not actually a fungus. Where did
<v Speaker 1>that that species that is problematic for American chestnuts in
<v Speaker 1>the East Coast? Where did that come from? Do you know?
<v Speaker 1>And what species? Is it? A phytostera?
<v Speaker 2>Yeah, it's Phytothra cinemoni.
<v Speaker 1>Was fucking everywhere. Man, that's in Australia, fucking up boll
<v Speaker 1>of my pines too, And and yeah, anyway, sorry, go ahead,
<v Speaker 1>where to come from?
<v Speaker 2>I believe it's one of the worst global plant pathogens.
<v Speaker 2>It can infect like thousands of different plant species, so
<v Speaker 2>it's like a really major pathogen. And I think they
<v Speaker 2>believe it was originally brought over on cinnamon trees and
<v Speaker 2>that's where it got its species name from the cinemomi.
<v Speaker 2>And it actually predates chestnut blade introduction. They were having
<v Speaker 2>phytaluther issues prior to chestnut blade.
<v Speaker 1>But unlike chestnut blighte, it doesn't knock out every tree right.
<v Speaker 2>It's actually a lot of people consider it worse than
<v Speaker 2>chestnut blake because it attacks the roots and so it
<v Speaker 2>will kill the whole root system and then the tree
<v Speaker 2>can't re sprout like it does when it gets chestnut.
<v Speaker 1>But does it I mean, is it because chestnut blight
<v Speaker 1>is nearly one hundred percent fatal, right, I mean there
<v Speaker 1>are no American chestnuts that seem resistant to it. Is
<v Speaker 1>that true? Or are there supposedly a couple outliers or what?
<v Speaker 2>There isn't any validated resistance to chestnut blight, but I
<v Speaker 2>don't think there's anything resistant to fightoth it either. The
<v Speaker 2>thing with phytopha, though, is that it's only in the south,
<v Speaker 2>on the south end of the range. It hasn't come
<v Speaker 2>north north. Yeah, it's not north of like Pennsylvania. So
<v Speaker 2>it's mainly an issue in the south right now, but
<v Speaker 2>as temperatures continue to increase, there's a pretty good chance
<v Speaker 2>it will make its way up here as well.
<v Speaker 1>Jesus. Okay, so this this idea, how did you guys
<v Speaker 1>catch this this mistake between them, this miss labeling? How
<v Speaker 1>do you guys catch this screw up between fifty Darling
<v Speaker 1>fifty eight and Darling fifty four.
<v Speaker 2>Yeah, So, like we were talking about before with this
<v Speaker 2>inheritance of the single gene, that's because we have when
<v Speaker 2>you start with the original transformant, there's just the one
<v Speaker 2>gene on one chromosome. But what you can do is
<v Speaker 2>actually cross two trees together that both have the gene,
<v Speaker 2>and then you can end up with offspring that have
<v Speaker 2>a copy of the gene on both chromosomes, so they
<v Speaker 2>would be considered homozygous. And so we were doing a
<v Speaker 2>breeding program for that, and we were screening the offspring
<v Speaker 2>for the inheritance of both of those genes.
<v Speaker 1>ALM not heterozygous YEP.
<v Speaker 2>And so we were trying to do PCRs on the
<v Speaker 2>region of the chromosome where Darling fifty eight's ox insertion
<v Speaker 2>is and we were having unsuccessful PCRs, and eventually through
<v Speaker 2>like sequencing, we figured out that it wasn't actually Darling
<v Speaker 2>fifty eight, but it was Darling fifty four.
<v Speaker 1>When you say unsuccessful PCRs, tell everybody what was going on.
<v Speaker 2>Yeah, So when you do a PCR, you design little
<v Speaker 2>pieces of DNA to bind to certain places in the
<v Speaker 2>genome to amplify and make lots of copies of DNA.
<v Speaker 2>And so we were trying to do a PCR of
<v Speaker 2>this place in the genome that didn't exist because it
<v Speaker 2>wasn't Darling fifty eight, so it was.
<v Speaker 1>It was located the oxogene was located in a different
<v Speaker 1>part of the genome. So you were basically putting it
<v Speaker 1>was was it primers? Yes, you were using so the
<v Speaker 1>primers were not matching because they were primers made for
<v Speaker 1>Darling fifty eight not Darling fifty four.
<v Speaker 2>Yep, exactly, And so.
<v Speaker 1>That was what made you guys then realize, Okay, we
<v Speaker 1>got Look, but Darling fifty eight and Darling fifty four,
<v Speaker 1>I mean, what are and I think I maybe already
<v Speaker 1>asked you this, but what are some of the differences
<v Speaker 1>between these two different phenotypes that both have the oxogene?
<v Speaker 2>They're completely indistinguishable. I mean, when if you look at
<v Speaker 2>them next to each other, they look exactly the same.
<v Speaker 1>Is there is there any difference in Bleich resistance?
<v Speaker 2>No, they're functionally equivalent. And also gene expression is almost
<v Speaker 2>exactly the same to the level of that oxoexpression.
<v Speaker 1>Jesus. So this really, it really isn't that big of
<v Speaker 1>a deal. It sounds like, I mean, it is for
<v Speaker 1>a fucking study, that's a huge fuck up. But in
<v Speaker 1>the long game, in terms of restoring an ecosystem, these
<v Speaker 1>are both would you guys, would Darling fifty four ever
<v Speaker 1>be released or was that just kind of a backup
<v Speaker 1>that is not planning to be released in the first place.
<v Speaker 2>I mean it could have potentially been released, but we
<v Speaker 2>were focusing more on Darling fifty eight because you kind
<v Speaker 2>of need to just like choot, we have limited operations
<v Speaker 2>so to like split the breeding between two different lines,
<v Speaker 2>which is result in like fewer numbers of one or
<v Speaker 2>the other. So we were just focusing on fifty eight
<v Speaker 2>or what we thought was fifty eight for all those years.
<v Speaker 2>But yeah, the yeah, I'm sorry train of thought.
<v Speaker 1>That's okay, Well this is a casual conversation anyway, don't
<v Speaker 1>worry about it.
<v Speaker 2>But I'm sorry, I just remembered. So it wasn't as
<v Speaker 2>if we weren't testing the offspring with PCRs and stuff
<v Speaker 2>the whole time. The problem was eight years ago and
<v Speaker 2>have that annotated genome like we have now, like we
<v Speaker 2>were talking about, that is something that came years after
<v Speaker 2>this labeling mistake happened, So we didn't actually have the
<v Speaker 2>resources to identify in the genome where these inserts were.
<v Speaker 2>So that was part of the issue early on, is
<v Speaker 2>that we didn't have the fully annotated genome at that point.
<v Speaker 1>So that and that's why this was that's why this
<v Speaker 1>wasn't caught before. Right, Well, I mean I still so
<v Speaker 1>what do you think is there a point in time
<v Speaker 1>at which this you know, OXO containing phenotypes of American
<v Speaker 1>chestnut trees, of pure American chestnut trees, not hybridized ones
<v Speaker 1>where parts of the Chinese chestnut are just shotgun throughout
<v Speaker 1>the genome. Is there a point in time at which
<v Speaker 1>American chestnuts with blight resistance might be available to the public.
<v Speaker 2>Yeah, that's what we're working towards right now now. Because
<v Speaker 2>they're genetically engineered, they're highly regulated by the US Biotechnology
<v Speaker 2>Regulatory Service, so that's the USDA, the EPA, and the FDA,
<v Speaker 2>and so we have to go through a regulatory process
<v Speaker 2>basically demonstrating that the trees are safe, and then once
<v Speaker 2>they're given approval, then we can begin distributing distributing them
<v Speaker 2>to the public.
<v Speaker 1>The companies that like when Monsanto does the round up
<v Speaker 1>ready crops and all the shit that they put I mean,
<v Speaker 1>they have to do the same thing, right or do they?
<v Speaker 1>They probably lobby. They're a huge organization, so they probably
<v Speaker 1>lobby to get you know, las you know, to have
<v Speaker 1>some of this stuff more lax with them, to get
<v Speaker 1>some of these regulations lacks, or to speed things up.
<v Speaker 2>Well, actually they benefit from having really strong regulations because
<v Speaker 2>they can afford to go through that process. They can
<v Speaker 2>hire lawyers and of scientists to generate data to go
<v Speaker 2>through this process, whereas smaller guys, say small university labs
<v Speaker 2>or something, don't have the resources to fund all the
<v Speaker 2>studies and the lawyers to put their products through this
<v Speaker 2>regulatory process. So people like Monsanto actually benefit from having
<v Speaker 2>really strong regulation, whereas the little guys are the ones
<v Speaker 2>that are pushed out of the market because I can't
<v Speaker 2>afford to do it.
<v Speaker 1>I don't see it. How do they benefit though.
<v Speaker 2>Because it reduces their competition because there are so that
<v Speaker 2>they can They can actually afford to go through that
<v Speaker 2>regulatory process, but they're the only ones who can. So
<v Speaker 2>there's little.
<v Speaker 1>Yeah hoops that they can jump through, but no one
<v Speaker 1>else can, right, So that's why they get a monopoly.
<v Speaker 1>The fucking round up readything is so insane, like the
<v Speaker 1>fact that they spray round up on crops, that's fucking insane,
<v Speaker 1>just even if you're feeding it the livestock. That's just nuts,
<v Speaker 1>you know, And then it's bad for people like me
<v Speaker 1>to want to you know, roundup gets all the bad
<v Speaker 1>press because it's you know, given everybody cancer because they're
<v Speaker 1>eating it, versus I just want to use it to
<v Speaker 1>get rid of some fucking bermuda grass in my yard
<v Speaker 1>and I can't. Now I'm getting Now I'm getting mean
<v Speaker 1>comments from these permaculture wackos. No offense to the ideology
<v Speaker 1>of permaculture. It's just the association that I've noticed you
<v Speaker 1>just you tend to meet, you get like a fifty
<v Speaker 1>to fifty Tell me someone who's in the permaculture, it's
<v Speaker 1>a fifty to fifty chance I might not be able
<v Speaker 1>to tolerate being around them for more than ten minutes.
<v Speaker 1>That okay, But anyway, sorry, So the the regulations, I mean,
<v Speaker 1>let's say I'm a corporates lease right, and I've got
<v Speaker 1>some sort of I've got some sort of ulterior motive
<v Speaker 1>with this. I want to make money off of it.
<v Speaker 1>I'm approaching you guys saying, hey, I've got this great idea.
<v Speaker 1>We can you know a way that we can generate
<v Speaker 1>so much revenue and corner of the market using this
<v Speaker 1>technology you guys invented. Is what the chances of that happening?
<v Speaker 1>Am I going to be successful or are you guys?
<v Speaker 1>Is there is there good intention, good solid work that's
<v Speaker 1>going to prevent the corruption of this work. You guys
<v Speaker 1>have done.
<v Speaker 2>Well with the distribution of these trees. We want it
<v Speaker 2>to be as open as possible so that anyone who
<v Speaker 2>wants to grow these trees are able to. So the
<v Speaker 2>way you do that is with things like non exclusive
<v Speaker 2>licenses without patents, and so it's easier for people to
<v Speaker 2>grow these trees and move them around and basically do
<v Speaker 2>whatever they want with them, versus having, you know, a patented,
<v Speaker 2>non propagatable piece of you know, plant material that that
<v Speaker 2>only gets in the way of restoration.
<v Speaker 1>So this will be non patented. Have you guys been
<v Speaker 1>approached by any money hungry villains out there. I'm not
<v Speaker 1>asking you to name names. I'm just wondering if any
<v Speaker 1>dirt bag, you know, corporate the you know what I mean,
<v Speaker 1>corporate interests have approached you and tried to, you know,
<v Speaker 1>tempt you. Has that happened.
<v Speaker 2>I don't think so. But I mean, even if that
<v Speaker 2>were to happen, I mean, it seems like it would
<v Speaker 2>be something better than what they're already doing. Right If
<v Speaker 2>a corporation wants to get behind something to restore the
<v Speaker 2>American trusts, not that that probably wouldn't be the worst
<v Speaker 2>thing in the world.
<v Speaker 1>Well, it's not with the intention of putting money behind.
<v Speaker 1>It's just there's a cost. And then when you privatize something,
<v Speaker 1>you always seem to lose efficacy for the sake of
<v Speaker 1>because the main motive is not it's no longer being
<v Speaker 1>you know, being you know, the purpose now becomes making
<v Speaker 1>money as opposed to actually achieving the goal in the
<v Speaker 1>first place, you know, and if you are achieving the goal,
<v Speaker 1>it's only as a stepping stone to make more money
<v Speaker 1>and please shareholders, et cetera. Which is kind of like
<v Speaker 1>why it's so ridiculous when people talk about, you know, well,
<v Speaker 1>private sector it's the post to the government. Well, yeah,
<v Speaker 1>the government can be tainted and fucked up and inefficient too.
<v Speaker 1>The private sector almost certainly is looking like the power company.
<v Speaker 1>Like you know, in Texas, there's like ten different sleezebag
<v Speaker 1>companies that are all going to equally rob you, well
<v Speaker 1>not equally. Some definitely shake you down more than others,
<v Speaker 1>you know, for the purges of electricity versus if it's
<v Speaker 1>like a public utility. Yeah, there might be problems, but
<v Speaker 1>privatizing is sometimes the worst thing you can you know,
<v Speaker 1>it's all this this nineteen eighties Reaganomics garbage. But I
<v Speaker 1>was just curious because it seems like that would be Okay,
<v Speaker 1>pardon it. We hit a technical difficulty. I'll put a
<v Speaker 1>commercial break in there. Got the fucking commercials on this
<v Speaker 1>podcast have gotten horrible. It used to do this auto
<v Speaker 1>select thing to put the commercials in, and they would
<v Speaker 1>put like nine commercial breaks, and then now I just
<v Speaker 1>fucking go in there. Have to put them in myself,
<v Speaker 1>because it's you know, you'll be listening to a podcast
<v Speaker 1>on American chestnuts and all of a sudden you get
<v Speaker 1>this just crass, horrific. Hey wait, come come out to
<v Speaker 1>Jumbu Casino. Just fills me with fucking hate. Anybody talks
<v Speaker 1>to me like that, I just Jesus Christ. Anyway, Okay,
<v Speaker 1>so we were we were just talking about you know,
<v Speaker 1>possible corruption of of uh you know, uh market opportunities
<v Speaker 1>for for marketing industry. But but again, there's going to
<v Speaker 1>be non patented. This is just open source basically, it's
<v Speaker 1>like a like an open source software.
<v Speaker 2>Yeah, essentially, Yeah, because we wanted a few barriers in
<v Speaker 2>the way of restoration as possible, and if rustoration is
<v Speaker 2>our goal, it's definitely not to make a bunch of
<v Speaker 2>money to be able to get these threatened trees back
<v Speaker 2>on the forests.
<v Speaker 1>And so how did you guys get funding for this?
<v Speaker 2>By the way, I guess there's a variety of different sources.
<v Speaker 2>Some came from the State of New York.
<v Speaker 1>Uh.
<v Speaker 2>Some from the New York chapter of the American Chess Foundation,
<v Speaker 2>which is basically our biggest partner, a biggest supporter. For
<v Speaker 2>a few years, we were funded by the Templeton World
<v Speaker 2>Charity Foundation. They actually funded us for three years, which
<v Speaker 2>was a huge help, really allowed us to grow and
<v Speaker 2>advanced as a project. And recently we actually partnered with
<v Speaker 2>this company called American Castania, which is a small B
<v Speaker 2>corporation which is a public interest corporation startup. It's just
<v Speaker 2>like a few guys and they're starting to plant some
<v Speaker 2>orchards with our trees for future distribution.
<v Speaker 1>God, that seems so great, man. I don't know, I
<v Speaker 1>would love to see it. I'd love to see it,
<v Speaker 1>you know. Comit wasn't there some project on abandoned coal
<v Speaker 1>mining sites like pit mines that got filled in in
<v Speaker 1>Appalachia where they were going to restore them. They were
<v Speaker 1>going to because to plant these things you would need
<v Speaker 1>you know, they need sun. They're not they're kind of
<v Speaker 1>shade intolerant. I guess maybe when not when they're young,
<v Speaker 1>but certainly when they get older, so you'd need open canopies.
<v Speaker 1>Was there, and you know, abandoned coal mining operations are
<v Speaker 1>a great spot to do it. Was there something about
<v Speaker 1>that going on from what you know?
<v Speaker 2>Yeah, I remember there was ideas to plant on coal
<v Speaker 2>reclamation sites. That was one of the ideas. There's other
<v Speaker 2>programs I've been discussed, such as converting foul farmland and
<v Speaker 2>stuff like that. There's quite a bit of that on
<v Speaker 2>the East Coast that was formerly forests that was converted
<v Speaker 2>into agriculture, you know, like one hundred and two hundred
<v Speaker 2>years ago and has kind of just been abandoned and
<v Speaker 2>is now not really productive land. Those would be good
<v Speaker 2>places to plant them as well.
<v Speaker 1>You're going to have some competition from shipbag developers though,
<v Speaker 1>because they want to put you know, ugly tracked housing
<v Speaker 1>and you know, big box stores in those all at
<v Speaker 1>abandoned farmland. That's the big thing that the United States
<v Speaker 1>is doing these days is that you see what is
<v Speaker 1>what's the cul de sac neighborhood Seene like around seat.
<v Speaker 1>What are the suburbs of Syracuse like.
<v Speaker 2>Pretty typical, I guess, I mean equally just.
<v Speaker 1>Good old standard American bleak like you know.
<v Speaker 2>Yeah, there's lots of trees though, So that's nice.
<v Speaker 1>That is cool. That is cool, Yeah, because we don't
<v Speaker 1>you know, it's it's a lot of lawns in Texas,
<v Speaker 1>like in the I thirty five corridor. Those are some
<v Speaker 1>of the bleakst fucking suburbs I've seen not a lot
<v Speaker 1>of trees.
<v Speaker 2>So we get lots of uh forestation around here from
<v Speaker 2>all the rain that we got.
<v Speaker 1>Did you have you guys sourced when you were looking
<v Speaker 1>for phenotypes of of Castania and tile, were you guys
<v Speaker 1>sourcing any trees from out west? Because there's a number
<v Speaker 1>of of trees that got planted, you know, in western
<v Speaker 1>North America where the blight is not as good at surviving,
<v Speaker 1>probably due to the aridity. Were you guys sourcing any
<v Speaker 1>phenotypes from out there they got planted before the blight
<v Speaker 1>you know, took hold in nineteen ten or whenever it was.
<v Speaker 2>Yeah, I think we did actually get a couple from
<v Speaker 2>out there. There is this one now the names it's
<v Speaker 2>like a light cemetery or something like that. There's a tree.
<v Speaker 2>I think it's either Oregon or Washington.
<v Speaker 1>Washington, Tumbwater, Washington. Yeah, there's two beasts there.
<v Speaker 2>Yeah, yeah, I think we got a clone from that,
<v Speaker 2>and that's part of our orchard. We have thousands of
<v Speaker 2>different genotypes from all across the range. We have southern trees,
<v Speaker 2>we have northern trees, stuff in the middle. You know,
<v Speaker 2>we tried as much diversity as possible.
<v Speaker 1>There was a I remember seeing a massive fucking American
<v Speaker 1>chestnut that I guess was planted in the late eighteen
<v Speaker 1>hundreds and the Klamath River in Hamburg, California, and I
<v Speaker 1>talked to the lady who lived there, so this kind
<v Speaker 1>of the fucking shack looks like it belongs to Appalachia.
<v Speaker 1>It's like this old house, and she's you know, I
<v Speaker 1>went and talked to him, like, is that a fuck
<v Speaker 1>that looks like an American chestnut. She's like, yeah, it
<v Speaker 1>was planted by a miner in eighteen fifty two that
<v Speaker 1>came out here from West Virginia. And I was like,
<v Speaker 1>that's a fucking one hundred and sixty year old American
<v Speaker 1>chest not holy shit, But it was only one, so
<v Speaker 1>there wasn't the you know, no good seed, but they
<v Speaker 1>fuck it was a beautiful I got a photo of it. Yeah,
<v Speaker 1>I was in Hamburg looking for Baker cypress trees. There's
<v Speaker 1>a cool grove that I guess burned wonderful fucking Hespero
<v Speaker 1>ciprus species.
<v Speaker 2>I'd love to get out there. I know there's this
<v Speaker 2>one farm that's near LA called Skyline Chestnuts, and they've
<v Speaker 2>got quite a few American chestnuts and American chestnut hybrids
<v Speaker 2>that are like fully mature and producing notes. I'd love
<v Speaker 2>to see those in person.
<v Speaker 1>What's the biggest American chestnut that you've seen or that
<v Speaker 1>you know of on the East Coast.
<v Speaker 2>Well, the one that I know of is the biggest
<v Speaker 2>is in Maine. It's like one hundred and fifteen feet tall.
<v Speaker 1>I think Jesus, and it didn't How did it not
<v Speaker 1>get black?
<v Speaker 2>It must have grown really fast and hassmissed it so far.
<v Speaker 2>But if I recall correctly, I think I heard that
<v Speaker 2>people have found some lesions on it, so it might
<v Speaker 2>not be there too much longer. But I myself, I've
<v Speaker 2>found a few large surviving American chestnuts in the Syracuse area.
<v Speaker 2>I actually found one that was probably around eighty or
<v Speaker 2>ninety feet tall, and I actually took cuttings from that
<v Speaker 2>and grafted it into our orchard, and you've done crosses
<v Speaker 2>with it.
<v Speaker 1>When you're grafting, you have to make sure that that
<v Speaker 1>ring of vascular cambia meets up perfectly or what.
<v Speaker 2>In theory. Yeah, I understand that's how it works, but
<v Speaker 2>I'm I just do cleft grafts. If you're familiar with that,
<v Speaker 2>it's basically the simplest form of grafting, and that seems
<v Speaker 2>to work pretty well. I've gotten a few successful grass
<v Speaker 2>using that type.
<v Speaker 1>I've only ever grafted cacti before, and it's easier because
<v Speaker 1>they've got cat I have cortical bundles, unlike I think
<v Speaker 1>every other family of plants made. There's probably a couple
<v Speaker 1>other families that have cortical bundles, but they've got vascular
<v Speaker 1>tissue in the cortex basically.
<v Speaker 2>So I think that's one of the great ways of
<v Speaker 2>preserving genetic diversity, because there's still like millions of trees
<v Speaker 2>out there that are just like these little stump sprouts
<v Speaker 2>that are stuck under the canopy and so they can't flower.
<v Speaker 2>If you locate those you can go and take cuttings
<v Speaker 2>from lows and graft them into orchards that get good sun,
<v Speaker 2>and then you can do crosses with those with those
<v Speaker 2>trees that otherwise would never be able to make nuts.
<v Speaker 1>Yeah, so you're preserving genetic diversity. But still if that
<v Speaker 1>stem I mean, you can graft an American onto a Chinese,
<v Speaker 1>I presume, but the American will still get blight, at
<v Speaker 1>least that branch will.
<v Speaker 2>Yeah, that's correct. So what you're doing is you're capturing
<v Speaker 2>its genetics in the offspring.
<v Speaker 1>Right, yeah, yeah, yeah, but still the Yeah, the American tree,
<v Speaker 1>the American, that American branch will still get blight, but
<v Speaker 1>but the whole tree that it's a part of will
<v Speaker 1>be fine. But still, I mean a branch, there's a
<v Speaker 1>lot less service area for blight the colonies compared to
<v Speaker 1>a whole giant tree trunk.
<v Speaker 2>So yeah, and most of them it is sorry.
<v Speaker 1>Blight germinates, I mean, the spores germinate on the kind
<v Speaker 1>of like the opening for them where they get in
<v Speaker 1>is where the bark is splitting as the tree grows.
<v Speaker 2>Right, yep, that's right. And so you see kinkers form
<v Speaker 2>on the graft unions pretty often.
<v Speaker 1>Yeah, so right on the graft Yeah, okay.
<v Speaker 2>Yeah, I've done quite a bit of grafting over the years,
<v Speaker 2>and a lot of it takes and grows for a
<v Speaker 2>couple of years, and then usually within like three or
<v Speaker 2>four years, it dies from the blight. We have some
<v Speaker 2>transgenic trees that I use as root sock, and I've
<v Speaker 2>been able to to keep more alive longer using those
<v Speaker 2>as the roots suck than just using non transgenic trees,
<v Speaker 2>and I've had the best success with that, and some
<v Speaker 2>of our biggest trees are actually from grass.
<v Speaker 1>So what happens to the transgenic trees when they get blight?
<v Speaker 2>Yeah, they get these gnarly looking cankers on them. They
<v Speaker 2>it kind of you could describe it as like a blocky,
<v Speaker 2>quirky kind of texture, and it gets swollen versus like
<v Speaker 2>a normal wild type canker. They actually look like they're
<v Speaker 2>sinking in. And what the oxod does is it slows
<v Speaker 2>the blight down enough that the trees' natural defenses can
<v Speaker 2>kick in and compartmentalize up blight infection so the tree
<v Speaker 2>can keep growing.
<v Speaker 1>So they still get infected and they'll get a canker,
<v Speaker 1>but the canker swells up instead of sinking in and
<v Speaker 1>then what happened you know, a year down the line,
<v Speaker 1>when the tree has sealed it off and recovered it.
<v Speaker 2>Uh, it just continues to grow and it just a scar.
<v Speaker 2>It does have a scar, yeah, it does. It gets
<v Speaker 2>a bit of a gnarly texture to it. Have you
<v Speaker 2>ever heard of the phenomenon of kruddy bark before?
<v Speaker 1>No cruddy bark?
<v Speaker 2>Yeah, So it's kind of this thing that was discovered
<v Speaker 2>in American chestnuts where these kinkers get this like really
<v Speaker 2>bizarre texture and uh, and but the trees are able
<v Speaker 2>to survive, and they think it's some kind of infection
<v Speaker 2>going on of the fungus. The fungus is getting infected
<v Speaker 2>either with another fungus or maybe a virus, and it
<v Speaker 2>seems to spread. But the tree it gets like this
<v Speaker 2>really nasty looking bark on the tree that's kind of
<v Speaker 2>like a qirky, blocky texture. But the tree is able
<v Speaker 2>to grow. And that's on non trans trees where that occurs,
<v Speaker 2>But on these tracks genic trees it looks really similar
<v Speaker 2>to that. It gets this like kind of this coirky,
<v Speaker 2>gnarly texture where the canker is and that allows the
<v Speaker 2>tree to survive.
<v Speaker 1>And so in Chinese chestnuts, I mean, is it known
<v Speaker 1>how the Chinese chestnuts resist the black because my how
<v Speaker 1>I presume this how chryphenectria got so pathogenic and so
<v Speaker 1>deadly was a series of baby steps. You know, ten
<v Speaker 1>million years ago the ancestor of chryphenectria or whatever lineage
<v Speaker 1>you know, if you go far ten million years back
<v Speaker 1>in that lineage, whenever this thing evolved or first interacted
<v Speaker 1>with the genus castania, you know, it probably wasn't that bad.
<v Speaker 1>But then then the genus I assume it's like an
<v Speaker 1>evolutionary arms race in other words, like the Chinese chestnuts
<v Speaker 1>get some resistance, all the ones that don't have it
<v Speaker 1>get culled out by the fungus. Now the fungus has
<v Speaker 1>to up its you know, enzymes to break down more
<v Speaker 1>chestnut tissue and get around this compartmentalization whatever. Now that
<v Speaker 1>then the Chinese. Now it's the Chinese, you know, just
<v Speaker 1>like the zigzag back and forth ping pong of you know,
<v Speaker 1>upping defenses versus upping pathogenicity. How does the Chinese chestnut
<v Speaker 1>resist resist chryphenectria?
<v Speaker 2>Yeah, that's right, And you described me pretty pretty well.
<v Speaker 2>That co evolutionary process. One one theory is called the
<v Speaker 2>red Queen theory, which is that you have to run
<v Speaker 2>as fast as you can just to stay in place.
<v Speaker 2>And there's there's gene gene for gene interactions, so you know,
<v Speaker 2>the the tree will evolve a gene for resistance to
<v Speaker 2>the blight, the blight will evolve a gene for resistance
<v Speaker 2>to that, and that just keeps going on and on forever.
<v Speaker 2>And so there's thousands of genes that are likely involved
<v Speaker 2>in blight resistance. In Chinese chestnut, they've found a disease
<v Speaker 2>resistant losai on almost all of the chromosomes in Chinese chestnut,
<v Speaker 2>So there's so election going on on all of those
<v Speaker 2>chromosomes for resistance. So they've got different mechanisms. One is
<v Speaker 2>the oxalic acid degradation. They have a multi step pathway
<v Speaker 2>for that which is hypothesized, it hasn't actually been fully
<v Speaker 2>elucidated yet. And then they have different types of tannins
<v Speaker 2>they use. They have a different enzyme inhibiting proteins that
<v Speaker 2>they express, and all that adds up to them compartmentalizing
<v Speaker 2>the fungus. They wall off the fungus and hold it
<v Speaker 2>in place and then basically outgrow the fungus.
<v Speaker 1>So they're just they're throwing a ton of shit at it. Basically, yes,
<v Speaker 1>and this was again the result of millions of years
<v Speaker 1>of coevolution and ecosystem and Chryfhenectara parasitica is an invasive fungus,
<v Speaker 1>which is another point to make. That's another thing I
<v Speaker 1>always hear about. Again, it's a fifty to fifty chance
<v Speaker 1>with the permaculture people, not all hashtag not all permaculture people,
<v Speaker 1>but they're always not always fifty to fifty percent of
<v Speaker 1>the time. I feel like they're just denying invasion biology,
<v Speaker 1>which is easy to do when you don't understand it.
<v Speaker 1>I used to be like that fifteen years ago. I
<v Speaker 1>get it, but it's this is this is invasive. What
<v Speaker 1>invasive means. It's not going back to some mythical period
<v Speaker 1>and time where you know everything was pure, or any
<v Speaker 1>of these goofy fucking you know, straw man arguments people make.
<v Speaker 1>It's about ecological function and coevolution in an ecosystem. And
<v Speaker 1>when you when you cross that ocean that's been separating
<v Speaker 1>North American ecosystem from the Asian ecosystem, and you're basically
<v Speaker 1>eradicating those ten million years of coevolution, and now you've
<v Speaker 1>got this pathogen that's a mild pain in the ass,
<v Speaker 1>you know, pragmatically speaking for the tree that it's co
<v Speaker 1>evolved with, and now it's this fucking nightmare that just
<v Speaker 1>basically causes an extinction. American justnut would be extinct one
<v Speaker 1>day were it not for hybridizing programs, and wow, that
<v Speaker 1>it's not even it would still be extinct. It's not
<v Speaker 1>a pure American or not for this transgenic process, it
<v Speaker 1>would be extinct. There's no natural resistance, there's no way
<v Speaker 1>around it. It would be gone. Give another century or two,
<v Speaker 1>the stump sprouts run out of juice, they're no longer functional.
<v Speaker 1>Boom that that tree is gone. It only exists and
<v Speaker 1>hopefully fossil record somewhere.
<v Speaker 2>Right And even though it's not like extinct right now,
<v Speaker 2>it's functionally extinct. It's completely lost its place in the
<v Speaker 2>ecosystem and it's not feeding the ecosystem the way it
<v Speaker 2>used to, and it's not reproducing and replacing itself.
<v Speaker 1>Yeah, it's one of those fucking blocks that a huge
<v Speaker 1>block has been taken out of that Jenga tower that
<v Speaker 1>is the North American East Coast ecosystem, and it's teetering.
<v Speaker 1>So that's why I applauded the work that you guys
<v Speaker 1>are doing. And I can't imagine why anybody would be
<v Speaker 1>against it. I certainly understand it, you know, exercising caution,
<v Speaker 1>but I don't understand why anybody would just would be
<v Speaker 1>against it, are quick to condemn it, or you know,
<v Speaker 1>it's the best we can do. We're affecting as a species,
<v Speaker 1>we're affecting life on earth, no matter what, we might
<v Speaker 1>as well try to do it in a positive fashion.
<v Speaker 1>You know, well studied, et cetera. What made you? What
<v Speaker 1>made you want to take this up? What got you
<v Speaker 1>interested in chestnuts? Because you're not from You didn't grow
<v Speaker 1>up in the native range of these things, did you.
<v Speaker 2>Yeah, that's right. I'm actually originally from North Dakota, and
<v Speaker 2>when I was in undergrad at the University of North Dakota,
<v Speaker 2>I worked on transgenic poplars. Poplars are actually another one
<v Speaker 2>of my things I'm really interested in, which is funny
<v Speaker 2>to some people. A lot of people think they're just weeds,
<v Speaker 2>but I'm really into poplars. But anyway, I was working
<v Speaker 2>on transgenic popular and undergrad and I was looking at
<v Speaker 2>what I wanted to do for grad school, and I
<v Speaker 2>learned about the American Trust and how they're using this technology,
<v Speaker 2>this these molecular biology tools to have like really positive change.
<v Speaker 2>You know. This is such a great example of what
<v Speaker 2>the technology can be used for for good, and that
<v Speaker 2>really appealed to me, you know, And so I came
<v Speaker 2>out here to Syracuse to work on a new line
<v Speaker 2>of chestnut that actually uses a promoter from Poplar. That
<v Speaker 2>was actually my master's degree project, and I'm actually still
<v Speaker 2>working on that project now, this new line of American
<v Speaker 2>chestnut that uses the same oxogene, but it uses a
<v Speaker 2>promoter from Poplar to regulate it.
<v Speaker 1>When you say promoter and terminator, can you explain to
<v Speaker 1>everybody what those are? Basically, they're just what begin and
<v Speaker 1>and a gene region yep.
<v Speaker 2>So, a promoter is a non coding region of DNA
<v Speaker 2>that regulates how a gene's expressed. It regulates where it's
<v Speaker 2>expressed in plant, so in the leaves or the stem
<v Speaker 2>or the roots it expressed. It determines when it's expressed,
<v Speaker 2>whether it's expressed all the time or only under certain conditions,
<v Speaker 2>and it controls how highly it's expressed, So if it
<v Speaker 2>expresses a lot of gene product or if maybe just
<v Speaker 2>a moderate or a low amount.
<v Speaker 1>Okay, So yeah, it's basically the start and stop. It's
<v Speaker 1>a non coded region, but they're still really important for
<v Speaker 1>how the gene is expressed.
<v Speaker 2>Yeah, it's like a genetic switch. So it's like the
<v Speaker 2>genetic switch that controls how a gene's expressed.
<v Speaker 1>Okay, So okay, Well, changing the subject briefly from DNA
<v Speaker 1>to ecological function. How does where does so? Cryphenectria obviously
<v Speaker 1>can exploit other food sources. It's not a specifist for
<v Speaker 1>castanas because it's got to be hiding out somewhere in
<v Speaker 1>the ecosystem since all of the all the chestnut are
<v Speaker 1>basically gone except for stump sprouts. Where does it? Where
<v Speaker 1>does it hide out?
<v Speaker 2>Yeah? So it's also a saffar troph. It can consume
<v Speaker 2>dead wood, and it also has a host range that
<v Speaker 2>goes beyond just chessnut. It can actually also go on
<v Speaker 2>several different oak species and a few different maples I believe,
<v Speaker 2>But yeah, it it can host on most of the
<v Speaker 2>oak species, and it can actually be an opportunistic pathogen
<v Speaker 2>on some of them. Like the oglethorpe oak and the
<v Speaker 2>scarlet oak. It can be pretty pathogenic.
<v Speaker 1>On what what species of quircus are those? Do you
<v Speaker 1>know of him?
<v Speaker 2>Uh? Yeah, I think that's Quirkus orgle oglethal bit and
<v Speaker 2>I believe. And this scarlet oak escapes me. I'm not
<v Speaker 2>sure what.
<v Speaker 1>One of the prime reasons for not naming plants after
<v Speaker 1>people is just that how fucking ridiculous some of these
<v Speaker 1>these names are when you try to latinize them and
<v Speaker 1>put them as a species epithet. Yeah.
<v Speaker 2>Yeah, it can be pretty silly when someone inserts the name.
<v Speaker 1>For its.
<v Speaker 2>Uh.
<v Speaker 1>Okay, so this thing's hiding out. I mean it's a
<v Speaker 1>sapartrough too. So this is like a wonderfungus.
<v Speaker 2>Fuck yeah, it's not going anywhere. It's here for good.
<v Speaker 2>It's the only way to get around is to create
<v Speaker 2>trees that are resistant to it.
<v Speaker 1>What was when it because there was a virus that
<v Speaker 1>was taking the fungus back or putting a hold on it, right,
<v Speaker 1>I mean the fungus has to have has to have
<v Speaker 1>some ecological checks and balances back home too. Do you
<v Speaker 1>know about any of those?
<v Speaker 2>Yeah, there's this type of micovirus that's called hypovirus and
<v Speaker 2>it can get into the fungus and decrease the fungus's pathogenicity.
<v Speaker 2>And this was originally discovered in Europe. They also have
<v Speaker 2>blight over there, and they discovered a few decades ago
<v Speaker 2>that there were some kinkers that were healing on their trees,
<v Speaker 2>and they discovered that there were these viruses in them
<v Speaker 2>that were causing the blight to kind of get turned around.
<v Speaker 2>But in the US they tried to apply that in
<v Speaker 2>the US and it didn't work as well. And the
<v Speaker 2>reason for that is in Europe there was very limited
<v Speaker 2>genetic diversity in their blight fungus, and so the virus
<v Speaker 2>is very easy to spread around. But in the United
<v Speaker 2>States there's a ton of genetic diversity in the blight.
<v Speaker 2>There was probably several different introductions, and so these different
<v Speaker 2>blight strains are not compatible and so the virus doesn't
<v Speaker 2>spread between them.
<v Speaker 1>And so this fungus was brought over on other chestnut trees,
<v Speaker 1>right on Chinese and Japanese chestnuts or.
<v Speaker 2>Yeah, it was originally brought over on Japanese chestnuts.
<v Speaker 1>Wow. Fuck, that's man, that's brutal. So and in Europe
<v Speaker 1>the species is Castania sativa. It's different chestnut species too.
<v Speaker 2>Yes, yep. And it's also a susceptible species.
<v Speaker 1>Is it slightly more resistant then tata?
<v Speaker 2>Yeah, it is slightly more, but not that much. Still susceptible,
<v Speaker 2>so it gets killed by it.
<v Speaker 1>Well, how do you think, I mean, when these species
<v Speaker 1>diverge from the same common ancestor, how did one end
<v Speaker 1>up in Asia and one end up in the eastern
<v Speaker 1>North America? Just probably because the higher latitudes were warmer,
<v Speaker 1>and that was probably because they weren't going across the ocean.
<v Speaker 1>There were probably ranges that extended up into the Arctic
<v Speaker 1>when it was a lot warmer up there, and then
<v Speaker 1>would make their way down, you know, shorter distance at
<v Speaker 1>the top of the globe versus at the lower latitudes.
<v Speaker 1>I mean, is that how it's surmised because there's a
<v Speaker 1>lot of disjuncts between eastern North America and Asia.
<v Speaker 2>Yeah. Millions of years ago, all the continents were connected
<v Speaker 2>in the northern hemisphere, and so the chesshunt was able
<v Speaker 2>to spread from its origin in Asia around to North
<v Speaker 2>America and into Europe before they were eventually separated by
<v Speaker 2>plate tech tonics.
<v Speaker 1>I mean, that was much longer though, that wasn't just
<v Speaker 1>ten or twenty million years ago, That was a lot
<v Speaker 1>much longer period of time, right, Or am I wrong?
<v Speaker 2>No? I believe you're right.
<v Speaker 1>But these things because these things separated not that long ago,
<v Speaker 1>only ten or twelve million years ago according to the
<v Speaker 1>molecular clock est mints. So I mean, but either way,
<v Speaker 1>it's still a shorter distance if you're going across bodies
<v Speaker 1>of water, migratory paths, you know, between birds or among
<v Speaker 1>birds are going to be again shorter at the top
<v Speaker 1>of the ball the globe than at the lower latitudes,
<v Speaker 1>you know. So I assume that's what it was. It
<v Speaker 1>was just the Arctic was warmer, and because there's a
<v Speaker 1>lot of fucking disjunctions, whereas you know, humid and the
<v Speaker 1>climates of eastern North America and Asia are similar enough
<v Speaker 1>in humidity and temperature, et cetera. But in western North America,
<v Speaker 1>had any of those species been there, you know, in
<v Speaker 1>the western North America you've got these orogenes, these mountains uplifting,
<v Speaker 1>creating rain shadows, whole western part of the continents drying out,
<v Speaker 1>and so a lot of stuff was probably there, but
<v Speaker 1>one extinct.
<v Speaker 2>That's exactly right. They've found chestnut fossils in like the
<v Speaker 2>Rocky Mountains and on the West coast, and chestnut used
<v Speaker 2>to be all across North America. But when the mountain
<v Speaker 2>ranges on the West coast were raised up, like you said,
<v Speaker 2>and tried out the middle of the continent, that's when
<v Speaker 2>most of those chestnuts were pushed to the East coast.
<v Speaker 1>Yeah. I guess the only place they survived was the
<v Speaker 1>East coast because it was warmer. Yeah, that's crazy. Yeah,
<v Speaker 1>they used to be fucking Don redwoods. I mean even
<v Speaker 1>like twelve million years ago. I've found huge fossilized permenized
<v Speaker 1>some with still fucking wood fibers in them, like the
<v Speaker 1>really like partially permenialized wood fibers in southeastern Oregon northwestern Nevada,
<v Speaker 1>and the layer of basalt above them is like twelve
<v Speaker 1>million years old. I think these these you know, dispetrified
<v Speaker 1>wood was estimated to be fourteen million years old. I
<v Speaker 1>looked at the layer of rock that it isn't entombed
<v Speaker 1>in white volcanic ash. But that's crazy.
<v Speaker 2>The comparison to the Don redwood as far as the
<v Speaker 2>migration goes, because you can find Don redwood across you know,
<v Speaker 2>North America and Asia and Europe the same way the
<v Speaker 2>chestnuts were because it was that area was all connected.
<v Speaker 1>Oh yeah, yeah, similar and similar, similar climate, similar humidity
<v Speaker 1>requirements between medisaquoia and American chestnut. Medisaquoia doesn't do well
<v Speaker 1>in a dry climate. I mean, even if it's growing
<v Speaker 1>in a fucking river, it's there's still just the that
<v Speaker 1>dry summer era just pulls moisture out of it too quick.
<v Speaker 1>But yeah, that's that's true. I didn't even think of
<v Speaker 1>that because that's where medisaquoia is still in existence today.
<v Speaker 1>Is is eastern China, right. I think there's like five
<v Speaker 1>thousand trees left. God, I would love to go there.
<v Speaker 1>Do you know anyone who's seen Medisaicoia in the wild.
<v Speaker 1>There's far more in cultivation than there are in left
<v Speaker 1>and habitat.
<v Speaker 2>I don't. That would be a really cool trip, though.
<v Speaker 2>I've always wanted to go to China for a botanical
<v Speaker 2>adventure because they got all the cool stuff over there.
<v Speaker 1>Yeah I have. I haven't even touched Asia yet, man,
<v Speaker 1>except like the fucking area right around the Hong Kong airport.
<v Speaker 1>I would love to go. I'm kind of saving it
<v Speaker 1>till I can start digesting it all at once, and
<v Speaker 1>I want to just dip my feet in and become
<v Speaker 1>familiar with these certain families and lineages and shit, and
<v Speaker 1>then leave and not come back for three years.
<v Speaker 2>You know. Yeah, I remember when I was taking undergrad
<v Speaker 2>body course and you're Washington documentary of the sky and
<v Speaker 2>mountains in China, and he's just going like down the list,
<v Speaker 2>like every flowering plant family was represented on this hillside.
<v Speaker 2>You know, he's going basically down the list and everything
<v Speaker 2>was there because you know that that's the center of
<v Speaker 2>radiation for all angiosprings.
<v Speaker 1>Really my recollection, Wow, Jesus God, damn. Yeah, man, I
<v Speaker 1>gotta get over there. And plus, it seems like China
<v Speaker 1>actually funds a lot of science, at least compared to
<v Speaker 1>the United States.
<v Speaker 2>You know, yeah they do. They've got really great forestry programs.
<v Speaker 1>Fucking Ronald Reagan quote in the eighties like we're not
<v Speaker 1>here to subsidize intellectual curiosity. And so fifty years later,
<v Speaker 1>you got a fucking population that's, you know, getting dumber
<v Speaker 1>by the with every passing year. Well I guess that.
<v Speaker 1>I guess that concludes. I mean, I got time left,
<v Speaker 1>but I don't.
<v Speaker 2>Talk about some newer stuff we've been working on.
<v Speaker 1>Sure, sure, Yeah, what's what's been going on?
<v Speaker 2>Yeah, so I mentioned briefly this, uh, this new line
<v Speaker 2>using the palslar promoter. We're pretty excited about it because
<v Speaker 2>this oxogene, it is highly regulated by that promoter where
<v Speaker 2>it's kept turned off most of the time. It's called
<v Speaker 2>an inducible promoter. And so basically what that promoter does
<v Speaker 2>is it keeps expression turned off until the trees are
<v Speaker 2>actually infected with the blight fungus, and then the gene
<v Speaker 2>gets turned on and then it can mount a defense.
<v Speaker 2>And so what that does is it preserves resources of
<v Speaker 2>the tree that would otherwise be going to the oxode
<v Speaker 2>expression to be used for other stuff like growth.
<v Speaker 1>And so instead of so, how does the oxygen work
<v Speaker 1>in It produces an enzyme that breaks down oxalic acid.
<v Speaker 2>Yep, So it produces this enzyme. It's a cell wall
<v Speaker 2>embedded enzyme. And yeah, it breaks down that acid into
<v Speaker 2>hydrogen peroxide and CO two and.
<v Speaker 1>So if but it's not in this case with this
<v Speaker 1>new promoter, it's not producing that enzyme unless unless there's
<v Speaker 1>an actual infection, Unless an infection is.
<v Speaker 2>Detected, right, it's induced by physical damage or by pathogen infection.
<v Speaker 2>That when a pathogen infects a plant, it releases all
<v Speaker 2>kinds of chemicals into the plant tissue and those chemicals
<v Speaker 2>will actually trigger the promoter to start producing expression.
<v Speaker 1>The oxidale like acid that's produced by chryphenectria. What what
<v Speaker 1>does that's a byproduct of the enzyme that the fungus produces.
<v Speaker 1>What can we get into that? What's going on there?
<v Speaker 2>Yeah, So oxalic acid is actually produced from oxalo acetate,
<v Speaker 2>which is a normal metabolic compound that's produced like during
<v Speaker 2>cellular respiration. And there's this pathway that ends with it's
<v Speaker 2>called oxaloacetate and hydrolyse and it basically converts that oxalo
<v Speaker 2>acetate into oxalic acid. And so that mechanism is really
<v Speaker 2>well understood because of that high overhelence research that we
<v Speaker 2>talked about.
<v Speaker 1>And this is we're talking about what cellular respiration of
<v Speaker 1>the fungus or what are we talking about here?
<v Speaker 2>Yeah, so just during normal like metabolism, you know, producing
<v Speaker 2>ATP in the citric acid cycle, oxal acid is.
<v Speaker 1>Just like the of the fungus you're talking about. In
<v Speaker 1>the fungus, we got to specify that for anybody listening
<v Speaker 1>to this. So this is occurring in the fungus.
<v Speaker 2>Yes, the fungus is drawing its own resources and creating
<v Speaker 2>the acid. And then the acid gets secreted out of
<v Speaker 2>the fungus into the tissue of the plant and that
<v Speaker 2>kills the tissue of the plant. And then the fungus
<v Speaker 2>will release this other enzyme called oxalate decarboxylates and it
<v Speaker 2>breaks down the acid and then the funst grows into
<v Speaker 2>that dead tissue and consumes it.
<v Speaker 1>Wow, so it's not really a byproduct. I mean it
<v Speaker 1>is initially oxalic acid is a byproduct of just cellular
<v Speaker 1>respiration in the fungus, but then it serves the fungus
<v Speaker 1>by killing the tissue first and then allowing other enzymes
<v Speaker 1>to move into the tree tissue.
<v Speaker 2>Yep, that's right. And the acid does a few things.
<v Speaker 2>It kind of breaks apart the cell walls of the
<v Speaker 2>tree tissue and it makes it more digestible, and then
<v Speaker 2>it secreased digestive enzymes into that dead tissue to break
<v Speaker 2>it down more before the fungus consumes it.
<v Speaker 1>And would those digestive enzymes by the fungus not be
<v Speaker 1>able to break down the tree tissue. If this oxalic
<v Speaker 1>acid hadn't been there first.
<v Speaker 2>They're greatly reduced. The advocacy of everything else is reduced
<v Speaker 2>without the oxalic acid. It's one of the main components.
<v Speaker 2>They've actually taken fungus, normal healthy fungus, and they've disrupted
<v Speaker 2>the gene that makes oxalic acid and then infected trees
<v Speaker 2>with that mutant fungus, and the pathogenicity is greatly reduced.
<v Speaker 2>So everything kind of revolves around that oxalic acid.
<v Speaker 1>We'll talk would there be a way using like gene
<v Speaker 1>drive technology to create oxalic acid impotent strains of fungus
<v Speaker 1>and then throw that into the the fungus itself, like
<v Speaker 1>throw that into the population. I mean, I'm not saying
<v Speaker 1>we should do this, but what is that theoretical is like?
<v Speaker 1>Is that is that Another approach is like basically what
<v Speaker 1>they do with mosquitoes. They create these bunk you know,
<v Speaker 1>shooting blanks mosquitoes. You know, that produced an entirely male
<v Speaker 1>population of mosquitos, and then it's slowly over time, you
<v Speaker 1>can have to keep putting it out there, these dud mosquitos,
<v Speaker 1>but eventually you reduce the mosquito population by like ninety
<v Speaker 1>six percent using gene drive.
<v Speaker 2>The main issue there would just be the sexual incompatibility
<v Speaker 2>between the strains like we were talking about before. But
<v Speaker 2>there there is actually a lab right now that's working
<v Speaker 2>on genetically engineering the fungus and what they're doing is
<v Speaker 2>creating what was called the super donor, And what they're
<v Speaker 2>doing is knocking out genes within the fungus that creates
<v Speaker 2>those barriers for the different strains. So basically that strain
<v Speaker 2>is able to spread virus between all strains.
<v Speaker 1>Wow shit, Okay, well that's good, is there? What would
<v Speaker 1>be the the thing about repercussions? Like back in the
<v Speaker 1>ecosystem in Asia where this fungus is native, what would
<v Speaker 1>be the repercussions of knocking out this fungus? Say it
<v Speaker 1>could happen of just the you know, we were trying
<v Speaker 1>to knock out the fungus in the American ecosystem and
<v Speaker 1>then you end up you know, the fund exploit. It's
<v Speaker 1>so it works so well that the fungus now becomes
<v Speaker 1>extinct in Asia. What would be that you could think
<v Speaker 1>of what would be a potential repercussion of that now
<v Speaker 1>that you remove this pathogen, Like what other function is
<v Speaker 1>there a beneficial function to other organisms that this fungus
<v Speaker 1>produces in the ecosystem of Asia where it's native Eastern Asia.
<v Speaker 2>That's a really interesting question and I don't know the answer,
<v Speaker 2>but I do know that this fungus, they weren't even
<v Speaker 2>aware that this fungus was a thing until it was
<v Speaker 2>brought to America and it began infecting the American chestnut trees.
<v Speaker 2>They later then went back to China and found it
<v Speaker 2>in the wild and like, oh, yeah, it is in
<v Speaker 2>the wild here. So it's so little of a problem
<v Speaker 2>there that they weren't even aware of it until it
<v Speaker 2>made the jump onto American chess.
<v Speaker 1>See, there was no fungal diversity survey in the early
<v Speaker 1>twentieth century in Asia. That's the problem. You know, if
<v Speaker 1>you had had if you had had just regular Joe's
<v Speaker 1>regular quote citizen scientists out there studying fungi in Asia
<v Speaker 1>in the early twentieth century, this all could have been prevented,
<v Speaker 1>right right.
<v Speaker 2>And the original infected trees that they think were brought
<v Speaker 2>over in the eighteen seventies, and that was before the
<v Speaker 2>germ theory was even widely accepted, so they really had
<v Speaker 2>no idea what they were doing.
<v Speaker 1>At that point, God as a species were such babies,
<v Speaker 1>like we're so young, like we haven't no clue what's
<v Speaker 1>going on. We're so oblivious to so many parts of
<v Speaker 1>this living world that we're a part of, you know,
<v Speaker 1>it's wild. And then you've got idiots like fucking Reagan saying, well,
<v Speaker 1>we're out here to fund intellectual curiosity. Wow, you're gonna get.
<v Speaker 1>That's what you're gonna get. You're gonna get, You're gonna
<v Speaker 1>get this and more shit every I mean, it's safe
<v Speaker 1>to say basically every tree pathogen in North America except
<v Speaker 1>for pine beetles is an invasive species that didn't exist
<v Speaker 1>in anywhere in the North American continent until it was
<v Speaker 1>brought over the last one hundred two hundred years. Right right, yep,
<v Speaker 1>What are some other bad t I mean, do you
<v Speaker 1>have any other do you have any interest in other
<v Speaker 1>tree pathogens?
<v Speaker 2>Yes, So one of the things that we're starting to
<v Speaker 2>work on now is beech trees. So beach has a
<v Speaker 2>couple of pathogens. The older one that's been around for
<v Speaker 2>a while is beach park disease. But there's actually a
<v Speaker 2>new one that's come out that's even worse and it's
<v Speaker 2>called beach leaf disease, and it's caused by a limatode
<v Speaker 2>and possibly complexes with other fungi or something like that,
<v Speaker 2>but it's primarily caused by nematode. And we're doing some
<v Speaker 2>of the earliest research on this disease, and we're beginning
<v Speaker 2>to do the beginning steps of tissue culture with beach
<v Speaker 2>and we could potentially apply the techniques that we've developed
<v Speaker 2>in American chests not to beach as well to hopefully
<v Speaker 2>address issues like the speechleief disease.
<v Speaker 1>I mean, yeah, that's what's going on, is like, you know,
<v Speaker 1>people are just swapping parts of ecosystems from one to
<v Speaker 1>another that have been separated for dozens of millions of years,
<v Speaker 1>and no shit, you know, not surprisingly things go haywire.
<v Speaker 1>Not every non native is an invasive species, but every
<v Speaker 1>invasive species is a non native primarily from another continent.
<v Speaker 1>There's a couple exceptions. Someone sent me an email about
<v Speaker 1>a loop and species from the West coast of North
<v Speaker 1>America that's causing you know, that's fucking with a loop
<v Speaker 1>and species in Maine. And then there's a you know, Rabinia,
<v Speaker 1>which is a you know, invasive legume tree that can
<v Speaker 1>become a little invasive out west, you know, especially because
<v Speaker 1>it can you know, clone it sends up runners, it
<v Speaker 1>sends up you know, cloned stems, so it basically forms
<v Speaker 1>a colony. But but it's these continental ecosystems when you
<v Speaker 1>start fucking around, you know, it's like it's like it's
<v Speaker 1>like taking a piece from a nineteen eighty nine Honda
<v Speaker 1>prelude and trying to put it in a in a
<v Speaker 1>fucking buick, you know, and being amazed when the trans
<v Speaker 1>transmission blows up. So it just it blows my mind.
<v Speaker 1>I mean, it's and all of these were presumably unintentional.
<v Speaker 1>I mean, white pine blister rust, same thing that's from
<v Speaker 1>Germany where you know where that's from.
<v Speaker 2>I don't remember where that one's from. That sounds right
<v Speaker 2>that I think it was originally imported on nursery stock
<v Speaker 2>from Europe.
<v Speaker 1>Yeah, and it's got that alternate host to that it
<v Speaker 1>needs because it's life cycle. It's on you know, members
<v Speaker 1>of rose ci No ribies.
<v Speaker 2>Yes, ribies, that's correct.
<v Speaker 1>Yeah. Are there there's ribies in New York Native ribies,
<v Speaker 1>aren't there?
<v Speaker 2>Yes?
<v Speaker 1>Jesus, yeah, man.
<v Speaker 2>Yeah, there was there was a an eradication program in
<v Speaker 2>the early twentieth century when they discovered that that was
<v Speaker 2>the intermediate host of the white pine blister us and
<v Speaker 2>they had bounties on them, and so people were out
<v Speaker 2>destroying those on the ecosystem, but there is just too
<v Speaker 2>many and eventually they gave.
<v Speaker 1>Up destroying ribies. Yeah, fucking idiot, They're going to take
<v Speaker 1>out this other native plant like fucking morons. What do
<v Speaker 1>you think again? You know, the field of ecology is
<v Speaker 1>so young, the whole you know, it's just it's crazy.
<v Speaker 1>You have such repercussions. It's like removing a predator from
<v Speaker 1>an ecosystem and now the fucking deer population goes goes
<v Speaker 1>up and they end up eating, nibbling everything down and
<v Speaker 1>then getting deer of course don't know how to regulate
<v Speaker 1>their own population those species really does humans included. I'm
<v Speaker 1>not advocating anything. I'm just saying we should be encouraging,
<v Speaker 1>let you know, reduced population. But that's all I'm saying.
<v Speaker 2>But anyway, there's should be less deer geh and.
<v Speaker 1>The deer get bit Well, that's like the Kaibab plateau.
<v Speaker 1>The population gets too big and the whole population crashes
<v Speaker 1>and now they're suffering among all the deer. They're starving
<v Speaker 1>and getting ridden with disease, and it's all because of people.
<v Speaker 2>People. It seems to be a problem across the entire country.
<v Speaker 2>Everywhere I hear there's these deer issues. There's definitely issues
<v Speaker 2>here on the East coast, but I know there's issues
<v Speaker 2>on the West coast, and I know they're taking out
<v Speaker 2>the aspen forests. Like I'm sure you've heard of Pando before.
<v Speaker 2>Pando's having a bad having issues because it can't regenerate
<v Speaker 2>because the deer comes in and they eat all the
<v Speaker 2>root suckers.
<v Speaker 1>Deer are horrible. They're horrible. They I mean really they
<v Speaker 1>where I live, they're really bad. They hang out in town,
<v Speaker 1>they nibble everything down, and they all look like shit too,
<v Speaker 1>Like they've all got it looks like they've got main
<v Speaker 1>or some sort of you could tell they've probably got
<v Speaker 1>the parasites on those things. I mean, not just the tics,
<v Speaker 1>but other disease. Yeah, yeah, and then there's probably some
<v Speaker 1>sort of mite that they get to because they all
<v Speaker 1>look scraggly and there's patches of fur that it's you know,
<v Speaker 1>we've fucked up this ecosystem, this civilization we have with
<v Speaker 1>with its mentality. You know, we haven't. We were able
<v Speaker 1>to separate ourselves from nature through agriculture and technology, and
<v Speaker 1>then you know, goat ourselves into thinking, fool ourselves into
<v Speaker 1>thinking we didn't need it anymore unless it directly benefited us.
<v Speaker 1>And so here we are with these ecological ruins. If
<v Speaker 1>you don't hold the life around you to be sacred
<v Speaker 1>and don't have a reverence for it, you end up
<v Speaker 1>with this, you know, and fucking Elon Muskus wants to
<v Speaker 1>take us to Mars now. And you know, you've got
<v Speaker 1>this man, this grown ass man, doing these horrible dances,
<v Speaker 1>and you could very clearly tell he just never got
<v Speaker 1>validation as a youngster. It probably doesn't go to therapy,
<v Speaker 1>and now he's desperate for it. He just wants to
<v Speaker 1>be cool. But it's he's going to take all of
<v Speaker 1>us downe It's going to take all of us down
<v Speaker 1>with him. Oh God, the lack of self love, it
<v Speaker 1>has such far reaching repercussions depending on the individual. So
<v Speaker 1>this beach leaf disease, I mean, you guys are working
<v Speaker 1>with that is what's the target of approach you have there?
<v Speaker 2>So right now we're trying to understand the disease and
<v Speaker 2>get a good understanding of the life cycle of the
<v Speaker 2>nematode and how it's transmitted between trees and how it
<v Speaker 2>overwinters and and things like that, and eventually, if we're
<v Speaker 2>able to develop a tissue culture and a transformation protocol
<v Speaker 2>for beach, we could potentially start testing genes for resistance
<v Speaker 2>against that disease.
<v Speaker 1>But again it'd be like a century before if this,
<v Speaker 1>you know, before you're able to actually restore an ecosystem,
<v Speaker 1>but you got to start somewhere and you'll at least
<v Speaker 1>be preserved being genetic diversity. So this nematode is it
<v Speaker 1>it's just the nematod that goes nuts. It's not the
<v Speaker 1>nematod introducing a fungus like you've got with you know,
<v Speaker 1>American elms, like the beetle is the vector for Dutch
<v Speaker 1>elm disease? Is it the case like that or is it?
<v Speaker 1>Is it just the nema toad.
<v Speaker 2>Right, that's something we're still working to understand. We don't
<v Speaker 2>know yet if there's anything else involved other than the nematod,
<v Speaker 2>But the nematod definitely is the causal agent that coaxed
<v Speaker 2>postulates have been completed with that. It's been you know,
<v Speaker 2>found on trees, isolated, inoculated onto healthy trees which then
<v Speaker 2>became disease. So it certainly is the nematoad causing the problem.
<v Speaker 1>What about hemlock wooly addle jide.
<v Speaker 2>Yeah, that's another big problem. We haven't started working with that,
<v Speaker 2>but I certainly would like to work with that because
<v Speaker 2>hemlocks are a really important species in our area, but
<v Speaker 2>obviously across the entire East Coast. It would be nice
<v Speaker 2>to be able to develop some lines that are resistant
<v Speaker 2>to that.
<v Speaker 1>But it's I mean, that's a similar case. It's they're
<v Speaker 1>probably I think that I didn't even look, but they're
<v Speaker 1>from Asia. Native hemlock over there that isn't driven to
<v Speaker 1>extinction by them, and then they they're brought over here
<v Speaker 1>and now they can they're taking apart hemlocks here.
<v Speaker 2>Yes, yep, what's the name?
<v Speaker 1>What's the do you know the native host of adult
<v Speaker 1>kids in Asia? I don't Probably another Suga species. I
<v Speaker 1>could look that up. But got the list of tree
<v Speaker 1>pathogens in North America that are just wrecking forests here
<v Speaker 1>as well. Emerald ashbor that's another one. Have you guys
<v Speaker 1>worked with those at all.
<v Speaker 2>We haven't worked with those, but I am aware that
<v Speaker 2>there are people in other labs that are working on that.
<v Speaker 2>I believe the US Forest Services working on approaches for
<v Speaker 2>emerald ashboard.
<v Speaker 1>Okay, what species of Suga this hemlock wooly addl jid
<v Speaker 1>hit in Asia, Suga cenensis, do moosa see boldy eye? Yeah,
<v Speaker 1>so those are all probably resistant, but uh, Suga caroliniana
<v Speaker 1>and Suga canadensis in eastern North America before nineteen fifty one,
<v Speaker 1>the adult jid was introduced, Like man, I bet it's
<v Speaker 1>pathogenic as hell on western hemlocks as well. Damn, what's
<v Speaker 1>with Do you know anything about Dutch elm disease and
<v Speaker 1>the the Beatles?
<v Speaker 2>Yeah, actually, we're working on ELM in our lab. We're
<v Speaker 2>not working on Dutch ELM, but we're actually working on
<v Speaker 2>elm yellows. Have you heard of that disease? No?
<v Speaker 1>What's that?
<v Speaker 2>It's so that's like the second disease of ELM that
<v Speaker 2>I think came after Dutch ELM. And it's a bacterial
<v Speaker 2>infection of the vascular tissue of the floam tubes. It's
<v Speaker 2>a it's a fytal plasma similar to like citrus greening
<v Speaker 2>if you're familiar with that, and it's spread by leaf hoppers.
<v Speaker 1>Native leaf hoppers or invasive ones.
<v Speaker 2>I believe they are native.
<v Speaker 1>Yeah, okay, do you know the beetles that hit the
<v Speaker 1>the elm trees and spread Dutch helm? Are they native?
<v Speaker 2>I'm not sure. I'm not sure about that one.
<v Speaker 1>I could, I could look that up. Okay, So, well,
<v Speaker 1>tell me about this, uh, this other pathoge in this bacteria.
<v Speaker 2>Yeah, so it's this bacteria is a phytal plasma, which
<v Speaker 2>is kind of like a strange type of bacteria that
<v Speaker 2>they're really hard to study because you can't culture them.
<v Speaker 2>They they they have to be within host tissue, and
<v Speaker 2>so that makes studying it very difficult because you can't
<v Speaker 2>culture it and do your normal types of studies you
<v Speaker 2>do on microbes. And so it's causing this flowing tube disorder.
<v Speaker 2>It causes the tree to turn yellow, so we call
<v Speaker 2>it elm yellows. First you'll see the leaves of the
<v Speaker 2>county to turn yellow and then eventually the tree dies.
<v Speaker 2>And so we're looking at creating transformation protocols and testing
<v Speaker 2>genes for both the bacteria and potentially for the vector
<v Speaker 2>as well. That that leaf hopper.
<v Speaker 1>Jesus, Okay, yeah, I just looked. Yeah, the bark beetles
<v Speaker 1>that are the beetles that's yeah, it's a bark beetle.
<v Speaker 1>The bark beetles that spread Dutch some of them are native,
<v Speaker 1>So that's gonna be that's gonna be another component of
<v Speaker 1>the ecosystem that's removed if elms goes a beetle that
<v Speaker 1>that you know, needs that that parasitizes it, but doesn't
<v Speaker 1>take it out unless this fungus is present. So God,
<v Speaker 1>that's fucking it's brutal. So what is the with this?
<v Speaker 1>How far? How widespread is this this bacteria that that
<v Speaker 1>hits elm trees? And what's it called? Yellow?
<v Speaker 2>What? Yeah? Elm? Yellows elm the yellow Okay, spread it
<v Speaker 2>on the East coast and I don't think it's spread
<v Speaker 2>across the entire range yet. But yeah, it's like an
<v Speaker 2>emerging disease.
<v Speaker 1>Do you know, are there is there any resistance and
<v Speaker 1>also was there any resistance to Dutch elm disease among
<v Speaker 1>American elms?
<v Speaker 2>Yeah? I don't think there's resistance to either of them
<v Speaker 2>in the pure species.
<v Speaker 1>Jesus God, that's fucking and probably a similar case, millions
<v Speaker 1>of years of co evolution in another ecosystem and then
<v Speaker 1>you bring that bacteria or that fungus here and it
<v Speaker 1>just runs wild. Hmm, Well, so you've.
<v Speaker 2>Something else I wanted to mention is that we're working
<v Speaker 2>with the two other American chests on species, the Ozark
<v Speaker 2>chinkapin and the Allegheny chinkapin. I guess there's one other
<v Speaker 2>species that we're not working.
<v Speaker 1>With, Alabamensis. Yes, there's an Alabama. Those have some resistance,
<v Speaker 1>don't they.
<v Speaker 2>I'm not aware of that, but that I haven't looked
<v Speaker 2>into that. They they've done testing on those for blight resistance. Yeah,
<v Speaker 2>for which one, the Alabama mensis?
<v Speaker 1>For any of them, they're they're all. I just know
<v Speaker 1>Ozark Kansas is not as endangered as as Dentata. And
<v Speaker 1>there's another one I saw in Florida, what a pumula
<v Speaker 1>is not as endangered. Do you know? Do you know
<v Speaker 1>why that might be? And is that is that even true?
<v Speaker 2>Yeah? So with Pumola, the Allegheny chinkapin, it's got quite
<v Speaker 2>a bit of a different growth habit than the American chestnut.
<v Speaker 2>American chess note has just one single stem. It grows
<v Speaker 2>in like a timber type tree, whereas the Alleghany chinkapin
<v Speaker 2>is like a shrub, a multi stem shrub. So if
<v Speaker 2>it gets infected one of its branches and it loses
<v Speaker 2>it to blight, it can be easily caught and saated
<v Speaker 2>for because it's already a shrub, so it can just
<v Speaker 2>send up more shoots.
<v Speaker 1>So that flowers earlier too and produces. Yeah.
<v Speaker 2>Yeah, and it's much shorter, so it's less of an
<v Speaker 2>issue there. But the ozark chincapin is, uh, just like
<v Speaker 2>the American chess. It has like the greet the timber
<v Speaker 2>type growth with this single stem. So it was pretty
<v Speaker 2>devastated when blade came through. And it is very limited
<v Speaker 2>in its range. So we've begun working with the ozark
<v Speaker 2>chinkapin to increase resistance in that tree as well.
<v Speaker 1>Where does that Where does that occur.
<v Speaker 2>The ozark chinkapin. Yeah, yeah, it's in the Ozark Mountains
<v Speaker 2>in like Arkansas, and it's it's a very limited range.
<v Speaker 2>I think Mississippi. It goes into a little bit in Missouri.
<v Speaker 1>Diverged from data probably last few million years.
<v Speaker 2>I would assume, yeah, that it's unclear. I remember reading
<v Speaker 2>a paper about that, and there's the the genomics are
<v Speaker 2>kind of strange because they were unsure how much hybridization
<v Speaker 2>had occurred, and uh, when when that diversions occurred, whether
<v Speaker 2>because they're very different with their their nut morphology, that
<v Speaker 2>chinkpin has just like that single nut, whereas the Dentata
<v Speaker 2>has the three nuts per bur and you see that
<v Speaker 2>in Asian species too. There's the chinkapin there that have
<v Speaker 2>like a single nut, and the Chinese shots sound has
<v Speaker 2>the three nuts. So it's unclear to me when that
<v Speaker 2>diversions happened, if they came here at the same time,
<v Speaker 2>or if they came as a single species and split.
<v Speaker 2>It's kind of an interesting question. Yeah, I don't think
<v Speaker 2>the definitive answer to.
<v Speaker 1>Yet producing one nut per burr. I'd assume that you
<v Speaker 1>could just produce more more burrs then, and it's maybe
<v Speaker 1>it's cost saving on that, you know, in terms of
<v Speaker 1>energy for the tree and probably faster, I would assume.
<v Speaker 1>I don't know, I don't know. I don't know what
<v Speaker 1>the adaptive benefits, if any there would be, you know,
<v Speaker 1>in seed size or seed number to fruit. But it's
<v Speaker 1>interesting stuff to think about. How big does the the
<v Speaker 1>ozark chikapin.
<v Speaker 2>Get it's quite a bit shorter than the American chests.
<v Speaker 2>Not I'm not actually sure what the maximum height is.
<v Speaker 2>I would say maybe like sixty feet sounds about right,
<v Speaker 2>so it's still a decent sized tree, but it's not
<v Speaker 2>massive like the American chess.
<v Speaker 1>How big were some of the American chestnuts, because those
<v Speaker 1>photos you find online they're huge. How tall can they get?
<v Speaker 2>I'm not sure what the max is, but I know
<v Speaker 2>the tallest one right now is one hundred and fifteen feet,
<v Speaker 2>so at least at large.
<v Speaker 1>Probably one hundred so it's safe to assume probably one
<v Speaker 1>hundred and forty hundred and fifty feet on some of
<v Speaker 1>those old growth ones that can probably lick. Because I
<v Speaker 1>wonder those trees in those photos they're I mean they're
<v Speaker 1>six feet diameter, breast tight, they're fucking huge. They must
<v Speaker 1>have been massive. I wonder how long they can live?
<v Speaker 1>How long? How what's the most East Coast oaks? How
<v Speaker 1>long can they live? You know, five six centuries or something.
<v Speaker 2>I know that the European chestnuts. There's European chestnuts that
<v Speaker 2>are like over a thousand years old over there, so
<v Speaker 2>I would.
<v Speaker 1>Reliably dated, not just estimated, but have been stem board
<v Speaker 1>like tree ring counted or what.
<v Speaker 2>Yeah, yeah, and uh, there's I believe it's in Sardinia.
<v Speaker 2>They have these very old chess on farms and that
<v Speaker 2>you know, they're growing trees there that are over eight
<v Speaker 2>hundred years old.
<v Speaker 1>We're Sardinia. Is that in Italy?
<v Speaker 2>Yeah, it's in the Mediterranean. It's an island.
<v Speaker 1>You ever want to go there?
<v Speaker 2>Oh, I would love to go there. Yeah, that'd be great.
<v Speaker 1>I'm going to go there sometime. I want to see
<v Speaker 1>if my if my fucking you know, because I'm not
<v Speaker 1>making any any you know, claims that I wonder if
<v Speaker 1>the you know, my fucking I just want something to
<v Speaker 1>blame my psychological flows on. They're they're not flashing laws
<v Speaker 1>I exploit. But I've heard I've heard a lot of these,
<v Speaker 1>you know, behavioral traits that I exhibit might be a
<v Speaker 1>result of ancestry. So I wonder, you know, reckless driving, uh,
<v Speaker 1>you know, wild gesticulations, fucking loudness, neurosis, but you know,
<v Speaker 1>sometimes over sensitivity. I wonder if those anyway, I'd love
<v Speaker 1>to go there.
<v Speaker 2>Anyway on your Italian ancestry, right, yeah.
<v Speaker 1>Okay, well listen, sorry, we were talking about Sardinia, I
<v Speaker 1>distracted us.
<v Speaker 2>Yeah. So they've got and I believe it's Sardinia. It's
<v Speaker 2>one of those Mediterranean islands. And they've got this old
<v Speaker 2>culture of chestnut agriculture there, and they've got these massive
<v Speaker 2>old European chestnut trees there that produce you know, tons
<v Speaker 2>of nuts every year. So and they've been lying on
<v Speaker 2>those for centuries.
<v Speaker 1>How do they not get the blight? Because it's dry there?
<v Speaker 1>Because it's dry.
<v Speaker 2>Air, yeah, and they might have never had an introduction.
<v Speaker 1>Do you think I mean arid climate where you have
<v Speaker 1>arid summers probably helps a lot, right, because for the
<v Speaker 1>fungus to produce spores, it's ever to produce the ascocarps,
<v Speaker 1>it's gotta the little mushrooms, it's gotta you have to
<v Speaker 1>have a reasonable amount of humidity. Or does the humidity
<v Speaker 1>help it travel? Do you any you know anything about
<v Speaker 1>that effects of aridity on on the ability to spread
<v Speaker 1>of this fungus.
<v Speaker 2>Yeah, this is kind of why they're able to grow
<v Speaker 2>well on the West coast. We believe is because the
<v Speaker 2>the dry air is not as conducive to the spores traveling.
<v Speaker 2>The knidia spores that the asexual spores are need to
<v Speaker 2>be moist otherwise they will die, and so I think
<v Speaker 2>that prevents a lot of its spreading.
<v Speaker 1>Yeah, because it's I mean it's you rarely get warmth
<v Speaker 1>and humidity on the West coast. I'm talking about like
<v Speaker 1>significant well like not like summer humidity like you get
<v Speaker 1>on the east, and so you put those little mushrooms
<v Speaker 1>out there and they just fucking they're going to have
<v Speaker 1>a hard time surviving when the air is just pulling
<v Speaker 1>moisture out of them.
<v Speaker 2>Right, So maybe that has something to do. And you
<v Speaker 2>know what, it wasn't Serginia. It was Corsica, That's what.
<v Speaker 1>Yeah, where's Corsca? Is that in Italy?
<v Speaker 2>It's in the French Mediterranean.
<v Speaker 1>I think you said, never been over the Mediterranean. The
<v Speaker 1>Mediterranean lifestyle should be over to just eating olives and
<v Speaker 1>sucking on tubes olive oil. And they make this tomato
<v Speaker 1>paste now in a tube. You can get it at
<v Speaker 1>Trader Joe's. You got a Trader Joe's and Syracuse.
<v Speaker 2>Yeah, we do.
<v Speaker 1>Fuck, I wish we had that out here. I have
<v Speaker 1>to go to like San Antonio to get it. But
<v Speaker 1>I'm telling you, man, that tube tomato paste. You know,
<v Speaker 1>it's like a little snack it it'll change your life.
<v Speaker 1>So there's groves of this stuff in Corsica of Castania.
<v Speaker 2>Sativa, yes, very old trees, and they have a whole
<v Speaker 2>culture there. You know, they've got their their mills and stuff.
<v Speaker 2>In the fall. During harvest time, they're making lots of
<v Speaker 2>chestnut beer and chestnut bread and chestnut desserts and all
<v Speaker 2>that stuff.
<v Speaker 1>Yeah, they're delicious pigs. There was a tree in a
<v Speaker 1>there was a tree in Mount Shasta and I used
<v Speaker 1>to I'd be up there, you know for work sometimes
<v Speaker 1>when I worked for the railroad, and there was a
<v Speaker 1>tree in two trees in someone's yard of Mount Shasta,
<v Speaker 1>and they would let me come get trees out of
<v Speaker 1>their backyard and or not get trees, get seeds out
<v Speaker 1>of their backyard because they would just fall and they
<v Speaker 1>didn't want them. And so I would get like bushels
<v Speaker 1>of these fucking and it was casting a stativa. I'm
<v Speaker 1>pretty sure not Melissa, and I would just get bushels
<v Speaker 1>of these these nuts. Was they were fucking delicious. And
<v Speaker 1>you stick a fork in and put them in the oven,
<v Speaker 1>spraying with some olive oil and then just crack them open.
<v Speaker 1>And snack. God, they were still good, man.
<v Speaker 2>I love roasted chestnuts.
<v Speaker 1>Are there are there any groves of Melissima in New
<v Speaker 1>York that you know of? A on the East Coast?
<v Speaker 1>I mean, because I would I presume they can grow
<v Speaker 1>those fine out there. Huh. Yeah.
<v Speaker 2>Actually, there is a an up and coming chestnut industry
<v Speaker 2>that's starting on the East Coast, and those are mostly
<v Speaker 2>Chinese seedling trees. There's this program at the University of
<v Speaker 2>I believe it's Missouri called HARK and They've got these
<v Speaker 2>very well tuned cultivars of Chinese and hybrids that they breed.
<v Speaker 2>They breed together, and then they send those seeds out
<v Speaker 2>to people to plant orchards with. And so these newer
<v Speaker 2>orchards that are coming online are mostly Melissima seedling trees.
<v Speaker 1>And how are How are the Chinese chests taste? They're
<v Speaker 1>still pretty good? Yeah.
<v Speaker 2>I actually like the Chinese chestnuts the best of all
<v Speaker 2>the nuts I've tasted. I think Chinese. The Chinese ones
<v Speaker 2>I've had are the best.
<v Speaker 1>What doesta taste like? Are they good?
<v Speaker 2>Yeah? They're good. The main issue with them is others
<v Speaker 2>was small, so you have to do a lot of
<v Speaker 2>work to get a little bit of food out of them.
<v Speaker 1>Yeah, but again we're not far who cares about, you know,
<v Speaker 1>anthropocentric use. We're carried, but we're concerned about restoring the ecosystem.
<v Speaker 1>And also I'm sure if you know, people would appreciate
<v Speaker 1>the lumber at some point too. Chestnut lumbers is supposed
<v Speaker 1>to be fucking great.
<v Speaker 2>Yeah, it's definitely preferable for wildlife to have those little nuts,
<v Speaker 2>so the blue jays, you know, can park their crops
<v Speaker 2>full of those little nuts, and all the other smaller creters.
<v Speaker 2>I think they prefer the smaller nut size versus the
<v Speaker 2>you know, the giant things that come off of Chinese
<v Speaker 2>chestnuts and Japanese chestnuts.
<v Speaker 1>But the wood, though, I mean, if you were trying
<v Speaker 1>to do like a like a couple of acre lumber
<v Speaker 1>plantation not my favorite thing in the world, but it's
<v Speaker 1>got to happen occasionally, and you would want to plant.
<v Speaker 1>I mean, chestnut wood is it's mostly for ornamental use.
<v Speaker 1>It's not really great for building or is it good
<v Speaker 1>for building? Do anything about the actual wood.
<v Speaker 2>Well, one of the best properties of it is that
<v Speaker 2>it's really wrought resistant. What they would use it for
<v Speaker 2>is stuff for outdoor exposures, so they would use it
<v Speaker 2>for like barns or like fence posts or telephone polls,
<v Speaker 2>stuff like that, because it's really route resistant and learn't
<v Speaker 2>have to treat it with chemicals, so it.
<v Speaker 1>Would be like a like an East Coast equivalent of
<v Speaker 1>red seeds. And yeah, damn well, Eric, I want to
<v Speaker 1>thank you for coming on again and for explaining all
<v Speaker 1>of this, and I'm really happy that it's still it's
<v Speaker 1>still happening. I'm really happy to I mean, what are
<v Speaker 1>we looking at in terms of time when these can,
<v Speaker 1>you know, be introduced to the ecosystem again, or when
<v Speaker 1>they can especially be available to the public, these transgendic
<v Speaker 1>American chestnuts.
<v Speaker 2>We're hoping pretty soon. We would have thought it would
<v Speaker 2>be done already, but there's been few delays along the way.
<v Speaker 2>And the current delay right now is that there is
<v Speaker 2>a change of administration and so all the those agencies
<v Speaker 2>are going through like a shuffling of employees and stuff
<v Speaker 2>like that, which which always happens during changes of administration.
<v Speaker 2>So that will probably delay things a little bit. But
<v Speaker 2>we're towards the end of the process. We've done all
<v Speaker 2>the paperwork and all that stuff, so we're we're hoping
<v Speaker 2>within the next you know, year or two.
<v Speaker 1>But you guys are regulated. This is going to be
<v Speaker 1>regulated by what federal agencies.
<v Speaker 2>By the USDA, the e p A, and the FDA.
<v Speaker 1>Oh well, the e p A and the FDA might
<v Speaker 1>not even exist in you know, a year, So I
<v Speaker 1>guess there's that going for you, right, four years of
<v Speaker 1>fucking chaos. Man. I'm I'm just like God, I don't care.
<v Speaker 1>You know, I don't get into politics people have. I
<v Speaker 1>don't need people to fucking agree with me one hundred percent.
<v Speaker 1>I don't care people get duped.
<v Speaker 2>You know.
<v Speaker 1>The fucking American left has its own share of wacos,
<v Speaker 1>of which I've certainly exposed, especially when I live in
<v Speaker 1>the bear. I just fucking like the mean girl syndrome too,
<v Speaker 1>just fucking but but still, I mean, it's just the
<v Speaker 1>fucking chaos. It's going to ensue chaos and incompetence and whatever.
<v Speaker 2>You know, I've learned it's really hard to predict what
<v Speaker 2>they're going to do. They'll We've been given all kinds
<v Speaker 2>of timelines of when to expect things to happen, and
<v Speaker 2>it's pretty rare when those timelines are adhered to. So
<v Speaker 2>it's hard because people will ask when this is going
<v Speaker 2>to happen, and I want to give a definitive answer,
<v Speaker 2>but it's just really hard for any of us to
<v Speaker 2>know what to expect because it's it's really out of
<v Speaker 2>our hands.
<v Speaker 1>I mean, but literally, the EPA might not exist, might
<v Speaker 1>not exist. But dude, he's got RFK, he's got RFK.
<v Speaker 1>Don't worry. What is So you're thinking three or four years?
<v Speaker 1>Maybe if you think it will take longer than that,
<v Speaker 1>what estimate? Just throw it out there and no one's
<v Speaker 1>going to hold you to it. No one's gonna put
<v Speaker 1>your feet in the fire if you're wrong.
<v Speaker 2>What do you think, Well, I would say probably closer
<v Speaker 2>to one to two because we're at the end of
<v Speaker 2>the process. We actually submitted our paperwork in twenty twenty,
<v Speaker 2>so it's been a full four years already and it's
<v Speaker 2>gone through most of the processes it needs to go through.
<v Speaker 2>So we're at the end. The question is how long
<v Speaker 2>is it going to take for like these final steps.
<v Speaker 2>Hopefully it shouldn't be too much longer. I'm mostly optimistic,
<v Speaker 2>but it's like I said, it's just really hard to
<v Speaker 2>predict what comes next.
<v Speaker 1>Yeah, but I mean I guess once it's out, then
<v Speaker 1>it's out. And then you can get people like backyard propagators,
<v Speaker 1>you know, buying a bunch of these and trying to
<v Speaker 1>do their you know, basically start their own chess American
<v Speaker 1>chestnut orchards. Yeah, and potentially doing more work. I mean,
<v Speaker 1>then it's kind of then it's out there. Once the
<v Speaker 1>cat's out the bag, then it's fucking I mean, that's
<v Speaker 1>kind of what I hope happens, because there's some there's
<v Speaker 1>some amazing people just doing crazy shit in their back
<v Speaker 1>are that it's pretty cool. I mean someone you might
<v Speaker 1>get someone like who's retired as you know, stable income
<v Speaker 1>and is like, I'm just going to fucking breathe thousands
<v Speaker 1>of these things now, you know.
<v Speaker 2>Yeah, Yeah, I'm really excited about that. And there's there
<v Speaker 2>are people who have been waiting a long time for
<v Speaker 2>these trees and are very enthusiastic about doing what you
<v Speaker 2>just said. So that's kind of something I'm the most
<v Speaker 2>excited about, is getting the trees into those people's hands,
<v Speaker 2>so you know, they can start going to town with
<v Speaker 2>these trees.
<v Speaker 1>Meanwhile, what is going on with the back cross breeding?
<v Speaker 1>I guess we should mention that.
<v Speaker 2>Yeah, their organization, they've done kind of like a change
<v Speaker 2>in strategy. They've been approaching back cross breeding from trying
<v Speaker 2>to get trees that are just like minimally Chinese and
<v Speaker 2>like mostly American. They were going for a fraction. They
<v Speaker 2>were going for like fifteenth sixteenth American and then like
<v Speaker 2>one sixteenth Chinese genetics. But that was before or they
<v Speaker 2>made their discovery that all these genes are on all
<v Speaker 2>the different chromosomes of the Chinese russ on. So they've
<v Speaker 2>readjusted their strategy a bit so that they're breeding for
<v Speaker 2>higher levels of Chinese genetics so they can get more
<v Speaker 2>Morval's resistance alleles in their hybrid population.
<v Speaker 1>Because it's not just one it's not just one genetic
<v Speaker 1>low side. It's not just one location on the genome.
<v Speaker 1>It's how many dozens, hundreds?
<v Speaker 2>What? Yeah, there's losai on every chromosome with unknown numbers
<v Speaker 2>of genes at each low side, so it could be
<v Speaker 2>as much as thousands.
<v Speaker 1>Jesus christ Man. That's that's good luck with that that's
<v Speaker 1>going to take for that's like that. It really is
<v Speaker 1>like the monkey and the typewriter analogy, just hoping that
<v Speaker 1>you end up. I mean, it's I don't know, but
<v Speaker 1>they're still I mean, they're still doing it. It's better
<v Speaker 1>than nothing, but it was there was the method that
<v Speaker 1>existed before we had genetic modification.
<v Speaker 2>I I don't think that any one method is going
<v Speaker 2>to be the solution. So I think that obviously I
<v Speaker 2>believe in what we're doing, but I think that it's
<v Speaker 2>gonna take a combination of breeding and genetic engineering and
<v Speaker 2>then maybe some of that biocontrol we were talking about
<v Speaker 2>with the super donor restrains. It's gonna it's gonna take
<v Speaker 2>a combination approach to be able to get to the
<v Speaker 2>restoration goal that we're all shooting for.
<v Speaker 1>Do you know what's up with are they are they
<v Speaker 1>like mapping the genomes? Are they mapping what they're doing?
<v Speaker 2>Like?
<v Speaker 1>Is there any way for them to have oversight over
<v Speaker 1>what different phenotypes they're creating, Like where they can actually
<v Speaker 1>map the genome and see, oh okay, I see when
<v Speaker 1>we move this pollen to this overary. Uh this this happened,
<v Speaker 1>We created this phenotype that's got these genes and et cetera.
<v Speaker 2>Yeah, that's actually exactly what they're doing. They're doing genomics
<v Speaker 2>based selection, so they they look at the the genotypes
<v Speaker 2>of their best trees, and so they can see where
<v Speaker 2>these low sie are and then when they do their
<v Speaker 2>next generations of breeding, they can go and do genotyping
<v Speaker 2>on their offspring and do a little bit of selection
<v Speaker 2>based just on the genomics versus you know, stem inoculation assets.
<v Speaker 2>So they're doing like pre screening using genetics.
<v Speaker 1>Yeah, because I remember I visited one of their fields
<v Speaker 1>in College Station. No it wasn't college Station, what is it,
<v Speaker 1>pennsyl Is it college Station, Pennsylvania?
<v Speaker 2>Yeah?
<v Speaker 1>Yeah, And I remember, God, the fucking field was loaded
<v Speaker 1>with ticks. That was my first experience with East Coast tics.
<v Speaker 1>God talk about the graded ecosystems. Got to bring fireback
<v Speaker 1>to those those ecosystems. But but I remember going out
<v Speaker 1>there and seeing, you know, they were inoculated all these
<v Speaker 1>trees planted, They were inoculating them, testing them for blight resistance.
<v Speaker 1>But now it seems like they may not even I mean,
<v Speaker 1>they still need to do that, but that would be
<v Speaker 1>a last step because now you can just look at
<v Speaker 1>the genome of the chest and see what genes it's inherited,
<v Speaker 1>and I mean you can you look at it. You
<v Speaker 1>can probably look at a chestnut genome and see whether
<v Speaker 1>it's whether that genome has light resistance or whether it's susceptible.
<v Speaker 2>Right, you can make predictions for sure by looking at
<v Speaker 2>some of these low si and whether or not they've
<v Speaker 2>been inherited. So you could look at a seedling that
<v Speaker 2>has inherited you know, low si you know, one, two, three, whatever,
<v Speaker 2>and say, okay, well there's probably a fifty percent chance
<v Speaker 2>that this is going to have increased resistance because it's
<v Speaker 2>inherited this. So that's that's the type of work that
<v Speaker 2>they're developing right now.
<v Speaker 1>Okay, cool, So there is Yeah, you can make somewhat
<v Speaker 1>accurate predictions. Well, I guess that's sums I guess that
<v Speaker 1>sums it up. Man. Thanks again for coming on. And
<v Speaker 1>I guess if you want to, you know, put anything
<v Speaker 1>out there for people who are interested in your work
<v Speaker 1>to read about or look up, and now's your chance.
<v Speaker 2>Sure. Our website is ESF dot E slash chestnut. We
<v Speaker 2>do regular updates on there, including publishing some of our
<v Speaker 2>data and other news around the project. If you're on Facebook,
<v Speaker 2>we have a Facebook group called the American Chestnut Research
<v Speaker 2>and Restoration Project, and if you join that group, we
<v Speaker 2>have discussions about chestnuts, people post pictures of their treaties
<v Speaker 2>and discuss topics around restoration, and we also post our
<v Speaker 2>updates there.
<v Speaker 1>Okay, well, great man, thanks a lot, appreciate you coming
<v Speaker 1>on and until next time, thanks a lot. Joy Okay,