Calibri;Consolas;Times New Roman;  I'm Tom Zinnen.  I work here at the University of Wisconsin-Madison Biotechnology Center.  I also work for UW Extension Cooperative Extension, and on behalf of the Science Alliance, Wisconsin Public Television, Wisconsin Alumni Association, thanks for coming to Wednesday Nite at the Lab.  We do this every Wednesday night, 50 times a year.  Tonight, it's my pleasure to introduce to you Dr. Ken Cameron.  He's a professor here at the UW Madison Department of Botany, and he's also the director of the Wisconsin State Herbarium, which is one of the largest botanical collections in North America.  His research includes the use of molecular data to address questions of flowering plant evolution, classification, taxonomy, distribution, and conservation.  I was a botany undergraduate at UW Platteville, and I took a course in every one of those.  In particular, he's recognized as a leading authority on the systematics of the orchid family, especially vanilla, pogonia, and the relatives. 

 Dr. Cameron earned his PhD from the University of North Carolina at Chapel Hill in 1996.  He was a curator at the New York Botanical Garden for nearly 10 years.  In 2008, he joined the UW faculty.  Particularly interesting night to me because he also, I think in June, your book came out called Vanilla Orchids: Natural History and Cultivation.  It's not every day that we get to talk about plants, vanilla, and evolution.  Please join me in welcoming Dr. Cameron to Wednesday Nite at the Lab.  [APPLAUSE]  >> Ken Cameron: Well, thank you very much.  It's a pleasure to be here.  Considering that next week's speaker is talking about beer, lager, I was thinking maybe the two of us ought to get together and create a new Wisconsin float.  Vanilla ice cream and lager, sounds pretty good to me. 

 [LAUGHTER] It's a pleasure to speak with you about three of my favorite topics: evolution, orchids in general, and then more specifically vanilla orchids.  And, although those topics may seem sort of unrelated, in fact this week is the week of orchids in evolution.  Not only do we celebrate Darwin Day next weekend here on campus, but at the same time the Madison Orchid Growers Guild is hosting their annual orchid quest at the Alliant Energy Center.  There's some outstanding speakers and plants on exhibit.  And I hope everyone can make an effort to attend at least one or both of those events.  So, before we get started, I've heard that there are some former botany majors amongst us.  Some of you may have had course in botany in the past.  I have to tell you for a moment to change your mind for a minute, clear your mind about everything you know concerning flowering plant structure.  So this typical lily flower is often used in textbooks to talk about the parts of a flower.  And the reason I say this is because as we look at the orchid family, you're going to see that orchids generally break the rules when it comes to flowering plants. 

 And as I move forward into vanilla, you'll see that vanilla, in particular, breaks a lot of the rules.  So let's just consider the orchid family for one moment.  This is a very large family.  At least 800 different genera and more than 27,000 different natural species.  We have quite a few here in Wisconsin, in fact.  Of course, they're popular with ornamental hobbyists, and there are well over 100,000 artificial hybrids and cultivars that have been named and registered.  So this is a very large and diverse group.  Unlike a lily flower or another stereotypical flowering plant, orchids only have one male reproductive organ.  Typically one stamen and that single stamen is united or entirely fused with the female pistil to form a special structure in the orchid flower called a column, and I'll show you this in just a minute.  Moreover, one of the petals is differentiated from all the others. 

 It's often called a lip or labellum.  And many orchids, as you know, especially in the tropics, are epiphytes.  They live in the canopies.  They're not parasites on trees, but they use trees as supports high up in the canopy to capture light.  And to do that, of course, they have to adapt to that unusual environment, and they have specialized roots with a structure called velamen that allows them to live high up in the canopy in the air.  Their pollen is different from most flowering plants in that it's typically packaged into a single cohesive unit called pollenia.  Their seeds are, essentially, microscopic, or at least dust-like.  I'm talking like tens of thousands of seeds in a given orchid capsule.  Unlike other flowering plants, the embryo within the seed is only a few cells at maturity, and they lack the characteristic feature of all angiosperms which is double fertilization leading to endosperm as a food reserve.  You may remember that from general biology. 

 Orchids don't have endosperm.  And then, finally, orchids require fungi in their life cycle, which makes them very unique in the plant kingdom.  In fact, so unique that there are actually parasites on fungi, and I'll talk more about that in a minute.  But we typically think of fungi, of course, parasitizing animals or causing disease or plant pathogens, but in this case the orchid turns the tables and is actually a parasite on the fungus.  Okay, so some of these structures that I mentioned, we look at a typical orchid flower here, beautiful, showy.  You can see the labellum very characteristic here as a landing platform for the insect pollinator.  Dorsal sepals, lateral sepals, lateral petals and so forth.  So there are six of these structures in total.  And then in the center of the flower, unlike the lily that we were looking down at, is the column, that white elongated structure.  And down here, in this rectangular box, I've profiled the column. 

 I've turned it upside down a little bit so that you can see the female stigma which is receptive for the pollen, and it's actually a depression filled with sticky fluid.  And then at the apex of that column is the male anther where the pollen is contained.  You can't see that there, but assuming that an insect, like this crane fly, perhaps had entered that orchid flower, searched for a reward.  Typically they don't find one.  Orchids are deceitful in that regard.  But if the insect withdraws from the flower, it often carries this large packet of pollen, the pollenia, along with it.  You can see, in this case, a yellow glob of pollenia attached to the proboscis of that insect.  So, really quite an unusual structure.  In terms of their reproduction, I mentioned the small seeds.  You can see here a typical orchid capsule. 

 If you tap it or blow on it in the wind, this dust seems to appear.  That dust, of course, are the seeds.  And under a microscope, under a light microscope, there you can see that the seed coat is only a single cell layer thick, so it's essentially transparent.  You can see the very small undifferentiated embryo within that seed.  There is no food reserve.  So this baby plant better find a partner in order to get food quickly.  And in nature they do that by allowing themselves to become infected by fungi.  So you can see some infected embryos, undifferentiated protocorms we call them.  Little globs of cells with fungal hyphae protruding from them.  Of course, the fungus thinks it's found a food source, but the orchid quickly turns the table and starts stealing carbohydrates from the fungus. 

 After a little bit of time and enough energy has been build up, these orchid seedlings, as you can see in a sterile flask here, are off to a good start.  They begin to differentiate root and shoot and eventually will become photosynthetic and can manufacture their own carbohydrates through photosynthesis.  On the other hand, there are quite a number of orchids that never bother to become photosynthetic at all.  They don't even make chlorophyl.  These are the so-called mycoheterotrophic orchids.  We have some here in Wisconsin.  For example, the coralroot orchid, Corallorhiza, lacks leaves, lacks green pigment, it lives its entire life as a parasite on the fungi.  One of the most famous in the world is from Australia, the underground orchid, Rhizanthella, which I show here on the slide.  It lives entirely subterranean.  It flowers below ground. 

 We assume that some sort of marsupial dig these plants up and ultimately disperses the fruits and seeds.  But orchids break the rules.  They're very strange plants.  In terms of their life in the canopy, if they are epiphytes, I mentioned that they have specialized roots.  If you think about it, your roots aren't growing in soil.  It's pretty hard to get minerals and even moisture if you're living in the air, drying winds.  So many orchids have pre-programmed cell death in their roots which, ultimately, results in a layer of dead cells at maturity.  So you can see that here in this electron micrograph.  A spongy layer known as the velamen.  If you grow orchids, you probably notice this white or silvery, spongy texture on some of the orchid roots coming out of the pots. 

 That's the velamen as well.  In fact, some of these orchids, like Polyradicion, many roots, Polyradicion lindenii from south Florida, the famous ghost orchid, never produces leaves.  It actually does all of its photosynthesis through its roots.  And you can even see here in some of these pictures some green pigment at the tips of the roots.  Plants aren't supposed to have photosynthetic roots.  Talk about breaking the rules.  So orchids are pretty strange in that regard as well.  Of course, I've mentioned that they're highly diverse.  And, in fact, if we consider that there are close to a quarter million flowering plants species known, the big families are the orchids, the sunflowers, and the legumes.  Orchidaceae, Asteraceae, Fabaceae. 

 Now, I have to admit that there's a bit of a competition between the Asteraceae people and those who study the Orchidaceae.  I think that there are more orchids than there are sunflowers, and you can see that here.  There's 27,001.  [LAUGHTER] In any respect, they're pretty close in terms of diversity, the sunflowers or the orchids, followed by the legumes, and then if you were just curious, the fourth largest family are the grasses with around 10,000 species.  So this is a hyper-diverse group of plants, the largest family of plants on Earth.  And, of course, people interested in diversity and the process of evolution that brings us that diversity have been fascinated by orchids for a very long time.  And that's where Charles Darwin comes in.  Some of you may have heard me speak about Darwin's interest in orchids at Darwin Days a couple years ago, but I'd like to spend a few minutes talking about Darwin's contribution to orchid evolution and the study of orchid diversity.  So, Darwin, we know, was fascinated by orchids.  We know that, in fact, because he collected a number of orchids on his voyage on the Beagle. 

 This is an actual herbarium specimen in London that Darwin collected while he was in Patagonia.  It's Gavalia patagonica.  And that sketch of the flower is actually in Darwin's own hand.  Darwin didn't do a whole lot of illustrations on his own, but he did sketch this orchid, obviously in the field, and I think he was a pretty darn good illustrator.  You can see the actual living flower above.  And I think he did a good job of capturing the three-dimensional structure of that flower.  In fact, I would argue that Darwin, despite the fact that we know his research on human origins and Galapagos finches and tortoises and all these other different animals, I would argue that Darwin is actually a botanist.  And I say that because following the publication of The Origin of Species in 1859, the eight subsequent books that Darwin published all had to do with botanical subjects.  He published, in 1876, a book on insectivorous plants like the Venus flytrap.  He was fascinated by plants that could move or plants that could climb. 

 But most important for our discussion this evening is the fact, and I've underlined it there at number one, the next book that Darwin published immediately after The Origin of Species was a book about orchids.  The Various Contrivances by Which Orchids Are Fertilized by Insects, 1862.  Now, this is important for a number of reasons.  Number one, Darwin received a great deal of criticism, as you can imagine, after publishing The Origin of Species.  I suppose he still does today.  And part of the criticism was that he hadn't given enough examples.  He hadn't substantiated or backed up his theory very well.  So he used the orchids to help substantiate and to prop up his theory of evolution by natural selection.  I also think there's a few other things that are interesting about this book.  You'll see here that the date at the bottom, 1862. 

 We're actually celebrating this year the 150th anniversary of this publication.  And so my students and I have actually organized a symposium at the annual botany meeting this summer to commemorate Darwin's contributions.  The other thing that I find very interesting in almost all of Darwin's books are not necessarily the titles, but the subtitles.  The subtitles really tell you what the book is all about.  And you know the publisher doesn't like the subtitle so they make them small.  But in the case of the orchid book, On the Various Contrivances of Which British and Foreign Ochids are Fertilized by Insects and on the Good Effects of Intercrossing.  Now, that's interesting.  And, in fact, a number of psychologists and historians of science have pointed out the fact that Charles Darwin had a little bit of a personal interest or obsession with inbreeding and outcrossing.  Why is that? One is that the wealthy Darwin family and the even wealthier Wedgwood families had intermarried for generations, and yes, indeed, this is the same Wedgwood of Wedgwood China, the famous blue and white pattern that you can still purchase today.  So two very prominent families that intermarried for generations. 

 Now, of the 62 aunts, uncles, and cousins born in the four generations founded by Josiah and Sarah Wedgwood, in other words, Darwin's grandparents, 38, or 61%, of those remained childless.  Now, you might think that has to do with disease or lack of medical care or nutrition, but, again, these were very wealthy, prominent families in England at the time.  And more specifically, a total of 73% of the 26 children born from the first cousin marriages in this Dar-Wood family did not reproduce.  So there's some interbreeding going on here, some suppression of reproductive success.  If you remember natural selection and how we measure fitness and so forth, it's all coming into play.  In fact, Charles Darwin himself married his own first cousin, Emma Wedgwood.  We know that you're not supposed to do that, right? And several of their children died young, three of the ten, or they suffered serious health problems such that only three of their seven living children produced grandchildren.  So Darwin was clearly very interested in inbreeding, inbreeding depression, fitness as measured by reproductive success.  So how does that come back to orchids? Well, again, remember the lily flower.  I told you not to think about that. 

 Instead, think about the orchid flower that I showed you a minute ago and the fact that you've got male and female sex organs fused together.  Now, wouldn't you think that would set the flower up for self-fertilization, for selfing? You can't get any closer than to put your male and female right next to each other.  So Darwin went through the book and gave many, many examples of how it is the orchids avoid selfing.  In fact, here's a quote from the book.  Unless we bear in mind the good effects which have been proved to follow in most cases from cross-fertilization, it is an astonishing fact that the flowers from orchids should not have been regularly self-fertilized.  It apparently demonstrates that there must be something injurious in this latter process, in other words, selfing, of which I've given elsewhere direct proof.  So lots of examples of that.  It is hardly an exaggeration to say that nature tells us, in the most emphatic manner, that she abhors perpetual self-fertilization.  Was he trying to tell something to the biologists or to his own family, we'll never know.  But, nevertheless, it was very important in helping to substantiate his theory of evolution. 

 In fact, Asa Gray, the great North American botanist, stated that if the orchid book, with a few trifling omissions, had appeared before The Origin, the author would have been canonized rather than anathematized by the natural theologians.  It's an amazing book even to this day.  Incredible detail.  And Darwin, himself, wrote to his publisher in 1861 saying, "I think this little volume will do good to The Origin, as it will show that I have worked hard at the details." So this orchid book was important then, it still is today.  Some other quotes, I love pulling out Darwin's quotes, and they're all online now.  "You cannot conceive how the orchids have delighted me.  I am sillily and very idly interested in them.  The orchids have been a splendid sport.  This subject is a passion with me." And even the portrait of the young Charles Darwin holding a potted orchid there.  "Orchids have interested me more than almost anything in my life." This guy was a botanist, and he had orchid fever big time. 

 The last quote that I'll put up here now as we work our way toward talking about vanilla is from 1861 to the director of Kew Gardens, Hooker.  "My dear Hooker, I was thinking of writing to you today when your note with the orchids came.  What frightful trouble you have taken about vanilla.  You really must not take an atom more, for the orchids are more play than real work." Don't tell my boss that.  [LAUGHTER] Okay, so Darwin wanted vanilla.  He was obsessed with getting his hands on some material of vanilla, some flowering material of vanilla because vanilla orchids are very special in the orchid family.  Now, you might not realize that if you just looked up the definition of vanilla.  Lacking distinction, plain, ordinary, conventional.  Well, I'm here to convince you, I hope, that nothing could be further from the truth.  As I mentioned a few minutes ago, there are more than 27,000 species of orchids found throughout the world, but only one, vanilla, is a major agricultural or commercial importance. 

 And that's what we're here to talk about today.  Now, before I continue on, I do have some ice cream for some taste-testing after the lecture, but while I'm speaking I also want to test your nose.  Got your schnoz cleared out? So, of course, vanilla is one of the world's most popular flavors and also fragrances.  And I'll be telling you more about that in just a minute, but what I have here with me today are four different samples.  The first is artificial synthetic vanilla, vanillin.  I have here a vanilla bean from a species called Tahitian vanilla from Tahiti.  And then two samples from a second species, vanilla planifolia, one grown and processed in Madagascar and the other from Mexico.  Now, most of us grew up with Madagascar vanilla, the McCormick's little brown bottle in your mom's cupboard.  So that's, I think, what you're most going to recognize, but I'd like you to sniff all four of these and appreciate the different aromas, profiles, characteristics.  Think of it as fine wine or tobacco in that regard. 

 So feel free to pass those around.  Okay, so I just got back from Singapore where I was at the World Orchid Congress, and I stood up there in front of hundreds of orchid specialists telling them that vanilla was the world's most popular orchid.  And, of course, they were all scratching their heads because very few of them actually grow vanilla in their greenhouses.  But the reason I think that we can say this is when we consider the economics of vanilla.  Worldwide imports are estimated around 2,100 metric tons per year of cured vanilla.  And these can fetch, on average, around $250 per kilo.  Now, if I've done my math right, and I'm not the world's greatest mathematician, but that amounts to $250,000 per metric ton or a worldwide annual value of $525 billion.  This is a huge market.  You may be surprised that almost everything you touched when you got up this morning and went to the bathroom, from your shampoo to your conditioner to your perfume to your toothpaste to your mouthwash, has vanilla in it.  Throughout the day you've probably encountered vanilla in Coca-Cola, in Pepsi, in all kinds of yogurts and cereals and so forth and so on. 

 So it's very important.  Just a couple of bits of trivia.  Madagascar alone exports 1,200 metric tons of cured vanilla each year.  The tiny little island nation of French Polynesia exports close to a billion Euros worth of vanilla.  That was from 2006 data.  Mexico has over a thousand hectares planted in vanilla.  And developing countries, especially in India and parts of Asia and Africa, are rapidly catching up.  So, for example, India now has almost 3,000 hectares in plantations of vanilla.  Here in the United States alone, if we just consider the vanilla ice cream sales and ice cream products, that's a $20 million industry just on its own.  So vanilla is big business, big stuff. 

 It's found in most of our food products and our fragrances, as I mentioned.  But this plant has a long and rich history dating back thousands of years to pre-Columbian times.  We believe that vanilla was first domesticated in Mexico, and the indigenous people in the state of Veracruz known as the Totonac people are the ones given credit for having discovered vanilla's magical properties.  This history is actually immortalized in a mural by Diego Rivera, the Mexican artist.  You can see this on the walls of the palace in the Zocalo in downtown Mexico City.  This is the ancient city of El Tajin.  You can still visit the ruins of the pyramids there.  And in the center of the mural you can see that Totonac Indians, in their traditional white garbs, bowing down to the Aztec contingency that's here on the right.  And what do we see at the center between the two cultures? A staff and a vanilla vine.  Can you see that? That vanilla vine there with the flower right there in white. 

 Now, if any of you have eaten dinner at Frida's down on State Street, this mural is actually replicated inside that restaurant.  Of course, Frida's recently closed so you won't be able to see this.  But I was a little bit miffed because whoever did the painting put the staff in the center but they left out the vanilla vine.  [LAUGHTER] That's the whole story.  It's right there in the middle.  So this plant, in fact, was used to flavor the ceremonial chocolate beverage that the Aztec court and the emperor was famous for consuming great quantities of raw chocolate which was barely sweetened, pretty nasty, bitter stuff with hot peppers and cornmeal but vanilla added to it.  We have archeological remains of these vessels with vanilla residue so that we know they were trading it.  They were using it almost as a form of currency.  Very important to pre-Columbian civilizations.  Of course, today in Mexico you still find farmers growing vanilla. 

 In fact, this gentleman that I visited in the city of Papantla told me that despite the fact that so much vanilla is grown in Africa and Asia, here in Mexico vanilla is in our blood.  We know vanilla.  It's part of our history.  And, in fact, I couldn't agree more.  So it's very, very interesting story.  Now, Europeans first encountered vanilla early in the 16th century.  The Spanish explorers we know must have encountered the plant, and we have some evidence in the form of these codices, these hand-drawn, often in pictographs, done by Mayans and indigenous people.  For example, the Florentine codex of the mid-1500s, that particular plant that's illustrated looks very much like a vanilla vine with beans hanging down.  And the one from 1552, in particular, which was labeled, I can't pronounce that -- the black flower, probably not in reference to the flower color but to the beans, to the vanilla fruits being long and black.  And sure enough, that looks like a vine with roots and pods hanging down and so forth. 

 So we do think that the Spanish explorers encountered vanilla, and ultimately took it to Europe to be cultivated.  But eventually the Spanish would lose interest in vanilla.  They would become more infatuated with hot spices like cinnamon and black pepper and nutmeg and allspice.  Instead, it was the French who really became fond of vanilla and took the cuttings of the vines to their tropical possessions and territories.  Places like Mauritius and La Reunion and French Polynesia and Madagascar.  In fact, that region of the world was known as the Bourbon Islands for the court, the Bourbon court, the family.  So when you see Bourbon vanilla on the label, it has nothing to do with sour mash whiskey or anything like that.  [LAUGHTER] It's in reference to its place of origin in the Bourbon Islands, Madagascar and Mauritius and Reunion.  But these plantations are pretty easy to set up.  So, vanilla is a vine, and it requires a support structure. 

 It prefers to grow in shady habitats and rich soils and so forth And ultimately, after nine months after pollination, the plants will, in fact, produce succulent long, cigar-shaped so-called beans, which of course are the fruit of the vanilla flower.  Now, one reason that vanilla is so expensive has to do with its processing.  The plant is native to Mexico but it's grown mostly outside of Mexico.  Now, as you can imagine, the natural insect pollinator, which is from Mexico, is not found in Madagascar or Reunion or Mauritius or so forth.  So in fact, every vanilla flower has to be hand-pollinated.  The flowers tend to open around 4:00 AM, in the middle of the night, and they are receptive for pollen between, like, daybreak and around 11:00.  By noon it's too late.  So, typically, young girls who have very dexterous fingers go out into the fields and they hand-pollinate.  Each plant only opens one flower each day, and they developed a very ingenious way of doing the hand-pollination which you can see here on the slide.  So it's a very laborious process. 

 As I mentioned, nine months for the beans to reach maturity.  And then, of course, they have to be hand-picked and ultimately cured and fermented and dried and processed into the beans that we're passing around now.  So, of course, in Mexico they kept doing this even after the Conquistadors took vanilla to the old world.  And you can see here an old historical photograph of vanilla beans spread out in the sun in Mexico during the day to dry and ferment and cure.  They typically are done, that's done on blankets or mats that get rolled up each night and carried indoors to allow for the fermenting, the chemical conversion.  Then the next day, brought out again, spread out, evenly spaced.  At night, rolled up again.  And this goes on for months.  This is typically at least a three-month process.  Incredible labor involved in bringing us those beans that we can buy at Penzeys or in the supermarket. 

 Now, I've mentioned that although Mexico continued to produce and supply some of the world's vanilla, today Madagascar leads the world far and above.  Half of the world's vanilla market is from Madagascar, and this represents 10% of their gross domestic product.  So important is vanilla to the Madagascar economy that they even depict vanilla vines on some of their bank notes, as you can see here.  So very important to their economy.  Vanilla did make its way, of course, to the United States.  This seems like the stuff of legend and lore, but believe it or not, Thomas Jefferson, our president, is given credit as one of the earliest known documented pieces of evidence for the use of vanilla in the United States.  This recipe for ice cream is written in Thomas Jefferson's own hand.  This is in the Library of Congress.  And I'm not sure how well you can read that in the center, but his recipe includes two bottles of good cream, I think it says five yolks of eggs, half a pound of sugar, blah, blah, blah.  Put this on a fire, first putting in a stick of vanilla. 

 You can see that right there.  You can barely read it.  So, Thomas Jefferson had access to vanilla beans.  We actually think that he probably brought them back, actually Benjamin Franklin I guess is thought, when he was ambassador to France, may have brought back the vanilla beans from France, introduced them to Thomas Jefferson.  Dolley Madison served vanilla ice cream at the president's inauguration at the White House, and from then on it was all full steam ahead.  So we love our vanilla ice cream.  Other parts of the world are only now beginning to appreciate vanilla.  For example, in Japan and China and other parts of Asia, vanilla has never been part of their cuisine.  But in the last couple of years, and I do quite a bit of field work in China so I go there at least once a year, and I have started seeing vanilla products and ice cream popping up all over China.  Last year I went to the southern island of Hainan and I saw this display garden here which was trying to introduce people to vanilla. 

 And they had plants being cultivated, and in their gift shop you could buy green tea scented with vanilla, and even black tea scented with vanilla.  So this plant that we have loved in our western cultures for so long is now becoming quite popular in Asian markets.  You can imagine that companies like Haagen-Dazs and Breyers are just salivating at the thought of tapping into that Asian ice cream market.  So vanilla looks like it's going to remain popular and continue to be so.  To recap, natural vanilla ranks as one of the most expensive commodities traded on the world market.  The price, obviously, has fluctuated up and down for various reasons.  Remember when Pepsi and Coke introduced vanilla Coke, and that was a real popular fad for a few years? Well, that pretty much corresponds right there to that spike in the chart.  And there's been some hurricanes that wiped out the crop, and the price has been fluctuating.  But natural vanilla is still very important.  And yet there's still so much that we don't know about these far from ordinary or plain orchids. 

 And as a biologist, of course this is what interests me.  I'll give you one example.  Nearly the entire vanilla market, the crop, is a monoculture.  The plants are taken as cuttings from original vines brought over from the new world to the old world, and now there are some serious fungal pathogens and other diseases that are affecting the crop.  You would think that there had been, somewhere along the way, an attempt to breed the plant or to improve the crop or to interbreed other species and improve the size or the fragrances, but that isn't that case.  Here's an example of where I think there is some potential.  This species, vanilla grandiflora, is growing in a palm swamp in Peru.  I don't know if you can see that there's standing water in that picture on the left, and there's some palm trees and a vanilla vine growing up the palm.  To the right are the fruits from that species.  Look at the size of these vanilla beans. 

 They're like bananas.  [LAUGHTER] And my colleagues, who spent some time in this region, said that if you allow those beans to ferment, even sitting in your hut in your field camp for a few weeks, the smell of vanilla is so powerful and so overwhelming that you actually have to toss them out of the house because it's too much, too perfumy.  Imagine if we bred this with our standard species.  We could really improve the yield and some interesting profiles, but that hasn't been done yet.  In fact, we know very little about vanilla biology, which is where my own research comes into play.  So, I've been fascinated by orchids since I was a kid.  Here I am at eight years old, when most boys are asking for a catcher's mitt or a basketball, I was begging my parents for an orchid.  [LAUGHTER] So my mom, there on the right, had to drive all over Detroit trying to find an orchid.  And she found one and you can see how happy I am about that.  [LAUGHTER] That ultimately led to research in graduate school in orchids, and I've been fortunate to travel all over the world. 

 Although I've added a little bit of extra pounds since I was eight years old, down there at the bottom right, I still enjoy getting out into the field.  That picture is in central Columbia in a rain forest next to a vanilla vine that I found.  So, these plants have become a passion for me as well.  And you've already heard that I recently published a book on vanilla orchids, and I have copies to sign and sell after the lecture if you're interested.  But what I've tried to do is capture some of the basic research that's taken place over the last decade by myself and some of my colleagues.  So I'm going to walk you through some of these research examples.  I won't have time to go into any of them in detail, but you can buy the book.  [LAUGHTER] Or you can send me an email or come visit.  So the first aspect, of course, I'm a plant systematist, which means that I'm interested in diversity.  And in terms of the diversity, vanilla has a number of close relatives that are equally remarkable. 

 For example, on the island of New Caledonia in the Pacific Ocean are two monotypic and endemic genera, Eriaxis and Clematepistephium.  One of these is a climbing vine just like vanilla is.  In South America, we have a genus called Epistephium which has spectacularly beautiful large, showy flowers, but they've resisted cultivation.  So few people are familiar with these.  I have a graduate student right now who's going to monograph this genus.  And you can see the difference in size between those two species Epistephium.  Another group of close relatives are completely mycoheterotrophic.  They lack chlorophyl.  They're fungal parasites.  They are the largest non-photosynthetic plants in the plant kingdom. 

 Some of these are enormous climbing vines.  They can be 10 feet or more tall.  Completely feeding on fungi in the soil.  Erythrorhiza, I photographed that in New South Wales.  I could actually see the Sydney Opera House in the distance from that picture.  I was really cool.  Lechanorchis I found on the island of Borneo.  Pseudo vanilla I photographed in Queensland.  Galeola in Singapore and then Cyrtosia is from Japan.  Look at these big, fat, fleshy, red fruits. 

 Now, you would imagine that a bird may be attracted to those and disperses the fruits and seeds, but we have no idea.  It's never been documented in the wild.  There's still so much we don't know.  Even here in North America we have close relatives of vanilla.  For example, the genus Cleistes is found in Florida and the southeastern US but doesn't quite make it up here to Wisconsin.  Isotra is a wildflower in northeast.  Found in pine forests and deciduous forests.  Doesn't quite make it to Wisconsin either.  It's in Michigan.  But one species that does make it to Wisconsin is this one right here. 

 This is Pogonia ophioglossoides.  And this is a native orchid found in bogs, found all the way up north in Wisconsin, even into Canada.  And it's a close relative of vanilla, surprisingly.  So, some of my research has focused on phylogenetics.  In other words, trying to understand the relationships of these orchids to one another and to other plant groups.  DNA, molecular phylogenetics has become an important tool in determining these relationships, and so a lot of my research has focused on vanilla phylogenetics.  Now, being a botanist, I have the fortune of being able to tap into three genomes in the plant cell of course.  There is DNA within the nucleus, as you know, in the form of chromosomes.  And like animals, there are mitochondria with their own DNA, but in the plant kingdom, botanists also, of course, have the plastid genome, chloroplasts with their own DNA.  And that's where a number of the gene sequences have come from working with the chloroplasts. 

 And so over the last few years a much clearer picture of orchid evolution and relationships has emerged.  For example, it was always assumed that orchids are related to lilies, but in fact, lilies are their own branch of the monocot tree of life, right here.  Instead, orchids, as it turns out, belong to an order of monocot flowering plants known as Asparagales, and, as the name implies, within in that order is asparagus but also agaves, onions, daffodils, the daylilies which aren't true lilies, irises, and at the base of the Asparagales are the orchids.  So this was quite a surprise when the DNA evidence revealed this relationship of orchids to asparagus and so forth.  My own work has taken that lineage of orchids and delved further into understanding relationships of the 27,000 orchid species.  Of course, I can't do that.  That's a lot of data.  But I've sampled most of the major lineages of orchids with multiple genes, and, as it turns out, we have a very good phylogeny now for the orchid family, so this family tree.  And the one thing I want to point out is that there appear to be five major groups of orchids.  We recognize these as subfamilies of the orchid family, and one of them, lo and behold, the reason I got interested in vanilla altogether, one of these subfamilies is the vanilla orchids. 

 They are so different that they actually deserve their own subfamily status, and we call that subfamily Vanilloideae.  And these are among the most primitive of the living orchids.  So, very unusual.  That was a real wake up call to orchid taxonomists because traditionally the Vanilloideae orchids had been classified higher up in the family tree with the advance orchids.  Within the Vanilloideae itself, within the subfamily, my colleagues and I have been reconstructing the phylogeny, the relationships of these different genera that I mentioned to you.  And I'll point out two things.  The two bold branches here represent the two major groups of Vanilloid orchids.  One of them includes the North American species, like Pogonia, so tempered orchids, but seem to be evolved from South American relatives.  And then vanilla itself is related to those mycoheterotrophic parasites, which seems quite odd, and these are worldwide in distribution but ultimately with a South American origin as well.  So we feel like we understand the relationships of these enigmatic orchids pretty well. 

 And it was quite surprising, in fact, to see the closest living relative of vanilla is a plant like pseudo vanilla, one of these climbing, parasitic, leafless plants from Australia.  I don't know if you can see that very well in the slide, but certainly this is a climbing vine.  It has aerial roots at each node.  It's lacking leaves but it does have the long, cylindrical fruits that you can see here high up in the canopy similar to a vanilla plant.  So now that we know the closest living relative or the out group of vanilla, my research has delved even deeper into the genus vanilla itself, and you can see here a preliminary phylogeny of vanilla species relationships.  And I'll just point out a couple of things.  First of all, some vanillas, although they are green and photosynthetic, have also lost their leaves.  They're leafless green vines.  And that leaflessness, the loss of leaves, seems to have occurred, I've marked them in green, at least three independent times in the evolution of the genus vanilla.  Furthermore, there's a pretty clear split in the phylogeny of vanilla between new world, meaning South America and North American, and old world, Asian and African species. 

 So there's a clear biogeographic pattern, but one anomaly is that all of the Caribbean species, which are all leafless, seem to be derived from African ancestors.  Now, that might seem a little odd except that in the Caribbean islands we have a number of African orchids.  It seems as though hurricanes occasionally pick up orchid seeds from Africa, carry them across the Atlantic, and deposit them in Puerto Rico, and even south Florida every now and then an African orchid will show up.  So we know that they can move around a little bit.  I also find it interesting that when they got to the Caribbean they lost their leaves.  And there are some other orchids that have done the same thing, so there seems to be a pattern there.  The orchids are so pretty you have to show pictures.  So let's just see some of the diversity within the genus vanilla.  These are the most primitive.  We call them the membranaceous leaved vanilla because they have very thin leaves. 

 Very unusual flowers.  They're quite primitive in structure.  I'll point out one in particular.  Vanilla --, this is the only non-climbing species in the genus vanilla.  This is a non-climbing species.  It was so special when it was first discovered that the botanist actually put it in its own genus, Dictyophyllaria.  It was only known from the type specimen in Brazil and hasn't been seen for almost a hundred years, and last year it was rediscovered.  So the botanist sent me this photograph, and we actually have a small DNA sample now.  And sure enough, it's a vanilla.  It doesn't deserve its own genus rank, but it is the only non-climbing member of the group which kind of maybe gives us a little clue about how the climbing habit might have evolved. 

 In the old world, some really spectacular plants.  Big flowers, showy, bright labella.  I love this one.  I photographed this in China.  Vanilla siamensis has these little whiskers on the labellum, and its close relative from Mount Kinabalu in Borneo down here, Vanilla kinabaluensis, also has whiskers on its chin.  Some of these are also leafless, like vanilla madagascariensis.  In the Caribbean, I mentioned, they're all leafless.  There's a few species, you can see them here.  And then, finally, the only species that actually produce the fragrant fruits are restricted to the new world.  So they include vanilla planifolia, the common vanilla of commerce and agriculture, but close relatives like vanilla odorata, vanilla --, photographed in Takahachi Swamp in south Florida, and a few other species here. 

 I should have mentioned that earlier.  It seems as though there's only one group of vanilla species that actually produce the fragrant aromatic capsules.  And I have a hard time convincing some people in Papua New Guinea and parts of Asia that you can't just go out into the forest and collect vanilla pods and cure them and sell them.  They don't have fragrance.  It's only the new world species that do that.  Anyway, now that we've reconstructed the phylogeny, we have a good sense of vanilla relationships, that has led to a very stable robust classification system that I've published along with some colleagues.  I won't go into that, but, again, vanilla is its own subfamily, tribe, and subtribe.  We also can use the phylogeny to do some pretty nifty things like molecular clock dating.  Only a few years ago the first fossil orchid was found.  It was a bee, extinct genus of bee, preserved in Dominican amber with orchid pollenia on its back, on its thorax. 

 And so based on morphological analysis, the people were able to place this fossil within the phylogeny that I proved to them and then dated the age of the orchids and the age of the vanilla group.  So you can see here that the common ancestor of orchids is at least somewhere here in the late Cretaceous.  So orchids were living with dinosaurs, basically.  And the vanilla orchids, in particular, in that blue box right there trace their origins back somewhere between 65 and 80 million years ago as well.  So this is an ancient group of plants.  They probably evolved in Gondwana, and there's some interesting biogeographic hypotheses that I've published with some colleagues considering plate tectonics and patterns of continental shift and comparing that to the phylogeny of these particular groups, and there's a very strong correlation there.  But even more recently, during the Pleistocene following Pleistocene glaciation, we know that the Vanilloid orchids, like Pogonia, were confined here to the glacial refugia in eastern United States but then also restricted to eastern Asia.  This is a classic example of continental disjunction.  We see it with trilliums and Jack-in-the-pulpits and all kinds of our typical spring wildflowers.  And so the phylogeny has allowed us to explore those kinds of questions as well. 

 Vanilla origins, of course, are always interesting.  I'm not an ethnobotanist but I got interested in the origin of this species in particular, the Tahitian vanilla.  So I passed that around.  You might have noticed that when you smelled the Tahitian vanilla that it was really quite different from the others.  Its chemical profile is more floral, fruity.  It contains heliotropin, which is a floral fragrance.  And that particular variety or species is restricted to the islands of French Polynesia.  Now, this is an enigma because we know that prior to European settlement there were no vanillas in French Polynesia.  And vanilla tahitensis isn't found in the wild.  It's only found in plantations in Tahiti. 

 So where did it come from? What's it origin? It's so different from all the others in terms of its flora morphology and its chemistry and so forth and so on.  Well, this is where DNA and a little bit of forensic paternity testing, I suppose, comes into play.  I got my hands on a number of accessions of Tahitian vanilla.  So here in the middle panel in green is the Tahitian vanilla.  And we have the fortune in molecular systematics the fact that the DNA of the organelles, the mitochondria and the plastids, are inherited maternally.  So they're only passed on from the mother.  Whereas, the nuclear DNA, of course, is biparentally inherited.  You get half from your mother and half from your father.  So we sequence some chloroplast genes, which should come from the mother, and we sequence some nuclear genes, which should pick up a paternal signal as well, and lo and behold we found that all of the maternal DNA sequences the chloroplast genes of the Tahitian vanilla were identical to vanilla planifolia, the common vanilla of agriculture.  This made us think that Tahitian vanilla might be a hybrid. 

 But then who's the father? Who's your daddy? And so we sampled a lot of species, and in red here, if you can read that, the paternal signal showed a wild plant from Guatemala, vanilla odorata, a wild species.  So we think vanilla tahitensis is not a species.  It's actually a hybrid of two neotropical species, planifolia and odorata.  So that was quite interesting.  Of course, there's actually a lot of legal ramifications because if you downgrade something from a species to a hybrid, it loses some protection, and the Tahitians claim this as part of their cultural heritage and we're telling them, well, actually it's not from Tahiti, it's from Mexico.  Now, the next question, of course, is where did the hybrid come from? Did humans make it? Did it occur in nature? Did it occur in Guatemala and the Spaniards collected the plants and took them to Tahiti? That we don't know.  We don't have a time machine, but certainly we've at least answered some interesting questions.  I want to just quickly run through a few more examples.  It's not all about DNA in my lab.  I'm an a traditional botanist as well as a modern gene jock. 

 And I've done some work on seed morphology.  When you buy vanilla, Breyers vanilla ice cream, the label says with real vanilla bean specks.  Those little black spots that are in the ice cream? Well, those specks are the microscopic seeds.  And here, under the electron microscope, is the seed of vanilla.  It's a big crusty globular thing.  Clearly, it can survive passing through the digestive tract of an animal, and it's quite likely that, whether bird or mammal, is attracted to the vanilla beans, they eat them and they pass the seeds out.  And that's really unusual in the orchid family because I showed you how they have a very thin seed coat.  But look at the other Vanilloid orchids, the close relatives of vanilla.  They've also got these crusty seeds.  Some of them have these big round wings. 

 They look like elm seeds to us or elm fruits.  This one has long capillary filaments on either side of the central core.  So a lot of interesting seed morphology.  Vanilla also breaks the rules when it comes to leaf architecture.  In general botany you learned that dicots have reticulate leaf venation and monocots always have parallel venation, right? Hm.  Well, vanilla kind of fits that pattern, and this -- does as well, but look at these guys.  Look at Epistephium and Clematepistephium.  Highly reticulate leaf venation, a very interesting adaptation for life in the understory, especially if you're a climbing vine.  Ovary structure is intriguing because most orchids have a single chamber within their ovary and vanilla does that.  But some of these others, the placenta of the ovary actually intrude to the central cavity and form a trilocular, a three-chambered structure. 

 Now, why would ovary structure be interesting if you're studying vanilla? Well, that's where the good stuff is.  The good stuff, and I'm passing around the synthetic version, is a molecule called vanillin.  We probably have some chemists in the audience so they'll probably correct me if I'm wrong, but vanillin is the byproduct of a reaction involving glucovanillin, which is a non-aromatic, has no odor, but when it comes into contact with the beta-glucosidase enzyme, there's a fermentation step, a chemical conversion to vanillin.  So the processors have known this for a long time.  They kill the beans, they chop them up, and then they let them sweat and go through this fermentation process.  It's only been in the last maybe five or six years that we've actually knew where this chemical reaction was occurring in the fruit structure.  As it turns out, and this graph here on the bottom shows the location of most of the glucovanillin.  So the precursor molecule, as it turns out, is mostly found in zone 4 with a little bit in zone 3 and zone 5.  Zone 4, in the fruit wall, is right here along the interior of that chamber.  In fact, there are little papillae, little hairs, on the inside of the fruit wall, and that's where the glucovanillin resides. 

 On the other hand, the enzyme, the beta-glucosidase, is mostly found in the outer green part of the wall, zones 1 and 2.  It's like being in a chemistry lab where you keep your acid over here in this glass beaker and your base over here in this glass beaker, and you better not let the two combine.  The orchid actually does that.  It keeps its substrate over here and its enzyme over here.  It's only when we crush them and we disrupt the cells that we bring that enzyme in contact with the substrate and the glucovanillin is produced.  So here's my thought.  I like to cook.  You know when you watch cooking shows and they tell you to take a vanilla bean and slice it up the long way and scrape out all the seeds because you don't want all that and put the pod in your custard or whatever.  You're scraping away all the good stuff.  All the glucovanillin is right there in zone 4, and you're scraping away zone 4. 

 That's where all the good material is.  So, anyway, there's something to be learned in plant anatomy kind of comes back to gourmet cooking.  And then the last bit of research, I haven't published this yet, but I find it a really kind of fun example.  It almost relates to this bird call app.  All kinds of new technology is coming into vogue to help us to better make identifications in the natural world.  And with vanilla, the example is known as DNA barcoding.  It's using DNA to help identify unknown plant samples.  So, for example, in addition to vanilla planifolia and vanilla tahitensis, there's a third species that one finds for sale occasionally, vanilla pompona.  Now, if you were just given beans or a non-flowering sterile vine, how could you really tell the difference between those three? You could do some chemical assays, I suppose, but that would be quite laborious.  Instead, DNA barcoding shows some promise as a cheap, easy, fast way of identifying species of life in a manner analogous to the commercially ubiquitous barcodes that we find on all products in the supermarket. 

 So, for example, let's assume we were asking the question is this cutting really vanilla tahitensis? If I'm going to pay top dollar for this plant, I want to make sure I'm getting what it says it is.  Well, I'm a vanilla expert and without flowers I don't know if that's tahitensis.  I'll have to take the seller's word for it.  Can the buyer be certain that these vanilla beans are from vanilla tahitensis? Well, they just look like long, brown vanilla beans to me.  Unless you smell them or do some chemical assays could you be certain.  This picture I took in Ecuador, and this particular nursery was propagating thousands of vanilla cuttings.  And I asked them what species are these? We don't know, we just went out into the forest.  Isn't there only one vanilla? I said, no, there's actually almost a hundred different species, so I may need to do some testing for you to tell you what you've got there.  When you're looking at the leafless vanillas, then you're really up a creek.  Which one of these is vanilla madagascariensis? Well, that one says it is according to the label, but can we really trust the plant tag? These all look the same to me. 

 So DNA barcoding, I think, has some promise.  So I'm going to show you a case study that I've done recently.  Here's leafless cutting a friend of mine gave me.  It was being cultivated in a greenhouse in Florida.  That's all I know about it.  So it's just vanilla species, unknown species.  I gave it my own number, and I made a note that it is being cultivated in Florida.  So how am I going to put a name on this thing? Well, I could wait 20 years for it to flower, but I don't have that much time.  So I would probably go to a standard key in the flora of Florida.  And assuming that this really is even a Florida native, which is a leap of faith, but if did that, I would work my way through the key and I would find that there are two species in that region that are leafless, but in order to distinguish vanilla dilloniana from vanilla barbellata, I need flowers. 

 All the parts here that distinguish them have to do with floral structure.  So that's not going to help me much.  So here's where DNA barcoding comes in.  The zoologists have rallied around a single gene, cytochrome oxidase 1, COX1 or CO1, from the mitochondria which seems to work quite well at distinguishing animal species from each other.  There's a few problems.  The reptiles have some trouble and some seagulls and a few other groups.  But it works quite well at distinguishing animals species.  But when it comes to plants, that particular gene doesn't work very well.  Instead, I'm part of a group called the Plant Working Group of the Consortium for the Barcode of Life, and in the proceedings of the National Academy of Sciences a couple years ago, we published some alternative candidate genes that may be useful for plant DNA barcoding.  And so I did a little case study with my unknown vanilla. 

 So I looked at one of these loci, one of these genes, RPOC1.  I hope you can read this.  Our unknown is in blue, and according to the DNA, if we just kind of look at DNA sequence similarities, that gene tells us some good news and bad news.  Out of 250,000 possible flowering plants, this gene, this tiny little 500-base pair snippet has narrowed my choices down to three.  It's either vanilla barbellata, vanilla claviculata, or vanilla dilloniana.  Those are Caribbean leafless vanillas.  So I'm in the right ballpark, and those are possible candidates.  But the DNA sequence is identical from my unknown and those three species.  So that gene alone isn't going to tell me the definitive answer.  What about this gene, RPOB? Well, in this case, I've actually narrowed it down to just two possibilities. 

 It's either vanilla dilloniana or vanilla barbellata.  Remember, those are the two that grow in south Florida so I have faith that this is probably at least in the right ballpark.  This particular gene region, PSBA to turn H spacer, actually gives us a pretty definitive answer.  The best match of my blue unknown here is with the two accessions of vanilla dilloniana that I've sampled and nothing else.  So with just a tiny little snippet of DNA and an unknown plant specimen, I can put a name on this unknown plant species.  So I actually feel confident in relabeling my collection as vanilla dilloniana, and I think DNA barcoding has great potential in this particular aspect.  I'll just close with two other thoughts.  Despite the fact that we love vanilla ice cream and it's the world's most popular orchid in flavor and fragrance, these plants are under great threat of extinction.  These are just three native species in Florida, and, as you can see here, they are either listed as endangered or in some cases they've even been extirpated.  They're extinct now in some parts of the wild. 

 These plants are difficult to cultivate because they're large, massive vines.  And they kind of get overlooked when it comes to orchid conservation, in favor of the more showy, beautiful flowers.  But I think if we can convince the next generation that when they're enjoying their vanilla ice cream that this flavor comes from an orchid and orchids deserve to be protected and so forth, we can make great headway.  My book is on sale.  [LAUGHTER] I do have copies I'm willing to sign.  And for you, the low, low price of just $30.  There's lots of interesting tidbits of information in there.  And then, finally, in conclusion, I hope I've convinced you that there's nothing plain about vanilla.  Thank you very much for your time.  [APPLAUSE]  Now, before we do ice cream tasting, I'm assuming there may be some questions. 

 Do we have time?