Gel of the Day: A Horny Little Bug That's Pissing Me Off

Just for the record, I would like it known that the 3-4mm long diaperine beetles lumped together into the species Neomida bicornis are stinky stupid-heads and I don't want to play with them anymore.

What can I say after that deeply professional and scientific appraisal? Well, I can offer up this Gel of the Day as evidence of my contentions:

The first two lanes are 28S and 5.8S nuclear ribosomal subunits and the third is cox1 from Neomida bicornis. The last three are the same genes from a positive control, Bolitotherus cornutus. Everything else is, of course, the same. The same aliquots of primers, the same Taq polymerase, the same dNTPs, the same thermocycler protocol.

Now, if this were the first time I'd gotten a result like this, it would be no problem. And, in fact, it wasn't a problem four attempts ago. This gel is my fifth attempt at getting these sequences! I've tried tweaking everything I can think of with my existing protocol, but nothing changes the result. My next step will be to use a forensic DNA extraction and amplification kit, which I've been trying not to do because of the expense involved.

If not for the fact that I have some evidence that N. bicornis is more than one species, I would have given up on this by now. The thing is, it's worth it to me because part of my research has to do with the evolution of host specificity and I've collected this beetle from four different fungi now. Not only is the coloration different in these populations, but there are more subtle morphological differences as well (for example, the proportions of the tarsomeres of the metatarsi). I've also used the large subunit sequences from two populations — one from Ganoderma lucidum in Florida and one from Pycnoporus cinnibarinus in Massachusetts — in a molecular phylogenetic analysis and guess what... they turn out to fall into two separate clades with 99% bootstrap support in a neighbor-joining tree after 500 replications. In other words, they're taxonomically classified as a single species, but considering how slowly this sequence evolves it's highly unlikely that they really are. I need to get the more rapidly-evolving cox1 sequence to clinch this, along with analyzing a few more populations and I can't get the damned thing.

What a tease.

I mean, just look...

If I had hair, I'd be pulling it out right now. I can take or leave the small subunit, but I must have the cox1 sequence.

OK, that's it, "Neomida bicornis," if that is your real name. You've pushed me too far. Next time you see me on the street, you'd better run. I'm gonna be packing Raid.

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Busy with Beetles: Collections and Hypotheses

I haven't had much free time since returning from Florida, hence the lack of updates here. My schedule this semester is rather packed. For instance, on Thursdays I have two hours of open time between 10 AM and 9 PM which I will be using to eat. It will all work out, but it will also take a couple of weeks for me to get settled into this new calendar. As it stands right now, though, I haven't even had the chance to edit my photos from the trip.

I have, however, finished identifying the beetle specimens to the extent I've been able without molecular work. It's not a bad collection for under a week's time, and a couple of the species are of particular interest for reasons I'll mention below.

Megalodacne heros is without doubt the most visually spectacular of the bunch. It's one of the largest (if not the largest) of the Erotylidae and quite a colorful insect. I collected four specimens from the pore surface of Ganoderma applanatum, which is not nearly so common in Florida as it is in cooler climates. In Florida, it seems to be replaced by species in the Ganoderma lucidum complex. I didn't find M. heros on any other fungi, including the numerous G. applanatum specimens I saw in the field. That might be chance, of course.

A teneb mimic of M. heros, Platydema ellipticus, was also collected from the pore surface of Phellinus gilvus. While this beetle is substantially smaller than Megalodacne, it has what I suspect is aposematic coloration. That is to say, natural selection has favored individuals with markings similar to the larger and more toxic erotylid in this particular teneb. Five specimens of this insect came back with me.

Platydema nigratum is a close relative of the above. It doesn't have much going for it in terms of coloration, but this is an exciting find for me for two reasons. First, the habitat of this beetle is uncertain. I can say definitively that I collected it from between fruiting bodies of Stereum ostrea, and I watched one specimen chewing on that fungus. I have no doubt at all that it is a fungivore and macrophagous at that. Second, the larva of this species isn't described in the literature. I recovered a specimen of what I believe will turn out to be a larval P. nigratum from within the fleshy base of a relatively large S. ostrea sporophore. Once I've sequenced both adult and larva I should be able to connect the two definitively.

I collected numerous specimens of Neomida bicornis from two fungi. I first found them inside the same G. applanatum sporophores that had Megalodacne on their surface. I found specimens again several days later living in the context of Trametes elegans, a particularly fleshy member of the same genus that contains the ubiquitous turkey tail, T. versicolor. An interesting thing happened when I collected N. bicornis from that fungus that I wish I had noticed in the field. When I first started collecting, I found six specimens and put them in a microtube. Then I noticed additional galleries and collected fourteen more specimens in a second tube. When I brought them all back to the lab, it turned out that all six specimens in the first tube were male and all fourteen in the second were female. The probability of this happening purely by chance is so small that it suggests to me that males and females were segregated for some reason at the moment that I found the insects. I know nothing about the behavior of N. bicornis; I doubt that anyone has really investigated it. Also, it's worth noting that the specimens from Florida are very, very different from what's called N. bicornis in New England, which I've collected from Pycnoporus cinnibarinus at Wachusett. The Floridian specimens have an orange prothorax and males have straight horns. The New England version is dark metallic green and the male's horns are slightly curved. There's a chance, I think, that the two are really two different species and, indeed, I don't find any sequences for Neomida in EntrezGene other than one for a basidiomycete yeast from its gut discovered by Suh and Blackwell. I'm looking forward to pulling sequences from the northern and southern "morphs." If they do turn out to be two different species, I think I get to name one of them. The two insects look so different that I find it hard to believe they could be the same species.

I also found a second Neomida, N. ferruginea, in the context of a second G. applanatum sporophore. I only recovered one specimen of this rather drab little teneb, though. When I eventually do the phylogeny from the genus, I suspect that this beetle will be more closely related to the southern "morph" of N. bicornis than it is to the one from New England. That's just a hunch, of course.

Of course, no teneb hunting expedition would be complete without the recovery of a couple of specimens of Bolitotherus cornutus. I collected specimens of the widely-distributed beetle from both G. applanatum (the same cluster of fruiting bodies above) and G. lucidum on this trip. B. cornutus seems to be the most successful of the larger fungus-dependent tenebs of North America based on my experience. I can hardly help but find the insects whenever I search polypores. The genus, and in fact the whole tribe, is quite small as compared to other teneb tribes, but Bolitotherus seems to be the champ in sheer numbers. Having some specimens from Florida will give me the chance to eventually compare sequences from specimens from three states. Perhaps I'll someday investigate population genetic structure in this insect, which doesn't get around much but seems to be everywhere.

I also have a couple of specimens that I haven't been able to identify beyond family yet, including a few of what are probably Nitidulidae but are potentially Leiodidae, too. I'm hoping for the former, though. Nitidulidae seems to be a family with taxa in transition between feeding on the sap from tree wounds and living on the polypores that grow from those wounds. Most beetle families have hind feet with five segments (tarsomeres). Tenebrionidae, which I've hypothesized to be primitively macrofungivorous, have only four. Nitidulidae still have five, but many species in the family have a markedly decreased fourth tarsomere, so one can imagine a scenario in which the transition from living on the surface of a fungus and eating the hymenium (Erotylidae) leaves the beetles with five tarsomeres, there's a transitional state in which the fourth (or other) tarsomere is reduced as the beetles begin to tunnel into the fungi (Nitidulidae) and, ultimately, the tarsomere is lost entirely over time as a fully fungus-exploiting lifestyle is adopted (Tenebrionidae).

One of the fungi I collected also turned out to be an unusual specimen. A glossy, brown, trametes-like, sessile thing with subdaedaloid pores turned out to be the tropical Nigroporus vinosus. Nobody in my lab has seen it before. One of the ongoing projects in the lab is the exploration of wood-decaying fungi (N. vinosus is a white-rotter) for use in bioremediation. It may be worthwhile to mount another trip to look for unusual species to be cultured in this regard, too. We'll see about that. I'm planning on going back again at some point, anyhow, and perhaps swinging around the Gulf Coast a bit in the future to sample both polypores and beetles from more climes. I've barely scratched the surface here, after all.

Clearly, I've got a ton of work to do, and I haven't even really started yet... hence my absence from the blogosphere of late. Rest assured, though, that situation is only temporary. I have no intention of disappearing just yet!

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A Brief Summary of the Florida Expedition

Today is the last day that I will be here in Florida. The field work has been productive for both myself and my colleague. Each day has begun with our getting up no later than 6 AM, eating breakfast and getting out to the field before 8 AM in hopes of escaping the heat for a little while. That last part hasn't really worked, of course, because this is North Florida in early September we're talking about. A bank sign that gave the temperature in Tallahassee read 100° by the time we came in from the field yesterday.

I've collected numerous specimens of Tenebrionidae, Tetratomidae and Erotylidae. Already winging their way back to Massachusetts are such critters as Platydema ellipticus, Megalodacne heros, Neomida bicornis (the Florida morph, which has an orange prothorax — and which I suspect isn't actually the same species as the Northeastern N. bicornis), and Penthe obliquata. All in all, I've collected at least seven species for which I previously had no specimens and possibly more. I'll be able to better sort this out once I've gotten back to the lab in Worcester. I suspect the final total will be closer to ten. The specimens should be getting back there on Wednesday, and I'll be getting back on Thursday.

As for today, we're going to take care of some housekeeping. My field companion needs to get some water from within a pitcher plant back to her lab in order to culture the yeasts therein. Then we're going to do laundry; sweaty clothes accumulate quickly in this part of the world. After that, we'll play tourist for a bit and see the Florida Caverns in Marianna.

Don't get the impression that this trip has been all work. We've had time in the evenings to hang around with our gracious hosts, toss back a few beers and consume more than our fair share of oysters. It's been quite nice to see a few friends here in Tallahassee for the first time in over a year and also quite remarkable how well my mycological compatriot has fit in with them. She's generally an agreeable sort, of course, as long as one doesn't get between her and her oysters. The woman is a whirlwind of cocktail sauce and spent shells when the things are set before her; one could easily lose an intrusive hand!

I've been studiously avoiding mention of all things non-biological this week and no doubt have a ton of catching up to do on the latest in religious lunacies and electoral nincompooperies, and I could try to write a more detailed entry about this trip. I may still do that over the next few days as time permits. There will soon be photos as well. For now, though, I have to admit that as much as I've enjoyed the company of fellow science geeks and friends here in Florida, I'm feeling just a little bit lazy today. I'll neither be catching up with news nor editing photos today.

I'm also ready to get back home to LL. She's one of the very few things in this world I find even more interesting than a beetle-infested polyporoid fungus, truth be told. Yes, that's actually a compliment, and she's probably the only woman who knows me well enough to understand just how big of one it is.

For now, I'm going to step outside and get my first few breaths of the muggy morning air.

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Bad Blogger: Finally, an Update from Florida

No, I have not been eaten by an alligator. I'm still in Florida; it has been a busy few days that have started quite early. We're up before dawn each day in an attempt to get our field work done before the sticky heat of the day begins. It hasn't worked every day, but it has for most of them. Generally speaking, we're on-site around 8:00 AM and done with collecting by about 11:00. Then it's a quick shower, lunch and off to the lab.

I've been more focused on collecting Tenebrionidae than fungi on this trip, and until yesterday hadn't been doing too well at that, either. I tried doing things a little differently during yesterday's outing to Torreya State Park, however, and it paid off. I recovered specimens from at least five species that I hadn't found before. I also had an encounter with a wild boar that could have turned out much worse than it did. So far, the greatest hazards of this trip have been the heat, mosquitoes, and Southern cooking — not necessarily in that order.

My traveling companion has been enjoying her first glimpses of the region's tremendous biodiversity, particularly the arachnids (and who wouldn't be impressed by Nephila clavipes and the numerous Micranthea species?) and carnivorous plants, although the alligator seen from a distance at Overstreet a couple of days ago was also appreciated by her. She's also been impressed by the local cuisine; I have never seen anyone eat so much bacon in my life. The loomingly phallic Tallahassee capital complex will no doubt haunt the dark corners of her mind forever, too.

Today is our last scheduled field day. We've left tomorrow as an open day so that we have the option to either do more collecting if necessary or to do housekeeping tasks such as shipping specimens home and laundry and whatever else pops up. Our flight back leaves on Wednesday. As much as I've enjoyed the trip, I'm about ready to get back to Worcester and to LL as well.

In the meanwhile, I'm not spending much time on the Internet. I'll get back to it as time permits... but time's not going to be too permissive for a couple more days. Nonetheless, all of my limbs are still intact.

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Moreland Woods Survey

The date has at last been set for local biologists and naturalists to begin the surveying of Moreland Woods in an effort to catalog species on behalf of the Greater Worcester Land Trust. Everyone involved in the effort can't make it there the same day, so &e and I will be taking a look at fungi and Coleoptera on Tuesday in the company of perhaps one or two other investigators. Others will be at the site on Wednesday and Thursday while I'm in Florida.

The site is in the northern part of the city (map), bounded by Moreland Street to the north and west and by Old Brook Drive to the east. I have only a vague notion of what might be in there and whether any of it will justify use of a grant to protect the place, but we'll find out soon.

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Wachusett Wednesday Cancelled: Fall Semester, Florida Expedition and Shifting Priorities

Due to a backlog of lab work in the face of the upcoming semester and the fact that I'll be doing field work in Florida all next week, there will be no Wachusett Wednesday this week. As perfect as the weather is and as much as I will miss this part of my weekly routine, I must put my priorities elsewhere over the next few days.

It's hard to believe that summer has come and gone so quickly, but the fact is that this is the last week before the fall semester begins. I will once again be teaching two lab sections for Introductory Biology this semester. As much as I enjoy teaching (although honesty compels me to admit that I'm not too fond of grading), this essentially means that there will be some combination of a reduction of my research time and late nights at the lab. The classroom portion of my teaching assistantship alone entails six hours each week. Add in grading, office hours, lab setup and miscellaneous issues that arise and a day disappears from my schedule. Additionally, I will be auditing an entomology course at another local university this semester; that adds up to about another six hours each week. It's going to be another very busy semester. With this in mind, I'm trying to spend the rest of this week tying up a bunch of loose ends.

A week from today, of course, I leave for the North Florida field expedition. The loss of my Wednesday on the mountain this week helps to relieve the pressure that a full week out of town will create. On the other hand, all of the planning for that trip has continued to come together very well. The biology faculty at Florida State has been exceedingly accommodating to this alumnus. My colleague and I have been furnished with laboratory space in the new biology building and the head of the university's herbarium has allowed us use of the dehydration equipment for the preservation of specimens. I still need to set up temporary accounts at FSU so that we'll have computer access from the lab.

Florida's DEP and parks management have also been incredibly helpful. In fact, I feel compelled to write a thank you to state biologist Harold Mitchell here. He has been a tremendous help in making the upcoming trip possible and interacting with him has been a pleasure. Moreover, the park managers at both Torreya and Lake Talquin State Parks have likewise been kind enough to provide my colleague and I with free and open access to their land anytime we wish, even going so far as to give us the combinations to open the gates to our sites before regular hours so that we can start as early as we like in order to beat the heat as much as possible in the Big Bend/Panhandle in early September.

Wachusett Wednesday has become one of this blog's most popular recurring features, so to readers who will miss it this week, all I can say is that it's a trade off. Wait until you see what's popping up in Florida this time of year. With the recent and unfortunate stormy weather, I expect that you'll be seeing a bumper crop of amazing and beautiful fungi in this space in the coming week.

On the other hand, Gel of the Day fans (because some people just can't get enough electrophoresis) will get a second, and quite possibly third, installment this week as I attempt to sequence a Ganoderma in the G. lucidum complex whose morphology alone hasn't been enough to provide a positive identification. I also intend to do the same with a sap beetle (Nitidulidae) collected earlier from Laetiporus sulphureus; while not in my core family, I've noticed something interesting about its foot that makes me think that having molecular data from these insects will be informative about one of the evolutionary hypotheses in my study. Markedly reduced fourth tarsomere, anyone? I have so far been unable to adequately resolve the genus of this beetle, though, and genetic sequencing should clarify the picture. Agarose lovers, rejoice!

In the words of Willie Wonka, "So much time, so little to do! Strike that. Reverse it."

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Gel of the Day: What the Heck Happened Here?

Sometimes, a perfectly good gel gets funkified in the process of handling. I think that's what happened with this gel, which was otherwise indicative of a near-perfect PCR:
The specimens in this case were three adult and one larval Bolitotherus cornutus which I collected at the MSA foray in Pennsylvania on August 10. The last eight lanes, 25-32, are from the larva. I haven't successfully sequenced a larva before; maybe my dissection technique is improving. The cox1 bands (31) look a bit weak, but the product should do for sequencing.

If you're familiar with electrophoresis gels or have at least been peeking at previous gels of the day, you're probably asking about now, "Mike, what the heck happened to lanes 21, 22 and 27?"

Well, I'll tell you. In order to take this photo using a UV light, I have to remove the gel from the casting tray, put it in a weigh boat and take it to a dark room upstairs from my lab. In the process of transferring the gel from tray to boat to camera, it gets a little scrape over an edge or two. This time, for whatever reason, that caused the bands in three lanes to get smeared. Nonetheless, there are bands there and since they are duplicates from two different dilutions (as far as specimen and gene go, 21=17, 22=18 and 27=31), it's not a problem.

Also, every fourth well is a control; there shouldn't be anything there. There seems to be a very faint band in lane 4, but it's nothing to worry about. I may just try and sequence it to see what the contamination is, since the position of that band is unlike that of any of the legitimate bands on this gel. I suspect that the culprit is a dirty microtube; if my primers, buffers, etc. had been contaminated, I would expect to see that phantom band in all of the control lanes.

The reason I'm running another batch of B. cornutus is that these are the first specimens I've collected from a site outside of Massachusetts. It's worth looking at them just to see if there are any polymorphisms present in introns in the sequences I'm analyzing.

I also read a very interesting paper today:

Leschen RAB and TR Buckley. 2007. Multistate characters and diet shifts: evolution of Erotylidae (Coleoptera). Systematic Biology 56(1):97-112. DOI: 10.1080/10635150701211844.

It's very much relevant to my own research. In this paper, Leschen and Buckley analyze the evolution of Erotylidae, a family of beetles only distantly related to the family I'm studying, and link their hyperdiversity to a shift to a diet of macrofungi. It sounds rather familiar, no? It's a very interesting read if you're into this sort of thing. It's based on morphology, not molecular biology, and finds a good link between diet and the external reproductive structure (gonocoxa and ovipositor) of Coleopteran females. Mycologists (at least those with an interest in basidiomycetes) and coleopterists alike will undoubtedly find something of interest in the paper. Enjoy.

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Neomida bicornis: in ur pycnoporus tunnelin ur context

While dissecting some galleried Pycnoporus cinnibarinus this morning, I found another teneb that I hadn't encountered before. It's a horned member of the tribe Diaperini called Neomida bicornis (more info here). It looks exactly like the photo shown here except, of course, much much smaller. The male I found was 3.25mm long, 1.75mm wide and 1.15mm deep. It's the first beetle in Tenebrionidae I've collected from a Pycnoporus. Previously, I've collected a few Anthribinae from that fungus. The sporocarp from which I collected this beetle came from a spot in Wachusett that I've been checking since April.

While examining the insect, taking measurements and such, I dropped it on the black-speckled linoleum floor. I then spent about 20 minutes crawling around looking for the thing. You have no idea how quickly a tiny, dark-colored beetle can disappear, and it's the only one I've recovered so far. Since it's in Diaperinae, is apparently a context tunneler and doesn't damage the hymenium, it may prove to be an important data point for my overall work. Next week, I'll sequence it and see if it has a mutation in common with it's relative, Diaperis maculata.

Now that I know where to find them, I'll keep going back to that spot for more specimens. As small as they are, it will probably take a few tries to get clean DNA sequences and build solid consensus contigs. We shall see.

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Gel of the Day: Half a Forked Fungus Beetle is Better Than None

Not a particularly successful PCR run today. For some reason, the ribosomal small subunit DNA didn't get amplified. I suspect that either I may have used an incorrect primer by mistake or else one of the pair may have gone bad. It happens. Luckily, I still have plenty of stock DNA and can have another crack at it. At least I got the large subunit and one of the cox1 genes from all of the specimens (the last eight wells are just a negative control; it's a good thing there's nothing there). I got nucLSU DNA, so it must be an error on my part that explains why the nucSSU didn't show up.

Oddly, the last two wells were taken from a 1:100 dilution and smeared; the previous two bands from a 1:50 dilution prepared from the same specimen didn't. Smearing usually indicates an excess of product. I'm not sure why that would have happened in this case. It's a duplicate, anyhow, so that's not important, just a bit strange.

The first sixteen wells in this gel are local specimens, collected at Wachusett Mountain. The third set of eight are from one of the specimens I picked up during the MSA foray in Pennsylvania last week. I have three more of those to sequence, too, once I get everything straight with these three.

As for tomorrow's field work, any B. cornutus I find will be shipped off live to Louisiana State for investigation of fungal symbionts in their gut. I don't really need to sequence anymore of them for the time being. Hopefully, I'll get lucky and find species other than this one and Diaperis maculata tomorrow. Those two seem to be dominant around here, but I'm going to try looking in a couple of different niches and see what turns up.

The North Florida field trip is two weeks from tomorrow. I know there will be more diversity down there, but I'd still like to come up with a few more local tenebs before winter sets in.

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Blog Logged by Worcester Magazine

An entry I wrote recently about my inclination to shake my fist and scream at the Asian longhorned beetle has been blog-logged by Worcester Magazine. It's becoming something of a hot topic around town, I guess. Fellow Worcester blogger Jeff of Wormtown Taxi has written several bits on it, including this newest one.

I've also just learned this morning of the Massachusetts Introduced Pests Outreach Blog. There's useful information there about how to recognize the damage done by the beetles even if you don't see the beetle itself. I suggest that any locals take a look at MIPO Blog and learn how they can help their state keep track of this infestation. It's always possible that the extra bit of information you provide could be the one that tips the scales against the big bad bug.

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Asian Longhorned Beetles Employ a Fungal Mutualist in Digesting Wood

I wrote recently about the appearance of the Asian longhorned beetle in the Worcester area. The insect is a highly problematic invasive species that's likely to do a great deal of damage to hardwood tree populations in the area before it's brought to heel. In fact, the unfortunate truth is that it is likely to be many years before it's stopped, and if it's like most invasives it's probably here to stay, maintaining at least small populations that will allow for outbreaks from time to time when conditions are favorable for elevated rates of reproduction.

It behooves us, therefore, to understand the biology of Anoplophora glabripennis as thoroughly as possible. One of the fundamental questions on the road to understanding the ecology of any organism is, what does it eat and how does it eat it? When it comes to beetles, a fungus or two (or twenty) is frequently involved. Fungi are everywhere in the biosphere, but there are few interkingdom mutualisms as old and as intimate as those between Coleoptera and fungi. A forthcoming paper in the Proceedings of the National Academy of Science has found evidence that a different sort of symbiosis is at work in the Asian longhorned beetle, giving it the ability to digest living wood — a very unusual ability among beetles and one that is key to Anoplophora's destructive potential.

Novel fungus helps beetles to digest hard wood

A little known fungus tucked away in the gut of Asian longhorned beetles helps the insect munch through the hardest of woods according to a team of entomologists and biochemists. Researchers say the discovery could lead to innovative methods of controlling the invasive pest, and potentially offer more efficient ways of breaking down plant biomass for generating biofuels.

Microbes in the gut of insects are known to break down cellulose, but little is known about how, or whether, insects degrade lignin. This natural polymer helps plants stay upright and protects them from most forms of microbial attack.

"Lignin is nature's plastic and any organism that wants to get to the sugars in a plant has to be able to get past this protective barrier," said Ming Tien, study co-author and Penn State professor of biochemistry and molecular biology. "We suspect that the fungus produces enzymes that help the beetles degrade lignin."

Before this report, it was thought that insects are unable to extensively break down lignin, and that they get around the problem either by feasting on wood that has already degraded, or by living close to fungi that can degrade the wood for them...

The researchers compared the chemical structure of non-degraded wood before and after it had passed through the gut of two wood-eating insects. To measure the degree of change in the lignin, they first fed pin oak wood to Asian longhorned beetles. Next they fed ponderosa pine wood to the Pacific dampwood termite, an insect that typically eats only dead wood.

Chemical analyses of feces from the two bugs indicated that they are able to alter the chemical structure of lignin by selectively adding or removing certain groups of molecules from the polymer...

"This fungus has genes that then make enzymes," explained Hoover, whose team's findings appear today (Aug. 18) in the Proceedings of the National Academy of Sciences. "We have been able to detect messages from the [fungal] DNA, which get translated into enzymes." [NOTE: I have checked the PNAS Early Edition and the paper is not available yet; this is typical for PNAS. It's a vexing problem that I really wish they would correct. It takes far too long for material to appear after authors are led to believe their publication will be made available on a particular date.]

While the researchers have identified the fungus residing in the gut of the Asian longhorned beetle, they have yet to find one in the gut of the termite.

"The types of chemical changes we see in the beetle are similar to those seen in the white-rot fungus," said Geib. "Changes that we see in the termite are similar to those in the brown-rot fungus. The chemical changes to the lignin are similar..."

"This type of fungus [in the Asian longhorned beetle] is known to cause disease in plants," said Hoover, whose work is funded by the Alphawood Foundation and the Penn State College of Agricultural Sciences. "But this particular strain appears to be unique. It looks like these insects somehow acquired the fungus to live in their gut and help them break down wood..."

Because of the way that this release has been written, a lot of information has to be inferred. I look forward to the day when the actual paper is made available, whenever that may come, so that we can learn more about what's been discovered — including the identity of the fungus, which is only referred to as a "novel fungus" in the release itself.

Still, it sounds from the release that the fungus is likely to be a basidiomycete. The reference to it as being like a white-rotter means that it likely produces two different varieties of peroxidases, perhaps both lignin peroxidase and manganese peroxidase if it's anything like saprophytic polypores like Ganoderma and Polyporus. As the release mentions that the fungus is capable of living freely, outside the guts of Asian longhorned beetles, as a pathogen, there's a strong likelihood that it's not a yeast, such as those found in the digestive tracts of Bolitotherus cornutus (see this paper by Suh et al. for a background on yeasts in beetle bellies). It would be very unusual for a yeast to produce lignin-degrading enzymes, particularly in large enough quantities to be useful to a wood-eating insect. Termites get around the problem of lignin's refractory nature with a battery of dinoflagellates and bacteria. The use of a mutualist fungus is an exciting new find in symbiology.

From the release, it also sounds like the researchers suspect that the longhorned beetles may have multiple allies working on their behalf. Gut flora is generally a complex thing, so that's likely, and the release itself doesn't make any mention of the relative importance of the novel fungus in the overall picture. Nonetheless, the discovery of the fungus could provide one more target in the attempt to control a particularly dangerous invasive species. In tinkering with ecology, whether on the scale of a digestive system or a forest, removing just one organism frequently has a ripple effect on total health. Whether this might turn out to be the case here — handcuff the fungus, kill the beetle — remains to be seen.

I anticipate finding out the identity of the new fungus and where it fits into the taxonomy of lignin degraders. In the meantime, I plan on being up on Wachusett on Wednesday to look for polypores, tenebs... and Asian longhorned beetles, unfortunately.

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Invasive Insects: This Time, It's Personal

The Asian longhorned beetle, Anoplophora glabripennis, has arrived in Massachusetts. In fact, it seems to have popped up in the Worcester area and begun to spread. The infestation zone now extends as far as West Boylston according to a story in the Worcester Telegram & Gazette. The map you see accompanying this update shows the quarantine area.

Because I am a rather sappy animal lover who can find admirable qualities in nearly every living thing, I admit to having a certain soft spot for certain invasive animal species. The Cuban tree frog, for instance, I found rather friendly and endearing when I used to encounter them in Florida. I could never bring myself to kill one. I once allowed one to ride around on my head, in fact.

If there are any Asian longhorned beetles reading this entry, however, be advised that I feel no such sentiments toward you. In particular, you are hereby warned to stay far away from Wachusett Mountain. If I find you in my study site on the south slope, I will show no mercy. You will be crushed and/or turned over to the authorities and/or fed to a large, spiny lizard.

Asian longhorned beetles kill hardwood trees. That, in turn, destroys polypore habitat which consequently destroys the favored living conditions for more benign Coleoptera like Diaperis maculata and Bolitotherus cornutus and that, my fine speckled arthropods, screws with my research.

You do not want to screw with my research. Anything that interferes with it is to be eliminated.

Were I Captain Kirk, I'd be shaking my fist and shouting "Khaaaaaaaan!" about now. Then again, shouting "Anoplophora!" just doesn't have the same ring to it.

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A Last Excellent Day at MSA

Today was my last day at the MSA conference. There's still one more day left, but for a variety of reasons I have to get up early tomorrow and make the long drive back to Worcester. Not the least of these is the fact that a petty, bureaucratic,politically-motivated dean at Florida State University has taken it upon himself to put up a massive and ridiculous roadblock in the way of LL's quest for a PhD. I won't get into the details; she'll write about it when and if she's ready. All I'll say is that I've never heard of such a moronic, pig-headed thing being done to someone who has been one of the few bright lights for a department that is now being dissolved largely because of the incompetence of another pig-headed bureaucrat in academic clothing. With people like these deans at the helm, I would have to advise anyone considering a PhD program at Florida State to put it as low on their list of possibilities as they can possibly can. If it were a boat, it would have the word "Titanic" painted at the prow.

Other than learning of LL being screwed over yet again, though, today was the kind of day that makes me regret having to leave a day early. Two lablings gave excellent presentations on their research as did my advisor. I take a good deal of pride in being part of this group even if I haven't begun contributing as much as I hope to... soon.

I had lunch today with Joey Spatafora and my advisor and so had the chance to listen in and/or talk about everything from plans for AFTOL2 to Wall-E and The Fantastic Planet. I learned that Joey reads this blog when he recognized me upon our introduction as Mike O'Risal. So did one of the postdocs from his lab. In fact, no fewer than four people I met today already knew me by my nom d'blog. I had best be careful not to mention my rampant crack addiction and penchant for devouring live kittens here. Someone might be looking.

Today was also the day of the Karling Lecture, delivered by Nick Talbot of the University of Exeter. The topic was "Investigating the biology of plant infection by the rice blast fungus Magnaporthe grisea using functional genomics." The rice blast fungus is a problem right up there with the potential of Puccinia graminis Ug99, wiping out up to 30% of the world's rice crops in any given year. Dr. Talbot's work on the mechanisms by which the fungus initiates and spreads infection seems to hold tremendous promise in addressing the problem. Some of the proteomics involved are well beyond the scope of my understanding; I'm not a proteins person by any stretch. Still, even I can grasp that if one can come to an understanding of how the subcellular mechanics of infection work, there's real hope of coming up with a solution.

After the Karling lecture, I met up with Meredith Blackwell and handed her three dead beetles. That might be perceived as an insult in some circles, but not in this one. I'll be sending some live specimens her way in the near future, too, and hopefully we'll be able to exchange some data and materials. I'm particularly looking forward to continued contact with the people I've met from her lab. Even though I don't think I'll be part of the next round of AFTOL research, others in my lab will be and her lab will also be involved, and we're all working on bugs and fungus in one way or another.

I had dinner at a little Indian place called Kaarma just around the corner from my hotel. I was joined by labling RiGa, Dan Durall from the University of British Columbia Okanagan, and Kentaro Hosaka from the National Museum of Nature and Science of Japan. It was a very nice cap to what has been an inspiring few days for me. If you should find yourself in State College, PA, I recommend checking out Kaarma.

I have indeed come away from this conference feeling inspired. I've got some new ideas and am itching to try them out, particularly when I get back from Florida laden with new specimens. Stephanie Gross and I talked today about there being a coleoptera/fungi symbiosis session at next year's MSA meeting. There are a bunch of us out there, and all we need to do is be in the same place at the same time with material to present. The place next year will be Snowbird in Utah and the time will be July 25-30, 2009. Now all I need is the material.

I'll get back to work on that on Thursday.

For now, my priorities are sleep and then the long drive back to Worcester. Sequences await.

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Pigs' Ears from Pennsylvania: MSA Foray Finds

These are a few of the fungi I collected during yesterday's Mycological Society of America foray in Central Pennsylvania. It's only about half. Other people made some even more interesting finds. One lucky collector came up with a beautiful Sparassis crispa, but to me the most unusual specimen I saw in the lab was Amanita jacksonii, which I had thought to be an exclusively southern species. In fact, David Geiser mentioned that he's never seen one in Pennsylvania in the years he's been collecting here. I'd only seen the species in Florida previously, where it's fairly common (I've collected them on the Florida State University campus in the quad in front of the College of Business).

I came up with some Gomphus clavatus, which until yesterday I had seen only on the west coast. Perhaps certain basidiomycetes need a GPS even more than I do. They seem to be getting lost!

		First up is a particularly beautiful Amanita close to A. brunnescens. I'm not sure of the exact species, but people far more experienced than I advised that it's likely the best identification we'll get for this one without sequencing it and I'm unlikely to have time to do that in the near future. Whatever the exact species might be, though, the pellis is a particularly rich brown, the stipe does bruise slightly brown, and the mushroom has a faint odor of raw potato. The bulb is cleft, too, so it could well just be a particularly dark-pigmented A. brunnescens.
		This is admittedly not going to be a fungus that most people will gasp over in astonishment. Coltricia montagnei is much like the Coltricia cinnamomea I found at Wachusett almost two weeks ago, but the one you're seeing here is the largest Coltricia I've ever seen. This specimen was just over four inches in diameter, making it more of a fairy sofa than a fairy stool. When I first flipped it over to see if it was yet another of the endless Phellodon sp. that were everywhere in the woods, I thought it might be some deformed Suillus fruiting body... until I looked at its spores. A drop of KOH turned the surface black instantly, too.
		I believed that Gomphus clavatus, also known as the pig's ear chanterelle, was a western species, but here it is in Pennsylvania. These are young specimens. The more aged fruiting bodies nearby had lost all violet tones and looked a god deal more like pigs' ears than do the ones in this photo.
		All Doctor Who fans should appreciate this little ascomycete, Leotia viscosa, also called the green jelly baby. It's not quite as common as its pallid yellow sibling species, Leotia lubrica, but I think it's far more esthetically appealing.
		The polypore Phaeolus schweinitzii, on the other hand, is beloved only by certain slightly mad mycologists. Wrap your kitchen sponge in felt, soak it in water and give it a good squeeze and you'll know what this fungus feels like in one's hand. It readily incorporates bits of twigs, leaves, grass and anything else that gets in its way as the fruiting body expands, and despite its squishy consistency these beasts can get quite large. I found another specimen in addition to the one in this photo; my second collection was a foot in diameter. P. schweinitzii is not considered edible, but some people do extract dye from it, hence its common name of "dye polypore." Not even beetles seem tempted by this one, although I suppose I'll need to collect more of these spongy, oozy polypores before knowing whether that's always true. I didn't find any on yesterday's collections, though.

Among the fungi I found yesterday but don't show here are Cantharellus cibarius and C. ignicolor (too few to bother bringing home, though), a green-brown Hygrophorus with both stipe and cap covered with slippery slime, a very large Paxillus atrotomentosus that smelled quite spicy after landing on the dryer (it's not supposed to, but even I could smell this one!), a couple more Elaphocordyceps ophioglossoides, a proliferation of Geoglossum sp., numerous Spathularia, and probably a number of things I'm forgetting entirely. I also came up with Ganoderma tsuga, but I only wanted the Bolitotherus cornutus that were living in it. Elisabet, who had never seen it outside of a herbarium before, took the fruiting bodies for further study.

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Gel of the Day: Meat the Beetles!

This Gel of the Day comes with a dedication to the unnamed UC Santa Cruz researcher whose car was blown up on Saturday by fruitcakes who want to liberate Drosophila.

The Fab Four in this case are four specimens of Diaperis maculata; John, Paul and George were collected together from a decomposing Piptoporus betulinus fruiting body. Ringo joined them later, having been recovered from a Ganoderma applanatum sporocarp. In each of the bosex, the first four lanes represent a 1:50 dilution and the second four a 1:100 dilution of the initial DNA solution. Lanes 1, 5, 9, 13, 17, 21, 25 and 29 are the nucLSU; 2, 6, 10, 14, 18, 22, 26 and 30 are nucSSU; 3, 7, 11, 15, 19, 23, 27 and 31 are cox1 amplified using the general Coleopteran PAT/JER primer set; 4, 8, 12, 16, 20, 24, 28 and 32 were an attempt to amplify cox1 using a custom-designed primer set that works on the other Tenebrionidae on which I've used it but apparently not on D. maculata. This is the same result I got last when I tried a competitive test of the cox1 primer sets on several species which also included another member of the same subfamily.

This might be a useful phylogenetic signal; it could very well indicate a mutation in the binding site for the new primer set which is conserved in all the other Tenebs for which I could find sequences (I could find none for D. maculata when I designed the primers). Moreover, an alignment from my previous attempt came up with an insertion and two transitions for D. maculata at a binding site that wasn't present in the other tenebs. I can't tell yet whether this is a species-specific situation or if the mutations are present in all members of the genus. I'm hoping to collect specimens of D. maculata' cousin, D. nigronotata, in Florida next month. If I can get sequences out of it, I will be able to say that much more about the evolution of genus Diaperis and perhaps about tribe Diaperini as well. That is, if Big Coleopterists don't Expel me over raising the slight possibility that the tribe might not be monophyletic and if no wackadoos with IEDs decide to set me on fire in my sleep.

Have I mentioned lately how cool it is to find something that nobody (as far as I can find in the literature) has seen before? This is the good stuff right here. This is why I get up in the morning and slave over a hot thermocycler.

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Wachusett Wednesday: Eat and Be Eaten Edition

I spent most of my energy on looking for beetle-infested polypores during yesterday's visit to my field site at Wachusett Mountain. Nonetheless, I saw a number of interesting specimens among the prolific mycofauna and collected a few. This included a couple of very interesting and rarely collected fungi and one truly excellent edible. Two of the fungi presented here, Nyctalis agaricoides and Elaphocordyceps ophioglossoides, specialize in eating other fungi. Cannibal fungi and tasty Lactarius are automatically interesting things, but that would leave out an oddball Cortinarius species known only from Massachusetts that I also present here. The flies were interesting, too, but I mean that in a different way. A species of brown fly with white-banded wings kept the foray particularly exciting as they bit chunks out of exposed skin, undeterred by repeated application of DEET. Yes, it hurt. The collections made yesterday are almost worth it, though.

		The genus Coltricia is collectively known in colloquial terms as the "fairy stools." They're stipitate-pileate members of the order Hymenochaetales. The majority of this order look more like polypores in general morphology and a number of them are resupinates (e.g., they form crust-like growths). This species, Coltricia cinnamomea, is unusual in being a hymenochaete that has a distinct cap-and-stem arrangement; it's probably an example of convergent evolution. The morphology has arisen independently several times in the Basidiomycetes. Despite the similarity of appearances, Coltricia are about as closely related to "true" polypores like Polyporus as humans are to shrews.
		As far as I've been able to find, Cortinarius dionysae has been collected only in Massachusetts. The purple color you see in these photos fades very quickly once the mushroom is removed from the substrate, changing to a pale brownish color within a few minutes as the mushroom dries. The largest pileus was very nearly 10 cm in diameter and the stipe is quite robust as well, terminating in a large bulb at the base. The universal veil leaves a sheath on the lower stipe that has a free collar, reminiscent of those commonly found in Amanita. Unlike most purple corts, C. dionysae has almost no bitter taste. The spores are shaped like lemons, also a bit of a departure from most other members of the genus. The flesh inside this mushroom is mostly white, but strong purple streaks can also be seen, particularly in the cap and top of the stipe. The edibility of this mushroom isn't known, but many corts are poisonous and some of the purple ones can be particularly nasty. I wouldn't suggest being the one to find out; it probably wouldn't be lethal, but it could do some serious damage to the digestive system.
		Elaphocordyceps ophioglossoides is one of the most interesting fungi in the world, and not just because its scientific name is such a mouthful. Most corcycepioid fungi are parasites on insects, killing them in rather spectacular and gruesome ways. Think Alien here. The spores infect a host, sometimes altering its behavior, and eventually the fruiting body emerges from the hapless arthropod's body. E. ophioglossoides, however, is one of a handful of these ascomycetes that have made the jump to some other host, in this case another fungus whose fruiting body is a false truffle in the genus Elaphomyces. Others parasites in the same genus haven't made that jump, though, and the closest relatives of the fungus you see in these photos are parasites of cicada nymphs. In an important sense, this host jumping is crystal clear evidence for an evolutionary mechanism called host-jumping. Researchers Naruo Nikoh and Takema Fukatsu published one of my favorite papers ever written about fungi on this very subject: Nikoh N and T Fukatsu. 2000. Interkingdom Host Jumping Underground: Phylogenetic Analysis of Entomoparasitic Fungi of the Genus Cordyceps. Molecular Biology and Evolution 17:629-638 In the time intervening, and based partly upon their work, the genus Cordyceps has been split up quite a bit. Check out Joey Spatafora's exhaustive website on the group of fascinating fungi, An Electronic Monograph of Cordyceps and Related Fungi, for more about the current state of knowledge about these intriguing parasites.
		It took me about an hour to figure out that this is Inocybe tahquamenonensis, a fungus that isn't at all rare but is seldom collected because it's small and nearly impossible to see against the forest floor. Unlike many members of its genus, this Inocybe is also odorless. Many of the Inocybes have odors variously described as crushed green corn, bleach or spermatic (which brings up some intersting questions of how mycologists might spend their free time that we won't get into here). Personally, I have a very poor sense of smell and in the past relied on former labling (and now associate professor at the University of Tennessee at Knoxville, though his faculty page isn't up yet) and Inocybe expert Brandon Matheny to help me out with collections of this genus. He can actually distinguish subtleties of these stinky mushroom's aromas. I can divide them only into "stinky" and "not quite so stinky" categories. Still, he's promised free beer at the MSA conference next month, so I'm sure he wouldn't mind my bringing along a bag or two full of these mushrooms for inspection by his far more educated nose. Eating any Inocybe is a bad idea. Most of them are poisonous. Some also contain hallucinogens, but the other compounds present insure that you'll have a very bad trip.
		Nyctalis agaricoides grows on dead Russula mushrooms; the slimy, mealy black corpse in these photos was probably Russula compacta as this collection was made from the area dominated by R. compacta that I was in two weeks ago and it is still largely overrun with that mushroom. There are only a few species of Nyctalis and they're a strange bunch. As you can see from these photos, they look like typical agaricoid fungi, but they're definitely not. In the second image, you can see that they lack either gills, pores or teeth. Where the gills should be there is instead a layer of translucent, very stiff gelatinous material that starts out white and turns a buff-brown color in older specimens. While it's technically a basidiomycete, N. agaricoides makes few or no basidia (I couldn't find any in the specimens I collected), and that's probably why it no longer bothers making a hymenium like most of its kin. When it want to reproduce, this mushroom simply clones itself. It walls off a hyphal tip and turns it into an asexual structure called a chlamydospore (here's a photo of the chlamydospores of this species), and these are produced not under the cap like in most basidiomycetes but on top< of the cap, which degenerates into a mass of beige fluff. The spores aren't forcibly discharged, either. They're dispersed by the wind and look for another dead Russula to germinate upon. Each of the offspring is an exact copy of the parent since no sexual recombination takes place. Other sexually-reproducing fungi can also form chlamydospores when circumstances force them to do so, but Nyctalis is unusual in having given up sexual reproduction in favor of parthenogenesis almost completely. A word of warning if you should happen to find any of these and think about bringing them home. They smell bad. Awful. They give off a powerful odor that combines garlic and phenolic compounds that can become nauseating in short order when confined to a room. The spore masses atop the fruiting bodies are very easily aerosolized, too. I learned this yesterday when I opened my collection box and was greeted by a puff of spores that I promptly inhaled. I could not only smell the things, I could taste them. Sucking down a lungful of asexual spores from an organism that lives as a scavenger, and has some mechanism by which it prevents the corpses upon which it thrives from drying out as they normally would, is likely a very bad idea. While it's unlikely that I'll wake one morning to find these pale little ghouls protruding from my ribcage, I would strongly suggest to others who might want to work with Nyctalis to do so under a hood or while wearing a pollen mask. The spores from these specimens were globular and 13 microns in diameter; use appropriate air filtration. I can't imagine anyone wanting to try eating something that grows on slimy mushroom corpses, but I can't imagine these critters would taste anything other than perfectly horrible. I have no idea as to whether they're poisonous. Despite their being fairly repulsive, I find them very interesting and would like to know more about how these things evolved. I'll likely sequence them and check into the literature to learn more.
		Moving away now from fungus that might eat you, here's a fungus that's good to eat. This is Lactarius volemus, which I consider a choice edible. I've collected and eaten these before; the last time was in Florida in 2002. Because of color variation I wound up collecting several of these yesterday, so half went into the dehydrator for archiving and the other half came home with me and wound up in a marsala sauce. They were excellent, far better than I remembered them from the last time I tried them. When any part of this mushroom is nicked, copious watery white latex will ooze out. This doesn't change color after more than ten minutes, but it will stain the flesh a light brown and the gills a darker brown. The odor can be fishy, but it isn't always. While authorities always mention the fishy scent, in my experience that may be a product of some small amount of decay. In any case, it's older specimens that take on the odor. Younger ones are close to odorless and as a rule of thumb mushrooms taste better before they get old. The cap color varies from an orange-brown in younger specimens to the pumpkin orange you see here in mature ones. The stipe is concolorous with the cap or a bit lighter and often fades to white at the apex. The gills are a pale cream-yellow color, often with brown bruises. The whole mushroom is finely pubescent; using a magnifying glass while make that feature more readily apparent. The spores are generally globose; the ones from the specimens I found yesterday were about 8 microns in diameter. Taste-testing the uncooked mushroom resulted in no acrid flavor at all. Cooking it in marsala wine and spooning it over pork chops resulted in a deeply appreciated deliciousness that almost made up for the bits of skin I'm now missing from the back of my head due to the nasty Diptera that think we humans far tastier than any Lactarius could ever hope to be.
		From the "There's a reason that they call it a variable Russula" department: Look at the mushroom in the photos to the left. Take a good, long look, noting coloration and morphology. Now take a look at this Russula, collected two weeks ago. They don't look at all alike, do they? According to current systematics, though, these are the same species, Russula variata. It's hard to believe (in fact, I'm not sure I believe it myself), but aside from the gross morphology, these two very different-looking fungi share all of the same traits. The repeatedly-forking gills, the lack of any color change in FeSO4, the size and shape of the spores and the presence of an amyloid reticulum joining the <0.5 micron-high warts on them... there's little doubt that this is also R. variata. I wonder, though, whether this is more a matter of convenience than a true phylogenetic hypothesis. Despite the taxonomic peculiarity, I have a hunch that what's called R. variata is actually a few species lumped together. Maybe mycologists are spending too much time thinking about spermatic Inocybes and Amanita with constricted volvae!

Flies eat people. People eat wild mushrooms that probably contain a few fly larvae. Fungi eating other fungi. Fungi eating insects. It's an eat-or-be-eaten world out there. Or more properly, it's eat-and-be-eaten.

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Natural Curiosity: An Inordinate Fondness for Fungi

My sister and her family are visiting this weekend. She and and my brother-in-law are both psychologists and my niece, Alex, is a rambunctious and extremely bright four year old with tremendous curiosity about nature. I don't know whether that's nature or nurture at work. They live in a rural area in the Hudson River Valley. Their home is the kind of place where deer wandering across the lawn is a daily occurrence and it's not too unusual to have a bear raid the garbage every so often. Nonetheless, as I've mentioned before, Alex seems to have been born with a particular interest in fungi. It's not something that anyone taught her about, though it's certainly been encouraged. I can't think of why...

We met up at Elm Park yesterday to let Alex blow off some steam at the playground. The plan was that the adults could check out Art in the Park at the same time, but there wasn't much art to see despite the banner proclaiming that the event runs through October. Practically the first thing that happened is that I was handed the note you see to the left. It was important that I see this right away!

When we started getting hungry, we left for Binh Duong, which still ranks as LL and my favorite Vietnamese restaurant in Worcester. I asked Alex if she'd like to see the lab where I work on the way and she was very interested in doing so.

It being Saturday night, nobody else was around. We did a quick tour of the clean lab; it's very hard to explain to a four year old what we do in there. She did want to wear the cool purple nitrile gloves, though. Alex was much more interested in the dirty lab. She wanted to see the mushrooms, of course, and she got to check out a whole preserved Bondarzewia berkeleyi and giant specimens of Ganoderma applanatum as well. She also went through some of the Russula specimens I've collected and she thought the big green ones were pretty impressive. I noticed that she was eying the microscopes, so I asked her if she wanted to look at a beetle and that was a cool idea. I dug out a big staphylinid from my bug box and set it up under a dissecting scope for her. Woah! Giant beetles... the kid was psyched.

I asked Alex if she'd like to look even closer and see what a beetle's foot looks like. Big surprise; let's do that! I dissected off a tarsus and described what I was doing. At each step, Alex replied with a "yeah" and implicit, "and then what," the pitch of her voice increasing a little each time. I showed her how to make a wet mount and each step brought a higher-pitched "yeah" dripping with curiosity. When I had the slide set up on the microscope, Alex climbed into my lap to look at the foot at 100X magnification and her eyes were bugging out enough that she could probably have seen it from a foot away. By the time she'd had her fill of those tarsomeres and tarsal claws, she was ready to do it again. "Let's do another one!"

It was time for dinner, though, so I let Alex use a pair of watchmaker's forceps to put the beetle back in its vial. I didn't tell her how to do it, and it struck me that she knew enough to be very delicate about the task. She started off trying to barely exert any pressure at all, dropping the insect a couple of times before she squeezed just hard enough to get the thing from the hack to the vial without crushing it in the slightest. The kid is a natural!

Our plan for today was to go up to Wachusett so that she could see the giant green mushrooms and whatever other kinds might be out. Alex wants to find beetles, too, so I let her pack a few vials of EtOH in my kit last night for use today. Unfortunately, we're having a wicked electrical storm at the moment and the forecast is calling for rain throughout the day, so I'm not sure whether we'll be able to follow through on the plans. We may need to come up with some kind of indoor alternative. I think I'd be as disappointed as Alex would about that. Most, if not all, kids are born with a curiosity about nature, I suppose. If the four year old keeps that same level of curiosity past adolescence, they become science geeks. When they keep it past the age of forty they become... well... me, I suppose. Doesn't every kid need a weird scientist uncle?

I hope the rain stops so I can go find mushrooms and bugs today! Am I a four year old dreaming that I'm a graduate student, or am I a graduate student in the company of a four year old?

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Gel of the Day: Moderate Success

Sound the trumpets! I actually got some PCR products out of today's experiment with COI primers.

It looks from the results like both primer sets work, although my new Tenebrionidae-specific primers (PATten and JERten) amplifies a smaller fragment than the traditional Coleoptera COI primers. Also, it seems that the traditional primer set worked for one species but not for another in the same tribe (Diaperini), and the new primers worked for the other member of that tribe (both are Diaperini; the new primers worked on Pentaphyllus but not Diaperis and the old ones on Diaperis but not Pentaphyllus). Why? I dunno. I still have my suspicions about how monophyletic tribes, and even subfamilies, really are in this family, and maybe that's part of what I'm seeing in these results. I may know more once I try aligning these sequences and especially when I add this data together with the large and small subunit stuff.

Otherwise, both primer sets worked on everything except one miss by the new set on a single Bolitotherus cornutus, but it worked on the other specimens of the same species, so I really have no idea with that one.

Still, here's the gel. There are a couple of faint bands in the lower half that may be hard to see in the reduced photo, but they're in there. Hopefully I'll still be able to get sequences out of those:

I used several different dilutions (1:100, 1:10, 1:5) and got some products from all of them. It all comes down to using a lower annealing temperature for the COI stuff. From here on in, I should be able to get those sequences reliably.

Now I'm just hanging around and waiting for my sequencing reactions to finish up, which they should do in another 2.5 hours. Thankfully, that Del Tenney DVD I have is a double feature; I can watch Curse of the Living Corpse while I'm waiting. I'll be here late on a Sunday, but I should be ready to go when tomorrow's sequencer run starts up at 11:00 AM. Go, me.

If anybody wants me, I'll be in the conference room.

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Reasons to Be Cheerful: DNA, Expeditions and Mycological Society Meeting

It's the little things in life sometimes that bring the greatest pleasures... at least when one revels in being a geek.

The first thing I need to do today is to clean up some DNA I have waiting in the freezer. Among them are the rDNA sequences from Diaperis maculata. Once I've cleaned the DNA I'll be able to load it onto a plate and will get the sequences back and be able to add them to my alignments. I'll know a tiny bit more about the evolution of fungus-associated Coleoptera. This is enough to make me look forward to a Monday. It's the little things... and D. maculata is only 5-6 mm long. That qualifies as "little."

I also have to start work on securing permits for the collecting expedition in September. Two months should be enough lead time, hypothetically, but one never knows with bureaucracies. We just need a couple of good sites to work at, though. I'll probably put feelers out on more than we'll actually use. That way if the process gets too long and we get turned down for a couple of sites at the last minute, we'll still have a couple of back-ups. Better to have it and not need it than to need it and not have it.

Speaking of science-related travel, it's only a month until I hit the road for Penn State and the annual Mycological Society of America meeting. It's the largest gathering of professional and amateur mycologists in the nation and promises to be a great time. I'm looking forward to the whole program, but I have to admit that I'm particularly tickled by the opportunity to collect in Pennsylvania, a state in which I've never forayed before. I'm even hauling my industrial-strength dehydrator on the trip so that folks can preserve specimens. The abstracts look like there will be some very interesting talks on fungal evolution and informatics tools. Folks from my lab are doing four of the presentations, I think. I'm not one of them yet (I've just started gathering data!) There are also a couple of talks by David Maddison that look like they'll be particularly interesting stuff. Days of full immersion... I can hardly wait.

Are any readers planning to attend MSA this year? If so, I'd love to meet up for a drink or two.

If you're new to this stuff and this is the first you're hearing about it and you'd like to attend, you can still register for the meeting. Even if you're a novice, you couldn't ask for a better opportunity to learn about fungi. Please consider joining us! If you're a newbie and can make it in time for the foray on August 10, I'd be more than happy to hang out with you in the field and help you get better acquainted with the various chitinous life-forms you'll encounter. I'd consider it a privilege to do so, in fact.

Today, and the next couple of months, are going to be great. Wonder-Geek Powers Activate!

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Florida Expedition: September 3-10

Guess who's coming to Florida?

A colleague from Harvard and I will be in North Florida to sample fungi and beetles from the area between September 3 and 10. The magnanimous Diego/James F/todabrilla has consented to put us up at his place for the week, insuring us of good company as well as saving two itinerant and impoverished graduate students the cost of a hotel. With our flight from Boston to Tampa now confirmed and lodging assured, I'll next be turning to securing the permits required to do our collecting legally. Flying into Tampa means that we'll have to make a three hour drive to Tallahassee, but it also saved us a ton of money on the cost of travel.

While our days will be filled with collecting, identifying and preserving specimens, I'm looking forward to seeing some old acquaintances in the evenings as well. I'll need to check into whether I can get access to some resources at Florida State (like a couple of microscopes), and in any case I expect to visit the campus and see a couple of my professors as well as attending a meeting of the Union of Freethinking Students. I'm certain I'll also pass at least one evening at Aladdin's Sheesha Café.

North Florida is one of the country's hottest biodiversity hotspots, renowned for its unique ecosystems and endemic species. Even though I spent five years investigating them on my own, it will be a whole new experience seeing them through eyes that have learned a great deal more about fungi and Coleoptera over the course of the past year. I must admit that it will be good to see the many venomous snakes that make field work in the area that much more exciting, too.

I could do without the Confederate flags, of course, but one doesn't see too many in the swamps and forests, anyhow.

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