Marine Microforay – foram and a thecofilosean party

Apologies for disappearing for a while – had an interview, finals and then my arm decided to temporarily rediscover RSI-like symptoms just when I had a term paper to write, so I had to lay off extraneous typing for a while. Then I realised just how much of my life depends on typing, and losing that ability would not only make me worthless and unemployable, but also unable to communicate with many of my friends who happen to be inconveniently dispersed around the globe. So yeah, I should probably stop casually dismissing ergonomics about now...as should you, if you haven't already!

I've accumulated another batch of microscopic findings, this time from marine samples. By the looks of it, I might be moving to the Midwest soon, and thus be deprived of my ocean (and mountains *sob*), so I figured that focusing on marine protists while I have the chance would be a good idea. Swampy pondwater is available pretty much anywhere anyway.

From time to time, you can be lucky enough to find a foram shell in the sediments around here. Live forams can be found too, but much more rarely – I have a couple, but still need to process the videos. This is not a snail:

Foram. Wreck Beach. 20x obj, DIC except for last image, which is in phase.

To save loading time, the rest are below the fold.

Sunday Protist - A sampling of Cercozoa Part I

This post grew out of proportion, so I'm splitting it into two or three parts, to cater to our ever-shortening attention spans (mine included)...

[Warning: Taxonomy. Of the harshest kind: involves Cavalier-Smith]

At the moment, among my favourite supergroups is Rhizaria (tree). Rhizaria is generally where all the obscure, interesting, and outright weird eukaryotes get sent by molecular data these days. The group itself is fairly recent, having been formally spewed out declared by Cavalier-Smith in 2002, as a fusion of Cercozoa and Retaria(=forams and 'radiolarians'), as well as Heliozoa and Apusozoa, apparently because they had "a centrosomal core or radiating microtubules and two microtubular roots and soft surface, typically with reticulopodia." (TC-S 2002 IJSEM:297) Don't worry, I don't really know what that means either. That is, those are fairly common traits in many non-Rhizarians, even according to the TC-S 2002 classification.

The name derives from the group's inclusion of many members of the then-defunct "Rhizopods" ('root-feet' - members typically had thin, branchy pseudopodia). Since then, Heliozoa died a horrible death with its limbs strewn all over the tree (Nikolaev et al. 2004 PNAS) and [many] Apusozoa now seem to enjoy their privileged life as the putative basal Opisthokonts (or their sisters). Ironically, many of the "Heliozoa" did return to Cercozoa later. Obligatory TC-S Diagram:

The birth of Rhizaria. As the young supergroup struggles to open its eyes to the world for the first time, it is confronted by the glaring faces of frustrated readers threatening to ban the author from ever birthing another taxon, for the sake of global sanity. Yet, despite its weak, fragile synapomorphies, the newborn supergroup, heavily-medicated by state-of-the-art molecular phylogenies, rises to become a bona fide citizen of the taxonomic world. For now. As all other life forms on earth, the higher taxa themselves are mortal. (diagram slightly modified (red box added) from Cavalier-Smith 2002 IJSEM)

"Radiolarians" (Acantharians+Taxopodids+Polycystines) and Forams (more generally, Granuloreticulosea) are massively diverse, complicated and awesome, but Cercozoa are more obscure to non-protistologists, and are a rather weird assemblage of stuff. I think the Amoebozoan taxon "Variosea" would have been quite fitting for them, were it not taken by amoebae instead. Cercozoa is older than Rhizaria, but not by much - it was formally established by Cavalier-Smith in 1998 (Biol Rev) as a modified successor of Rhizopoda:

"The recently revised phylum Rhizopoda is modified further by adding more flagellates and removing some ‘ rhizopods ’ and is therefore renamed Cercozoa" (TC-S 1998 BiolRev:203)

Of course, that was Tom's version of Rhizopoda to begin with. Taxonomy gets very fun when different people at different times mean different things by the same name. Can't seem to find the etymology of Cercozoa, but the formal description reads pretty much like 'miscellaneous eukaryotes with thin pseudopodia'. And that they are.

While Cercozoa was initially based loosely on morphology and sketchy data from the dawn of molecular phylogenetics, it mostly survived intact over the years, and grew further (with various things shaved off too, of course). The original members were Phytomyxids (incl. the plant pathogen Plasmodiophora), Reticulofilosa (basically, Chlorarachniophytes) and Monadofilosa (Cercomonas, Gymnophrys, Euglypha and Spongomonas are given as original examples). Curiously, all of them survived the onslaught of molecular reality (or so we hope...). Stuff has been added, like Ascetosporea (paramyxids and haplosporidia; added in TC-S 2002 IJSEM) and the gromiids, as well as various obscure incertae sedis orphans and a few refugees from 'Heliozoa'.

Eventually, the Cercozoa got 'sistered' to the forams (Keeling 2001 MBE) by ACTIN phylogenies, which got taxonomically recognised in the TC-S 2002 IJSEM revision of The Book of Tom by declaring the holy union of Retaria (forams and rads) and Cercozoa as Rhizaria. Going overboard as usual by adding in Heliozoa and Apusozoa, of course. We're talking about the mad taxonomist here ;-) (now someone needs to make that into a pop culture phenomenon to rival mad scientists..."And along comes the evil mad taxonomist...and RENAMES EVERYTHING!" *cue spooky music*) The group still lacks any solid synapomorphies (shared derived characters); the situation is such that even the use of obscure ultrastructural elements has been attempted, such as Cavalier-Smith's "transitional nonagonal fibre" (TC-S 2008 Protist) – even one of his own past students has no idea what he meant there!

And a whole bunch of other stuff happened but I think that was enough Historical Taxonomy (would make the most popular course evar, srsly) for...the month. Ok, so have we lost everyone yet? Or have the wiser ones employed the high art of The Scrollbar and skimmed accordingly? In any case, I'd like to very briefly and shallowly run over a few of the major cercozoans to give you a taste of the phylum, and just how diverse and varied it is. Things will be skipped, including, quite possibly, The Most Interesting Thing Ever Because You Studied it for the Past Ten Years. Apologies in advance. TMITEBYSiftPTY will get its chance, someday.

Some phylogeny and taxonomy sources: TC-S & Chao 2003; Bass & TC-S 2004; Bass et al. 2005; Pawlowski & Burki 2009; Chantangsi et al. 2010.

(L: Pawlowski & Burki 2009; R: Chantangsi et al. 2010)

To be continued in Part II – Endomyxa.

References
Bass D, & Cavalier-Smith T (2004). Phylum-specific environmental DNA analysis reveals remarkably high global biodiversity of Cercozoa (Protozoa). International journal of systematic and evolutionary microbiology, 54 (Pt 6), 2393-404 PMID: 15545489

BASS, D. (2005). Polyubiquitin Insertions and the Phylogeny of Cercozoa and Rhizaria Protist, 156 (2), 149-161 DOI: 10.1016/j.protis.2005.03.001

CAVALIER-SMITH, T. (1998). A revised six-kingdom system of life Biological Reviews of the Cambridge Philosophical Society, 73 (3), 203-266 DOI: 10.1017/S0006323198005167

Cavalier-Smith T (2002). The phagotrophic origin of eukaryotes and phylogenetic classification of Protozoa. International journal of systematic and evolutionary microbiology, 52 (Pt 2), 297-354 PMID: 11931142

Cavalier-Smith, T., & Chao, E. (2003). Phylogeny of Choanozoa, Apusozoa, and Other Protozoa and Early Eukaryote Megaevolution Journal of Molecular Evolution, 56 (5), 540-563 DOI: 10.1007/s00239-002-2424-z

CAVALIERSMITH, T., LEWIS, R., CHAO, E., OATES, B., & BASS, D. (2008). Morphology and Phylogeny of Sainouron acronematica sp. n. and the Ultrastructural Unity of Cercozoa Protist, 159 (4), 591-620 DOI: 10.1016/j.protis.2008.04.002

Chantangsi, C., Hoppenrath, M., & Leander, B. (2010). Evolutionary relationships among marine cercozoans as inferred from combined SSU and LSU rDNA sequences and polyubiquitin insertions Molecular Phylogenetics and Evolution, 57 (2), 518-527 DOI: 10.1016/j.ympev.2010.07.007

Keeling PJ (2001). Foraminifera and Cercozoa are related in actin phylogeny: two orphans find a home? Molecular biology and evolution, 18 (8), 1551-7 PMID: 11470846

Nikolaev, S. (2004). From the Cover: The twilight of Heliozoa and rise of Rhizaria, an emerging supergroup of amoeboid eukaryotes Proceedings of the National Academy of Sciences, 101 (21), 8066-8071 DOI: 10.1073/pnas.0308602101

PAWLOWSKI, J., & BURKI, F. (2009). Untangling the Phylogeny of Amoeboid Protists Journal of Eukaryotic Microbiology, 56 (1), 16-25 DOI: 10.1111/j.1550-7408.2008.00379.x

Sunday Protist -- Auranticordis: "Orange Heart"

(Also the answer to MM12, which nobody got, although I haven't particularly followed up on the hints either...)

If you're ever low on ideas for your next taxonomic adventure, Cercozoa is one endlessly fascinating clade that never fails to offer surpises. In fact, the more you explore them, the less familiar they become. Cercozoa (TC-S 1998) can offer pretty much anything -- from photosynthetic amoebae to the utterly bizarre Phaeodaria to beautiful testate Euglyphids (including a case of separate primary plastid endosymbiosis!) to a wide variety of random semi-amoeboid-semi-flagellate things that glide about on their flagella and make faces at anyone trying to classify these things by morphology. Cercozoa are a mess, but a beautiful mess.

SEM from looking down at the anterior apex of the cell. Bar = 10um; Chantangsi et al. 2008 BMC Microbiol (open access)

The baby Nautilus looking thing that was so mischievously confusing is Auranticordis quadriverbis, a rare benthic tetraflagellate of a distinctive orange colour.

A - L: lobe, arrows: orange muciferous bodies; C - arrowheads: putative endosymbionts. D - arrowhead points to ventral groove. E - phase contrast showing flagella protruding from the ventral groove. Bar = 10um. (Chantangsi et al. 2008 BMC Microbiol)

The orange colour is caused by the muciferous bodies lining the ridges along the cortex. These bodies secrete mucilage upon disturbance, and may participate in adhesion to the surface. The membranous ridges on the surface are supported by bundles of microtubules, in the absense of any cell wall. An SEM of the ridges reveals the mucilage being secreted by the pores that line them:

Sem close-up of the ridges with mucilage. Scalebars: C - 1um, D,E - 0.5um
(Chantangsi et al. 2008 BMC Microbiol)

Curiously enough, Auranticordis posesses potential endosymbionts -- thylakoid-bearing orange bodies inside the cytoplasm. While those could be food particles being digested, the lack of different extents of digestion and their intact appearance suggests these endosymbionts may be long-term (as in kleptoplasty) or permanent (endosymbiosis). The similarity of the ultrastructure to free-living cyanobacteria actually suggests this may be yet another case of primary endosymbiosis, as in Paulinella! (cyanobacteria don't have to be blue-green, by the way)

TEM of putative endosymbionts. The lines at the edges are thylakoids, surrounding a prominent invagination of the inner membrane. Bar = 2um (Chantangsi et al. 2008 BMC Microbiol)

Inside the endosymbionts one can find viruses that look similar to the ones that infect free-living cyanobacteria. The idea of a virus attacking your organelles, while not actually surprising, is still kind of cool!

Higher magnification TEM of the putative endosymbiont. B shows detail of the thylakoids C - cleavage furrow (arrowheads) supporting the idea of this body being an endosymbiont rather than a food particle, SC - sac-like vesicles. D - Viral particles at the thylakoid-free core. E - detail of the prominent inner membrane invagination. G - close up of more viral particles, (bar=0.5um) with an inset showing a complete one (bar=0.2um) (Chantangsi et al. 2008 BMC Microbiol)

Unfortunately these organisms are rare, so it's difficult to establish whether these pigmented bodies are in fact a third known case of primary plastid endosymbiosis. Wouldn't be so shocking after Paulinella, but still kind of cool. Perhaps primary endosymbiosis isn't as rare as previously thought, and that, were it true, could have a profound impact on our understanding of eukaryotic evolution. Conventionally, plastid gain is used as a very unlikely feature, and weighed heavily in analyses of parsimony. But the real picture could actually be much messier.

I hope you've enjoyed this brief encounter with an enigmatic, obscure Cercozoan. Warning: There's more where that came from, so expect more Cercozoa in the future. I really love that group...

Reference:
Chantangsi, C., Esson, H., & Leander, B. (2008). Morphology and molecular phylogeny of a marine interstitial tetraflagellate with putative endosymbionts: Auranticordis quadriverberis n. gen. et sp. (Cercozoa) BMC Microbiology, 8 (1) DOI: 10.1186/1471-2180-8-123