Choanoflagellates are a group of free-living unicellular and colonial flagellateeukaryotes considered to be the closest living relatives of animals. The name refers to the characteristic funnel-shaped "collar" of interconnected microvilli and the presence of a flagellum. Choanoflagellates are found globally in aquatic environments, and they are of particular interest to evolutionary biologists studying the origins of multicellularity in animals.
The flagellum of choanoflagellates is surrounded by microvilli at its base. Movement of the flagellum creates water currents that can propel free-swimming choanoflagellates through the water column and trap bacteria and detritus against the microvilli, where these foodstuffs are engulfed. This feeding plays an ecological role in the carbon cycle by linking different trophic levels.
Choanoflagellates bear morphological similarities to the choanocyte, a type of cell in sponges. As the proposed sister group to Animalia, choanoflagellates serve as a useful model for reconstructions of the last unicellular ancestor of animals. According to a 2021 study, crown group craspedids (and perhaps crown group choanoflagellates if Acanthoecida arose within Craspedida<ref>Template:Cite journal</ref>) appeared 422.78 million years ago,<ref>Template:Cite journal</ref> although a previous study from 2017 recovered the divergence of the crown group choanoflagellates (craspedids) at 786.62 million years.<ref>Template:Cite journal</ref>
Each choanoflagellate has a single flagellum, surrounded by a ring of actin-filled protrusions called microvilli, forming a cylindrical or conical "collar" (Template:Lang in Greek). Movement of the flagellum draws water through the collar, and bacteria and detritus are captured by the microvilli and ingested.<ref name=King2008>Template:Cite journal</ref> Water currents generated by the flagellum also push free-swimming cells along, as in animalsperm. In contrast, most other flagellates are pulled by their flagella.<ref>Klindt, G. S., & Friedrich, B. M. (2015). Flagellar swimmers oscillate between pusher- and puller-type swimming. Physical Review E, 92(6), 063019. https://doi.org/10.1103/PhysRevE.92.063019</ref>
In addition to the single apical flagellum surrounded by actin-filled microvilli that characterizes choanoflagellates, the internal organization of organelles in the cytoplasm is constant.<ref name=Leadbeater2000(1)>Template:Cite journal</ref> A flagellar basal body sits at the base of the apical flagellum, and a second, non-flagellar basal body rests at a right angle to the flagellar base. The nucleus occupies an apical-to-central position in the cell, and food vacuoles are positioned in the basal region of the cytoplasm.<ref name=Leadbeater2000(1)/><ref name=Karpov1998>Template:Cite journal</ref> Additionally, the cell body of many choanoflagellates is surrounded by a distinguishing extracellular matrix or periplast. These cell coverings vary greatly in structure and composition and are used by taxonomists for classification purposes. Many choanoflagellates build complex basket-shaped "houses", called lorica, from several silica strips cemented together.<ref name=Leadbeater2000(1)/> The functional significance of the periplast is unknown, but in sessile organisms, it is thought to aid attachment to the substrate. In planktonic organisms, there is speculation that the periplast increases drag, thereby counteracting the force generated by the flagellum and increasing feeding efficiency.<ref name=Leadbeater2001>Template:Cite journal</ref>
Choanoflagellates are either free-swimming in the water column or sessile, adhering to the substrate directly or through either the periplast or a thin pedicel.<ref name="Leadbeater1983">Template:Cite journal</ref> Although choanoflagellates are thought to be strictly free-living and heterotrophic, several choanoflagellate relatives, such as members of Ichthyosporea or Mesomycetozoa, follow a parasitic or pathogenic lifestyle.<ref name="Mendoza2002">Template:Cite journal</ref> The life histories of choanoflagellates are poorly understood. Many species are thought to be solitary; however, coloniality seems to have arisen independently several times within the group, and colonial species still retain a solitary stage.<ref name="Leadbeater1983" />
Over 125 extant species of choanoflagellates<ref name=King2008/> are known, distributed globally in marine, brackish and freshwater environments from the Arctic to the tropics, occupying both pelagic and benthic zones. Although most sampling of choanoflagellates has occurred between Template:Convert, they have been recovered from as deep as Template:Convert in open water<ref name=Thomsen1982>Template:Cite book</ref> and Template:Convert under Antarctic ice sheets.<ref name="Buck1988">Template:Cite journal</ref>
Many species are hypothesized to be cosmopolitan on a global scale [e.g., Diaphanoeca grandis has been reported from North America, Europe and Australia (OBIS)], while other species are reported to have restricted regional distributions.<ref name=Thomsen1991>Template:Cite journal</ref> Co-distributed choanoflagellate species can occupy quite different microenvironments, but in general, the factors that influence the distribution and dispersion of choanoflagellates remain to be elucidated.<ref>Template:Cite journal</ref>
A number of species, such as those in the genusProterospongia, form simple colonies,<ref name=King2008/> planktonic clumps that resemble a miniature cluster of grapes in which each cell in the colony is flagellated or clusters of cells on a single stalk.<ref name=Leadbeater2000(1)/><ref name=Carr2008>Template:Cite journal</ref> A colonial species from Mono Lake, Barroeca monosierra, forms spheres filled with a branched network of an extracellular matrix where a microbiome of different species of symbiotic bacteria live.<ref>Template:Cite journal</ref><ref>Template:Cite journal</ref> In October 2019, scientists found a new band behaviour of choanoflagellates: they apparently can coordinate to respond to light.<ref>Template:Cite web</ref>
The choanoflagellates feed on bacteria and link otherwise inaccessible forms of carbon to organisms higher in the trophic chain.<ref name=Butterfield1997>Template:Cite journal</ref> Even today, they are important in the carbon cycle and microbialfood web.<ref name=King2008/> There is some evidence that choanoflagellates feast on viruses as well.<ref>Template:Cite news</ref>
Choanoflagellates grow vegetatively, with multiple species undergoing longitudinal fission;<ref name="Karpov1998" /> however, the reproductive life cycle of choanoflagellates remains to be elucidated. A paper released in August 2017 showed that environmental changes, including the presence of certain bacteria, trigger the swarming and subsequent sexual reproduction of choanoflagellates.<ref name="Woznica2017">Template:Cite journal</ref> The ploidy level is unknown;<ref>Claus Nielsen. Animal Evolution: Interrelationships of the Living Phyla. 3rd ed. Claus Nielsen. Oxford, UK: Oxford University Press, 2012, p. 14.</ref> however, the discovery of both retrotransposons and key genes involved in meiosis<ref name="Carr2010">Template:Cite journal</ref> previously suggested that they used sexual reproduction as part of their life cycle. Some choanoflagellates can undergo encystment, which involves the retraction of the flagellum and collar and encasement in an electron dense fibrillar wall. On transfer to fresh media, excystment occurs; though it remains to be directly observed.<ref name="Leadbeater2000(2)">Template:Cite journal</ref>
Evidence for sexual reproduction has been reported in the choanoflagellate species Salpingoeca rosetta.<ref name="pmid28867285">Template:Cite journal</ref><ref name="pmid24139741">Template:Cite journal</ref> Evidence has also been reported for the presence of conserved meiotic genes in the choanoflagellates Monosiga brevicollis and Monosiga ovata.<ref name="pmid20015185"/>
The Acanthoecid choanoflagellates produce an extracellular basket structure known as a lorica. The lorica is composed of individual costal strips, made of a silica-protein biocomposite. Each costal strip is formed within the choanoflagellate cell and is then secreted to the cell surface. In nudiform choanoflagellates, lorica assembly takes place using a number of tentacles once sufficient costal strips have been produced to comprise a full lorica. In tectiform choanoflagellates, costal strips are accumulated in a set arrangement below the collar. During cell division, the new cell takes these costal strips as part of cytokinesis and assembles its own lorica using only these previously produced strips.<ref name="Leadbeater et al. 2009">Template:Cite journal</ref>
Choanoflagellate biosilicification requires the concentration of silicic acid within the cell. This is carried out by silicon transporter (SiT) proteins. Analysis of choanoflagellate SiTs shows that they are similar to the SiT-type silicon transporters of diatoms and other silica-forming stramenopiles. The SiT gene family shows little or no homology to any other genes, even to genes in non-siliceous choanoflagellates or stramenopiles. This suggests that the SiT gene family evolved via a lateral gene transfer event between Acanthoecids and Stramenopiles. This is a remarkable case of horizontal gene transfer between two distantly related eukaryotic groups, and has provided clues to the biochemistry and silicon-protein interactions of the unique SiT gene family.<ref name=Marron2013>Template:Cite journal</ref>
Félix Dujardin, a French biologist interested in protozoan evolution, recorded the morphological similarities of choanoflagellates and sponge choanocytes and proposed the possibility of a close relationship as early as 1841.<ref name=Leadbeater2001/> Over the past decade, this hypothesized relationship between choanoflagellates and animals has been upheld by independent analyses of multiple unlinked genetic sequences: 18S rDNA, nuclear protein-coding genes, and mitochondrial genomes (Steenkamp, et al., 2006; Burger, et al., 2003;<ref name=Mendoza2002/> Wainright, et al., 1993). Importantly, comparisons of mitochondrial genome sequences from a choanoflagellate and three sponges confirm the placement of choanoflagellates as an outgroup to Metazoa (animals, also known as Animalia) and negate the possibility that choanoflagellates evolved from metazoans (Lavrov, et al., 2005). Finally, a 2001 study of genes expressed in choanoflagellates has revealed that choanoflagellates synthesize homologues of metazoan cell signaling and adhesion genes.<ref name=King2001>Template:Cite journal</ref> Genome sequencing shows that, among living organisms, the choanoflagellates are most closely related to animals.<ref name=King2008/>
Because choanoflagellates and metazoans are closely related, comparisons between the two groups promise to provide insights into the biology of their last common ancestor and the earliest events in metazoan evolution. The choanocytes (also known as "collared cells") of sponges (considered among the most basal metazoa) have the same basic structure as choanoflagellates. Collared cells are found in other animal groups, such as ribbon worms,<ref>Template:Cite journal</ref> suggesting this was the morphology of their last common ancestor. The last common ancestor of animals and choanoflagellates was unicellular, perhaps forming simple colonies; in contrast, the last common ancestor of all eumetazoan animals was a multicellular organism, with differentiated tissues, a definite "body plan", and embryonic development (including gastrulation).<ref name=King2008/> The timing of the splitting of these lineages is difficult to constrain, but was probably in the late Precambrian, >Template:Ma.<ref name=King2008/>
The choanoflagellates were included in Chrysophyceae until Hibberd, 1975.<ref>Reviers, B. de. (2006). Biologia e Filogenia das Algas. Editora Artmed, Porto Alegre, p. 156.</ref> Recent molecular phylogenetic reconstruction of the internal relationships of choanoflagellates allows the polarization of character evolution within the clade. Large fragments of the nuclear SSU and LSUribosomal RNA, alpha tubulin, and heat-shock protein 90 coding genes were used to resolve the internal relationships and character polarity within choanoflagellates.<ref name=Carr2008/> Each of the four genes showed similar results independently and analysis of the combined data set (concatenated) along with sequences from other closely related species (animals and fungi) demonstrate that choanoflagellates are strongly supported as monophyletic and confirm their position as the closest known unicellular living relative of animals.
Previously, Choanoflagellida was divided into these three families based on the composition and structure of their periplast: Codonosigidae, Salpingoecidae and Acanthoecidae. Members of the family Codonosigidae appear to lack a periplast when examined by light microscopy, but may have a fine outer coat visible only by electron microscopy. The family Salpingoecidae consists of species whose cells are encased in a firm theca that is visible by both light and electron microscopy. The theca is a secreted covering predominately composed of cellulose or other polysaccharides.<ref>(Adl, et al., 2005)</ref> These divisions are now known to be paraphyletic, with convergent evolution of these forms widespread. The third family of choanoflagellates, the Acanthoecidae, has been supported as a monophyletic group. This clade possess a synapomorphy of the cells being found within a basket-like lorica, providing the alternative name of "Loricate Choanoflagellates". The Acanthoecid lorica is composed of a series of siliceous costal strips arranged into a species-specific lorica pattern."<ref name=Leadbeater2000(1)/><ref name=Leadbeater2001/>
The choanoflagellate tree based on molecular phylogenetics divides into three well supported clades.<ref name=Carr2008/> Clade 1 and Clade 2 each consist of a combination of species traditionally attributed to the Codonosigidae and Salpingoecidae, while Clade 3 comprises species from the group taxonomically classified as Acanthoecidae.<ref name=Carr2008/> The mapping of character traits on to this phylogeny indicates that the last common ancestor of choanoflagellates was a marine organism with a differentiated life cycle with sedentary and motile stages.<ref name=Carr2008/>
The genome of Monosiga brevicollis, with 41.6 million base pairs,<ref name=King2008/> is similar in size to filamentous fungi and other free-living unicellular eukaryotes, but far smaller than that of typical animals.<ref name=King2008/> In 2010, a phylogenomic study revealed that several algal genes are present in the genome of Monosiga brevicollis. This could be due to the fact that, in early evolutionary history, choanoflagellates consumed algae as food through phagocytosis.<ref>Template:Cite journal</ref> Carr et al. (2010)<ref name="pmid20015185">Template:Cite journal</ref> screened the M. brevicollis genome for known eukaryotic meiosis genes. Of 19 known eukaryotic meiotic genes tested (including 8 that function in no other process than meiosis), 18 were identified in M. brevicollis. The presence of meiotic genes, including meiosis specific genes, indicates that meiosis, and by implication, sex is present within the choanoflagellates.
Salpingoeca rosetta genome
The genome of Salpingoeca rosetta is 55 megabases in size.<ref name=Fairclough2013>Template:Cite journal</ref> Homologs of cell adhesion, neuropeptide and glycosphingolipid metabolism genes are present in the genome.
S. rosetta has a sexual life cycle and transitions between haploid and diploid stages.<ref name="pmid24139741" /> In response to nutrient limitation, haploid cultures of S. rosetta become diploid. This ploidy shift coincides with mating during which small, flagellated cells fuse with larger flagellated cells. There is also evidence of historical mating and recombination in S. rosetta.
S. rosetta is induced to undergo sexual reproduction by the marine bacterium Vibrio fischeri.<ref name="pmid28867285" /> A single V. fischeri protein, EroS fully recapitulates the aphrodisiac-like activity of live V. fisheri.
Other genomes
The single-cell amplified genomes of four uncultured marine choanoflagellates, tentatively called UC1–UC4, were sequenced in 2019. The genomes of UC1 and UC4 are relatively complete.<ref>Template:Cite journal</ref>
Transcriptomes
Template:Anchor
An EST dataset from Monosiga ovata was published in 2006.<ref name="Snell et al. 2006">Template:Cite journal</ref> The major finding of this transcriptome was the choanoflagellate Hoglet domain and shed light on the role of domain shuffling in the evolution of the Hedgehog signaling pathway. M. ovata has at least four eukaryotic meiotic genes.<ref name="pmid20015185" />
Template:Anchor
The transcriptome of Stephanoeca diplocostata was published in 2013. This first transcriptome of a loricate choanoflagellate<ref name=Marron2013/> led to the discovery of choanoflagellate silicon transporters. Subsequently, similar genes were identified in a second loricate species, Diaphanoeca grandis. Analysis of these genes found that the choanoflagellate silicon transporters show homology to the SIT-type silicon transporters of diatoms and have evolved through horizontal gene transfer.
An additional 19 transcriptomes were published in 2018. A large number of gene families previously thought to be animal-only were found.<ref>Template:Cite journal</ref>
.png){kind=link}
