Cryptophyte Algae Research Articles

Quantum design for a light trap

The photosynthetic apparatus of cryptophyte algae is odd — its pigments are farther apart than is expected for efficient functioning. A study into how this apparatus works so well finds quantum effects at play.

Transcriptome analysis reveals nuclear-encoded proteins for the maintenance of temporary plastids in the dinoflagellate Dinophysis acuminata

BackgroundDinophysis is exceptional among dinoflagellates, possessing plastids derived from cryptophyte algae. Although Dinophysis can be maintained in pure culture for several months, the genus is mixotrophic and needs to feed either to acquire plastids (a process known as kleptoplastidy) or obtain growth factors necessary for plastid maintenance. Dinophysis does not feed directly on cryptophyte algae, but rather on a ciliate (Myrionecta rubra) that has consumed the cryptophytes and retained their plastids. Despite the apparent absence of cryptophyte nuclear genes required for plastid function, Dinophysis can retain cryptophyte plastids for months without feeding.ResultsTo determine if this dinoflagellate has nuclear-encoded genes for plastid function, we sequenced cDNA from Dinophysis acuminata, its ciliate prey M. rubra, and the cryptophyte source of the plastid Geminigera cryophila. We identified five proteins complete with plastid-targeting peptides encoded in the nuclear genome of D. acuminata that function in photosystem stabilization and metabolite transport. Phylogenetic analyses show that the genes are derived from multiple algal sources indicating some were acquired through horizontal gene transfer.ConclusionsThese findings suggest that D. acuminata has some functional control of its plastid, and may be able to extend the useful life of the plastid by replacing damaged transporters and protecting components of the photosystem from stress. However, the dearth of plastid-related genes compared to other fully phototrophic algae suggests that D. acuminata does not have the nuclear repertoire necessary to maintain the plastid permanently.

Physical origins and models of energy transfer in photosynthetic light-harvesting

We perform a quantitative comparison of different energy transfer theories, i.e. modified Redfield, standard and generalized Förster theories, as well as combined Redfield-Förster approach. Physical limitations of these approaches are illustrated and critical values of the key parameters indicating their validity are found. We model at a quantitative level the spectra and dynamics in two photosynthetic antenna complexes: in phycoerythrin 545 from cryptophyte algae and in trimeric LHCII complex from higher plants. These two examples show how the structural organization determines a directed energy transfer and how equilibration within antenna subunits and migration between subunits are superimposed.

Nucleomorph Genomes

Nucleomorphs are the remnant nuclei of algal endosymbionts in cryptophytes and chlorarachniophytes, two evolutionarily distinct unicellular eukaryotic lineages that acquired photosynthesis secondarily by the engulfment of red and green algae, respectively. At less than one million base pairs in size, nucleomorph genomes are the most highly reduced nuclear genomes known, with three small linear chromosomes and a gene density similar to that seen in prokaryotes. The independent origin of nucleomorphs in cryptophytes and chlorarachniophytes presents an interesting opportunity to study the reductive evolutionary forces that have led to their remarkable convergence upon similar genome architectures and coding capacities. In this article, we review the current state of knowledge with respect to the structure, function, origin, and evolution of nucleomorph genomes across the known diversity of cryptophyte and chlorarachniophyte algae.

Heme oxygenase 2 of the cyanobacterium Synechocystis sp. PCC 6803 is induced under a microaerobic atmosphere and is required for microaerobic growth at high light intensity

Cyanobacteria, red algae, and cryptomonad algae utilize phycobilin chromophores that are attached to phycobiliproteins to harvest solar energy. Heme oxygenase (HO) in these organisms catalyzes the first step in phycobilin formation through the conversion of heme to biliverdin IXalpha, CO, and iron. The Synechocystis sp. PCC 6803 genome contains two open reading frames, ho1 (sll1184) and ho2 (sll1875), whose products have in vitro HO activity. We report that HO2, the protein encoded by ho2, was induced in the cells growing under a microaerobic atmosphere [0.2% (v/v) O(2)], whereas HO1 was constitutively expressed under both aerobic and microaerobic atmospheres. Light intensity did not have an effect on the expression of both the HOs. Cells, in which ho2 was disrupted, were unable to grow microaerobically at a light intensity of 40 micromol m(-2) s(-1), but did grow microaerobically at 10 micromol m(-2) s(-1) light intensity. These cells grew normally aerobically at both light intensities. Comparative analysis of complete cyanobacterial genomes revealed that possession of two HOs is common in cyanobacteria. In phylogenetic analysis of their amino acid sequences, cyanobacterial HO1 and HO2 homologs formed distinct clades. HO sequences of cyanobacteria that have only one isoform were most similar to HO1 sequences. We propose that HO2 might be the more ancient HO homolog that functioned under low O(2) tension, whereas the derived HO1 can better accommodate increased O(2) tension in the environment.

Zebra mussels mediate benthic–pelagic coupling by biodeposition and changing detrital stoichiometry

Summary1. The zebra mussel (Dreissena polymorpha) is one of the most successful invasive species; it has colonised many aquatic systems in Europe and North America with strong impacts on various ecosystem processes. The effect of D. polymorpha filtration on pelagic seston concentrations has been quantified in several studies, but the magnitude and stoichiometry of the transfer of sestonic biomass into benthic detritus by D. polymorpha and the accompanying enrichment of the benthic habitat is still under‐investigated.2. We studied biodeposition by zebra mussels in two series of laboratory experiments with the food algae Cryptomonas erosa and Scenedesmus obliquus. We also measured the year‐round biodeposition rate under natural conditions in the oligotrophic Lake Constance.3. In all experiments, zebra mussel biodeposition was linearly related to seston concentration. In the field, the relationship changed with a seasonal shift in algal composition and lower biodeposition rates during the spring algal bloom.4. For both algal species in laboratory experiments, biodeposited material was depleted in phosphorous at an algal concentration ≤0.6 mg ash‐free dry mass L−1, but not at higher concentrations. This effect was not observed in the field, probably because of high variation in C : N : P stoichiometry.5. By mediating the transfer of pelagic resources into the benthos zebra mussels provide a sufficient amount of detritus for benthic invertebrates, especially during summer. Thus, material biodeposited by the mussels might increase benthic secondary production from pelagic resources, and zebra mussels are important mediators of this flux of organic matter from the pelagic zone into the benthos.

Phycobiliprotein diffusion in chloroplasts of cryptophyte Rhodomonas CS24

Unicellular cryptophyte algae employ antenna proteins with phycobilin chromophores in their photosynthetic machinery. The mechanism of light harvesting in these organisms is significantly different than the energy funneling processes in phycobilisomes utilized by cyanobacteria and red algae. One of the most striking features of cryptophytes is the location of the water-soluble phycobiliproteins, which are contained within the intrathylakoid spaces and are not on the stromal side of the lamellae as in the red algae and cyanobacteria. Studies of mobility of phycobiliproteins at the lumenal side of the thylakoid membranes and how their diffusional behavior may influence the energy funneling steps in light harvesting are reported. Confocal microscopy and fluorescence recovery after photobleaching (FRAP) are used to measure the diffusion coefficient of phycoerythrin 545 (PE545), the primary light harvesting protein of Rhodomonas CS24, in vivo. It is concluded that the diffusion of PE545 in the lumen is inhibited, suggesting possible membrane association or aggregation as a potential source of mobility hindrance.

Immobility of phycobilins in the thylakoid lumen of a cryptophyte suggests that protein diffusion in the lumen is very restricted

The thylakoid lumen is an important photosynthetic compartment which is the site of key steps in photosynthetic electron transport. The fluidity of the lumen could be a major constraint on photosynthetic electron transport rates. We used Fluorescence Recovery After Photobleaching in cells of the cryptophyte alga Rhodomonas salina to probe the diffusion of phycoerythrin in the lumen and chlorophyll complexes in the thylakoid membrane. In neither case was there any detectable diffusion over a timescale of several minutes. This indicates very restricted phycoerythrin mobility. This may be a general feature of protein diffusion in the thylakoid lumen.

The Complete Plastid Genome Sequence of the Secondarily Nonphotosynthetic Alga Cryptomonas paramecium: Reduction, Compaction, and Accelerated Evolutionary Rate

The cryptomonads are a group of unicellular algae that acquired photosynthesis through the engulfment of a red algal cell, a process called secondary endosymbiosis. Here, we present the complete plastid genome sequence of the secondarily nonphotosynthetic species Cryptomonas paramecium CCAP977/2a. The ∼78 kilobase pair (Kbp) C. paramecium genome contains 82 predicted protein genes, 29 transfer RNA genes, and a single pseudogene (atpF). The C. paramecium plastid genome is approximately 50 Kbp smaller than those of the photosynthetic cryptomonads Guillardia theta and Rhodomonas salina; 71 genes present in the G. theta and/or R. salina plastid genomes are missing in C. paramecium. The pet, psa, and psb photosynthetic gene families are almost entirely absent. Interestingly, the ribosomal RNA operon, present as inverted repeats in most plastid genomes (including G. theta and R. salina), exists as a single copy in C. paramecium. The G + C content (38%) is higher in C. paramecium than in other cryptomonad plastid genomes, and C. paramecium plastid genes are characterized by significantly different codon usage patterns and increased evolutionary rates. The content and structure of the C. paramecium plastid genome provides insight into the changes associated with recent loss of photosynthesis in a predominantly photosynthetic group of algae and reveals features shared with the plastid genomes of other secondarily nonphotosynthetic eukaryotes.

Evolutionary Dynamics of Light-Independent Protochlorophyllide Oxidoreductase Genes in the Secondary Plastids of Cryptophyte Algae

Plastid genes encoding light-independent protochlorophyllide oxidoreductase (LIPOR) subunits were isolated from cryptophyte algae, the first example of such genes in plastids of secondary endosymbiotic origin. The presence of functional and nonfunctional copies of LIPOR genes in cryptophytes suggests that light-independent chlorophyll biosynthesis is a nonessential pathway in these organisms.

Multiple Gene Phylogenies Support the Monophyly of Cryptomonad and Haptophyte Host Lineages

Cryptomonad algae acquired their plastids by the secondary endosymbiotic uptake of a eukaryotic red alga. Several other algal lineages acquired plastids through such an event [1], but cryptomonads are distinguished by the retention of a relic red algal nucleus, the nucleomorph [2]. The nucleomorph (and its absence in other lineages) can reveal a great deal about the process and history of endosymbiosis, but only if we know the relationship between cryptomonads and other algae, and this has been controversial. Several recent analyses have suggested a relationship between plastids of cryptomonads and some or all other red alga-containing lineages [3-6], but we must also know whether host nuclear genes mirror this relationship to determine the number of endosymbiotic events, and this has not been demonstrated. We have carried out an expressed sequence tag (EST) survey of the cryptomonad Guillardia theta. Phylogenetic analyses of 102 orthologous nucleus-encoded proteins (18,425 amino acid alignment positions) show a robust sister-group relationship between cryptomonads and the haptophyte algae, which also have a red secondary plastid. This relationship demonstrates that loss of nucleomorphs must have taken place in haptophytes independently of any other red alga-containing lineages and that the ancestor of both already contained a red algal endosymbiont.

Competition between two planktonic rotifer species at different temperatures: an experimental test

Summary1. The effect of temperature on the outcome of resource competition between two planktonic rotifers (Synchaeta pectinata and Brachionus calyciflorus) was investigated in laboratory experiments. In addition to the competition experiments, several physiological variables and their temperature‐dependence were characterised, including ingestion rate and starvation tolerance.2. Because of a lower threshold food level (TFL) for population growth for the food algae Cryptomonas erosa, Synchaeta was predicted to be the superior competitor at low temperatures (12 °C). In contrast, Brachionus had a lower TFL at 20 °C and was predicted to be competitively superior at this temperature.3. In both rotifer species, ingestion rates increased with temperature, but the increase was much more pronounced in Brachionus. Ingestion rates of Brachionus at temperatures from 8 to 24 °C were always higher than in Synchaeta (up to 4.6‐fold).4. Starvation resistance reduced with temperature in both rotifer species. At all temperatures investigated (12, 16 and 20 °C) Brachionus could survive starvation for longer than Synchaeta. This difference was strongest at 12 °C (5.8 days versus 2.5 days).5. In the first competition experiment, food was supplied at 48 h‐intervals. Brachionus displaced Synchaeta at both experimental temperatures (12 and 20 °C). Competitive exclusion of Synchaeta at the lower temperature was probably because of large fluctuations in algal densities that resulted from the long intervals between feeding, a condition that favoured Brachionus because of its higher starvation resistance.6. In the second competition experiment, one third of the food suspension was renewed every 8 h, resulting in a much better approximation to a continuous resource supply. At 12 °C Synchaeta and Brachionus coexisted for more than 1 month and the densities of both rotifer species were significantly lower in the presence of their competitor. In contrast to expectations, Brachionus was able to persist even when Cryptomonas concentrations fell below its TFL. This was probably because Brachionus was using detritus and associated bacteria as additional food sources, which were present in the cultures during the later phase of the experiment.7. Autocorrelation analysis of the temporal changes in egg ratios revealed significant periodic cycles in Synchaeta during the second competition experiment. A possible explanation for this is the fecundity schedule of Synchaeta, in which reproduction is highly concentrated in a few age classes. According to demographic theory, such a life cycle feature can cause slower convergence to a stable age distribution.

Novel Nucleomorph Genome Architecture in the Cryptomonad Genus Hemiselmis

Cryptomonads are ubiquitous aquatic unicellular eukaryotes that acquired photosynthesis through the uptake and retention of a red algal endosymbiont. The nuclear genome of the red alga persists in a highly reduced form termed a nucleomorph. The nucleomorph genome of the model cryptomonad Guillardia theta has been completely sequenced and is a mere 551 kilobases (kb) in size, spread over three chromosomes. The presence of three chromosomes appears to be a universal characteristic of nucleomorph genomes in cryptomonad algae as well as in the chlorarachniophytes, an unrelated algal lineage with a nucleomorph and plastid genome derived from a green algal endosymbiont. Another feature of nucleomorph genomes in all cryptomonads and chlorarachniophytes examined thus far is the presence of subtelomeric ribosomal DNA (rDNA) repeats at the ends of each chromosome. Here we describe the first exception to this canonical nucleomorph genome architecture in the cryptomonad Hemiselmis rufescens CCMP644. Using pulsed-field gel electrophoresis (PFGE), we estimate the size of the H. rufescens nucleomorph genome to be approximately 580 kb, slightly larger than the G. theta genome. Unlike the situation in G. theta and all other known cryptomonads, sub-telomeric repeats of the rDNA cistron appear to be absent on both ends of the second largest chromosome in H. rufescens and two other members of this genus. Southern hybridizations using a variety of nucleomorph protein gene probes against PFGE-separated H. rufescens chromosomes indicate that recombination has been a major factor in shaping the karyotype and genomic structure of cryptomonad nucleomorphs.

The photophysics of cryptophyte light-harvesting

Recent studies of the optical properties and the critical role of phycobiliproteins in the absorption of green light for photosynthesis in cryptophyte algae ( Rhodomonas CS24 and Chroomonas CCMP270) are reviewed. Investigations of two different isolated proteins, phycoerythrin 545 (PE545) and phycocyanin 645 (PC645), whose crystal structures are known to 0.97 and 1.4 Å resolution respectively, are described. Steady-state spectroscopic measurements, including polarization anisotropy and circular dichroism, are used in combination with ultrafast transient grating and transient absorption techniques to elucidate a detailed picture of resonance energy transfer within the light-harvesting proteins. Quantum chemical calculations are employed to estimate phycobilin excited states, and generate transition density cubes which are used to calculate accurately the electronic coupling between the chromophores in PE545 and PC645. Energy transfer dynamics are examined using the generalized Förster theory. Kinetic models for energy transfer dynamics in both proteins are presented for comparison. Investigations of energy transfer from phycoerythrin 545 to chlorophyll-containing light harvesting complexes and photosystems in the intact algae Rhodomonas CS24 and Chroomonas CCMP270 are also reported.

Insight into the Diversity and Evolution of the Cryptomonad Nucleomorph Genome

The cryptomonads are an enigmatic group of marine and freshwater unicellular algae that acquired their plastids through the engulfment and retention of a eukaryotic ("secondary") endosymbiont. Together with the chlorarachniophyte algae, the cryptomonads are unusual in that they have retained the nucleus of their endosymbiont in a miniaturized form called a nucleomorph. The nucleomorph genome of the cryptomonad Guillardia theta has been completely sequenced and with only three chromosomes and a total size of 551 kb, is a model of nuclear genome compaction. Using this genome as a reference, we have investigated the structure and content of nucleomorph genomes in a wide range of cryptomonad algae. In this study, we have sequenced nine new cryptomonad nucleomorph 18S ribosomal DNA (rDNA) genes and four heat shock protein 90 (hsp90) gene fragments, and using pulsed-field gel electrophoresis and Southern hybridizations, have obtained nucleomorph genome size estimates for nine different species. We also used long-range polymerase chain reaction to obtain nucleomorph genomic fragments from Hanusia phi CCMP325 and Proteomonas sulcata CCMP704 that are syntenic with the subtelomeric region of nucleomorph chromosome I in G. theta. Our results indicate that (1) the presence of three chromosomes is a common feature of the nucleomorph genomes of these organisms, (2) nucleomorph genome size varies dramatically in the cryptomonads examined, (3) unidentified cryptomonad species CCMP1178 has the largest nucleomorph genome identified to date at approximately 845 kb, (4) nucleomorph genome size reductions appear to have occurred multiple times independently during cryptomonad evolution, (5) the relative positions of the 18S rDNA, ubc4, and hsp90 genes are conserved in three different cryptomonad genera, and (6) interchromosomal recombination appears to be rapidly changing the size and sequence of a repetitive subtelomeric region of the nucleomorph genome between the 18S rDNA and ubc4 loci. These results provide a glimpse into the genetic diversity of nucleomorph genomes in cryptomonads and set the stage for more comprehensive sequence-based studies in closely and distantly related taxa.

Distinctive organization of genes for light-harvesting proteins in the cryptophyte alga Rhodomonas

Cryptophyte algae contain two kinds of light-harvesting protein, phycobiliproteins and chlorophyll a,c-binding proteins. The β subunit of the phycobiliprotein phycoerythrin (PE) is encoded in the chloroplast. Genes for the other PE polypeptides are located in the nucleus but little is known of their organization. We cloned and sequenced six cpeA genes encoding the phycoerythrin α subunit from a genomic library of the cryptophyte Rhodomonas CS24. Derived peptide sequences of the cpeA genes show that α subunits occur in at least two forms, a longer α 1 form and a shorter α 2 form. Remarkably, all six cpeA genes occur in divergent pairs encoding one α 1 and one α 2 subunit. Four cac genes encoding chlorophyll a,c-binding proteins were cloned and sequenced and also found to occur in divergent pairs comprising one cac1 and one cac2 gene. Inspection of the predicted targeting sequences of the α 1 and α 2 phycoerythrin polypeptides shows that only the α 1 polypeptides have a thylakoid lumen targeting sequence, corresponding to the TAT pathway. Given the previously reported lack of a lumen-targeting sequence on the β subunit, we propose a novel import mechanism in which the entire α 1α 2ββ phycoerythrin complex is assembled in the stroma and transported into the thylakoid under the direction of the single targeting sequence on the α 1 protein. The FAP motif implicated in plastid targeting in diatoms appears to be conserved in this cryptophyte.

CONTROLS OF ALGAL ABUNDANCE AND COMMUNITY COMPOSITION DURING ECOSYSTEM STATE CHANGE

Shallow lake ecosystems can shift between clear-water, macrophyte-rich conditions and turbid states with abundant phytoplankton. However, little is known about the controls of algal community composition and primary production before, during, and after ecosystem state change, because long time series that monitor biological change through the transition are scarce. Using proxy data sets derived from sediment cores from two shallow hypertrophic lakes in Denmark, variance-partitioning analysis (VPA) was used to determine the relative importance of changes in total phosphorus (diatom inferred), planktivorous fish density (zooplankton inferred), and submerged macrophyte communities (as macrofossil abundance) as determinants of algal abundance and community composition (as sedimentary pigments) over ecosystem state transitions since 1750 (CE) for Lake Lading and 1900 for Lake Søbygaard. Past variation in densities of planktivorous fish explained 12.3% and 18.2% of historical algal community change in lakes Lading and Søbygaard, respectively, while a further 22.3% and 6% of algal variability was explained by variation in macrophyte abundance. Total phosphorus (TP) alone explained nonsignificant amounts of variance (1.5%, 3.6%) but had a significant effect in combination with macrophytes and fish (27%, 13.4%). State transitions occurred ca. 1940 but were preceded by increases in benthic diatoms and macrophytes, suggesting that transitions were gradual rather than instantaneous. In contrast, green, colonial blue-green, and cryptophyte algae were abundant only during turbid states after ca. 1960 and were correlated to changes in planktivorous fish or fish–TP interactions. Contrary to expectations, the shift from predominantly benthic to pelagic algal production represented only a change in habitat and did not result in an increase in total abundance of primary producers.

Phagotrophy in chlorarachniophyte algae: implications for eukaryotic genome evolution

The chlorarachniophytes are a group of marine amoeboflagellate algae that acquired photosynthesis by secondary endosymbiosis. Together with cryptomonad algae, the chlorarachniophytes are unusual in that they have retained the nucleus of their eukaryotic endosymbiont. The host component of chlorarachniophyte cells is derived from a member of the Cercozoa while the chlorarachnion endosymbiont is green algal in origin. In addition to being able to photosynthesize, chlorarachniophytes have been shown to ingest other algae and bacteria, although the exact nature of engulfment, and the frequency with which this occurs in nature is unknown. Here, I present a light micrographic study of phagotrophy in three chlorarachniophytes, Chlorarachnion reptans, Lotharella sp. 240 and Gymnochlora stellata. These organisms readily ingest a wide variety of algae, including cryptomonads, haptophytes and green algae. The mechanism of capture and ingestion involves adhesion to, fusion with, and digestion within the pseudopodia formed by chlorarachniophyte cells. The implications of phagotrophy on the chlorarachniophyte nuclear genome are discussed.

Role of feeding in growth and photophysiology of Myrionecta rubra

Myrionecta rubra is a cosmopolitan estuarine and neritic ciliate, known to cause 'red- water' blooms. The current study was conducted to achieve a better understanding of the relationship of photosynthetic performance and growth with feeding on cryptophyte algae in M. rubra. During the experiment the cryptophyte Teleaulax acuta was introduced for 2 consecutive growth periods (14 d each) and the cultures were then starved during 4 additional periods. In both high light (HL) and low light (LL) treatments, a significant decrease in per capita growth rates (µ) was observed over time (p < 0.05) in the absence of new prey. In the LL treatment, chlorophyll a content (chl a cell -1 ), photosyn- thetic capacity (Pmax (cell)), and photosynthetic efficiency (αcell) increased after feeding and then declined during starvation. In the HL treatment, chl cell -1 and αcell also increased and then declined after feeding; however, Pmax (cell) showed only a slight decrease with starvation. In both treatments, M. rubra appeared to undergo an acclimation-like response following declines in chl cell -1 , with increases in Pmax (chl) and the light saturation parameter (Ik). While photosynthetic efficiency declined during starvation, overall photosynthetic capacity appeared to become uncoupled with growth over time. M. rubra demonstrated a high capacity for chl production (7 to 10 µg chl a ml -1 d -1 ) after feed- ing, which decreased during starvation. Declines in growth and photosynthetic parameters coincided with the loss of prey nuclei from M. rubra cells, implicating a possible functional role for retained prey nuclei. These data show that M. rubra can function phototrophically for extended periods without ingesting prey, but that feeding is periodically required for optimal growth and photosynthe- sis, especially in high light.

A molecular timeline for the origin of photosynthetic eukaryotes.

The appearance of photosynthetic eukaryotes (algae and plants) dramatically altered the Earth's ecosystem, making possible all vertebrate life on land, including humans. Dating algal origin is, however, frustrated by a meager fossil record. We generated a plastid multi-gene phylogeny with Bayesian inference and then used maximum likelihood molecular clock methods to estimate algal divergence times. The plastid tree was used as a surrogate for algal host evolution because of recent phylogenetic evidence supporting the vertical ancestry of the plastid in the red, green, and glaucophyte algae. Nodes in the plastid tree were constrained with six reliable fossil dates and a maximum age of 3,500 MYA based on the earliest known eubacterial fossil. Our analyses support an ancient (late Paleoproterozoic) origin of photosynthetic eukaryotes with the primary endosymbiosis that gave rise to the first alga having occurred after the split of the Plantae (i.e., red, green, and glaucophyte algae plus land plants) from the opisthokonts sometime before 1,558 MYA. The split of the red and green algae is calculated to have occurred about 1,500 MYA, and the putative single red algal secondary endosymbiosis that gave rise to the plastid in the cryptophyte, haptophyte, and stramenopile algae (chromists) occurred about 1,300 MYA. These dates, which are consistent with fossil evidence for putative marine algae (i.e., acritarchs) from the early Mesoproterozoic (1,500 MYA) and with a major eukaryotic diversification in the very late Mesoproterozoic and Neoproterozoic, provide a molecular timeline for understanding algal evolution.