Chromalveolates and the Evolution of Plastids by Secondary Endosymbiosis1

ABSTRACT. The establishment of a new plastid organelle by secondary endosymbiosis represents a series of events of massive complexity, and yet we know it has taken place multiple times because both green and red algae have been taken up by other eukaryotic lineages. Exactly how many times these events have succeeded, however, has been a matter of debate that significantly impacts how we view plastid evolution, protein targeting, and eukaryotic relationships. On the green side it is now largely accepted that two independent events led to plastids of euglenids and chlorarachniophytes. How many times red algae have been taken up is less clear, because there are many more lineages with red alga‐derived plastids (cryptomonads, haptophytes, heterokonts, dinoflagellates and apicomplexa) and the relationships between these lineages are less clear. Ten years ago, Cavalier‐Smith proposed that these plastids were all derived from a single endosymbiosis, an idea that was dubbed the chromalveolate hypothesis. No one observation has yet supported the chromalveolate hypothesis as a whole, but molecular data from plastid‐encoded and plastid‐targeted proteins have provided strong support for several components of the overall hypothesis, and evidence for cryptic plastids and new photosynthetic lineages (e.g. Chromera) have transformed our view of plastid distribution within the group. Collectively, these data are most easily reconciled with a single origin of the chromalveolate plastids, although the phylogeny of chromalveolate host lineages (and potentially Rhizaria) remain to be reconciled with this plastid data.     

A cryptic algal group unveiled: a plastid biosynthesis pathway in the oyster parasite Perkinsus marinus

Plastids are widespread in plant and algal lineages. They are also exploited by some nonphotosynthetic protists, including malarial parasites, to support their diverse modes of life. However, cryptic plastids may exist in other nonphotosynthetic protists, which could be important in studies on the diversity and evolution of plastids. The parasite Perkinsus marinus, which causes mass mortality in oyster farms, is a nonphotosynthetic protist that is phylogenetically related to plastid-bearing dinoflagellates and apicomplexans. In this study, we searched for P. marinus methylerythritol phosphate (MEP) pathway genes, responsible for de novo isoprenoid synthesis in plastids, and determined the full-length gene sequences for 6 of 7 of these genes. Phylogenetic analyses revealed that each P. marinus gene clusters with orthologs from plastid-bearing eukaryotes, which have MEP pathway genes with essentially the same mosaic pattern of evolutionary origin. A new analytical method called sliding-window iteration of TargetP was developed to examine the distribution of targeting preferences. This analysis revealed that the sequenced genes encode bipartite targeting peptides that are characteristic of proteins targeted to secondary plastids originating from endosymbiosis of eukaryotic algae. These results support our idea that Perkinsus is a cryptic algal group containing nonphotosynthetic secondary plastids. In fact, immunofluorescent microscopy indicated that 1 of the MEP pathway enzymes, 1-deoxy-D-xylulose 5-phosphate reductoisomerase, was localized to small compartments near mitochondrion, which are possibly plastids. This tiny organelle seems to contain very low quantities of DNA or may even lack DNA entirely. The MEP pathway genes are a useful tool for investigating plastid evolution in both of the photosynthetic and nonphotosynthetic eukaryotes and led us to propose the hypothesis that ancestral "chromalveolates" harbored plastids before a secondary endosymbiotic event.     

Gene replacement of fructose-1,6-bisphosphate aldolase supports the hypothesis of a single photosynthetic ancestor of chromalveolates

Plastids (photosynthetic organelles of plants and algae) are known to have spread between eukaryotic lineages by secondary endosymbiosis, that is, by the uptake of a eukaryotic alga by another eukaryote. But the number of times this has taken place is controversial. This is particularly so in the case of eukaryotes with plastids derived from red algae, which are numerous and diverse. Despite their diversity, it has been suggested that all these eukaryotes share a recent common ancestor and that their plastids originated in a single endosymbiosis, the so-called "chromalveolate hypothesis." Here we describe a novel molecular character that supports the chromalveolate hypothesis. Fructose-1,6-bisphosphate aldolase (FBA) is a glycolytic and Calvin cycle enzyme that exists as two nonhomologous types, class I and class II. Red algal plastid-targeted FBA is a class I enzyme related to homologues from plants and green algae, and it would be predicted that the plastid-targeted FBA from algae with red algal secondary endosymbionts should be related to this class I enzyme. However, we show that plastid-targeted FBA of heterokonts, cryptomonads, haptophytes, and dinoflagellates (all photosynthetic chromalveolates) are class II plastid-targeted enzymes, completely unlike those of red algal plastids. The chromalveolate enzymes form a strongly supported group in FBA phylogeny, and their common possession of this unexpected plastid characteristic provides new evidence for their close relationship and a common origin for their plastids.     

Nucleus-encoded,plastid-targeted glyceraldehyde-3-phosphate dehydrogenase (GAPDH) indicates a single origin for chromalveolate plastids

Plastids (the photosynthetic organelles of plants and algae) originated through endosymbiosis between a cyanobacterium and a eukaryote and subsequently spread to other eukaryotes by secondary endosymbioses between two eukaryotes. Mounting evidence favors a single origin for plastids of apicomplexans, cryptophytes, dinoflagellates, haptophytes, and heterokonts (together with their nonphotosynthetic relatives, termed chromalveolates), but so far, no single molecular marker has been described that supports this common origin. One piece of evidence comes from plastid-targeted glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which originated by a gene duplication of the cytosolic form. However, no plastid GAPDH has been characterized from haptophytes, leaving an important piece of the puzzle missing. We have sequenced genes encoding cytosolic, mitochondrion-targeted, and plastid-targeted GAPDH proteins from a number of haptophytes and heterokonts and found haptophyte homologs that branch within a strongly supported clade of chromalveolate plastid-targeted genes, being more closely related to an apicomplexan homolog than was expected. The evolution of plastid-targeted GAPDH supports red algal ancestry of apicomplexan plastids and raises a number of questions about the importance of plastid loss and the possibility of cryptic plastids in nonphotosynthetic lineages such as ciliates.     

PHYLOGENOMICS AND SECONDARY PLASTIDS: A LOOK BACK AND A LOOK AHEAD1

Despite their importance to evolution, ecology, and cell biology, eukaryotes that acquired plastids through secondary endosymbiosis remain poorly studied from a genomic standpoint. Chromalveolata, a eukaryotic supergroup proposed to have descended from a heterotrophic eukaryote that acquired a red algal plastid by secondary endosymbiosis, includes four major lineages (alveolates, cryptophytes, haptophytes, and heterokonts). The chromalveolates exhibit remarkable diversity of cellular organization, and the available data suggest that they exhibit equal diversity in their genome organization. One of the most obvious differences in cellular organization is the retention of a highly reduced red algal nucleus in cryptophytes (also known as cryptomonads), but there are other major differences among chromalveolate lineages, including the loss of photosynthesis in multiple lineages. Although the hypothesis of chromalveolate monophyly is appealing, there is limited support for the hypothesis from nuclear genes, and questions have even been raised about the monophyly of chromalveolate plastids. Evidence for the chromalveolate hypothesis from large‐scale nuclear data sets is reviewed, and alternative hypotheses are described. The potential for integrating information from chromalveolate genomics into functional genomics is described, emphasizing both the methodological challenges and the opportunities for future phylogenomic analyses of these groups.     

The endosymbiotic origin,diversification and fate of plastids

Plastids and mitochondria each arose from a single endosymbiotic event and share many similarities in how they were reduced and integrated with their host. However, the subsequent evolution of the two organelles could hardly be more different: mitochondria are a stable fixture of eukaryotic cells that are neither lost nor shuffled between lineages, whereas plastid evolution has been a complex mix of movement, loss and replacement. Molecular data from the past decade have substantially untangled this complex history, and we now know that plastids are derived from a single endosymbiotic event in the ancestor of glaucophytes, red algae and green algae (including plants). The plastids of both red algae and green algae were subsequently transferred to other lineages by secondary endosymbiosis. Green algal plastids were taken up by euglenids and chlorarachniophytes, as well as one small group of dinoflagellates. Red algae appear to have been taken up only once, giving rise to a diverse group called chromalveolates. Additional layers of complexity come from plastid loss, which has happened at least once and probably many times, and replacement. Plastid loss is difficult to prove, and cryptic, non-photosynthetic plastids are being found in many non-photosynthetic lineages. In other cases, photosynthetic lineages are now understood to have evolved from ancestors with a plastid of different origin, so an ancestral plastid has been replaced with a new one. Such replacement has taken place in several dinoflagellates (by tertiary endosymbiosis with other chromalveolates or serial secondary endosymbiosis with a green alga), and apparently also in two rhizarian lineages: chlorarachniophytes and Paulinella (which appear to have evolved from chromalveolate ancestors). The many twists and turns of plastid evolution each represent major evolutionary transitions, and each offers a glimpse into how genomes evolve and how cells integrate through gene transfers and protein trafficking.     

Rethinking plastid evolution

How easy is it to acquire an organelle? How easy is it to lose one? Michael Gray considers the latest evidence in this regard concerning the chromalveolates.How easy is it to acquire an organelle? How easy is it to lose one? These questions underpin the current debate about the evolution of the plastid—that is, chloroplast—the organelle of photosynthesis in eukaryotic cells.The origin of the plastid has been traced to an endosymbiosis between a eukaryotic host cell and a cyanobacterial symbiont, the latter gradually ceding genetic control to the former through endosymbiotic gene transfer (EGT). The resulting organelle now relies for its biogenesis and function on the expression of a small set of genes retained in the shrunken plastid genome, as well as a much larger set of transferred nuclear genes encoding proteins synthesized in the cytosol and imported into the organelle.This scenario accounts for the so-called primary plastids in green algae and their land plant relatives, in red algae and in glaucophytes, which together comprise Plantae (or Archaeplastida)—one of five or six recognized eukaryotic supergroups (Adl et al, 2005). In other algal types, plastids are ‘second-hand''—they have been acquired not by taking up a cyanobacterium, but by taking up a primary-plastid-containing eukaryote (sometimes a green alga, sometimes a red alga) to produce secondary plastids. In most of these cases, all that remains of the eukaryotic symbiont is its plastid; the genes coding for plastid proteins have moved from the endosymbiont to the host nucleus. A eukaryotic host—which may or may not itself have a plastid—might also take up a secondary-plastid symbiont (generating tertiary plastids), or a secondary-plastid host might take up a primary-plastid symbiont. You get the picture: plastid evolution is complicated!Several excellent recent reviews present expanded accounts of plastid evolution (Reyes-Prieto et al, 2007; Gould et al, 2008; Archibald, 2009; Keeling, 2009). Here, I focus on one particular aspect of plastid evolutionary theory, the ‘chromalveolate hypothesis'', proposed in 1999 by Tom Cavalier-Smith (1999).The chromalveolate hypothesis seeks to explain the origin of chlorophyll c-containing plastids in several eukaryotic groups, notably cryptophytes, alveolates (ciliates, dinoflagellates and apicomplexans), stramenopiles (heterokonts) and haptophytes—together dubbed the ‘chromalveolates''. The plastid-containing members of this assemblage are mainly eukaryotic algae with secondary plastids that were acquired through endosymbiosis with a red alga. The question is: how many times did such an endosymbiosis occur within the chromalveolate grouping?A basic tenet of the chromalveolate hypothesis is that the evolutionary conversion of an endosymbiont to an organelle should be an exceedingly rare event, and a hard task for a biological system to accomplish, because the organism has to ‘learn'' how to target a large number of nucleus-encoded proteins—the genes of many of which were acquired by EGT—back into the organelle. Our current understanding of this targeting process is detailed in the reviews cited earlier. Suffice it to say that the evolutionary requirements appear numerous and complex—sufficiently so that the chromalveolate hypothesis posits that secondary endosymbiosis involving a red alga happened only once, in a common ancestor of the various groups comprising the chromalveolates.Considerable molecular and phylogenetic data have been marshalled over the past decade in support of the chromalveolate hypothesis; however, no single data set specifically unites all chromalveolates, even though there is compelling evidence for various subgroup relationships (Keeling, 2009). Moreover, within the proposed chromalveolate assemblage, plastid-containing lineages are interspersed with plastid-lacking ones—for example, ciliates in the alveolates, and oomycetes such as Phytophthora in the stramenopiles. The chromalveolate hypothesis rationalizes such interspersion by assuming that the plastid was lost at some point during the evolution of the aplastidic lineages. The discovery in such aplastidic lineages of genes of putatively red algal origin, and in some cases suggestive evidence of a non-photosynthetic plastid remnant, would seem to be consistent with this thesis, although these instances are still few and far between.In this context, two recent papers are notable in that the authors seek to falsify, through rigorous testing, several explicit predictions of the chromalveolate hypothesis—and in both cases they succeed in doing so. Because molecular phylogenies have failed to either robustly support or robustly disprove the chromalveolate hypothesis, Baurain et al (2010) devised a phylogenomic falsification of the chromalveolate hypothesis that does not depend on full resolution of the eukaryotic tree. They argued that if the chlorophyll c-containing chromalveolate lineages all derive from a single red algal ancestor, then similar amounts of sequence from the three compartments should allow them to recover chromalveolate monophyly in all cases. The statistical support levels in their analysis refuted this prediction, leading them to “reject the chromalveolate hypothesis as falsified in favour of more complex evolutionary scenarios involving multiple higher order eukaryote–eukaryote endosymbioses”.In another study, Stiller et al (2009) applied statistical tests to several a priori assumptions relating to the finding of genes of supposed algal origin in the aplastidic chromalveolate taxon Phytophthora. These authors determined that the signal from these genes “is inconsistent with the chromalveolate hypothesis, and better explained by alternative models of sequence and genome evolution”.So, is the chromalveolate hypothesis dead? These new studies are certainly the most serious challenge yet. Additional data, including genome sequences of poorly characterized chromalveolate lineages, will no doubt augment comparative phylogenomic studies aimed at evaluating the chromalveolate hypothesis—which these days is looking decidedly shaky.     

Multiple losses of photosynthesis in Nitzschia (Bacillariophyceae)

In order to obtain insights into the evolution of colorless (apochlorotic) diatoms, we investigated newly established apochlorotic strains of Nitzschia spp. using light and electron microscopy and molecular phylogenetic analyses. Fluorescence microscopic observations demonstrated that the apochlorotic diatoms lack chlorophylls. Transmission electron microscopy of two apochlorotic strains also demonstrated that their plastids lacked thylakoids; instead, having four‐membrane‐bound organelles without thylakoids, similar to nonphotosynthetic plastid remnants. From the apochlorotic strains, we also found plastid small subunit rRNA genes that were unusually long branched in phylogenetic analyses, as observed in other nonphotosynthetic plastids. Molecular phylogenetic analysis of the nucleus‐encoded large subunit rRNA genes showed eight distinct lineages for apochlorotic diatoms. The eight apochlorotic lineages were not monophyletic, suggesting that the loss of photosynthesis took place multiple times independently within Nitzschia. Several diatoms, including Nitzschia spp., are mixotrophic, which is an expected mode of nutrition that would help explain the evolutionary switch from a photosynthetic lifestyle to a heterotrophic lifestyle.     

Review

Most photosynthetic dinoflagellates harbour the peridinin plastid. This plastid is surrounded by three membranes and its characteristic pigments are chlorophyll c and the carotenoid peridinin. The evolutionary origin of this peculiar plastid remains controversial and is hotly debated. On the recently published tree of concatenated plastid-encoded proteins, dinoflagellates emerge from within the Chromista (clade containing cryptophytes, heterokonts, and haptophytes) and cluster specifically with Heterokonta. These data inspired a new version of the ‘chromalveolate’ model, according to which the peridinin plastid evolved by ‘descent with modification’ from a heterokont-like plastid that had been acquired from a rhodophyte by an ancestral chromalveolate. However, this model of plastid evolution encounters serious obstacles. Firstly, the heterokont plastid is surrounded by four membranes, which means that the ancestral peridinin plastid must have lost one of these primary membranes. However, such a loss could be traumatic, because it could potentially disturb protein import into and/or within the plastid. Secondly, on the phylogenetic tree of Dinoflagellata and Heterokonta, the first to diverge are not plastid, but heterotrophic, aplastidal taxa. Thus, when accepting the single origin of the heterokont and peridinin plastids, we would have to postulate multiple plastid losses, but such a scenario is highly doubtful when the numerous non-photosynthetic functions of plastids and their existence in heterotrophic protists, including parasitic lineages, are considered. Taking these obstacles into account, we suggest an alternative interpretation of the concatenated tree of plastid-encoded proteins. According to our hypothesis, the peridinin plastid evolved from a heterokont alga through tertiary endosymbiosis.     

What Happened before Losses of Photosynthesis in Cryptophyte Algae?

In many lineages of algae and land plants, photosynthesis was lost multiple times independently. Comparative analyses of photosynthetic and secondary nonphotosynthetic relatives have revealed the essential functions of plastids, beyond photosynthesis. However, evolutionary triggers and processes that drive the loss of photosynthesis remain unknown. Cryptophytes are microalgae with complex plastids derived from a red alga. They include several secondary nonphotosynthetic species with closely related photosynthetic taxa. In this study, we found that a cryptophyte, Cryptomonas borealis, is in a stage just prior to the loss of photosynthesis. Cryptomonas borealis was mixotrophic, possessed photosynthetic activity, and grew independent of light. The plastid genome of C. borealis had distinct features, including increases of group II introns with mobility, frequent genome rearrangements, incomplete loss of inverted repeats, and abundant small/medium/large-sized structural variants. These features provide insight into the evolutionary process leading to the loss of photosynthesis.     

Chlorophyll c-containing plastid relationships based on analyses of a multigene data set with all four chromalveolate lineages

The chlorophyll c-containing algae comprise four major lineages: dinoflagellates, haptophytes, heterokonts, and cryptophytes. These four lineages have sometimes been grouped together based on their pigmentation, but cytological and rRNA data had suggested that they were not a monophyletic lineage. Some molecular data support monophyly of the plastids, while other plastid and host data suggest different relationships. It is uncontroversial that these groups have all acquired plastids from another eukaryote, probably from the red algal lineage, in a secondary endosymbiotic event, but the number and sequence of such event(s) remain controversial. Understanding chlorophyll c-containing plastid relationships is a first step towards determining the number of endosymbiotic events within the chromalveolates. We report here phylogenetic analyses using 10 plastid genes with representatives of all four chromalveolate lineages. This is the first organellar genome-scale analysis to include both haptophytes and dinoflagellates. Concatenated analyses support the monophyly of the chlorophyll c-containing plastids and suggest that cryptophyte plastids are the basal member of the chlorophyll c-containing plastid lineage. The gene psbA, which has at times been used for phylogenetic purposes, was found to differ from the other genes in its placement of the dinoflagellates and the haptophytes, and in its lack of support for monophyly of the green and red plastid lineages. Overall, the concatenated data are consistent with a single origin of chlorophyll c-containing plastids from red algae. However, these data cannot test several key hypothesis concerning chromalveolate host monophyly, and do not preclude the possibility of serial transfer of chlorophyll c-containing plastids among distantly related hosts.     

DO PLASTID‐RELATED CHARACTERS SUPPORT THE CHROMALVEOLATE HYPOTHESIS?1

According to the idea of secondary endosymbiosis, plastids with three and four envelope membranes have evolved from either red or green algal endosymbionts engulfed by phagotrophic protozoans. Although this hypothesis is nowadays commonly accepted, the number of secondary endosymbioses still remains controversial. One of the models, known as the ”chromalveolate” hypothesis, postulates that the 4 membrane‐bound plastids of Chromista and the 3 or 4 membrane‐bound plastids of Alveolata result from a single secondary endosymbiosis involving a rhodophyte as the endosymbiont. Although this model has found many followers, a variety of data clearly contradict it. The ideas that became the direct inspiration for formulation of the “chromalveolate” hypothesis are also now questionable. In this comment, I discuss all these problems in the light of the most recent phylogenetic, cytological, and genomic data.     

NADPH thioredoxin reductase C is localized in plastids of photosynthetic and nonphotosynthetic tissues and is involved in lateral root formation in Arabidopsis

Plastids are organelles present in photosynthetic and nonphotosynthetic plant tissues. While it is well known that thioredoxin-dependent redox regulation is essential for leaf chloroplast function, little is known of the redox regulation in plastids of nonphotosynthetic tissues, which cannot use light as a direct source of reducing power. Thus, the question remains whether redox regulation operates in nonphotosynthetic plastid function and how it is integrated with chloroplasts for plant growth. Here, we show that NADPH-thioredoxin reductase C (NTRC), previously reported as exclusive to green tissues, is also expressed in nonphotosynthetic tissues of Arabidopsis thaliana, where it is localized to plastids. Moreover, we show that NTRC is involved in maintaining the redox homeostasis of plastids also in nonphotosynthetic organs. To test the relationship between plastids of photosynthetic and nonphotosynthetic tissues, transgenic plants were obtained with redox homeostasis restituted exclusively in leaves or in roots, through the expression of NTRC under the control of organ-specific promoters in the ntrc mutant. Our results show that fully functional root amyloplasts are not sufficient for root, or leaf, growth, but fully functional chloroplasts are necessary and sufficient to support wild-type rates of root growth and lateral root formation.     

After the primary endosymbiosis: an update on the chromalveolate hypothesis and the origins of algae with Chl <Emphasis Type="Italic">c</Emphasis>

The chromalveolate hypothesis proposed by Cavalier-Smith (J Euk Microbiol 46:347–366, 1999) suggested that all the algae with chlorophyll c (heterokonts, haptophytes, cryptophytes, and dinoflagellates), as well as the ciliates, apicomplexans, oomycetes, and other non-photosynthetic relatives, shared a common ancestor that acquired a chloroplast by secondary endosymbiosis of a red alga. Much of the evidence from plastid and nuclear genomes supports a red algal origin for plastids of the photosynthetic lineages, but the number of secondary endosymbioses and the number of plastid losses have not been resolved. The issue is complicated by the fact that nuclear genomes are mosaics of genes acquired over a very long time period, not only by vertical descent but also by endosymbiotic and horizontal gene transfer. Phylogenomic analysis of the available whole-genome data has suggested major alterations to our view of eukaryotic evolution, and given rise to alternative models. The next few years may see even more changes once a more representative collection of sequenced genomes becomes available.     

Multiple genes of apparent algal origin suggest ciliates may once have been photosynthetic

Plantae (as defined by Cavalier-Smith, 1981) plastids evolved via primary endosymbiosis whereby a heterotrophic protist enslaved a photosynthetic cyanobacterium. This "primary" plastid spread into other eukaryotes via secondary endosymbiosis. An important but contentious theory in algal evolution is the chromalveolate hypothesis that posits chromists (cryptophytes, haptophytes, and stramenopiles) and alveolates (ciliates, apicomplexans, and dinoflagellates) share a common ancestor that contained a red-algal-derived "secondary" plastid. Under this view, the existence of several later-diverging plastid-lacking chromalveolates such as ciliates and oomycetes would be explained by plastid loss in these lineages. To test the idea of a photosynthetic ancestry for ciliates, we used the 27,446 predicted proteins from the macronuclear genome of Tetrahymena thermophila to query prokaryotic and eukaryotic genomes. We identified 16 proteins of possible algal origin in the ciliates Tetrahymena and Paramecium tetraurelia. Fourteen of these are present in other chromalveolates. Here we compare and contrast the likely scenarios for algal-gene origin in ciliates either via multiple rounds of horizontal gene transfer (HGT) from algal prey or symbionts, or through endosymbiotic gene transfer (EGT) during a putative photosynthetic phase in their evolution.     

The evolutionary history of haptophytes and cryptophytes: phylogenomic evidence for separate origins

An important missing piece in the puzzle of how plastids spread across the eukaryotic tree of life is a robust evolutionary framework for the host lineages. Four assemblages are known to harbour plastids derived from red algae and, according to the controversial chromalveolate hypothesis, these all share a common ancestry. Phylogenomic analyses have consistently shown that stramenopiles and alveolates are closely related, but haptophytes and cryptophytes remain contentious; they have been proposed to branch together with several heterotrophic groups in the newly erected Hacrobia. Here, we tested this question by producing a large expressed sequence tag dataset for the katablepharid Roombia truncata, one of the last hacrobian lineages for which genome-level data are unavailable, and combined this dataset with the recently completed genome of the cryptophyte Guillardia theta to build an alignment composed of 258 genes. Our analyses strongly support haptophytes as sister to the SAR group, possibly together with telonemids and centrohelids. We also confirmed the common origin of katablepharids and cryptophytes, but these lineages were not related to other hacrobians; instead, they branch with plants. Our study resolves the evolutionary position of haptophytes, an ecologically critical component of the oceans, and proposes a new hypothesis for the origin of cryptophytes.     

64 Nucleus-encoded, plastid-targeted glyceraldehyde-3-phosphate dehydrogenase (GAPDH) indicates a single origin for chromalveolate plastids

Plastids (the photosynthetic organelles of plants and algae) ultimately originated through an endosymbiosis between a cyanobacterium and a eukaryote. Subsequently, plastids spread to other eukaryotes by secondary endosymbioses that took place between a eukaryotic alga and a second eukaryote. Recently, evidence has mounted in favour of a single origin for plastids of apicomplexans, cryptophytes, dinoflagellates, haptophytes, and heterokonts (together with their non-photosynthetic relatives, collectively termed chromalveolates). As of yet, however, no single molecular marker has been described which supports a common origin for all of these plastids. One piece of the evidence for a single origin of chromalveolate plastids came from plastid-targeted glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which originated by a gene duplication of the cytosolic form. However, no plastid GAPDH has been characterized from haptophytes, leaving an important piece of the puzzle missing. We have sequenced genes encoding cytosolic, mitochondrial-targeted, and plastid-targeted GAPDH proteins from a number of haptophytes and heterokonts, and found the haptophyte homologues to branch within the strongly supported clade of chromalveolate plastid-targeted GAPDH genes. Interestingly, plastid-targeted GAPDH genes from the haptophytes were more closely related to apicomplexan genes than was expected. Overall, the evolution of plastid-targeted GAPDH reinforces other data for a red algal ancestry of apicomplexan plastids, and raises a number of questions about the importance of plastid loss and the possibility of cryptic plastids in non-photosynthetic lineages such as ciliates.     

Evolution of glutamine synthetase in heterokonts: evidence for endosymbiotic gene transfer and the early evolution of photosynthesis

Although the endosymbiotic evolution of chloroplasts through primary and secondary associations is well established, the evolutionary timing and stability of the secondary endosymbiotic events is less well resolved. Heterokonts include both photosynthetic and nonphotosynthetic members and the nonphotosynthetic lineages branch basally in phylogenetic reconstructions. Molecular and morphological data indicate that heterokont chloroplasts evolved via a secondary endosymbiosis, involving a heterotrophic host cell and a photosynthetic ancestor of the red algae and this endosymbiotic event may have preceded the divergence of heterokonts and alveolates. If photosynthesis evolved early in this lineage, nuclear genomes of the nonphotosynthetic groups may contain genes that are not essential to photosynthesis but were derived from the endosymbiont genome through gene transfer. These genes offer the potential to trace the evolutionary history of chloroplast gains and losses within these lineages. Glutamine synthetase (GS) is essential for ammonium assimilation and glutamine biosynthesis in all organisms. Three paralogous gene families (GSI, GSII, and GSIII) have been identified and are broadly distributed among prokaryotic and eukaryotic lineages. In diatoms (Heterokonta), the nuclear-encoded chloroplast and cytosolic-localized GS isoforms are encoded by members of the GSII and GSIII family, respectively. Here, we explore the evolutionary history of GSII in both photosynthetic and nonphotosynthetic heterokonts, red algae, and other eukaryotes. GSII cDNA sequences were obtained from two species of oomycetes by polymerase chain reaction amplification. Additional GSII sequences from eukaryotes and bacteria were obtained from publicly available databases and genome projects. Bayesian inference and maximum likelihood phylogenetic analyses of GSII provided strong support for the monophyly of heterokonts, rhodophytes, chlorophytes, and plants and strong to moderate support for the Opisthokonts. Although the phylogeny is reflective of the unikont/bikont division of eukaryotes, we propose based on the robustness of the phylogenetic analyses that the heterokont GSII gene evolved via endosymbiotic gene transfer from the nucleus of the red-algal endosymbiont to the nucleus of the host. The lack of GSIII sequences in the oomycetes examined here further suggests that the GSIII gene that functions in the cytosol of photosynthetic heterokonts was replaced by the endosymbiont-derived GSII gene.