THE MESOZOIC RADIATION OF EUKARYOTIC ALGAE: THE PORTABLE PLASTID HYPOTHESIS1

Although all chloroplasts appear to have been derived from a common ancestor, a major schism occurred early in the evolution of eukaryotic algae that gave rise to red and green photoautotrophic lineages. In Paleozoic and earlier times, the fossil record suggests that oceanic eukaryotic phytoplankton were dominated by the green (chl b‐containing) algal line. However, following the end‐Permian extinction, a diverse group of eukaryotic phytoplankton evolved from secondary symbiotic associations in the red (chl c‐containing) line and subsequently rose to ecological prominence. In the contemporary oceans, red eukaryotic phytoplankton taxa continue to dominate marine pelagic food webs, whereas the green line is relegated to comparatively minor ecological and biogeochemical roles. To help elucidate why the oceans are not dominated by green taxa, we analyzed and compared whole plastid genomes in both the red and green lineages. Our results suggest that whereas all algal plastids retain a core set of genes, red plastids retain a complementary set of genes that potentially confer more capacity to autonomously express proteins regulating oxygenic photosynthetic and energy transduction pathways. We hypothesize that specific gene losses in the primary endosymbiotic green plastid reduced its portability for subsequent symbiotic associations. This corollary of the plastid “enslavement” hypothesis may have limited subsequent evolutionary advances in the green lineage while simultaneously providing a competitive advantage to the red lineage.     

Cytoplasmic inheritance in green algae: patterns,mechanisms and relation to sex type

Cytological and genetic investigations of two major groups of green algae, chlorophyte and streptophyte green algae, show a predominance of uniparental inheritance of the plastid and mitochondrial genomes in most species. However, in some crosses of isogamous species of Ulva compressa, these genomes are transmitted from mt⁺, mt⁻, and both parents. In species with uniparental organelle inheritance, various mechanisms can eliminate organelles and their DNA during male gametogenesis or after fertilization. Concerning plastid inheritance, two major mechanisms are widespread in green algae: (1) digestion of plastid DNA during male gametogenesis, during fertilization, or after fertilization; and (2) disintegration or fusion of the plastid in the zygote. The first mechanism also eliminates the mitochondrial DNA in anisogamous and oogamous species. These mechanisms would ensure the predominantly uniparental inheritance of organelle genomes in green algae. To trace the evolutionary history of cytoplasmic inheritance in green algae, the relations between uniparental inheritance and sex type were considered in isogamous, anisogamous, and oogamous species using sex-specific features that might be nearly universal among Chlorophyta.     

Direct and indirect influence of sulfur availability on phytoplankton evolutionary trajectories

The sulfate facilitation hypothesis suggests that changes in ocean sulfate concentration influenced the rise to dominance of phytoplankton species of the red lineage. The mechanistic reasons for this phenomenon are not yet understood. We started to address this question by investigating the differences in S utilization by algae of the green and red lineages and in cyanobacteria cultured in the presence of either 5 mmol · L^?1 (approximately equivalent to Paleozoic ocean concentrations) or 30 mmol · L^?1 (corresponding to post‐Mesozoic/extant concentrations) sulfate. The activities of the main enzymes involved in SO₄^2? assimilation changed in response to changes in growth sulfate concentration. ATP sulfurylase showed different kinetics in the various taxa, with an especially odd behavior for the dinoflagellate. Sulfate availability had a modest effect on cell organic composition. Species‐specific differences in the use of some elements were instead obvious in algae grown in the presence of different sulfate concentrations, overall confirming that algae of the red lineage do better at high sulfate than algae of the green lineage. The increase in sulfate concentration may thus have had an impact on phytoplankton radiation both through changes in their enzymatic machinery and through indirect repercussion on elemental usage.     

Is cell composition related to the phylogenesis of microalgae? An investigation using Hierarchical Cluster Analysis of Fourier Transform Infrared spectra of whole cells

Fourier Transform Infra-Red (FTIR) spectroscopy was used to investigate whether algae belonging to the green evolutionary lineage and algae belonging to the red evolutionary lineage consistently differ for their biomass composition and for the way their composition responds to changes in NO₃⁻ availability. For this purpose, the FTIR spectra of eleven algal species cultured in media containing different amount of NO₃⁻ were subjected to Hierarchical Cluster Analysis (HCA). The same analysis was conducted on the differential FTIR spectra of cells grown at different NO₃⁻ concentrations. Although the organic composition of cells was affected by the availability of NO₃⁻, the analysis of its change did not elicit a clear relationship with the phylogenetic position of the species used. However, when the two evolutionary lineages, and not the individual species, are compared, some differences emerge: the species belonging to the green lineage reorganize their biomass composition similarly in response to variations in N availability, whereas the species of the red lineage modulate their allocation patterns more heterogeneously. This difference may contribute to the ecological fitness of microalgae and may have had an influence on the evolutionary trajectories of phytoplankton groups.     

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.     

Phylogenetic relationships within Korthalsella (Viscaceae) based on nuclear ITS and plastid trnL-F sequence

The nuclear encoded internal transcribed spacer (ITS) region and the plastid encoded trnL-F region were sequenced for 25 populations of Korthalsella, a genus of reduced, monoecious, Old World misletoes. The molecular study confirms the hypothesis that branch shape and cladotaxy (the arrangement of branches with respect to their parent axis) are unreliable indicators of relationship in the genus and demonstrates that many of the taxa previously recognized are not monophyletic. Both gene regions identify three major subgroups within the genus and find lower level relationships within these subgroups highly correlated with geographic distance. An analysis based upon 18S and rbcL sequences identifies Ginalloa as the sister group to Korthalsella, which together with the branching order within the genus, indicates that Korthalsella originated in Papuasia and aids in elucidating evolution of the peculiar inflorescence structure. There are problems associated with species delimitation when evolutionary units are more restricted than morphological lineages, and justification is offered for recognizing only morphologically diagnosable monophyletic lineages as species. Varying substitution rates and differing modes of inheritance in ITS and trnL-F result in complementary utility of the two regions for elucidating infrageneric relationships in Korthalsella.     

Did some red alga‐derived plastids evolve via kleptoplastidy? A hypothesis

The evolution of plastids has a complex and still unresolved history. These organelles originated from a cyanobacterium via primary endosymbiosis, resulting in three eukaryotic lineages: glaucophytes, red algae, and green plants. The red and green algal plastids then spread via eukaryote–eukaryote endosymbioses, known as secondary and tertiary symbioses, to numerous heterotrophic protist lineages. The number of these horizontal plastid transfers, especially in the case of red alga‐derived plastids, remains controversial. Some authors argue that the number of plastid origins should be minimal due to perceived difficulties in the transformation of a eukaryotic algal endosymbiont into a multimembrane plastid, but increasingly the available data contradict this argument. I suggest that obstacles in solving this dilemma result from the acceptance of a single evolutionary scenario for the endosymbiont‐to‐plastid transformation formulated by Cavalier‐Smith & Lee (1985). Herein I discuss data that challenge this evolutionary scenario. Moreover, I propose a new model for the origin of multimembrane plastids belonging to the red lineage and apply it to the dinoflagellate peridinin plastid. The new model has several general and practical implications, such as the requirement for a new definition of cell organelles and in the construction of chimeric organisms.     

Evolution and functional diversification of fructose bisphosphate aldolase genes in photosynthetic marine diatoms

Diatoms and other chlorophyll-c containing, or chromalveolate, algae are among the most productive and diverse phytoplankton in the ocean. Evolutionarily, chlorophyll-c algae are linked through common, although not necessarily monophyletic, acquisition of plastid endosymbionts of red as well as most likely green algal origin. There is also strong evidence for a relatively high level of lineage-specific bacterial gene acquisition within chromalveolates. Therefore, analyses of gene content and derivation in chromalveolate taxa have indicated particularly diverse origins of their overall gene repertoire. As a single group of functionally related enzymes spanning two distinct gene families, fructose 1,6-bisphosphate aldolases (FBAs) illustrate the influence on core biochemical pathways of specific evolutionary associations among diatoms and other chromalveolates with various plastid-bearing and bacterial endosymbionts. Protein localization and activity, gene expression, and phylogenetic analyses indicate that the pennate diatom Phaeodactylum tricornutum contains five FBA genes with very little overall functional overlap. Three P. tricornutum FBAs, one class I and two class II, are plastid localized, and each appears to have a distinct evolutionary origin as well as function. Class I plastid FBA appears to have been acquired by chromalveolates from a red algal endosymbiont, whereas one copy of class II plastid FBA is likely to have originated from an ancient green algal endosymbiont. The other copy appears to be the result of a chromalveolate-specific gene duplication. Plastid FBA I and chromalveolate-specific class II plastid FBA are localized in the pyrenoid region of the chloroplast where they are associated with β-carbonic anhydrase, which is known to play a significant role in regulation of the diatom carbon concentrating mechanism. The two pyrenoid-associated FBAs are distinguished by contrasting gene expression profiles under nutrient limiting compared with optimal CO2 fixation conditions, suggestive of a distinct specialized function for each. Cytosolically localized FBAs in P. tricornutum likely play a role in glycolysis and cytoskeleton function and seem to have originated from the stramenopile host cell and from diatom-specific bacterial gene transfer, respectively.     

Testing the ecological consequences of evolutionary change using elements

Understanding the ecological consequences of evolutionary change is a central challenge in contemporary biology. We propose a framework based on the ~25 elements represented in biology, which can serve as a conduit for a general exploration of poorly understood evolution‐to‐ecology links. In this framework, known as ecological stoichiometry, the quantity of elements in the inorganic realm is a fundamental environment, while the flow of elements from the abiotic to the biotic realm is due to the action of genomes, with the unused elements excreted back into the inorganic realm affecting ecological processes at higher levels of organization. Ecological stoichiometry purposefully assumes distinct elemental composition of species, enabling powerful predictions about the ecological functions of species. However, this assumption results in a simplified view of the evolutionary mechanisms underlying diversification in the elemental composition of species. Recent research indicates substantial intraspecific variation in elemental composition and associated ecological functions such as nutrient excretion. We posit that attention to intraspecific variation in elemental composition will facilitate a synthesis of stoichiometric information in light of population genetics theory for a rigorous exploration of the ecological consequences of evolutionary change.     

UNIVERSAL PRIMERS AMPLIFY A 23S rDNA PLASTID MARKER IN EUKARYOTIC ALGAE AND CYANOBACTERIA1

The challenge in the development of universal algal primers lies in the genetic diversity contained within the vast array of evolutionary lineages present in this informally named group of organisms. A comparative genomics approach was used previously to identify conserved primers flanking a region of the plastid genome. Our present research illustrates the feasibility of amplifying and sequencing this marker across multiple algal lineages. We present a preliminary framework of 107 novel sequences of this region from 62 red algae, 19 green algae, 14 brown algae, 8 cyanobacteria, 2 diatoms, 1 xanthophyte, and 1 euglenoid, and illustrate levels of divergence of the marker for well‐represented groups in a neighbor‐joining analysis. This ～410 nt region distinguishes most species included in the analysis. The remarkable universality of these primers suggests potential for their use in assays of environmental samples in which they could be used to simultaneously detect a number of different algal lineages.     

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.     

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.     

The evolution of ecosystem processes: growth rate and elemental stoichiometry of a key herbivore in temperate and arctic habitats

Understanding the reciprocal interactions between the evolved characteristics of species and the environment in which each species is embedded is a major priority for evolutionary ecology. Here we use the perspective of ecological stoichiometry to test the hypothesis that natural selection on body growth rate affects consumer body stoichiometry. As body elemental composition (nitrogen, phosphorus) of consumers influences nutrient cycling and trophic dynamics in food webs, such differences should also affect biogeochemical processes and trophic dynamics. Consistent with the growth rate hypothesis, body growth rate and phosphorus content of individuals of the Daphnia pulex species complex were lower in Wisconsin compared to Alaska, where the brevity of the growing season places a premium on growth rate. Consistent with stoichiometric theory, we also show that, relative to animals sampled in Wisconsin, animals sampled in Alaska were poor recyclers of P and suffered greater declines in growth when fed low‐quality, P‐deficient food. These results highlight the importance of evolutionary context in establishing the reciprocal relationships between single species and ecosystem processes such as trophic dynamics and consumer‐driven nutrient recycling.     

Photosynthetic eukaryotes unite: endosymbiosis connects the dots

The photosynthetic organelle of algae and plants (the plastid) traces its origin to a primary endosymbiotic event in which a previously non-photosynthetic protist engulfed and enslaved a cyanobacterium. This eukaryote then gave rise to the red, green and glaucophyte algae. However, many algal lineages, such as the chlorophyll c-containing chromists, have a more complicated evolutionary history involving a secondary endosymbiotic event, in which a protist engulfed an existing eukaryotic alga (in this case, a red alga). Chromists such as diatoms and kelps then rose to great importance in aquatic habitats. Another algal group, the dinoflagellates, has undergone tertiary (engulfment of a secondary plastid) and even quaternary endosymbioses. In this review, we examine algal diversity and show endosymbiosis to be a major force in algal evolution. This area of research has advanced rapidly and long-standing issues such as the chromalveolate hypothesis and the extent of endosymbiotic gene transfer have recently been clarified.     

Paradoxically, prior acquisition of antioxidant activity enhances oxidative stress-induced cell death

Oxidative stress has been implicated in the induction of programmed cell death in a wide variety of organisms. Acquiring antioxidant capacity is thought to enhance the viability of cells challenged by a subsequent oxidative stress. Counter-intuitively, we show that in two phytoplankton species, Chlamydomonas reinhardtii and Peridinium gatunense , representing the green and red plastid lineages, oxidative stress induced cell death in cultures that already possessed high antioxidant activity but not in cells that exhibited low activity. Cell death of low antioxidant possessing cultures was markedly enhanced by the addition of dehydroascorbate, a product of ascorbate peroxidase (APX), but not of ascorbate or reduced glutathione, and was preceded by increased metacaspase expression and activity. These data suggested that the level of APX and its products, strongly upregulated by oxidative stress, serves as a possible surveillance signal, reporting that the cells already experienced an earlier oxidative stress. Our data presents a novel role of APX in antioxidant activity and response to oxidative stress in photosynthetic microorganisms. Elimination of cysts production by phytoplankton cells that were already damaged by oxidative stress (indicated by the rise in oxidized proteins) as the inoculum for the following year's population may be the evolutionary trigger for this phenomenon.     

Exploring patterns and mechanisms of interspecific and intraspecific variation in body elemental composition of desert consumers

Key processes such as trophic interactions and nutrient cycling are often influenced by the element content of organisms. Previous analyses have led to some preliminary understanding of the relative importance of evolutionary and ecological factors determining animal stoichiometry. However, to date, the patterns and underlying mechanisms of consumer stoichiometry at interspecific and intraspecific levels within natural ecosystems remain poorly investigated. Here, we examine the association between phylogeny, trophic level, body size, and ontogeny and the elemental composition of 22 arthropod as well as two lizard species from the coastal zone of the Atacama Desert in Chile. We found that, in general, whole‐body P content was more variable than body N content both among and within species. Body P content showed a significant phylogenetic signal; however, phylogeny explained only 4% of the variation in body P content across arthropod species. We also found a significant association between trophic level and the element content of arthropods, with carnivores having 15% greater N and 70% greater P contents than herbivores. Elemental scaling relationships across species were only significant for body P content, and even the P content scaling relationship was not significant after controlling for phylogeny. P content did decrease significantly with body size within most arthropod species, which may reflect the size dependence of RNA content in invertebrates. In contrast, larger lizards had higher P contents and lower N:P ratios than smaller lizards, which may be explained by size‐associated differences in bone and scale investments. Our results suggests that structural differences in material allocation, trophic level and phylogeny can all contribute to variation in the stoichiometry of desert consumers, and they indicate that the elemental composition of animals can be useful information for identifying broad‐scale linkages between nutrient cycling and trophic interactions in terrestrial food webs.     

Twenty-fold difference in evolutionary rates between the mitochondrial and plastid genomes of species with secondary red plastids

Within plastid-bearing species, the relative rates of evolution between mitochondrial and plastid genomes are poorly studied, but for the few lineages in which they have been explored, including land plants and green algae, the mitochondrial DNA mutation rate is nearly always estimated to be lower than or equal to that of the plastid DNA. Here, we show that in protists from three distinct lineages with secondary, red algal-derived plastids, the opposite is true: their mitochondrial genomes are evolving 5-30 times faster than their plastid genomes, even when the plastid is nonphotosynthetic. These findings have implications for understanding the origins and evolution of organelle genome architecture and the genes they encode.     

MULTIPLE CRYPTIC SPECIES: MOLECULAR DIVERSITY AND REPRODUCTIVE ISOLATION IN THE BOSTRYCHIA RADICANS/B. MORITZIANA COMPLEX (RHODOMELACEAE, RHODOPHYTA) WITH FOCUS ON NORTH AMERICAN ISOLATES

Red algae of the Bostrychia radicans/B. moritziana complex are common in warm temperate areas of North America. Phylogenetic analysis of both plastid and mitochondrial DNA sequence data revealed seven distinct evolutionary lineages among worldwide samples. Although only two haplotypes (plastid and mitochondrial) were found in Pacific Mexico, four plastid and 11 mitochondrial haplotypes were found in a similar latitudinal spread along the Atlantic coast of the United States. On the U.S. Atlantic coast only one plastid haplotype was found in northern samples (Connecticut to North Carolina), whereas further south several plastid haplotypes were found. Phylogenetic analyses suggested that this single plastid haplotype found among northern samples could be the result of a northward range expansion possibly since the last glacial maximum. Crossing data of samples within the same evolutionary lineage showed that samples with the same plastid haplotypes were generally sexually compatible; samples with different plastid haplotypes were reproductively isolated. Samples from Pacific Mexico were partially reproductively compatible with some samples from the Atlantic USA (plastid haplotype C) and were more closely related to these samples than these U.S. samples were to other U.S. Atlantic samples. Compatible solute types mirrored the plastid haplotype, with plastid haplotype B having only sorbitol, whereas all other haplotypes also contained dulcitol. Samples from Atlantic USA, with different plastid haplotypes (e.g. B vs. C), but within the same evolutionary lineage, were reproductively isolated from each other. Data indicate that reproductive isolation occurs between and within supported evolutionary lineages and that the number of cryptic species is high.     

Phylogeny of Plastids Based on Cladistic Analysis of Gene Loss Inferred from Complete Plastid Genome Sequences

Based on the recent hypothesis on the origin of eukaryotic phototrophs, red algae, green plants, and glaucophytes constitute the primary photosynthetic eukaryotes (whose plastids may have originated directly from a cyanobacterium-like prokaryote via primary endosymbiosis), whereas the plastids of other lineages of eukaryotic phototrophs appear to be the result of secondary or tertiary endosymbiotic events (involving a phototrophic eukaryote and a host cell). Although phylogenetic analyses using multiple plastid genes from a wide range of eukaryotic lineages have been carried out, some of the major phylogenetic relationships of plastids remain ambiguous or conflict between different phylogenetic methods used for nucleotide or amino acid substitutions. Therefore, an alternative methodology to infer the plastid phylogeny is needed. Here, we carried out a cladistic analysis of the loss of plastid genes after primary endosymbiosis using complete plastid genome sequences from a wide range of eukaryotic phototrophs. Since it is extremely unlikely that plastid genes are regained during plastid evolution, we used the irreversible Camin-Sokal model for our cladistic analysis of the loss of plastid genes. The cladistic analysis of the 274 plastid protein-coding genes resolved the 20 operational taxonomic units representing a wide range of eukaryotic lineages (including three secondary plastid-containing groups) into two large monophyletic groups with high bootstrap values: one corresponded to the red lineage and the other consisted of a large clade composed of the green lineage (green plants and Euglena) and the basal glaucophyte plastid. Although the sister relationship between the green lineage and the Glaucophyta was not resolved in recent phylogenetic studies using amino acid substitutions from multiple plastid genes, it is consistent with the rbcL gene phylogeny and with a recent phylogenetic study using multiple nuclear genes. In addition, our analysis robustly resolved the conflicting/ambiguous phylogenetic positions of secondary plastids in previous phylogenetic studies: the Euglena plastid was sister to the chlorophycean (Chlamydomonas) lineage, and the secondary plastids from the diatom (Odontiella) and cryptophyte (Guillardia) were monophyletic within the red lineage.     

Babesia bovis: A comprehensive phylogenetic analysis of plastid-encoded genes supports green algal origin of apicoplasts

Apicomplexan parasites commonly contain a unique, non-photosynthetic plastid-like organelle termed the apicoplast. Previous analyses of other plastid-containing organisms suggest that apicoplasts were derived from a red algal ancestor. In this report, we present an extensive phylogenetic study of apicoplast origins using multiple previously reported apicoplast sequences as well as several sequences recently reported. Phylogenetic analysis of amino acid sequences was used to determine the evolutionary origin of the organelle. A total of nine plastid genes from 37 species were incorporated in our study. The data strongly support a green algal origin for apicoplasts and Euglenozoan plastids. Further, the nearest green algae lineage to the Apicomplexans is the parasite Helicosporidium, suggesting that apicoplasts may have originated by lateral transfer from green algal parasite lineages. The results also substantiate earlier findings that plastids found in Heterokonts such as Odontella and Thalassiosira were derived from a separate secondary endosymbiotic event likely originating from a red algal lineage.