Christopher E. Lane

The New Higher Level Classification of Eukaryotes with Emphasis on the Taxonomy of Protists

Abstract. This revision of the classification of unicellular eukaryotes updates that of Levine et al. (1980) for the protozoa and expands it to include other protists. Whereas the previous revision was primarily to incorporate the results of ultrastructural studies, this revision incorporates results from both ultrastructural research since 1980 and molecular phylogenetic studies. We propose a scheme that is based on nameless ranked systematics. The vocabulary of the taxonomy is updated, particularly to clarify the naming of groups that have been repositioned. We recognize six clusters of eukaryotes that may represent the basic groupings similar to traditional “kingdoms.” The multicellular lineages emerged from within monophyletic protist lineages: animals and fungi from Opisthokonta, plants from Archaeplastida, and brown algae from Stramenopiles.

The revised classification of eukaryotes.

This revision of the classification of eukaryotes, which updates that of Adl et al. [J. Eukaryot. Microbiol. 52 (2005) 399], retains an emphasis on the protists and incorporates changes since 2005 that have resolved nodes and branches in phylogenetic trees. Whereas the previous revision was successful in re‐introducing name stability to the classification, this revision provides a classification for lineages that were then still unresolved. The supergroups have withstood phylogenetic hypothesis testing with some modifications, but despite some progress, problematic nodes at the base of the eukaryotic tree still remain to be statistically resolved. Looking forward, subsequent transformations to our understanding of the diversity of life will be from the discovery of novel lineages in previously under‐sampled areas and from environmental genomic information.

Algal genomes reveal evolutionary mosaicism and the fate of nucleomorphs

Cryptophyte and chlorarachniophyte algae are transitional forms in the widespread secondary endosymbiotic acquisition of photosynthesis by engulfment of eukaryotic algae. Unlike most secondary plastid-bearing algae, miniaturized versions of the endosymbiont nuclei (nucleomorphs) persist in cryptophytes and chlorarachniophytes. To determine why, and to address other fundamental questions about eukaryote–eukaryote endosymbiosis, we sequenced the nuclear genomes of the cryptophyte Guillardia theta and the chlorarachniophyte Bigelowiella natans. Both genomes have >21,000 protein genes and are intron rich, and B. natans exhibits unprecedented alternative splicing for a single-celled organism. Phylogenomic analyses and subcellular targeting predictions reveal extensive genetic and biochemical mosaicism, with both host- and endosymbiont-derived genes servicing the mitochondrion, the host cell cytosol, the plastid and the remnant endosymbiont cytosol of both algae. Mitochondrion-to-nucleus gene transfer still occurs in both organisms but plastid-to-nucleus and nucleomorph-to-nucleus transfers do not, which explains why a small residue of essential genes remains locked in each nucleomorph.

A MULTI-GENE MOLECULAR INVESTIGATION OF THE KELP (LAMINARIALES, PHAEOPHYCEAE) SUPPORTS SUBSTANTIAL TAXONOMIC RE-ORGANIZATION1

Every year numerous ecological, biochemical, and physiological studies are performed using members of the order Laminariales. Despite the fact that kelp are some of the most intensely studied macroalgae in the world, there is significant debate over the classification within and among the three “derived” families, the Alariaceae, Laminariaceae, and Lessoniaceae (ALL). Molecular phylogenies published for the ALL families have generated hypotheses strongly at odds with the current morphological taxonomy; however, conflicting phylogenetic hypotheses and consistently low levels of support realized in all of these studies have resulted in conservative approaches to taxonomic revisions. In order to resolve relationships within this group we have sequenced over 6000 bp from regions in the nuclear, chloroplast, and mitochondrial genomes and included 42 taxa in Bayesian, neighbor‐joining, and parsimony analyses. The result is the first comprehensive and well‐supported molecular phylogeny for the ALL complex of the Laminariales. We maintain the three recognized families (Alariaceae, Laminariaceae, and Lessoniaceae), but with vastly different compositions, as well as propose the Costariaceae fam. nov. for Agarum, Costaria, Dictyoneurum, and Thalassiophyllum, the only genera in the Laminariales with flattened, occasionally terete, stipes and either a perforate or reticulate blade. In addition, our data strongly support a split of the genus Laminaria. We resurrect the genus Saccharina Stackhouse for the Laminaria clade that does not contain L. digitata (Hudson) J.V. Lamouroux, the type of the genus.

Diversity, Nomenclature, and Taxonomy of Protists

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Nucleomorph genome of Hemiselmis andersenii reveals complete intron loss and compaction as a driver of protein structure and function

Nucleomorphs are the remnant nuclei of algal endosymbionts that took up residence inside a nonphotosynthetic eukaryotic host. The nucleomorphs of cryptophytes and chlorarachniophytes are derived from red and green algal endosymbionts, respectively, and represent a stunning example of convergent evolution: their genomes have independently been reduced and compacted to <1 megabase pairs (Mbp) in size (the smallest nuclear genomes known) and to a similar three-chromosome architecture. The molecular processes underlying genome reduction and compaction in eukaryotes are largely unknown, as is the impact of reduction/compaction on protein structure and function. Here, we present the complete 0.572-Mbp nucleomorph genome of the cryptophyte Hemiselmis andersenii and show that it is completely devoid of spliceosomal introns and genes for splicing RNAs—a case of complete intron loss in a nuclear genome. Comparison of H. andersenii proteins to those encoded in the slightly smaller (0.551-Mbp) nucleomorph genome of another cryptophyte, Guillardia theta, and to their homologs in the unicellular red alga Cyanidioschyzon merolae reveal that (i) cryptophyte nucleomorph genomes encode proteins that are significantly smaller than those in their free-living algal ancestors, and (ii) the smaller, more compact G. theta nucleomorph genome encodes significantly smaller proteins than that of H. andersenii. These results indicate that genome compaction can eliminate both coding and noncoding DNA and, consequently, drive the evolution of protein structure and function. Nucleomorph proteins have the potential to reveal the minimal functional units required for basic eukaryotic cellular processes.

A revised classification of the dictyoteae (dictyotales, phaeophyceae) based on rbcL and 26S ribosomal DNA sequence analyses

Dictyota is a genus of tropical to warm temperate brown algae characterized by parenchymatous, flattened thalli that grow from a single, transversely oriented apical cell. Dictyota is currently distinguished from allied genera of the tribe Dictyoteae (Dilophus, Glossophora, Glossophorella, and Pachydictyon) by the structure of the cortical and medullary layers, as well as the relative abundance of surface proliferations. Even though the traditional classification of the Dictyoteae has repeatedly been criticized in the past, the absence of sound molecular data has so far discouraged any new taxonomic proposals apart from a merger of Dilophus with Dictyota, which has been accepted by only part of the phycological community. Phylogenetic analysis of rbcL gene, partial 26S rDNA sequence, and combined data sets, including four of five generitypes, demonstrates that the traditional classification does not accurately reflect the evolutionary history of the group. None of the genera are resolved as a monophyletic clade. Hence, a merger of Glossophora, Glossophorella, and Pachydictyon in Dictyota is proposed. Two new genera, Canistrocarpus (incorporating D. cervicornis, D. crispata, and D. magneana) and Rugulopteryx (accommodating D. radicans, Dil. suhrii, and Dil. marginata), are proposed. Both genera are supported by molecular indications and a combination of reproductive and vegetative characters. The position of Dil. fastigiatus as a clade sister to Dictyota s.l. and the absence of Dil. gunnianus, the generitype of Dilophus, from the analyses, prevented us from making a more definite statement on the status of the latter genus.

Insights into the evolutionary origin and genome architecture of the unicellular opisthokonts Capsaspora owczarzaki and Sphaeroforma arctica

ABSTRACT. Molecular phylogenetic analyses have recently shown that the unicellular amoeboid protist Capsaspora owczarzaki is unlikely to be a nucleariid or an ichthyosporean as previously described, but is more closely related to Metazoa, Choanoflagellata, and Ichthyosporea. However, the specific phylogenetic relationship of Capsaspora to other protist opisthokont lineages was poorly resolved. To test these earlier results we have expanded both the taxonomic sampling and the number of genes from opisthokont unicellular lineages. We have sequenced the protein‐coding genes elongation factor 1‐α (EF1‐α) and heat shock protein 70 (Hsp70) from C. owczarzaki and the ichthyosporean Sphaeroforma arctica. Our maximum likelihood (ML) and Bayesian analyses of a concatenated alignment of EF1‐α, Hsp70, and actin protein sequences with a better sampling of opisthokont‐related protist lineages indicate that C. owczarzaki is not clearly allied with any of the unicellular opisthokonts, but represents an independent unicellular lineage closely related to animals and choanoflagellates. Moreover, we have found that the ichthyosporean S. arctica possesses an EF‐like (EFL) gene copy instead of the canonical EF1‐α, the first so far described in an ichthyosporean. A maximum likelihood phylogenetic analysis shows that the EF‐like gene of S. arctica strongly groups with the EF‐like genes from choanoflagellates. Finally, to begin characterizing the Capsaspora genome, we have performed pulsed‐field gel electrophoresis (PFGE) analyses, which indicate that its genome has at least 12 chromosomes with a total genome size in the range of 22–25 Mb.

Red Algae Lose Key Mitochondrial Genes in Response to Becoming Parasitic

Red algal parasites are unusual because the vast majority of them parasitize species with which they share a recent common ancestor. This strategy has earned them the name “adelphoparasites,” from the Greek, adelpho, meaning “kin.” Intracellular adelphoparasites are very rare in nature, yet have independently evolved hundreds of times among the floridiophyte red algae. Much is known about the life history and infection cycle of these parasites but nearly nothing in known about their genomes. We sequenced the mitochondrial genomes of the free-living Gracilariopsis andersonii and its closely related parasite Gracilariophila oryzoides to determine what effect a parasitic lifestyle has on the genomes of red algal parasites. Whereas the parasite genome is similar to the host in many ways, the genes encoding essential proteins ATP8 and SDHC are pseudogenes in the parasite. The mitochondrial genome of parasite from a different class of red algae, Plocamiocolax puvinata, has lost the atp8 gene entirely, indicating that this gene is no longer critical in red algal parasite mitochondria.

Transcriptome of American Oysters, Crassostrea virginica, in Response to Bacterial Challenge: Insights into Potential Mechanisms of Disease Resistance

The American oyster Crassostrea virginica, an ecologically and economically important estuarine organism, can suffer high mortalities in areas in the Northeast United States due to Roseovarius Oyster Disease (ROD), caused by the gram-negative bacterial pathogen Roseovarius crassostreae. The goals of this research were to provide insights into: 1) the responses of American oysters to R. crassostreae, and 2) potential mechanisms of resistance or susceptibility to ROD. The responses of oysters to bacterial challenge were characterized by exposing oysters from ROD-resistant and susceptible families to R. crassostreae, followed by high-throughput sequencing of cDNA samples from various timepoints after disease challenge. Sequence data was assembled into a reference transcriptome and analyzed through differential gene expression and functional enrichment to uncover genes and processes potentially involved in responses to ROD in the American oyster. While susceptible oysters experienced constant levels of mortality when challenged with R. crassostreae, resistant oysters showed levels of mortality similar to non-challenged oysters. Oysters exposed to R. crassostreae showed differential expression of transcripts involved in immune recognition, signaling, protease inhibition, detoxification, and apoptosis. Transcripts involved in metabolism were enriched in susceptible oysters, suggesting that bacterial infection places a large metabolic demand on these oysters. Transcripts differentially expressed in resistant oysters in response to infection included the immune modulators IL-17 and arginase, as well as several genes involved in extracellular matrix remodeling. The identification of potential genes and processes responsible for defense against R. crassostreae in the American oyster provides insights into potential mechanisms of disease resistance.