Sung Min Boo

Differential Gene Retention in Plastids of Common Recent Origin

The cyanobacterium-derived plastids of algae and plants have supported the diversification of much of extant eukaryotic life. Inferences about early events in plastid evolution must rely on reconstructing events that occurred over a billion years ago. In contrast, the photosynthetic amoeba Paulinella chromatophora provides an exceptional model to study organelle evolution in a prokaryote-eukaryote (primary) endosymbiosis that occurred approximately 60 mya. Here we sequenced the plastid genome (0.977 Mb) from the recently described Paulinella FK01 and compared the sequence with the existing data from the sister taxon Paulinella M0880/a. Alignment of the two plastid genomes shows significant conservation of gene order and only a handful of minor gene rearrangements. Analysis of gene content reveals 66 differential gene losses that appear to be outright gene deletions rather than endosymbiotic gene transfers to the host nuclear genome. Phylogenomic analysis validates the plastid ancestor as a member of the Synechococcus-Prochlorococcus group, and the cyanobacterial provenance of all plastid genes suggests that these organelles were not targets of interphylum gene transfers after endosymbiosis. Inspection of 681 DNA alignments of protein-encoding genes shows that the vast majority have dN/dS ratios <<1, providing evidence for purifying selection. Our study demonstrates that plastid genomes in sister taxa are strongly constrained by selection but follow distinct trajectories during the earlier phases of organelle evolution.

Genetic diversity in Undaria pinnatifida (Laminariales, Phaeophyceae) deduced from mitochondria genes – origins and succession of introduced populations

S. Uwai, W. Nelson, K. Neill, W.D. Wang, L.E. Aguilar-Rosas, S.M. Boo, T. Kitayama and H. Kawai. 2006. Genetic diversity in Undaria pinnatifida (Laminariales, Phaeophyceae) deduced from mitochondria genes – origins and succession of introduced populations. Phycologia 45: 687–695. DOI: 10.2216/05-66.1 To elucidate the genetic diversity of the brown alga Undaria pinnatifida in native and introduced populations worldwide, and to discuss the transoceanic introduction processes, we investigated the haplotype divergence of the mitochondrial loci of the coding region of cox3 and noncoding region between tatC and tLeu genes. In its native range (Japan, Korea and China), we found 27 haplotypes, which were classified into 4 genetic and biogeographical groups: (1) Northern Japan type, distributed in Hokkaido and Pacific northern Honshu; (2) Continental type, found in Korea and China; (3) Pacific central Japan type; and (4) Sea of Japan type. Among the introduced populations, European and Mexican populations agreed with the Northern Japan type. In Australia, the Tasmanian population agreed with the Sea of Japan type, whereas the Victorian population was of the Continental type. Very high diversities were found in New Zealand: 10 haplotypes were found (including 2 only in old herbarium specimens), including both the Northern Japan type and the Continental type. The haplotype found in California agreed with a component of the Central Japan type collected at Kanagawa Prefecture. The samples from Argentina agreed with the Continental type. The alignment of the European populations with the Northern Japan type is consistent with the notion that the Undaria in Europe was first introduced with oyster spat. It is speculated that Californian and Mexican populations were recently introduced by shipping vectors. There have been many introduction events to New Zealand since the late 1980s, and the dominant haplotypes in the local populations appear to have changed over time. Introduction to Argentina/Australia (Victoria) could have resulted from secondary introductions from New Zealand populations, because transport within the same latitudinal range is considered to be easier than transport by shipping across the equator. Within Japan, the occurrence of both the Continental and the Northern Japan types in the Osaka Bay area is considered to be the result of recent intentional introduction for fisheries purposes.

Complex phylogeographic patterns in the freshwater alga Synura provide new insights into ubiquity vs. endemism in microbial eukaryotes

The global distribution, abundance, and diversity of microscopic freshwater algae demonstrate an ability to overcome significant barriers such as dry land and oceans by exploiting a range of biotic and abiotic colonization vectors. If these vectors are considered unlimited and colonization occurs in proportion to population size, then globally ubiquitous distributions are predicted to arise. This model contrasts with observations that many freshwater microalgal taxa possess true biogeographies. Here, using a concatenated multigene data set, we study the phylogeography of the freshwater heterokont alga Synura petersenii sensu lato. Our results suggest that this Synura morphotaxon contains both cosmopolitan and regionally endemic cryptic species, co‐occurring in some cases, and masked by a common ultrastructural morphology. Phylogenies based on both proteins (seven protein‐coding plastid and mitochondrial genes) and DNA (nine genes including ITS and 18S rDNA) reveal pronounced biogeographic delineations within phylotypes of this cryptic species complex while retaining one clade that is globally distributed. Relaxed molecular clock calculations, constrained by fossil records, suggest that the genus Synura is considerably older than currently proposed. The availability of tectonically relevant geological time (107–108 years) has enabled the development of the observed, complex biogeographic patterns. Our comprehensive analysis of freshwater algal biogeography suggests that neither ubiquity nor endemism wholly explains global patterns of microbial eukaryote distribution and that processes of dispersal remain poorly understood.

Supermatrix data highlight the phylogenetic relationships of photosynthetic stramenopiles.

Molecular data had consistently recovered monophyletic classes for the heterokont algae, however, the relationships among the classes had remained only partially resolved. Furthermore, earlier studies did not include representatives from all taxonomic classes. We used a five-gene (nuclear encoded SSU rRNA; plastid encoded rbcL, psaA, psbA, psbC) analysis with a subset of 89 taxa representing all 16 heterokont classes to infer a phylogenetic tree. There were three major clades. The Aurearenophyceae, Chrysomerophyceae, Phaeophyceae, Phaeothamniophyceae, Raphidophyceae, Schizocladiophyceae and Xanthophyceae formed the SI clade. The Chrysophyceae, Eustigmatophyceae, Pinguiophyceae, Synchromophyceae and Synurophyceae formed the SII clade. The Bacillariophyceae, Bolidophyceae, Dictyochophyceae and Pelagophyceae formed the SIII clade. These three clades were also found in a ten-gene analysis. The approximately unbiased test rejected alternative hypotheses that forced each class into either of the other two clades. Morphological and biochemical data were not available for all 89 taxa, however, existing data were consistent with the molecular phylogenetic tree, especially for the SIII clade.

A NEW BROWN ALGAL ORDER, ISHIGEALES (PHAEOPHYCEAE), ESTABLISHED ON THE BASIS OF PLASTID PROTEIN‐CODING rbcL, psaA, AND psbA REGION COMPARISONS1

The brown algal family Ishigeaceae currently includes a single genus, Ishige Yendo, with two species. The relationship of the family to other brown algal lineages is less studied in terms of their plastid ultrastructure and molecular phylogeny. We determined the sequences of rbcL from four samples of the two Ishige species and nine putative relatives and the psaA and psbA sequences from 37 representatives of the brown algae. Analyses of individual and combined data sets resulted in similar trees; however, the concatenated data gave greater resolution and clade support than each individual gene. In all the phylogenies, the Phaeophyceae was well resolved, the Ectocarpales being placed in a terminal position and the Ishigeaceae ending up in a basal position. From our ultrastructural study, we concluded that the pyrenoid is absent in the Ishigeaceae, despite the presence of a rudimentary pyrenoid in I. okamurae. These results suggest that the Ishigeaceae is an early diverging brown lineage. Our molecular and morphological data, therefore, lead us to exclude the Ishigeaceae from the Ectocarpales s.l., which have an elaborate pyrenoid, and to propose its own order Ishigeales ord. nov. The Ishigeales is distinguished by oligostichous structure of thalli, phaeophycean hairs formed within cryptostomata, unilocular sporangia transformed from terminal cortical cells, and plurilocular sporangia lacking sterile terminal cells. This study is the first to document the utility of the psaA and psbA sequences for brown algae and also the first report on the multigene phylogeny of the Phaeophyceae based on three protein‐coding plastid genes.

Genetic data hint at a common donor region for invasive Atlantic and Pacific populations of Gracilaria vermiculophylla (Gracilariales, Rhodophyta).

Gracilaria vermiculophylla (Ohmi) Papenf., an agar‐producing red alga introduced from northeast Asia to Europe and North America, is often highly abundant in invaded areas. To assay its genetic diversity and identify the putative source of invasive populations, we analyzed the mitochondrial cytochrome c oxidase subunit I (cox1) gene from 312 individuals of G. vermiculophylla collected in 37 native and 32 introduced locations. A total of 19 haplotypes were detected: 17 in northeast Asia and three in Europe and eastern and western North America, with only one shared among all regions. The shared haplotype was present in all introduced populations and in ∼99% of individuals in the introduced areas. This haplotype was also found at three native locations in east Korea, west Japan, and eastern Russia. Both haplotype and nucleotide diversities were extremely low in Europe and North America compared to northeast Asia. Our study indicates that the East Sea/Sea of Japan is a likely donor region of the invasive populations of G. vermiculophylla in the east and west Atlantic and the east Pacific.

Reinstatement of Ectocarpus crouaniorum Thuret in Le Jolis as a third common species of Ectocarpus (Ectocarpales, Phaeophyceae) in Western Europe, and its phenology at Roscoff, Brittany

Based on morphological characters, cross‐fertility and molecular systematics, two species are currently recognized in the ubiquitous temperate brown algal genus Ectocarpus: the type species E. siliculosus (Dillwyn) Lyngbye and E. fasciculatus Harvey. We studied diversity, cross‐fertility and ecology of Ectocarpus in megatidal areas in northwest France (Western Europe) and propose to reinstate a third species, E. crouaniorum Thuret in Le Jolis. Genotyping of 67 individuals from five localities, including the type locality of E. crouaniorum, using internal transcribed spacer 1 (ITS1) length as a marker, showed that the three species co‐occurred whenever the habitat was suitable. Our survey also revealed a single putative field hybrid between E. crouaniorum and E. siliculosus, and a single individual of a further Ectocarpus genotype. In laboratory experiments, E. crouaniorum was crossed with E. siliculosus and E. fasciculatus. In 12 of 13 crosses, the zygotes did not develop (postzygotic sterility); in one experiment a viable hybrid was produced after crossing a female E. crouaniorum with a male E. siliculosus, but this hybrid was unable to form meiospores. Phylogenetic analysis of five molecular markers from the nuclear, mitochondrial and plastid genomes (in total 1818 bp) confirmed genetic separation of the three species. Ecologically, E. crouaniorum was confined to high intertidal pools and run‐offs, where the gametophyte was common from spring to summer. Another characteristic was that it usually occurred as an epiphyte of up to 12 cm in length on erect thalli of Scytosiphon lomentaria. Sporophytes of E. crouaniorum were found all year long; they were <3 cm in size or microscopic and were epilithic in the same habitat. The presence of a third species of Ectocarpus in Western Europe suggests that species diversity in this genus is larger than recognized during the last 40 years.

Divergence time estimates and the evolution of major lineages in the florideophyte red algae

The Florideophyceae is the most abundant and taxonomically diverse class of red algae (Rhodophyta). However, many aspects of the systematics and divergence times of the group remain unresolved. Using a seven-gene concatenated dataset (nuclear EF2, LSU and SSU rRNAs, mitochondrial cox1, and plastid rbcL, psaA and psbA genes), we generated a robust phylogeny of red algae to provide an evolutionary timeline for florideophyte diversification. Our relaxed molecular clock analysis suggests that the Florideophyceae diverged approximately 943 (817–1,049) million years ago (Ma). The major divergences in this class involved the emergence of Hildenbrandiophycidae [ca. 781 (681–879) Ma], Nemaliophycidae [ca. 661 (597–736) Ma], Corallinophycidae [ca. 579 (543–617) Ma], and the split of Ahnfeltiophycidae and Rhodymeniophycidae [ca. 508 (442–580) Ma]. Within these clades, extant diversity reflects largely Phanerozoic diversification. Divergences within Florideophyceae were accompanied by evolutionary changes in the carposporophyte stage, leading to a successful strategy for maximizing spore production from each fertilization event. Our research provides robust estimates for the divergence times of major lineages within the Florideophyceae. This timeline was used to interpret the emergence of key morphological innovations that characterize these multicellular red algae.

Recent introduction of Polysiphonia morrowii (Ceramiales, Rhodophyta) to Punta Arenas, Chile

Abstract Polysiphonia morrowii (Rhodomelaceae, Rhodophyta) is abundant in intertidal zones of the northwestern Pacific Ocean in spring, and has been introduced to the Mediterranean Sea. We collected specimens of this species from intertidal rocks in February in Punta Arenas, in southernmost Chile. Thalli were densely tufted, slender, and elongate, with four pericentral cells, no cortication and short tetrasporangial ultimate branchlets. The plastid protein-coding rbcL was analyzed from specimens collected in Chile and Korea, as well as from putative relatives. The rbcL sequences of the Chilean specimens were almost identical to those from Korea, and were clearly separated from other related taxa from Chile and other areas. These results suggest that P. morrowii has been introduced recently from the northwestern Pacific Ocean to Chile.

Highly conserved mitochondrial genomes among multicellular red algae of the Florideophyceae

Two red algal classes, the Florideophyceae (approximately 7,100 spp.) and Bangiophyceae (approximately 193 spp.), comprise 98% of red algal diversity in marine and freshwater habitats. These two classes form well-supported monophyletic groups in most phylogenetic analyses. Nonetheless, the interordinal relationships remain largely unresolved, in particular in the largest subclass Rhodymeniophycidae that includes 70% of all species. To elucidate red algal phylogenetic relationships and study organelle evolution, we determined the sequence of 11 mitochondrial genomes (mtDNA) from 5 florideophycean subclasses. These mtDNAs were combined with existing data, resulting in a database of 25 florideophytes and 12 bangiophytes (including cyanidiophycean species). A concatenated alignment of mt proteins was used to resolve ordinal relationships in the Rhodymeniophycidae. Red algal mtDNA genome comparisons showed 47 instances of gene rearrangement including 12 that distinguish Bangiophyceae from Hildenbrandiophycidae, and 5 that distinguish Hildenbrandiophycidae from Nemaliophycidae. These organelle data support a rapid radiation and surprisingly high conservation of mtDNA gene syntheny among the morphologically divergent multicellular lineages of Rhodymeniophycidae. In contrast, we find extensive mitochondrial gene rearrangements when comparing Bangiophyceae and Florideophyceae and multiple examples of gene loss among the different red algal lineages.