Akira F. Peters

Oligoalginate Recognition and Oxidative Burst Play a Key Role in Natural and Induced Resistance of Sporophytes of Laminariales

Forty-five species of brown algae (Phaeophyceae) were surveyed for their capacity to respond by an oxidative burst to challenges with alginate oligosaccharides. Intertidal frondose brown algae (Fucales) constitutively released high quantities of peroxide. The capacity to recognize oligoguluronates and to react with an oxidative burst was confined to alginate-rich taxa with complex thallus morphology, epitomized by the sporophytes of Laminariales. When kelp sporophytes were impaired in their capacity to perform an oxidative burst by the NAD(P)Hoxidase inhibitor diphenylene iodonium, they were readily degraded by their bacterial epiflora. Thus, in these algae, the oxidative response is an essential element of natural resistance. We also report on the establishment of a well-defined experimental system for investigations on kelp immunity, with Laminaria digitata as the host and its phaeophycean endophyte, Laminariocolax tomentosoides, as the pathogen. We found that an alginate-triggered oxidative burst significantly induces resistance in Laminaria digitata against infection. From these findings we infer that oligoalginate signals are important cues in the interaction between laminarialean kelps and potential pathogens.

Development and physiology of the brown alga Ectocarpus siliculosus: two centuries of research.

Brown algae share several important features with land plants, such as their photoautotrophic nature and their cellulose-containing wall, but the two groups are distantly related from an evolutionary point of view. The heterokont phylum, to which the brown algae belong, is a eukaryotic crown group that is phylogenetically distinct not only from the green lineage, but also from the red algae and the opisthokont phylum (fungi and animals). As a result of this independent evolutionary history, the brown algae exhibit many novel features and, moreover, have evolved complex multicellular development independently of the other major groups already mentioned. In 2004, a consortium of laboratories, including the Station Biologique in Roscoff and Genoscope, initiated a project to sequence the genome of Ectocarpus siliculosus, a small filamentous brown alga that is found in temperate, coastal environments throughout the globe. The E. siliculosus genome, which is currently being annotated, is expected to be the first completely characterized genome of a multicellular alga. In this review we look back over two centuries of work on this brown alga and highlight the advances that have led to the choice of E. siliculosus as a genomic and genetic model organism for the brown algae.

Plastid genomes of two brown algae, Ectocarpus siliculosus and Fucus vesiculosus: further insights on the evolution of red-algal derived plastids

BackgroundHeterokont algae, together with cryptophytes, haptophytes and some alveolates, possess red-algal derived plastids. The chromalveolate hypothesis proposes that the red-algal derived plastids of all four groups have a monophyletic origin resulting from a single secondary endosymbiotic event. However, due to incongruence between nuclear and plastid phylogenies, this controversial hypothesis remains under debate. Large-scale genomic analyses have shown to be a powerful tool for phylogenetic reconstruction but insufficient sequence data have been available for red-algal derived plastid genomes.ResultsThe chloroplast genomes of two brown algae, Ectocarpus siliculosus and Fucus vesiculosus, have been fully sequenced. These species represent two distinct orders of the Phaeophyceae, which is a major group within the heterokont lineage. The sizes of the circular plastid genomes are 139,954 and 124,986 base pairs, respectively, the size difference being due principally to the presence of longer inverted repeat and intergenic regions in E. siliculosus. Gene contents of the two plastids are similar with 139-148 protein-coding genes, 28-31 tRNA genes, and 3 ribosomal RNA genes. The two genomes also exhibit very similar rearrangements compared to other sequenced plastid genomes. The tRNA-Leu gene of E. siliculosus lacks an intron, in contrast to the F. vesiculosus and other heterokont plastid homologues, suggesting its recent loss in the Ectocarpales. Most of the brown algal plastid genes are shared with other red-algal derived plastid genomes, but a few are absent from raphidophyte or diatom plastid genomes. One of these regions is most similar to an apicomplexan nuclear sequence. The phylogenetic relationship between heterokonts, cryptophytes and haptophytes (collectively referred to as chromists) plastids was investigated using several datasets of concatenated proteins from two cyanobacterial genomes and 18 plastid genomes, including most of the available red algal and chromist plastid genomes.ConclusionThe phylogenetic studies using concatenated plastid proteins still do not resolve the question of the monophyly of all chromist plastids. However, these results support both the monophyly of heterokont plastids and that of cryptophyte and haptophyte plastids, in agreement with nuclear phylogenies.

Inheritance of organelles in artificial hybrids of the isogamous multicellular chromist alga Ectocarpus siliculosus (Phaeophyceae)

Our knowledge about the sexual transmission of mitochondria and plastids (hereafter organelles) in isogamous eukaryotes comes mostly from studies of the yeast Saccharomyces cerevisiae and the green alga Chlamydomonas reinhardtii which are both unicellular species. To investigate organelle inheritance in a multicellular organism with morphological isogamy, we studied the filamentous brown alga Ectocarpus siliculosus, in which each gamete contributes one plastid and at least one mitochondrion to the zygote. We crossed strains whose organelle genotypes were distinguishable by PCR. Hybrids contained only maternal mitochondrial genotypes, indicating uniparental inheritance of this organelle. In contrast, hybrids were chimerical for the plastid genome. In 64 to 75% of cases examined, the two zygotic plastids were partitioned into the two halves of the sporophyte, which developed from the zygote. A smaller number of hybrids deviated from this pattern, suggesting more complex mechanisms such as irregular division, segregation, or exchange of genetic material between plastids. E. siliculosus is the first isogamous heterokont eukaryote in which inheritance of organelles has been determined with molecular markers. The deterministic pseudo-Mendelian mechanism of plastid inheritance in E. siliculosus is unusual among eukaryotes.

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.

EARLY DEVELOPMENT PATTERN OF THE BROWN ALGA ECTOCARPUS SILICULOSUS (ECTOCARPALES, PHAEOPHYCEAE) SPOROPHYTE1

The distant phylogenetic position of brown macroalgae from the other multicellular phyla offers the opportunity to study novel and alternative developmental processes involved in the establishment of multicellularity. At present, however, very little information is available about developmental patterning in this group. Ectocarpus siliculosus (Dillwyn) Lyngb. has uniseriate filaments and displays one of the simplest architectures in the Phaeophyceae. The aim of this study was to decipher the morphogenetic steps that lead to the development of the Ectocarpus sporophyte. We carried out a detailed morphometric study of the events that occurred between gamete germination and the 100‐cell stage. This analysis was performed on two ecologically distant isolates to assess plasticity in developmental patterning within this species. Cell sizes were measured in both isolates, allowing the definition of two main cell types based on their shape (round and elongated). On average, the filament is composed of about 40% round cells, which are present in the central region of the filament, but different combinations of the two cell types within filaments were observed and quantified. Young sporophytes grew apically, with elongated cells progressively differentiating into round cells. Secondary filaments emerged preferentially on round cells, primarily from the older central cells. Statistical analyses showed that the pattern of branching was regulated to ensure a stereotyped architecture. This description of the developmental patterning during the growth of the E. siliculosus sporophyte will serve as a base for more detailed studies of development, in this species and in brown algae in general.

The Potential Impact of Climate Change on Endophyte Infections in Kelp Sporophytes

There is a strong scientific consensus that coastal marine ecosystems are threatened by global climate change. These ecosystems are particularly vulnerable as many disturbances act at the terrestrial–marine interface and are predicted to increase, such as increased land run-off after floods or higher wave energies owing to increased storm frequency (Helmuth et al., 2006; IPCC, 2007). An alarming decrease in the density and biomass of canopy-forming kelps has been reported worldwide (Dayton et al., 1999; Steneck et al., 2002; Connell et al., 2008) and recent European monitoring programs indicate substantial losses of Laminaria digitata in France (Morizur, 2001) and of Saccharina latissima (formerly L. saccharina) along the Southwest coast of Norway and Sweden (survey in 1996–2006, Norwegian Institute for Water Research, 2007) and on the German island Helgoland (Pehlke and Bartsch, 2008). For instance, the losses of S. latissima at the Norwegian West and Skagerrak coasts are estimated to be 50% and 90%, respectively. Here, the decline in kelp abundance is most pronounced in sheltered waters, where the kelp forest in large areas has been replaced by a silty turf community dominated by filamentous algae. Anthropogenic influences, such as eutrophication and global climate change, have been postulated as possible causes for the loss of canopy-forming kelps. However, substantial scientific evidence is still lacking.

Examining the bank of microscopic stages in kelps using culturing and barcoding

This paper describes a method to study the diversity of young kelp sporophytes that are recruited from the bank of microscopic stages. Small samples of rocky substratum (0.5 cm2) were collected from the low intertidal zone, which was dominated by the kelp Laminaria digitata. Samples were cultivated in the laboratory under conditions permitting gametogenesis. Sporophyte recruits in the cultures were isolated and identified at the species level using the barcoding mitochondrial marker rpl31–rns. Sixty per cent of the collected samples had at least one to a maximum of 30 kelp recruits, belonging to five different species (L. digitata, L. hyperborea, L. ochroleuca, Saccharina latissima and Sacchorhiza polyschides). As the examination of freshly collected rocky samples under a stereo microscope did not reveal any kelp sporophytes, the recruitment in these samples after culture probably occurred from the bank of microscopic forms present on the substratum. Despite the dominance of L. digitata in the field, the young sporophytes obtained after culturing were mainly S. polyschides. This study illustrates the suitability of culturing in combination with molecular identification of young sporophytes to address several key aspects of kelp ecology related to the existence of a bank of microscopic stages in the field.