Jessica Kegel

Pan genome of the phytoplankton Emiliania underpins its global distribution

Coccolithophores have influenced the global climate for over 200 million years. These marine phytoplankton can account for 20 per cent of total carbon fixation in some systems. They form blooms that can occupy hundreds of thousands of square kilometres and are distinguished by their elegantly sculpted calcium carbonate exoskeletons (coccoliths), rendering them visible from space. Although coccolithophores export carbon in the form of organic matter and calcite to the sea floor, they also release CO2 in the calcification process. Hence, they have a complex influence on the carbon cycle, driving either CO2 production or uptake, sequestration and export to the deep ocean. Here we report the first haptophyte reference genome, from the coccolithophore Emiliania huxleyi strain CCMP1516, and sequences from 13 additional isolates. Our analyses reveal a pan genome (core genes plus genes distributed variably between strains) probably supported by an atypical complement of repetitive sequence in the genome. Comparisons across strains demonstrate that E. huxleyi, which has long been considered a single species, harbours extensive genome variability reflected in different metabolic repertoires. Genome variability within this species complex seems to underpin its capacity both to thrive in habitats ranging from the equator to the subarctic and to form large-scale episodic blooms under a wide variety of environmental conditions.

Life-cycle modification in open oceans accounts for genome variability in a cosmopolitan phytoplankton.

Emiliania huxleyi is the most abundant calcifying plankton in modern oceans with substantial intraspecific genome variability and a biphasic life cycle involving sexual alternation between calcified 2N and flagellated 1N cells. We show that high genome content variability in Emiliania relates to erosion of 1N-specific genes and loss of the ability to form flagellated cells. Analysis of 185 E. huxleyi strains isolated from world oceans suggests that loss of flagella occurred independently in lineages inhabiting oligotrophic open oceans over short evolutionary timescales. This environmentally linked physiogenomic change suggests life cycling is not advantageous in very large/diluted populations experiencing low biotic pressure and low ecological variability. Gene loss did not appear to reflect pressure for genome streamlining in oligotrophic oceans as previously observed in picoplankton. Life-cycle modifications might be common in plankton and cause major functional variability to be hidden from traditional taxonomic or molecular markers.

Genome Variations Associated with Viral Susceptibility and Calcification in Emiliania huxleyi

Emiliania huxleyi, a key player in the global carbon cycle is one of the best studied coccolithophores with respect to biogeochemical cycles, climatology, and host-virus interactions. Strains of E. huxleyi show phenotypic plasticity regarding growth behaviour, light-response, calcification, acidification, and virus susceptibility. This phenomenon is likely a consequence of genomic differences, or transcriptomic responses, to environmental conditions or threats such as viral infections. We used an E. huxleyi genome microarray based on the sequenced strain CCMP1516 (reference strain) to perform comparative genomic hybridizations (CGH) of 16 E. huxleyi strains of different geographic origin. We investigated the genomic diversity and plasticity and focused on the identification of genes related to virus susceptibility and coccolith production (calcification). Among the tested 31940 gene models a core genome of 14628 genes was identified by hybridization among 16 E. huxleyi strains. 224 probes were characterized as specific for the reference strain CCMP1516. Compared to the sequenced E. huxleyi strain CCMP1516 variation in gene content of up to 30 percent among strains was observed. Comparison of core and non-core transcripts sets in terms of annotated functions reveals a broad, almost equal functional coverage over all KOG-categories of both transcript sets within the whole annotated genome. Within the variable (non-core) genome we identified genes associated with virus susceptibility and calcification. Genes associated with virus susceptibility include a Bax inhibitor-1 protein, three LRR receptor-like protein kinases, and mitogen-activated protein kinase. Our list of transcripts associated with coccolith production will stimulate further research, e.g. by genetic manipulation. In particular, the V-type proton ATPase 16 kDa proteolipid subunit is proposed to be a plausible target gene for further calcification studies.

Transcriptional host–virus interaction of Emiliania huxleyi (Haptophyceae) and EhV-86 deduced from combined analysis of expressed sequence tags and microarrays

The cosmopolitan coccolithophore Emiliania huxleyi forms frequent massive blooms and thus is important for global climate and the carbon cycle. Lytic viral infection of this alga leads to termination of blooms and therefore influences global climate. To understand the host-virus interaction of E. huxleyi an expressed sequence tag (EST) approach was used to determine changes in gene expression during viral infection. Three cDNA libraries, generated 6, 12 and 24 h after viral infection, were compared to a library from an uninfected culture by sequencing, clustering and manual annotation of 1100–1500 ESTs per library. To verify the gene expression results of the ESTs we used two-colour oligonucleotide microarrays. A total of 4480 ESTs were assembled into 1871 clusters, of which, 223 are of viral origin. Microarray expression analysis indicated that 231 out of 565 oligonucleotides of E. huxleyi changed their expression level for at least at one time point in response to viral infection. Results suggest that viral infection affects the following processes: photosynthesis, transcription and translation, carbohydrate and lipid metabolism (particularly glycolysis), metabolism, and signal transduction. Results of this study provide insights into the gene expression of E. huxleyi during infection by the virus EhV-86.

In situ expression of eukaryotic ice-binding proteins in microbial communities of Arctic and Antarctic sea ice

Ice-binding proteins (IBPs) have been isolated from various sea-ice organisms. Their characterisation points to a crucial role in protecting the organisms in sub-zero environments. However, their in situ abundance and diversity in natural sea-ice microbial communities is largely unknown. In this study, we analysed the expression and phylogenetic diversity of eukaryotic IBP transcripts from microbial communities of Arctic and Antarctic sea ice. IBP transcripts were found in abundances similar to those of proteins involved in core cellular processes such as photosynthesis. Eighty-nine percent of the IBP transcripts grouped with known IBP sequences from diatoms, haptophytes and crustaceans, but the majority represented novel sequences not previously characterized in cultured organisms. The observed high eukaryotic IBP expression in natural eukaryotic sea ice communities underlines the essential role of IBPs for survival of many microorganisms in communities living under the extreme conditions of polar sea ice.

Genomics of Algal Host–Virus Interactions

Viruses in Earths aquatic environment outnumber all other forms of life and carry a vast reservoir of genetic information. A large proportion of the characterized viruses infecting eukaryotic algae are large double-stranded DNA viruses, each of their genomes carrying more than a hundred genes, but only a minority of their genes resemble genes with known biological functionalities. Unusual forms of single-stranded DNA and single- and double-stranded RNA viral genomes have been characterized over the last 10 years, and the number of novel taxa of viruses being discovered continues to increase. Although viral infections are usually specific to certain host strains in a species, lytic viral infections nevertheless affect a large proportion of algae and have a global impact, for example in the termination of blooms. Resistance to viruses is thus subject to strong selection, but little is known about its mechanism. Lateral gene transfer between host and virus has been shown by comparisons between their complete genomes and must play an important role in coevolution in the microbial world. Recent advances in bioinformatics and the possibility of amplifying complete genomes from single cells promise to revolutionize analyses of viral genomes from environmental samples.