images.nature.com€¦ · Web viewSupplementary Information. Supplementary information is provided in three files: a word file containing materials and methods, phylogenetic analysis

Supplementary Information

Supplementary information is provided in three files: a word file containing materials and methods,

phylogenetic analysis results, legend to supplementary figures 1, 2, 3, 4, 5, 6, 7 and 8;

supplementary tables 1, 2 and 3, sequences of Skeletonema marinoi transcripts of the prostaglandin

pathway reported in the main text; a word file with the sequences used in the phylogenetic analysis;

eight pdf file containing the supplementary figures 1, 2, 3, 4, 5, 6, 7 and 8.

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Supplementary information: file 1.

Content: materials and methods, phylogenetic analysis results, legend to supplementary figures 1, 2, 3, 4, 5, 6, 7 and 8; supplementary tables 1, 2 and 3; sequences of Skeletonema marinoi transcripts of the prostaglandin pathway reported in the main text.

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Materials and Methods

Bioinformatic identification of the Prostaglandin pathway

Transcriptome sequencing of Skeletonema marinoi stain FE7 and FE60 (MMETSP1039 and

MMETSP1040 respectively) were obtained thanks to the Marine Microbial Eukaryote

Transcriptome Sequencing Project initiative, by the National Centre for Genome resources (NGR)

(Santa Fe, New Mexico, USA), supported by the Betty and Gordon Moore Foundation

(www.moore.org). Sequences are now freely available in the public database CAMERA

(http://camera.crbs.ucsd.edu/mmetsp/list.php) and iMicrobes interactive query tool for microbial

data (http://data.imicrobe.us/sample/view/1867 http://data.imicrobe.us/sample/view/1868).

To identify expressed metabolic pathways, Skeletonema marinoi FE7 and FE60 contigs were

submitted to the Kyoto Encyclopaedia of Genes and Genome (KEGG) Automatic Annotation

Server (KAAS, http://www.genome.jp/tools/kaas/) as “Complete or Draft Genome” using bi-

directional best hit assignment method and adding to the default organisms the following others:

Phaeodactylum tricornutum, Thalassiosira pseudonana, Ostreococcus lucimarinus, Ostreococcus

tauri and Cyanidioschyzon merolae. Enzymes involved in prostaglandin synthesis were found

annotated in map 0590 relative to arachidonate metabolism (Supplementary Figure S1). The protein

sequence annotated as prostaglandin transporter was found exploring the annotation list obtained

using Blast2go PRO software (http://www.blast2go.com/b2ghome) with default parameters.

Phytoplankton cultures

The two Skeletonema marinoi strains, FE7 and FE60, were isolated from phytoplankton samples

collected during diatom blooms in the northern Adriatic Sea in 1997 and 2005, respectively

(Gerecht et al., 2011). Three replicate cultures of each clone, with initial cell concentrations of 5000

cells/mL, were grown in f⁄2-medium (Guillard, 1975) in 10 lt polycarbonate bottles under gentle

bubbling with sterile-filtered ambient air and kept in a climate chamber at 20°C on a 12:12 light:

dark (L:D) cycle at 100 µmol photons m-2 s-1. Culture growth was followed in each replicates by

daily cell enumeration in a Sedgewick-Rafter counting chamber (Hausser Scientific, Horsham, PA, 3

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USA), up to the 11th day from the inoculation. All cultures were harvested always at the same time

of the day in order to avoid interference due to circadian variability. Samples were collected on day

4, 7 and 11 representing exponential, stationary and senescent phases of the growth curve,

respectively (Supplementary figure S2). Cultures were concentrated by centrifugation in a cooled

centrifuge at 2753g for 15 min with a swing-out rotor (DR 15P, Braun Biotechnology International,

Allentown, PA, USA), immediately frozen in liquid nitrogen, and kept at -80°C until analysis.

Silica starvation experiment were conducted in triplicates as described in Orefice et al. (Orefice et

al., 2015).

RNA extraction and reverse transcription

Total RNA was extracted from 500 ml of cultures harvested at day 4 and 300 ml of cultures

harvested at days 7 and 11. Each pellet was treated following the procedure described in Barra et al.

(Barra et al., 2013). DNase treatment was carried out using DNAse I recombinant, RNAse-free

(Roche, Basel, Switzerland) method according to manufacturer's protocol to eliminate potential

genomic DNA contamination. Total RNA sample was purified and concentrated using RNeasy

MinElute Cleanup Kit (Qiagen, Venlo, Netherlands) and eluted in 20 μl RNase-free water.

Concentration of the resulting RNA samples was assessed by absorbance at 260 nm (ND-1000

Spectrophotometer; NanoDrop Technologies, Wilmington, DE, USA). The integrity of total RNA

was checked with agarose gel electrophoresis. From each RNA sample 1g was retro-transcribed in

the T100 Thermal cycler (Bio-Rad Laboratories, Hercules, CA, USA) following the manufacturer's

instructions of the sensiFAST™ cDNA synthesis kit (Bioline, Cat. No. BIO-65054).

Primer design and real time quantitative PCR

Transcript of interest were selected considering the annotation of the putative protein sequences

reported in the transcriptome of S. marinoi strain FE7 from CAMERA: MMETSP1039-20121108|

4563_1 Prostaglandin G/H synthase 1/2 (Ptgs 1/2) or cyclooxygenase-1 (COX-1), MMETSP1039-

20121108|5744_1 Prostaglandin E synthase 2 (PtgEs), MMETSP1039-20121108|1431_1 Glutatione

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S-transferase o Prostaglandin H2 D-isomerase (PtgHi or PtgDs), MMETSP1039-20121108|

17278_1 Prostaglandin Transport (PtgT).

Primers, designed using Primer3 program V. 0.4.0 (Untergasser et al., 2012; Koressaar and Remm,

2007), are:

COX-1: Ptgs_F: GGGTGGGATGATGAACGTGT; Ptgs_R: AAGGACCTGGCTCGACTTTG;

PtgEs: PtgEs_F: GAGTTGAAACCATGGTCAGGAG; PtgEs_R:

TGTGTATCAAGTAGCGCATCG; PtgDs or PtgHi: PtgHi_F: AAAAGAATGGCGGAGTGGGA;

PtgHi_R: GTCGCCACAATACCCGCATA; for PtgT: PtgTr_F:

GGAGAGCATTGGAGTCAGCA; PtgTr_R: TTCGCCACCCTCTGAAATGT.

Each sequence was initially tested by standard PCR. Reactions were carried out in 25 μl volume

with 2,5 μl of 10× PCR reaction buffer (Roche, Basel, Switzerland), 2,5 μl of 10 × 2 mM dNTP, 0.2

μl of 5 U/μl Taq (Roche, Basel, Switzerland), 1,25μl 10 of each oligo, 1 μl of cDNA templates

and nuclease-free water up to 25 μl. The PCR program consisted of a denaturation step at 95 °C for

3 min, 40 cycles at 95 °C for 30 s, 53 °C 30s, 72 °C for 30s, and a final extension step at 72 °C for 7

min. Amplified PCR products were analysed by agarose gel electrophoresis. The resulting bands

were excised from the gel and extracted according to the GenElute Gel Extraction Kit protocol

(Sigma-Aldrich, St. Louis, MO, USA). Sequences were obtained by BigDye Terminator Cycle

Sequencing Technology (Applied Biosystems, Foster City, CA, USA) and purified using the

Agencourt CleanSEQ Dye terminator removal Kit (Agencourt Bioscience Corporation, Beverly,

MA, USA) in automated robotic station Biomek FX (Beckman Coulter, Pasadena, CA, USA).

Products were analysed on the Automated Capillary Electrophoresis Sequencer 3730 DNA

Analyser (Applied Biosystems, Foster City, CA, USA). Alignments were performed with BioEdit

software V. 7.0.5.3 (http://www.mbio.ncsu.edu/BioEdit/bioedit.html).

Selected reference genes were ribosomal protein small subunit 30S (RPS), actin (ACT) and

ubiquitin (UB) which were the best reference genes in Orefice et al. (Orefice et al., 2015).

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Reverse transcription-quantitative PCR (RT-qPCR) experiments were performed in MicroAmp

Optical 384-Well reaction plate (Applied Biosystems, Foster City, CA, USA) with Optical

Adhesive Covers (Applied Biosystems, Foster City, CA, USA) in a Viia7 Real Time PCR System

(Applied Biosystem, Foster City, CA, USA). Five serial dilutions of mixed cDNAs were used to

determine primer reaction efficiency (E= 10−1/slope) that for each oligo couple was two. The PCR

volume for each sample was 10μl, with 5μl of Fast Start SYBR Green Master Mix with ROX

(Roche, Basel, Switzerland), 1μl of cDNA template (1 to 5 dilution each template) and 1μl of 10M

oligo mix (forward and reverse). Program reaction used was: 95°C for 20s, 40 cycles of 95°C for 1s

and 60°C for 20s. The program was set to reveal the melting curve of each amplicon from 60°C to

95°C, and read every 0.5°C. Single peaks for all genes confirmed gene-specific amplification and

the absence of primer-dimers. All RT-qPCR reactions were carried out in triplicate to capture intra-

assay variability. Each assay included three no-template negative controls for each primer pair.

The expression levels of each gene of interest relative to the most stable reference genes were

calculated by using the REST tool (Relative Expression Software Tool) (Pfaffl et al., 2002). The

evaluation of gene expression along the growth curve of S. marinoi, was calculated considering the

exponential phase at the fourth day of the growth curve as the reference condition.

Data are represented in graph as log2 x-fold difference in the expression level of the analysed genes

in the stationary (Day7) and senescent (Day11) phases compared to the control exponential phase

(Day4). Results of silica starvation experiment are reported as log2 x-fold difference in the

expression level of the analysed genes for each growth phase under silica starvation with respect to

the corresponding phase in standard culturing conditions (control: complete medium). Relative

expression ratios above two fold were considered significant. Statistical analysis was performed

using the Pair Wise Fixed Reallocation Randomisation test by REST.

Prostaglandin extraction

500 ml of cell culture from day 4 and 300 ml of cell culture from days 7 and 11 were harvested by

centrifugation at 2753g for 15 minutes. The pellets were re-suspended in 1 ml PBS per gr wet

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weight, added with 1µg of Prostaglandin E2-d4 (CAYMAN CHEMICAL, Michigan, USA) as

internal standard and sonicated with 1-minute pulse, 30 seconds on ice and another 30 seconds of

pulse at 20% of maximum intensity. After sonication, an aliquot of 50 l was taken to measure

proteins concentration by Bradford assay (PanReac AppliChem, product code A6932). Samples

were left 10 minutes at room temperature. To precipitate proteins, samples were added with 4

volumes of ethanol and left 5 minutes at 4°C. Precipitated proteins were removed by centrifugation

at 3000g for 10 minutes. After ethanol evaporation under nitrogen, samples were acidified with 4

volumes of 1M acetate buffer pH 4 and centrifuged 15 minutes at 4200g. Clean supernatant was

fractionated by using an automated procedure on a GX-271 ASPEC Gilson

apparatus equipped with a TRILUTION® LH Software (Gilson, Middleton, WI,

USA). C18 columns (Aspec C18 6 mL/500 mg. Gilson, Middleton, WI, USA) were

activated with 5 ml of methanol and equilibrated with 10 mL of distilled water. Elution

was performed after a first washing step with 5 mL 100% H2O using 5ml ethyl

acetate-1% methanol. The ethyl acetate-methanol fraction was dried under nitrogen and saved until

the chemical analysis.

LC/MSMS analysis

Samples were resuspended in 1ml methanol and analysed by using a 4000QTrap mass spectrometer

(Applied Biosystems, Toronto , Canada), a QqLIT instrument, working in negative ion mode and

coupled to a 1100 nanoHPLC system (Agilent Technologies, Waldbronn, Germany). The

prostaglandins were separated by using a micro C18 column (10 cm x 1,0mm, 5µm). The mobile

phase was generated by mixing eluent A (0.1% Acetic Acid) and eluent B (acetonitrile/isopropanol

50/50) and the flow rate was 30 nl/min. Starting condition was 20% to 95% B in 15 minutes.

Tandem mass spectrometry was performed using a turbo ion spray source operated in negative

mode, and the multiple reaction monitoring (MRM) mode was used for the selected analytes. For

the detection of metabolites, the Turbo ion spray source was operated as follows: curtain gas 20psi,

GS 1/2 50/50psi, ion spray voltage -5500V , DP, -50 V; Dwell 25ms and temperature of 550°C. MS

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were carried out on a quadrupole MRM mode for identification and quantification. MRM was

performed by the detection of a unique product ion arising from collision-induced fragmentation of

the deprotonated selected parent compounds (Wang et al., 2014; Yang et al., 2009) after proper

optimization of mass spectral parameters as reported in Supplementary Table 1 and Supplementary

Figure 3 and 4.

The amount of prostaglandins quantified as described above, was normalized by protein content of

each sample.

Phylogenetic analyses

The COX-1, PTGE Synthase, PTGH Isomerase and PTG Transporter sequences from the other

diatom species were obtained as follow: the file with the peptide sequences of each diatom species

was downloaded from the MMETSP website (http://data.imicrobe.us/project/view/104). Each

peptide file was indexed (following NCBI instructions:

http://www.ncbi.nlm.nih.gov/books/NBK1763/) and searched using the corresponding Skeletonema

marinoi clone FE7 sequence as a query in a blastp search with default parameters. Sequences with

highest similarity were blasted against the non-redundant (NR) NCBI database to verify the identity

and checked for their annotation in the corresponding MMETSP project swiss-prot table. Outgroup

sequences were selected from the blastp results of each Skeletonema marinoi strain FE7 protein

analyzed in the prostaglandin pathway against the NCBI non redundant sequences database

(https://blast.ncbi.nlm.nih.gov/Blast.cgi?PAGE=Proteins ).

Sequences were visualized and aligned using BioEdit Sequence Alignment Editor (Hall, 1999). A

first automatic alignment was performed using the ClustalW algorithm implemented in BioEdit,

then the alignment was manually refined. Bayesian analyses were performed on the alignment using

MrBayes (Ronquist and Huelsenbeck, 2003) computer program. All the settings for each analysis

are listed in the figure 2 legend. For Bayesian analyses, two parallel and totally independent

Markov Chain Monte Carlo (MCMC) runs were carried out on data matrices. Four chains (three hot

and one cold) drove each analysis. The number of generations and the split frequency standard

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deviation for each analysis are reported in figure 2 legend. Sampling frequency was set every 500

generations. The analyses were forced to jump among the evolutionary models for protein sequence

alignments implemented in the software. The first 25% of the samples from the cold chain were

discarded in order to stabilize the algorithm, reduce the variability among results and have a more

robust analysis. Consensus tree with posterior probability (PP) of each node and branch length are

reported after a 50% majority-rule consensus. Phylogenetic trees were visualized and edited in the

FigTree (Tree Figure Drawing Tool Version 1.4.2) software (http://tree.bio.ed.ac.uk/).The complete

list of sequences used in the present work for phylogenetic analyses are listed in Supplementary

table 3. The sequences from strain SkelB that were annotated as Skeletonema dohrnii in MMETSP

database were changed to S. marinoi after ribosomal sequence identification. Sequences ID were

left unchanged in the fasta sequences list attached at the end of this supplemental information file.

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http://tree.bio.ed.ac.uk/

Results

Effect of nutrient limitation on prostaglandin enzymes gene expression

Silica starvation experiment (36 μM Si(OH)4) revealed a large variability among replicates and no

significant differences were evident in the expression levels of prostaglandin synthesizing enzymes

(Supplementary Figure S5).

The MMETSP database includes transcriptomic data of different diatom species and clones grown

in many different conditions: nutrient limitation (Silica, Iron, Nitrate, Phosphorus), high or low

CO2, changes in light/dark cycle, freezing temperature, nocodazole treatment, different nitrogen

sources such as glutamate,urea, ammonium or nitrate. We used this information to analyze the

relative presence of the three enzymes of the prostaglandin pathway (Supplementary Table 2).

Results highlight a large variability of COX-1 expression (reported as Fragments Per Kilobase of

transcript per Million mapped reads, FPKM) among species and among different growth conditions

for the same species. For example, COX-1 expression in Thalassiosira gravida, clone GMp14c1,

seems to be affected by nitrogen limitation but not by phosphorus with respect to the control

condition. Skeletonema marinoi clone SkelA, contrary to our clones, seems to be affected by silica

deprivation, with COX-1 expression being almost doubled compared to the control condition. On

the other hand, COX-1 expression seems to be affected by silica limitation in Chaetoceros

neogracile, by iron deprivation in Chaetoceros debilis and by nitrogen, phosphorus and CO2 in

Chaetoceros affinis.

Detailed results of the phylogenetic analyses on Prostaglandin D Synthase (PTGDS) and Prostaglandin transporter (PTGT)

In addition to information given in the main text, here we report detailed description of the

phylogenetic analysis performed on two of the protein sequences involved in the prostaglandin

pathway.

PTGD Synthase

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The Bayesian phylogeny produced with PTGD Synthase alignment contained 230 sequences from

diatoms and other organisms (Supplementary Figure S7). The protein sequences were very variable

and in fact after 10 million generations, the split frequency standard deviation did not go below 0.04

and the log likelihood did not change over at least five million generations. This tree showed at least

nine variably supported big clades (PP from 0.53 to 1.00); five out of them contained only diatom

sequences. The remaining four clades contained at least one outgroup sequence, some of which

annotated as Glutathione S-transferase 4, Glutathione S-transferase-like protein, Glutathione-

requiring prostaglandin D synthase or Glutathione S-transferase. The other outgroup sequences in

fact cluster in unresolved branches or in supported but not resolved small clades. What is worth of a

note is that all but one clades contained both centric and pennate diatoms; this can corroborate the

hypothesis that the nine clades in fact cluster different proteins, present in both diatom classes. This,

together with the low level of resolution of the phylogeny, suggests that the sequences retrieved by

BLAST in MMETSP and in NCBI databases for diatoms and other organisms in fact share a certain

identity and contain similar domains, but can belong to different protein families. Conversely,

PTGD Synthase can have evolved quite differently in different organisms and also in diatoms

different proteins can serve the same function or be involved in different conditions. More

functional studies are required to disentangle this issue.

PTG transporter

PTGT sequence was annotated in the transcriptomes of our strains as prostaglandin transporter.

However, blastp of its protein sequence in the NCBI database retrieved similarity with the broad

ABC transporter family which include less specific transporters. For this reason, Bayesian analysis

of the PTGT sequence was restricted only to diatom sequences that showed a low e-value indicating

high rate of similarity. The analyses showed four main clades indicated by different shades in

Supplementary Figure 8. In this tree, some diatom genera are represented in all the clades, while

some others appear in only one (Cyclotella, Amphora, Coscinodiscus, Astrosyne, Cyclophora,

Nitzschia, Attheya, Striatella), two (Staurosira, Grammatophora, Dactyliosolen, Odontella,

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Aulacoseira, Thalassiotrix, Minutocellulus) or three clades (Amphiprora, Rhizosolenia,

Asterionellopsis, Licmophora, Thalassionema, Stephanopyxis, Proboscia, Letpocylindrus). Among

the genera that appear in only one clade, no clear taxonomic pattern can be identified because in

clade A two centric and three pennate diatoms are recorded as single appearances; while in clade D,

one centric and two pennates were recorded. What is worth noting is that single occurrences were

recorded only in clades A and D (that are the widest in the tree) and that pennate genera are the

most represented with five genera against 3 centric ones. Among the double and triple occurrences,

centric diatoms are the most represented, with five genera against two pennate for the former; and

six centric against two pennate genera for the latter. All the double and triple occurrences have at

least one representative in clades A and D (apart from Minutocellulus with sequences clustering in

clades A and C and Asterionellopsis not recorded in clade D). From this analysis of clustering

pattern it emerged that likely transporter proteins clustering in clades A and D are the most spread

among diatoms (no single occurrences were recorded in clades B and C) possibly revealing a

subordinate hierarchical level of transporter proteins B and C. No identification of subcellular

localization was performed on the sequences because transcripts usually miss the specific signal

peptide or were truncated.

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References

Barra L, Ruggiero MV, Sarno D, Montresor M, Kooistra WHCF. (2013). Strengths and weaknesses of microarray approaches to detect Pseudo-nitzschia species in the field. Environ Sci Pollut Res Int 20: 6705–6718.

Gerecht A, Romano G, Ianora A, d’Ippolito G, Cutignano A, Fontana A. (2011). Plasticity of Oxylipin Metabolism Among Clones of the Marine Diatom Skeletonema Marinoi (bacillariophyceae)1. J Phycol 47: 1050–1056.

Guillard RRL. (1975). Culture of Phytoplankton for Feeding Marine Invertebrates. In: Smith WL, Chanley MH (eds). Culture of Marine Invertebrate Animals. Springer US, pp 29–60.

Koressaar T, Remm M. (2007). Enhancements and modifications of primer design program Primer3. Bioinforma Oxf Engl 23: 1289–1291.

Orefice I, Lauritano C, Procaccini G, Ianora A, Romano G. (2015). Insights into possible cell-death markers in the diatom Skeletonema marinoi in response to senescence and silica starvation. Mar Genomics 24 Pt 1: 81–88.

Pfaffl MW, Horgan GW, Dempfle L. (2002). Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30: e36.

Ronquist F, Huelsenbeck JP. (2003). MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19: 1572–1574.

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucleic Acids Symposium Series 41: 95–98.

Untergasser A, Cutcutache I, Koressaar T, Ye J, Faircloth BC, Remm M, et al. (2012). Primer3--new capabilities and interfaces. Nucleic Acids Res 40: e115.

Wang Y, Armando AM, Quehenberger O, Yan C, Dennis EA. (2014). Comprehensive ultra-performance liquid chromatographic separation and mass spectrometric analysis of eicosanoid metabolites in human samples. J Chromatogr A 1359: 60–69.

Yang J, Schmelzer K, Georgi K, Hammock BD. (2009). Quantitative Profiling Method for Oxylipin Metabolome by Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry. Anal Chem 81: 8085–8093.

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Legend to Supplementary Figure 1, 2, 3, 4, 5, 6, 7 and 8

Supplementary Figure 1: Arachidonic acid metabolism reference pathway (Map 0590).

The map shows the metabolic pathways originating from the arachidonic acid, including the

prostaglandin pathway. Enzymes are identified by their code numbers (EC) enclosed in rectangular

boxes. Red arrows highlight the prostaglandin pathway. Green ovals indicate gene product

annotated in the Skeletonema marinoi strain FE7 transcriptome and blue rectangles the gene product

annotated in FE60 strain.

Supplementary Figure 2: Skeletonema marinoi growth curves.

Growth curve of Skeletonema marinoi strain Fe7 (A) and strain FE60 (B) from day 0 to day 11.

Day 4, 7 and 11, have been taken as representative point of the exponential, stationary and

senescent phases of the growth curve, respectively. Day 4 was considered the control phase, as it

represents the health phase of a diatom culture.

Supplementary Figure 3: MRM chromatogram.

MRM chromatogram for diatom sample displaying transitions for all targeted prostaglandins.

Supplementary Figure 4: MRM chromatogram of selected analytes.

MRM chromatogram only showing some detected PGs. The specific MRM transitions for

monitoring the different analytes are indicated as an example. The deuterated PG used as internal

standard is also reported.

Supplementary Figure 5: Expression levels of the prostaglandin enzymes in Skeletonema

marinoi (Clone FE7) under silica starvation.

Expression levels of enzymes responsible for the PGs synthesis and of the prostaglandin transporter

measured by Real Time qPCR in exponential, stationary and senescent phases of growth under

silica starvation. Results were analysed with the REST software and reported as 2log expression

ratio on the standard growth conditions (control) ±SD. Statistical analysis (N=3) was performed

using the Pair Wise Fixed Reallocation Randomization test by REST. Relative expression ratios

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above two fold were considered significant. Results are means of three technical replicates for each

biological replicate (N=3).

Supplementary Figure 6: Skeletonema marinoi-COX1 alignment.

Skeletonema marinoi COX1 sequence (highlighted in yellow) alignment with representative COX

sequences from vertebrate and invertebrate organisms. The alignment of Sm_COX-1 showed a

percent of identity ranging from about 36% to 24% having the highest similarity with cyanobacteria

followed by mammals, corals, teleosts and finally red algae. Amminoacidic residues Gln203 and

His388 involved in peroxidase activity and residue Tyr385, crucial to initiate cyclooxygenase

activity, were conserved, similarly to the red algae, while the region containing the amminoacidic

residue Arg120, which is the site that interacts with the substrate, was poor conserved.

Representative domains are highlighted in red boxes following the Ovis aries (highlighted in pink)

COX sequence positions. Important amminoacidic residues are highlighted in blue.

Supplementary Figure 7: PTGD Synthase phylogenetic tree.

Bayesian mid-point routed phylogenetic trees of PTGD Synthase proteins (10,000,000 generations,

split frequency standard deviation = 0.04) from pennate (blue) and centric (red) diatoms. Non-

diatom sequences are reported in black. Sequences used for phylogenetic analyses are listed in

Supplementary Table S3. The tree was built on scale in the branch length (scale bars reported).

Sequences are identified by the species name and annotation (where applicable), followed by their

MMETSP ID deprived of the taxon ID for simplicity or GenBank accession number. Posterior

probability (PP) values are reported at the nodes. Branch coloring refers to PP, color code is

reported in figure. The nine main clades are identified by letters (see Figure 2c in the main text)

highlighted with vertical bars on the right hand side of the picture. Extended clusters 10 and 11 are

also highlighted.

Supplementary Figure 8: PTG transporter phylogenetic tree.

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Bayesian mid-point routed phylogenetic tree (5,000,000 generations, split frequency standard

deviation = 0.01) of proteins from pennate (blue) and centric (red) diatoms. The tree was built on

scale in the branch length (scale bar reported). Sequences are identified by species name and

MMESP ID deprived of the taxon ID for simplicity. Four main clades (see text) are highlighted in

different colors. Posterior probability (PP) values are reported at the nodes. Branch coloring refers

to PP, color code is reported in figure.

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3132

Supplementary Table 1: Prostaglandin metabolites mass spectral parameters (LC/MSMS data)

Abbreviation Name Pathway Precursor fatty acid

Parent ion

Product Ions

collision energy (V)

PGE1 Prostaglandin E1 COX ETE 353 191 -28235 -29

PGD1 Prostaglandin D1 COX ETE 353 317 -30163 -29

PGF1 Prostaglandin F1 COX ETE 355 311 -24293 -36

6k PGE1 6-keto-prostaglandin E1 COX ETE 367 163 -30331 -36

PGE2 Prostaglandin E2 COX AA 351 235 -25271 -35

PGD2 Prostaglandin D2 COX AA 351 271 -24233 -35

PGF2 Prostaglandin F2 COX AA 353 309 -32193 -35

PGA2 Prostaglandin A2 COX AA 333 189 -30271 -20

PGB2 Prostaglandin B2 COX AA 333 175 -25235 -30

PGJ2 Prostaglandin J2 COX AA 333 251 -28189 -30

PGK2 Prostaglandin K2 COX AA 349 287 -31249 -28

11 PGF2 11 Prostaglandin F2 COX AA 353 299 -25335 -30

15d PGA2 15-deoxy-Prostaglandin A2 COX AA 315 233 -23255 -30

15d PGD2 15-deoxy-Prostaglandin D2 COX AA 333 203 -28271 -26

15d PGJ2 15-deoxy-Prostaglandin J2 COX AA 318 217 -30203 -30

15k PGD2 15-keto-Prostaglandin D2 COX AA 349 287 -24235 -18

15k PGE2 15-keto-Prostaglandin E2 COX AA 349 283 -27235 -22

15k PGF2 15-keto-Prostaglandin F2 COX AA 351 289 -30219 -30

dhk PGE2 13,14-dihydro-15-keto-Prostaglandin E2 COX AA 351 315 -26

207 -28

dhk-PGD2 13,14-dihydro-15-keto-Prostaglandin D2 COX AA 351 333 -24

175 -35

dh-PGF2 13,14-dihydro Prostaglandin F2 COX AA 355 275 -32

283 -22

dhomo-PGF2 dihomo-Prostaglandin F2 COX AA 381 301 -24337 -24

dihomo_PGJ2 dihomo-Prostaglandin J2 COX AA 361 284 -17299 -28

PGE3 Prostaglandin E3 COX EPA 349 313 -15269 -24

PGD3 Prostaglandin D3 COX EPA 349 233 -28269 -24

PGF3 Prostaglandin F3 COX EPA 351 147 -24193 -26

17

357

358

359

3334

Supplementary table 2: presence/absence of complete or partial prostaglandin pathway in all available MMETSP transcriptomes of diatom species grown in different conditions.

MMETSP ID Genus Species Clone Treatment COX1

FPKM

PtgE Synthas

e

PtgH isomerase

Ptg Transporte

r

1065 Amphiprora paludosa CCMP125 - - + + +

0724 Amphiprora sp CCMP467 Exponential - + + +

0727 Amphiprora sp CCMP467 Nocodazole - - - +

0725 Amphiprora sp CCMP467 SiLim - + + +

0316 Amphora coffeaeformis CCMP127

f/2 + silicate,

adjusted to 1 mM

nitrate and amended with 200

µM acetate

- - - +


f/2 + silicate, but

without nitrate

- - - -


f/2 + silicate,

adjusted to 1 mM nitrate

- - + -

1394 Asterionellopsis glacialis CCMP1581 - + + + +

0713 Asterionellopsis glacialis - - - + + +

0705 Asterionellopsis glacialis CCMP134 Exponential - - - -

0708 Asterionellopsis glacialis CCMP134 Nocodazole - - - -

0707 Asterionellopsis glacialis CCMP134 N-limited + 7305,03 - + +

0706 Asterionellopsis glacialis CCMP134 Si-limited + 8052,7 - + -

0418 Astrosyne radiata 13vi08-1A - - - + +

1449 Attheya septentrionalis CCMP2084 - - + + +

1064 Aulacoseira subarctica CCAP1002/5 - + + + +

0785 Bolidomonas pacifica CCMP1866 - - + + -

0088 Chaetoceros affinis CCMP159 Replete-ModernCO2 + 22383 - + -

0090 Chaetoceros affinis CCMP159 -N-ModernCO2 + 43066 - + +

0091 Chaetoceros affinis CCMP159 -P-ModernCO2 + 7994,5 + + +

18

360361

3536

0092 Chaetoceros affinis CCMP159 Replete-FutureCO2 + 2526,8 - + -

1435 Chaetoceros brevis CCMP164 - - - + -

0716 Chaetoceros curvisetus Exponential - - - -

0717 Chaetoceros curvisetus Exponential + + + +

0718 Chaetoceros curvisetus Exponential - + + +

0719 Chaetoceros curvisetus Exponential - - - -

0149 Chaetoceros debilis MM31A-1 CTR +21207,5

- + +

0150 Chaetoceros debilis MM31A-1 -Fe +10718,8

+ + +

1447 Chaetoceros dichaeta CCMP1751 - - + + +

1336 Chaetoceros neogracile RCC1993 - + + + +

0751 Chaetoceros neogracile CCMP1317 Exponential + 2887 - + +

0753 Chaetoceros neogracile CCMP1317 NLim + 2311,38 + - -

0754 Chaetoceros neogracile CCMP1317 Nocodazole + 5420,1 - + +

0752 Chaetoceros neogracile CCMP1317 SiLim + 4620 - + +

0200 Chaetoceros sp. clone GSL56 - - + + +

1429 Chaetoceros sp. clone UNC1202 - - + + +

0010 Corethron hystrix 308 - - + + +

0169 Corethron pennatum L29A3 CTR - - + +

0171 Corethron pennatum L29A3 -Fe - - + +

1066 Coscinodiscus wailesii CCMP2513 - - + + +

1442 Craspedostauros australis CCMP3328 - - + + +

0397 Cyclophora tenuis ECT3854 - - + + +

1057 Cyclotella meneghiniana CCMP 338 - + - + +

0017 Cylindrotheca closterium KMMCC:B-181 - - + + +

0580 Dactyliosolen fragilissimus - - + + +

1058 Detonula confervacea CCMP 353 - + + + +

1443 Entomoneis sp CCMP2396 - - + + -

193738

1437 Eucampia antarctica CCMP1452 - - + + +

0733 Fragilariopsis kerguelensis L26-C5 Fe LL - - + +

0734 Fragilariopsis kerguelensis L26-C5 Fe SL - - + +

0735 Fragilariopsis kerguelensis L26-C5 -Fe LL - + + -

0736 Fragilariopsis kerguelensis L26-C5 -Fe SL - - + +

0906 Fragilariopsis kerguelensis L2-C3

freezing_temperature (T=-2°C, light=45)

- - - -


control growth (T=5°C,

light=100)

+ - - +


iron limitation,

T=4°C, light=80

- - + +

0909 Fragilariopsis kerguelensis L2-C3 Darkness, T=5°C - - - -

0009 Grammatophora oceanica CCMP 410 - + - + +

0322 Leptocylindrus apora B651 - - + + +

0321 Leptocylindrus danicus B650 - - - - -

1362 Leptocylindrus danicus CCMP1856 - - + + +

1360 Licmophora paradoxa CCMP2313 - - + + +

1070 Minutocellus polymorphus NH13 - - - + +

1434 Minutocellus polymorphus CCMP3303 - - - - +

1322 Minutocellus polymorphus RCC2270 - - + + +

0744 Nitzschia punctata CCMP561 Exponential - - + +

0745 Nitzschia puncatata CCMP561 SiLim - + - +

0746 Nitzschia puncatata CCMP561 N-limit - - - -

0747 Nitzschia punctata CCMP561 Nocodazole - - + +

0014 Nitzschia sp RCC80 - + + + +

0015 Odontella aurita 1302-5 - + + + +

0160 Odontella sinensis Grunow1884 - + + + +

203940

0174 Proboscia alata PI-D3 CTR - - + +

0176 Proboscia alata PI-D3 -Fe - + + +

0816 Proboscia inermis CCAP1064/1 - - + + +

0329 Pseudonitzschia arenisensys B593 - + + + +

0142_2 Pseudonitzschia australis 10ABGLUTAMA

TE as N source

- - + +

0140 Pseudonitzschia australis 10AB NH4 as N source - - + -

0139 Pseudonitzschia australis 10AB NO3 as N source + - - +

0141 Pseudonitzschia australis 10AB UREA as N source - + + +

0327 Pseudonitzschia delicatissima B596 - - + + +

0850 Pseudonitzschia fraudolenta WWA7 Si Replete 390ppm - - - +

0851 Pseudonitzschia fraudolenta WWA7 Si Lim 390ppm - - + +

0852 Pseudonitzschia fraudolenta WWA7 Si Replete 750ppm - + + -

0853 Pseudonitzschia fraudolenta WWA7 Si Lim 750ppm - - - -

1423 Pseudonitzschia heimii UNC1101 - - + + +

- Pseudonitzschia multistriata female at little size - + + +

1061 Pseudonitzschia pungens - + - + -

1060 Pseudonitzschia pungens f cingulata - - + - +

0789 Rhizosolenia setigera CCMP 1694 - - + + +

0013_2 Skeletonema costatum 1716 - + - + +

0562 Skeletonema marinoi SkelB Replete + - - +

0563 Skeletonema marinoi SkelB N-limit - - - +

0578 Skeletonema grethea CCMP 1804 - + - + +

0593 Skeletonema japonicum CCMP2506 - - - + +

1040 Skeletonema marinoi FE60 - - + + +

1039 Skeletonema marinoi FE7 - + + + +

0918 Skeletonema marinoi SkelA Replete + 10270 - - -

214142

0920 Skeletonema marinoi SkelA SiLim + 24515 - - -

0320 Skeletonema marinoi SM1012Den-03 NoCuNoSi +

+ + +

0319 Skeletonema marinoi SM1012Hels-07 NoCuNoSi + - - +

1428 Skeletonema marinoi UNC1201 - + - - -

0603 Skeletonema menzelii CCMP793 Replete - - + +

0604 Skeletonema menzelii CCMP793 NLim - - - +

1361 Staurosira complex CCMP2646 - + - - +

0794 Stephanophyxis turris CCMP 815 - - + + +

0800 Striatella unipunctata CCMP2910 - - - + +

1176 Synedropsis recta CCMP1620 - - - + -

0786 Thalassionema frauenfeldii CCMP 1798 - + + + +

0156 Thalassionema nitzschioides L26-B CTR 1 - + + +

0158 Thalassionema nitzschioides L26-B -Fe - - + +

0905 Thalassiosira antarctica CCMP982 control growth - + - +

0902 Thalassiosira antarctica CCMP982 freezing temperature - - + +

0903 Thalassiosira antarctica CCMP982 iron limitation - - + -

0904 Thalassiosira antarctica CCMP982 silica limitation - - + -

0492 Thalassiosira gravida GMp14c1 Replete + 5168,2 - + -

0493 Thalassiosira gravida GMp14c1 NLim + 2093 - - +

0494 Thalassiosira gravida GMp14c1 PLim + 5953,6 - - +

0737 Thalassiosira minuscola CCMP1093 Exponential - - + +

0738 Thalassiosira minuscola CCMP1093 SiLim - - - -

0739 Thalassiosira minuscola CCMP1093 NLim - + + +

0740 Thalassiosira minuscola CCMP1093 Nocodazole - - - -

0970 Thalassiosira oceanica CCMP1005 Replete - + + +

0971 Thalassiosira oceanica CCMP1005 iron limitation - + - +

0972 Thalassiosira oceanica CCMP1005 Silicate Limitation - - - +

224344

0973 Thalassiosira oceanica CCMP1005 Nitrate Limitation - - - -

1067 Thalassiosira puntigera Tpunct2005C2 - - - + +

0404 Thalassiosira rotula CCMP3096 darkcycle - - + -

0403 Thalassiosira rotula CCMP3096 lightCycle - + - +

0910 Thalassiosira rotula GSO102 PLim - - + +

0911 Thalassiosira rotula GSO102 FeLim - - - -

0912 Thalassiosira rotula GSO102 FeLim_2 - - - -

0910 Thalassiosira rotula GSO102 Replete - - - -

1059 Thalassiosira sp. FreshWater - - + + +

1071 Thalassiosira sp. NH16 - - + + +

0878 Thalassiosira weissflogii CCMP1336 20nM Fe - - - -

0879 Thalassiosira weissflogii CCMP1336 60nM Fe - - - -

0880 Thalassiosira weissflogii CCMP1336 20nM Fe_2 - - - -

0881 Thalassiosira weissflogii CCMP1336 60nM Fe_2 - - - +

0898 Thalassiosira weissflogii CCMP1010 Replete - - - -

0899 Thalassiosira weissflogii CCMP1010 iron limitation - + + -

0900 Thalassiosira weissflogii CCMP1010 silica limitation - - - -

0901 Thalassiosira weissflogii CCMP1010 Nitrate Limitation - + + -

1405 Thalassiosira weissflogii CCMP1010 RepleteA - + - -

1406 Thalassiosira weissflogii CCMP1010 RepleteB - - - -

1407 Thalassiosira weissflogii CCMP1010 RepleteC - - - +

1408 Thalassiosira weissflogii CCMP1010 SiLimA - - - -

1409 Thalassiosira weissflogii CCMP1010 SiLimB - - - -

1410 Thalassiosira weissflogii CCMP1010 SiLimC - - - -

1411 Thalassiosira weissflogii CCMP1010 FeLimA - - - -

1412 Thalassiosira weissflogii CCMP1010 FeLimB - - - +

1413 Thalassiosira weissflogii CCMP1010 FeLimC - - - +

234546

1414 Thalassiosira weissflogii CCMP1010 +2HrFeAddA - - - -

1415 Thalassiosira weissflogii CCMP1010 +2HrFeAddB - - + -

1416 Thalassiosira weissflogii CCMP1010 +2HrFeAddC - - - +


1418 Thalassiosira weissflogii CCMP1010 +8HrFeAddB - - - -

1419 Thalassiosira weissflogii CCMP1010 +8HrFeAddC - - - +



1422 Thalassiosira weissflogii CCMP1010 +24HrFeAddB - - + +

0152 Thalassiothrix antarctica L6-D1 CTR1 - - + +

0154 Thalassiothrix antarctica L6-D1 Fe1 - + + +

24

362

363

4748

Supplementary table 3: list of sequences used in the phylogenetic analyses

Prostaglandin Enzyme Sequence ID Strain Specie Organism

COX1 MMETSP0707-20130614|6459 CCMP134 Asterionellopsis glacialis Microalga-Diatom

COX1 MMETSP1064-20121228|17645 CCAP1002/5 Aulacoseira subarctica Microalga-Diatom

COX1 MMETSP1064-20121228|57453 CCAP1002/5 Aulacoseira subarctica Microalga-Diatom

COX1 MMETSP0090-20130426|9955 CCMP159 Chaetoceros affinis Microalga-Diatom

COX1 MMETSP0717-20131115|51459 Chaetoceros curvisetus Microalga-Diatom

COX1 MMETSP0149-20130528|3320 MM31A-1 Chaetoceros debilis Microalga-Diatom

COX1 MMETSP0150-20130528|3844 MM31A-1 Chaetoceros debilis Microalga-Diatom

COX1 MMETSP0751-20121128|24195 CCMP1317 Chaetoceros neogracile Microalga-Diatom

COX1 MMETSP0751-20121128|3624 CCMP1317 Chaetoceros neogracile Microalga-Diatom

COX1 MMETSP1336-20130426|1470 RCC1993 Chaetoceros neogracile Microalga-Diatom

COX1 MMETSP1057-20121228|12622 CCMP338 Cyclotella meneghiniana Microalga-Diatom

COX1 MMETSP1057-20121228|4220 CCMP338 Cyclotella meneghiniana Microalga-Diatom

COX1 MMETSP1058-20130122|27510 CCMP353 Detonula confervacea Microalga-Diatom

COX1 MMETSP0907-20130614|13332 L2-C3 Fragilariopsis kerguelensis

Microalga-Diatom

COX1 MMETSP0009_2-20130614|5553 CCMP410 Grammatophora oceanica

Microalga-Diatom

COX1 MMETSP0014_2-20120614|6887 RCC80 Nitzschia sp Microalga-Diatom

COX1 MMETSP0015_2-20120614|17751 1302-5 Odontella aurita Microalga-Diatom

COX1 MMETSP0329-20121206|15499 B593 Pseudo-nitzschia arenysensis

Microalga-Diatom

COX1 MMETSP1061-20121228|74475 Pseudo-nitzschia pungens Microalga-Diatom

COX1 MMETSP0013_2-20120614|556 1716 Skeletonema costatum Microalga-Diatom

COX1 MMETSP0562-20121206|12376 SkelB Skeletonema dohrnii Microalga-Diatom

COX1 MMETSP0578-20130828|1168 CCMP1804 Skeletonema grethea Microalga-Diatom

COX1 MMETSP0319-20121206|1511 SM1012Hels-07 Skeletonema marinoi Microalga-Diatom

COX1 MMETSP0319-20121206|26582 SM1012Hels-07 Skeletonema marinoi Microalga-Diatom

COX1 MMETSP0320-20121206|19182 SM1012Den-03 Skeletonema marinoi Microalga-Diatom

COX1 MMETSP0920-20130426|33289 skelA Skeletonema marinoi Microalga-Diatom

COX1 MMETSP1039-20121108|4563 FE7 Skeletonema marinoi Microalga-Diatom

COX1 MMETSP1428-20130617|31509 UNC1201 Skeletonema marinoi Microalga-Diatom

COX1 MMETSP1428-20130617|783 UNC1201 Skeletonema marinoi Microalga-Diatom

COX1 MMETSP1361-20130828|23824 CCMP2646 Staurosira complex Microalga-Diatom

COX1 MMETSP1176-20130426|10672 CCMP1620 Synedropsis recta Microalga-Diatom

COX1 MMETSP0786-20121207|3776 CCMP1798 Thalassionema frauenfeldii

Microalga-Diatom

COX1 ADB65785 Caprella sp Animalia-shrimp

COXa XP_002127674 Ciona intestinalis Animalia-ascidia

25

364

4950

COXb XP_002123273 Ciona intestinalis Animalia-ascidia

COX1 AFN20596 Coccotylus truncatus Macroalga-read alga

COX1 ADB65786 Gammarus sp Animalia-shrimp

COXa AAF93168 Gersemia fruticosa Animalia-soft coral

COXb AAS48061 Gersemia fruticosa Animalia-soft coral

COX1 NP000953 Homo sapiens Animalia-mammalia

COX2 NP000954 Homo sapiens Animalia-mammalia

COX1 WP015113127 Nostoc sp Bacteria-cyanobacteria

COX1 CAC10360.1 Oncorhynchus mykiss Animalia-rainbow trout

COX1 NP001009476 Ovis aries Animalia-mammalia

15S-specific-cyclooxygenase AAU87497 Plexaura homomalla Animalia-gorgonian

type coral

COX1 AAF93169 Plexaura homomalla Animalia-gorgonian type coral

PtgE synthase MMETSP1065-20121228_57710 CCMP125 Amphiprora paludosa Microalga-Diatom

PtgE synthase MMETSP0724-20121128_6497 CCMP467 Amphiprora sp Microalga-Diatom

PtgE synthase MMETSP0713-20130426_43092 Asterionellopsis glacialis Microalga-Diatom

PtgE synthase MMETSP1394-20130617_3078 CCMP1581 Asterionellopsis glacialis Microalga-Diatom

PtgE synthase MMETSP1449-20131203_4452 CCMP2084 Attheya septentrionalis Microalga-Diatom

PtgE synthase MMETSP1064-20121228_59152 CCAP1002/5 Aulacoseira subarctica Microalga-Diatom

PtgE synthase MMETSP0785-20121206_16150 CCMP1866 Bolidomonas pacifica Microalga-Diatom

PtgE synthase MMETSP0091-20130426_66594 CCMP159 Chaetoceros affinis Microalga-Diatom

PtgE synthase MMETSP0717-20131115_41249 Chaetoceros curvisetus Microalga-Diatom

PtgE synthase MMETSP0150-20130528_6846 MM31A-1 Chaetoceros debilis Microalga-Diatom

PtgE synthase MMETSP0150-20130528_6846 MM31A-1 Chaetoceros debilis Microalga-Diatom

PtgE synthase MMETSP1447-20131203_65193 CCMP1751 Chaetoceros dichaeta Microalga-Diatom

PtgE synthase MMETSP0753-20130528_8419 CCMP1317 Chaetoceros neogracile Microalga-Diatom

PtgE synthase MMETSP1336-20130426_5187 RCC1993 Chaetoceros neogracile Microalga-Diatom

PtgE synthase MMETSP1336-20130426_5187 RCC1993 Chaetoceros neogracile Microalga-Diatom

PtgE synthase MMETSP0200_2-20121128_3741 GSL56 Chaetoceros sp Microalga-Diatom

PtgE synthase MMETSP0010_2-20120614_2064 308 Corethron hystrix Microalga-Diatom

PtgE synthase MMETSP1066-20121228_203475 CCMP2513 Coscinodiscus wailesii Microalga-Diatom

PtgE synthase MMETSP1442-20131203_60396 CCMP3328 Craspedostauros australis

Microalga-Diatom

PtgE synthase MMETSP0397-20121206_3519 ECT3854 Cyclophora tenuis Microalga-Diatom

PtgE synthase MMETSP0017_2-20120614_4529 KMMCC:B-181 Cylindrotheca closterium Microalga-Diatom

PtgE synthase MMETSP0580-20130426_33201 Dactyliosolen fragilissimus

Microalga-Diatom

265152

PtgE synthase MMETSP1058-20130122_59290 CCMP353 Detonula confervacea Microalga-Diatom

PtgE synthase MMETSP1443-20131203_66539 CCMP2396 Entomoneis sp Microalga-Diatom

PtgE synthase MMETSP0735-20130129_6806 L26-C5 Fragilariopsis kerguelensis

Microalga-Diatom

PtgE synthase MMETSP1360-20130828_7305 CCMP2313 Licmophora paradoxa Microalga-Diatom

PtgE synthase MMETSP1322-20130828_5091 RCC2270 Minutocellus polymorphus

Microalga-Diatom

PtgE synthase MMETSP0014_2-20120614_25505 RCC80 Nitzschia sp Microalga-Diatom

PtgE synthase MMETSP0014_2-20120614_43557 RCC80 Nitzschia sp Microalga-Diatom

PtgE synthase MMETSP0015_2-20120614_4973 isolate 1302-5 Odontella aurita Microalga-Diatom

PtgE synthase MMETSP0160_2-20130417_17287 Grunow 1884 Odontella sinensis Microalga-Diatom

PtgE synthase MMETSP0160_2-20130417_2021 Grunow 1884 Odontella sinensis Microalga-Diatom

PtgE synthase MMETSP0176-20130528_3501 PI-D3 Proboscia alata Microalga-Diatom

PtgE synthase MMETSP0816-20121228_18404 CCAP1064/1 Proboscia inermis Microalga-Diatom

PtgE synthase MMETSP0329-20121206_22727 B593 Pseudo-nitzschia arenysensis

Microalga-Diatom

PtgE synthase MMETSP0327-20121206_12465 B596 Pseudo-nitzschia delicatissima

Microalga-Diatom

PtgE synthase MMETSP0852-20130820_66215 WWA7 Pseudo-nitzschia fraudulenta

Microalga-Diatom

PtgE synthase MMETSP1423-20130617_783 UNC1101 Pseudo-nitzschia heimii Microalga-Diatom

PtgE synthase comp15013_c0 Small size female

Pseudo-nitzschia multistriata

Microalga-Diatom

PtgE synthase MMETSP1060-20121228_69939 Pseudo-nitzschia pungens Microalga-Diatom

PtgE synthase MMETSP0141_2-20121125_8366 10249 10 AB Pseudo-nitzschia_australis

Microalga-Diatom

PtgE synthase MMETSP0789-20121207_286 CCMP1694 Rhizosolenia setigera Microalga-Diatom

PtgE synthase MMETSP1039-20121108_5744 FE7 Skeletonema marinoi Microalga-Diatom

PtgE synthase MMETSP0794_2-20130614_15797 CCMP815 Stephanopyxis turris Microalga-Diatom

PtgE synthase MMETSP0786-20121207_2746 CCMP1798 Thalassionema frauenfeldii

Microalga-Diatom

PtgE synthase MMETSP0156-20130528_14046 L26-B Thalassionema nitzschioides

Microalga-Diatom

PtgE synthase MMETSP0905-20130828_2730 CCMP982 Thalassiosira antarctica Microalga-Diatom

PtgE synthase MMETSP0739-20130205_36184 CCMP1093 Thalassiosira miniscula Microalga-Diatom

PtgE synthase MMETSP0970-20121206_26001 CCMP1005 Thalassiosira oceanica Microalga-Diatom

PtgE synthase MMETSP0971-20121206_7568 CCMP1005 Thalassiosira oceanica Microalga-Diatom

PtgE synthase MMETSP0403-20120918_4534 CCMP3096 Thalassiosira rotula Microalga-Diatom

PtgE synthase MMETSP1059-20121228_69676 FW Thalassiosira sp Microalga-Diatom

PtgE synthase MMETSP1071-20121207_12737 NH16 Thalassiosira sp Microalga-Diatom

PtgE synthase MMETSP0899_2-20130528_50683 CCMP1010 Thalassiosira weissflogii Microalga-Diatom

PtgE synthase MMETSP0901_2-20130828_2659 CCMP1010 Thalassiosira weissflogii Microalga-Diatom

PtgE synthase MMETSP1405-20130829_715 CCMP1010 Thalassiosira weissflogii Microalga-Diatom

PtgE synthase MMETSP0154-20130528_3647 L6-D1 Thalassiothrix antarctica Microalga-Diatom

275354

PtgE synthase2 XP_006846913 Amborella trichopoda Plant

PtgE synthase2 OAY77991 Ananas comosus Plant

PtgE synthase2 like XP_015933989 Arachis duranensis Plant

PtgE synthase2 KYP65502 Cajanus cajan Plant

PtgE synthase2 like XP_010496074 Camelina sativa Plant

PtgE synthase2 like XP_004512741 Cicer arietinum Plant

PtgE synthase2 like XP_008453485 Cucumis melo Plant

PtgE synthase2 like XP_004137346 Cucumis sativus Plant

Electron carrier/proteindisulfide

oxidoreductaseXP_005785506 Emiliania huxleyi Microalga-

coccolithophore

PtgE synthase2 like XP_010069798 Eucalyptus grandis Plant

PtgE synthase2 KHN44626 Glycine soja Plant

PtgE synthase2 XP_012447492 Gossypium raimondii Plant

PtgE synthase2 EFN78959 Harpegnathos saltator Animalia-insect

PtgE synthase2 like XP_002161497 Hydra vulgaris Animalia-polyp

Glutathione_S-transferase XP_002406052 Ixodes scapularis Animalia-arthropod

PtgE synthase2 like XP_008393428 Malus domestica Plant

Glutathione_S-transferase XP_003619823 Medicago truncatula Plant

PtgE synthase2 XP_009400344 Musa acuminata Plant

Glutaredoxin EWM25410 Nannochloropsis gaditana Microalga

PtgE synthase2 like XP_010273922 Nelumbo nucifera Plant

PtgE synthase2 XP_009772831 Nicotiana sylvestris Plant

PtgE synthase2 CAH67930 Oryza sativa Plant

PtgE synthase2 like XP_015924269 Parasteatoda tepidariorum

Animalia-arthropode

PtgE synthase2 XP_011003060 Populus euphratica Plant

PtgE synthase2 like XP_007209338 Prunus persica Plant

PtgE synthase2 like XP_009356033 Pyrus bretschneideri Plant

Glutathione-S-transferase/glutaredoxin XP_845973 Trypanosoma brucei Excavata-protozoa

PtgE synthase2 XP_002282513 Vitis vinifera Plant

PtgE synthase2 XP_015882743 Ziziphus jujuba Plant

PtgE synthase KMZ69256 Zostera marina Plant

PtgH isomerase MMETSP1065-20121228_10087 CCMP125 Amphiprora paludosa Microalga-Diatom



PtgH isomerase MMETSP0725-20121227_3824 CCMP467 Amphiprora sp Microalga-Diatom

285556

PtgH isomerase MMETSP0724-20121128_16256 CCMP467 Amphiprora sp. Microalga-Diatom



PtgH isomerase MMETSP0318_2-20130129_1568 CCMP127 Amphora coffeaeformis Microalga-Diatom

PtgH isomerase MMETSP0318_2-20130129_6994 CCMP127 Amphora coffeaeformis Microalga-Diatom

PtgH isomerase MMETSP0706-20130129_10541 CCMP134 Asterionellopsis glacialis Microalga-Diatom






PtgH isomerase MMETSP0713-20130426_21585 Asterionellopsis glacialis Microalga-Diatom

PtgH isomerase MMETSP0713-20130426_4592 Asterionellopsis glacialis Microalga-Diatom





PtgH isomerase MMETSP0418-20121206_7922 13vi08-1A Astrosyne radiata Microalga-Diatom

PtgH isomerase MMETSP1449-20131203_1075 CCMP2084 Attheya septentrionalis Microalga-Diatom




PtgH isomerase MMETSP1064-20121228_1534 CCAP1002/5 Aulacoseira subarctica Microalga-Diatom

PtgH isomerase MMETSP1064-20121228_17710 CCAP1002/5 Aulacoseira subarctica Microalga-Diatom

PtgH isomerase MMETSP0785-20121206_15367 CCMP1866 Bolidomonas pacifica Microalga-Diatom

PtgH isomerase MMETSP0785-20121206_4282 CCMP1866 Bolidomonas pacifica Microalga-Diatom

PtgH isomerase MMETSP0088-20131115_39681 CCMP159 Chaetoceros affinis Microalga-Diatom




PtgH isomerase MMETSP0717-20131115_17446 Chaetoceros curvisetus Microalga-Diatom



PtgH isomerase MMETSP0149-20130528_3694 MM31A-1 Chaetoceros debilis Microalga-Diatom


295758



PtgH isomerase MMETSP1447-20131203_4865 CCMP1751 Chaetoceros dichaeta Microalga-Diatom

PtgH isomerase MMETSP1447-20131203_7059 CCMP1751 Chaetoceros dichaeta Microalga-Diatom

PtgH isomerase MMETSP0751-20121128_12958 CCMP1317 Chaetoceros neogracile Microalga-Diatom



PtgH isomerase MMETSP1336-20130426_27716 RCC1993 Chaetoceros neogracile Microalga-Diatom




PtgH isomerase MMETSP0200_2-20121128_14879 GSL56 Chaetoceros sp. Microalga-Diatom

PtgH isomerase MMETSP0200_2-20121128_2287 GSL56 Chaetoceros sp. Microalga-Diatom

PtgH isomerase MMETSP1429-20130617_66082 UNC1202 Chaetoceros sp. Microalga-Diatom



PtgH isomerase MMETSP0010_2-20120614_11223 308 Corethron hystrix Microalga-Diatom



PtgH isomerase MMETSP0169-20130528_22966 L29A3 Corethron pennatum Microalga-Diatom











PtgH isomerase MMETSP1066-20121228_16401 CCMP2513 Coscinodiscus wailesii Microalga-Diatom



PtgH isomerase MMETSP1442-20131203_40613 CCMP3328 Craspedostauros australis

Microalga-Diatom


Microalga-Diatom

PtgH isomerase MMETSP1442-20131203_5597 CCMP3328 Craspedostauros Microalga-Diatom

305960

australis


Microalga-Diatom

PtgH isomerase MMETSP0397-20121206_19965 ECT3854 Cyclophora tenuis Microalga-Diatom



PtgH isomerase MMETSP1057-20121228_314 CCMP 338 Cyclotella meneghiniana Microalga-Diatom

PtgH isomerase MMETSP0017_2-20120614_18451 KMMCC:B-181 Cylindrotheca closterium Microalga-Diatom




PtgH isomerase MMETSP0580-20130426_8153 Dactyliosolen fragilissimus

Microalga-Diatom

PtgH isomerase MMETSP0580-20130426_9785 Dactyliosolen fragilissimus

Microalga-Diatom

PtgH isomerase MMETSP1058-20130122_15807 CCMP 353 Detonula confervacea Microalga-Diatom



PtgH isomerase MMETSP1443-20131203_181 CCMP2396 Entomoneis sp. Microalga-Diatom



PtgH isomerase MMETSP1437-20131217_32895 CCMP1452 Eucampia antarctica Microalga-Diatom

PtgH isomerase MMETSP0733-20121128_4596 L26-C5 Fragilariopsis kerguelensis

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

PtgH isomerase MMETSP0009_2-20130614_10662 CCMP 410 Grammatophora oceanica

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

PtgH isomerase MMETSP0322-20121206_14221 B651 Leptocylindrus apora Microalga-Diatom

PtgH isomerase MMETSP0322-20121206_7581 B651 Leptocylindrus apora Microalga-Diatom

PtgH isomerase MMETSP1362-20130617_1296 CCMP1856 Leptocylindrus danicus Microalga-Diatom



316162

PtgH isomerase MMETSP1360-20130828_11548 CCMP2313 Licmophora paradoxa Microalga-Diatom

PtgH isomerase MMETSP1360-20130828_3247 CCMP2313 Licmophora paradoxa Microalga-Diatom

PtgH isomerase MMETSP1070-20121207_3283 NH13 Minutocellulus polymorphus

Microalga-Diatom

PtgH isomerase MMETSP1070-20121207_7688 NH13 Minutocellulus polymorphus

Microalga-Diatom

PtgH isomerase MMETSP1322-20130828_31186 RCC2270 Minutocellulus polymorphus

Microalga-Diatom

PtgH isomerase MMETSP1322-20130828_3156 RCC2270 Minutocellulus polymorphus

Microalga-Diatom

PtgH isomerase MMETSP0744-20121128_10137 CCMP561 Nitzschia punctata Microalga-Diatom

PtgH isomerase MMETSP0747-20121128_14861 CCMP561 Nitzschia punctata Microalga-Diatom

PtgH isomerase MMETSP0014_2-20120614_15951 RCC80 Nitzschia sp Microalga-Diatom



PtgH isomerase MMETSP0015_2-20120614_28361 isolate 1302-5 Odontella aurita Microalga-Diatom



PtgH isomerase MMETSP0160_2-20130417_1657 Grunow 1884 Odontella sinensis Microalga-Diatom






PtgH isomerase MMETSP0174-20130528_18662 PI-D3 Proboscia alata Microalga-Diatom






PtgH isomerase MMETSP0816-20121228_114861 CCAP1064/1 Proboscia inermis Microalga-Diatom




PtgH isomerase MMETSP0329-20121206_12053 B593 Pseudo-nitzschia arenysensis

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

PtgH isomerase MMETSP0140_2-20121125_19224 10249 10 AB Pseudo-nitzschia australis

Microalga-Diatom


Microalga-Diatom

326364


Microalga-Diatom

PtgH isomerase MMETSP0327-20121206_4279 B596 Pseudo-nitzschia delicatissima

Microalga-Diatom

PtgH isomerase MMETSP0327-20121206_8227 B596 Pseudo-nitzschia delicatissima

Microalga-Diatom

PtgH isomerase MMETSP0851-20130426_113318 WWA7 Pseudo-nitzschia fraudulenta

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

PtgH isomerase MMETSP1423-20130617_153 UNC1101 Pseudo-nitzschia heimii Microalga-Diatom



PtgH isomerase comp10947_c0_seq1 Female small size


Microalga-Diatom



Microalga-Diatom



Microalga-Diatom

PtgH isomerase MMETSP1061-20121228_2541 Pseudo-nitzschia pungens Microalga-Diatom



PtgH isomerase MMETSP0789-20121207_12219 CCMP1694 Rhizosolenia setigera Microalga-Diatom



PtgH isomerase MMETSP0013_2-20120614_1402 1716 Skeletonema costatum Microalga-Diatom

PtgH isomerase MMETSP0013_2-20120614_7554 1716 Skeletonema costatum Microalga-Diatom

PtgH isomerase MMETSP0578-20130828_3923 CCMP1804 Skeletonema grethae Microalga-Diatom

PtgH isomerase MMETSP0593-20130828_9896 CCMP2506 Skeletonema japonicum Microalga-Diatom

PtgH isomerase MMETSP0320-20121206_14352 SM1012Den-03 Skeletonema marinoi Microalga-Diatom

PtgH isomerase MMETSP1039-20121108_1431 FE7 Skeletonema marinoi Microalga-Diatom

PtgH isomerase MMETSP0603-20130417_1199 CCMP793 Skeletonema menzelii Microalga-Diatom

PtgH isomerase MMETSP0603-20130417_164 CCMP793 Skeletonema menzelii Microalga-Diatom

PtgH isomerase MMETSP0794_2-20130614_10777 CCMP815 Stephanopyxis turris Microalga-Diatom



PtgH isomerase MMETSP0800-20121206_5993 CCMP2910 Striatella unipunctata Microalga-Diatom

PtgH isomerase MMETSP1176-20130426_1126 CCMP1620 Synedropsis recta Microalga-Diatom

336566






PtgH isomerase MMETSP0786-20121207_16667 CCMP1798 Thalassionema frauenfeldii

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

PtgH isomerase MMETSP0156-20130528_11107 L26-B Thalassionema nitzschioides

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

PtgH isomerase MMETSP0902-20130614_41792 CCMP982 Thalassiosira antarctica Microalga-Diatom



PtgH isomerase MMETSP0492-20130828_47214 GMp14c1 Thalassiosira gravida Microalga-Diatom

PtgH isomerase MMETSP0737-20130205_6022 CCMP1093 Thalassiosira miniscula Microalga-Diatom

PtgH isomerase MMETSP0739-20130205_39664 CCMP1093 Thalassiosira miniscula Microalga-Diatom

PtgH isomerase MMETSP0970-20121206_1919 CCMP1005 Thalassiosira oceanica Microalga-Diatom

PtgH isomerase MMETSP0970-20121206_25055 CCMP1005 Thalassiosira oceanica Microalga-Diatom

PtgH isomerase MMETSP1067-20121228_17924 Tpunct2005C2 Thalassiosira puntigera Microalga-Diatom

PtgH isomerase MMETSP1067-20121228_6006 Tpunct2005C2 Thalassiosira puntigera Microalga-Diatom

PtgH isomerase MMETSP0404_2-20130426_37981 CCMP3096 Thalassiosira rotula Microalga-Diatom

PtgH isomerase MMETSP0910-20130528_8791 GSO102 Thalassiosira rotula Microalga-Diatom

PtgH isomerase MMETSP1059-20121228_75945 FW Thalassiosira sp Microalga-Diatom

PtgH isomerase MMETSP1071-20121207_10010 NH16 Thalassiosira sp Microalga-Diatom

PtgH isomerase MMETSP1071-20121207_6566 NH16 Thalassiosira sp Microalga-Diatom

PtgH isomerase MMETSP0899_2-20130528_8000 CCMP1010 Thalassiosira weissflogii Microalga-Diatom

PtgH isomerase MMETSP0901_2-20130828_34040 CCMP1010 Thalassiosira weissflogii Microalga-Diatom

PtgH isomerase MMETSP1415-20130829_7831 CCMP1010 Thalassiosira weissflogii Microalga-Diatom

PtgH isomerase MMETSP1422-20130829_36592 CCMP1010 Thalassiosira weissflogii Microalga-Diatom

PtgH isomerase MMETSP0152-20130528_2239 L6-D1 Thalassiothrix antarctica Microalga-Diatom



346768





Glutathione S-transferase B1N8I3 _CHOCR Chondrus crispus Macroalga-red alga

Glutathione s-transferase-like protein KOO22099 CCMP291 Chrysochromulina sp. Microalga

Glutathione-requiring prostaglandin D synthase EKC38075 Crassostrea gigas Animalia-oyster

Glutathione S-transferase CBJ30560 Ectocarpus siliculosus Macroalga-brown alga

Glutathione S-transferase ABR09274 Laminaria digitata Macroalga-brown alga

PtgH isomerase EGZ23680 Phytophthora sojae Plant

Glutathione S-transferase WP006973551 Plesiocystis pacifica Bacteria-marine micro-bacterium

Glutathione S-transferase ACV31082 Pyropia yezoensis Macroalga-red alga

PtgH isomerase XP004998907 Salpingoeca rosetta Microeukaryotes-choanoflagellate

Glutathione S-transferase-like protein AFV00540 Simiduia agarivorans Bacteria-marine

bacterium

ProstaglandinD synthase XP014151970 Sphaeroforma arctica Bacteria-marine bacterium

Glutathione S-transferase WP004371111 Thauera phenylacetica Bacteria

Ptg transporter MMETSP1065-20121228_11996 CCMP125 Amphiprora paludosa Microalga-Diatom

Ptg transporter MMETSP1065-20121228_67738 CCMP125 Amphiprora paludosa Microalga-Diatom

Ptg transporter MMETSP0724-20121128_19505 CCMP467 Amphiprora sp Microalga-Diatom







Ptg transporter MMETSP0316-20121206_6107 CCMP127 Amphora coffeaeformis Microalga-Diatom

Ptg transporter MMETSP0707-20130614_37570 CCMP134 Asterionellopsis glacialis Microalga-Diatom






356970


Ptg transporter MMETSP0418-20121206_1955 13vi08-1A Astrosyne radiata Microalga-Diatom

Ptg transporter MMETSP1449-20131203_57789 CCMP2084 Attheya septentrionalis Microalga-Diatom

Ptg transporter MMETSP1064-20121228_70784 CCAP 1002/5 Aulacoseira subarctica Microalga-Diatom

Ptg transporter MMETSP1064-20121228_9085 CCAP 1002/5 Aulacoseira subarctica Microalga-Diatom

Ptg transporter MMETSP0090-20130426_47634 CCMP159 Chaetoceros affinis Microalga-Diatom






Ptg transporter MMETSP0717-20131115_847 Chaetoceros curvisetus Microalga-Diatom

Ptg transporter MMETSP0149-20130528_2569 MM31A-1 Chaetoceros debilis Microalga-Diatom



Ptg transporter MMETSP1447-20131203_66063 CCMP1751 Chaetoceros dichaeta Microalga-Diatom

Ptg transporter MMETSP0751-20121128_20690 CCMP1317 Chaetoceros neogracile Microalga-Diatom





Ptg transporter MMETSP0200_2-20121128_13609 GSL56 Chaetoceros sp Microalga-Diatom



Ptg transporter MMETSP1429-20130617_78154 UNC1202 Chaetoceros sp Microalga-Diatom

Ptg transporter MMETSP1429-20130617_8616 UNC1202 Chaetoceros sp Microalga-Diatom

Ptg transporter MMETSP0010_2-20120614_11436 308 Corethron hystrix Microalga-Diatom



Ptg transporter MMETSP0169-20130528_5424 L29A3 Corethron pennatum Microalga-Diatom



Ptg transporter MMETSP1066-20121228_134602 CCMP2513 Coscinodiscus wailesii Microalga-Diatom

Ptg transporter MMETSP0397-20121206_3438 ECT3854 Cyclophora tenuis Microalga-Diatom

Ptg transporter MMETSP1057-20121228_10109 CCMP338 Cyclotella meneghiniana Microalga-Diatom


367172


Ptg transporter MMETSP0017_2-20120614_25628 KMMCC:B-181 Cylindrotheca closterium Microalga-Diatom



Ptg transporter MMETSP0580-20130426_41845 Dactyliosolen fragilissimus

Microalga-Diatom

Ptg transporter MMETSP0580-20130426_6520 Dactyliosolen fragilissimus

Microalga-Diatom

Ptg transporter MMETSP1058-20130122_5998 CCMP353 Detonula confervacea Microalga-Diatom

Ptg transporter MMETSP1058-20130122_60219 CCMP353 Detonula confervacea Microalga-Diatom

Ptg transporter MMETSP1437-20131217_3447 CCMP1452 Eucampia antarctica Microalga-Diatom



Ptg transporter MMETSP0733-20121128_115 L26-C5 Fragilariopsis kerguelensis

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

Ptg transporter MMETSP0009_2-20130614_86438 CCMP410 Grammatophora oceanica

Microalga-Diatom

Ptg transporter MMETSP0009_2-20130614_9365 CCMP410 Grammatophora oceanica

Microalga-Diatom

Ptg transporter MMETSP0322-20121206_16142 B651 Leptocylindrus apora Microalga-Diatom

Ptg transporter MMETSP0322-20121206_6063 B651 Leptocylindrus apora Microalga-Diatom

Ptg transporter MMETSP1362-20130617_58757 CCMP1856 Leptocylindrus danicus Microalga-Diatom

Ptg transporter MMETSP1362-20130617_9502 CCMP1856 Leptocylindrus danicus Microalga-Diatom

Ptg transporter MMETSP1360-20130828_11970 CCMP2313 Licmophora paradoxa Microalga-Diatom



Ptg transporter MMETSP1070-20121207_135 NH13 Minutocellus polymorphus

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

Ptg transporter MMETSP1322-20130828_30754 RCC2270 Minutocellus polymorphus

Microalga-Diatom

Ptg transporter MMETSP1322-20130828_31343 RCC2270 Minutocellus polymorphus

Microalga-Diatom

Ptg transporter MMETSP0744-20121128_13503 CCMP561 Nitzschia punctata Microalga-Diatom



Ptg transporter MMETSP0014_2-20120614_15610 RCC80 Nitzschia sp Microalga-Diatom

377374



Ptg transporter MMETSP0015_2-20120614_39172 isolate 1302-5 Odontella aurita Microalga-Diatom

Ptg transporter MMETSP0015_2-20120614_6159 isolate 1302-5 Odontella aurita Microalga-Diatom

Ptg transporter MMETSP0160_2-20130417_9596 Grunow 1884 Odontellasinensis Microalga-Diatom

Ptg transporter MMETSP0174-20130528_123001 PI-D3 Proboscia alata Microalga-Diatom




Ptg transporter MMETSP0816-20121228_116198 CCAP1064/1 Proboscia inermis Microalga-Diatom



Ptg transporter MMETSP0329-20121206_18166 B593 Pseudo-nitzschia arenysensis

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

Ptg transporter MMETSP0139_2-20121125_8846 10249 10 AB Pseudo-nitzschia australis

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

Ptg transporter MMETSP1060-20121228_6609 Pseudo-nitzschia cingulata

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

Ptg transporter MMETSP0327-20121206_2559 B596 Pseudo-nitzschia delicatissima

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

Ptg transporter MMETSP0850-20130426_54709 WWA7 Pseudo-nitzschia fraudulenta

Microalga-Diatom


Microalga-Diatom


Microalga-Diatom


Microalga-Diatom

Ptg transporter MMETSP1423-20130617_26482 UNC1101 Pseudo-nitzschia heimii Microalga-Diatom



Ptg transporter comp10879_c0_seq1 Female small size


Microalga-Diatom



Microalga-Diatom



Microalga-Diatom

387576



Microalga-Diatom



Microalga-Diatom

Ptg transporter MMETSP0789-20121207_14424 CCMP1694 Rhizosolenia setigera Microalga-Diatom





Ptg transporter MMETSP0013_2-20120614_20568 1716 Skeletonema costatum Microalga-Diatom




Ptg transporter MMETSP0562-20121206_3835 SkelB Skeletonema dohrnii Microalga-Diatom



Ptg transporter MMETSP0578-20130828_59445 CCMP1804 Skeletonema grethea Microalga-Diatom

Ptg transporter MMETSP0593-20130828_6310 CCMP2506 Skeletonema japonicum Microalga-Diatom

Ptg transporter MMETSP0319-20121206_7134 SM1012Hels-07 Skeletonema marinoi Microalga-Diatom

Ptg transporter MMETSP0320-20121206_143 SM1012Den-03 Skeletonema marinoi Microalga-Diatom



Ptg transporter MMETSP1039-20121108_17278 FE7 Skeletonema marinoi Microalga-Diatom

Ptg transporter MMETSP0603-20130417_70287 CCMP793 Skeletonema menzelii Microalga-Diatom




Ptg transporter MMETSP1361-20130828_34096 CCMP2646 Staurosira complex Microalga-Diatom

Ptg transporter MMETSP1361-20130828_4670 CCMP2646 Staurosira complex Microalga-Diatom

Ptg transporter MMETSP0794_2-20130614_50572 CCMP 815 Stephanopyxis turris Microalga-Diatom



Ptg transporter MMETSP0800-20121206_177 CCMP2910 Striatella unipunctata Microalga-Diatom

Ptg transporter MMETSP0786-20121207_3412 CCMP1798 Thalassionema frauenfeldii

Microalga-Diatom

Ptg transporter MMETSP0786-20121207_9273 CCMP1798 Thalassionema frauenfeldii

Microalga-Diatom

Ptg transporter MMETSP0156-20130528_26157 L26-B Thalassionema nitzschioides

Microalga-Diatom


Microalga-Diatom

397778


Microalga-Diatom

Ptg transporter MMETSP0902-20130614_56002 CCMP982 Thalassiosira antarctica Microalga-Diatom



Ptg transporter MMETSP0493-20130528_38175 GMp14c1 Thalassiosira gravida Microalga-Diatom




Ptg transporter MMETSP0737-20130205_1557 CCMP1093 Thalassiosira miniscula Microalga-Diatom



Ptg transporter MMETSP0970-20121206_8045 CCMP1005 Thalassiosira oceanica Microalga-Diatom




Ptg transporter MMETSP1067-20121228_139476 Tpunct2005C2 Thalassiosira puntigera Microalga-Diatom

Ptg transporter MMETSP1067-20121228_14711 Tpunct2005C2 Thalassiosira puntigera Microalga-Diatom

Ptg transporter MMETSP0403-20120918_14596 CCMP3096 Thalassiosira rotula Microalga-Diatom

Ptg transporter MMETSP0403-20120918_2990 CCMP3096 Thalassiosira rotula Microalga-Diatom

Ptg transporter MMETSP0910-20130528_2098 GSO102 Thalassiosira rotula Microalga-Diatom

Ptg transporter MMETSP0910-20130528_34273 GSO102 Thalassiosira rotula Microalga-Diatom

Ptg transporter MMETSP1059-20121228_3048 FW Thalassiosira sp Microalga-Diatom

Ptg transporter MMETSP1059-20121228_75934 FW Thalassiosira sp Microalga-Diatom

Ptg transporter MMETSP1071-20121207_17545 NH16 Thalassiosira sp Microalga-Diatom



Ptg transporter MMETSP0881-20121228_4845 CCMP1336 Thalassiosira weissflogii Microalga-Diatom







Ptg transporter MMETSP0152-20130528_78003 L6-D1 Thalassiothrix antarctica Microalga-Diatom



407980

41

365

366

8182

Transcripts sequences of the Skeletonema marinoi prostaglandin pathway reported in the main text.

>COX-1_Skel_mar_MMETSP1039-20121108|4563_1 len=1758

ATGAAGGCAGCAGCCAGCATCATCGTAGCCGTCATCGGCATCGTAATCTTTTACCTCAAACAAGCCAGTCTTCCATCCACCGCGCCCACACCACCAGGCTACCCCCTCCCACCCGACAGCATTGAATGGACGAGCCCACCCTATCCCTTTACAGAATGGCTCATCGACACTTTAAAGGCTCCAGTCATCAACTTCGGGTTGAGTCTGCCCGTCATATCTAGTTTTGTGAACCGTCGTCTACAGAACAGGGCAAGCACCAGCGGTTCCAATCGTCCATACAATCTATCCTGTAAATCGGACTACACCTCGTGGGATTCATTGAACGACCGTTCCTACTTTGGTAGACACTTGCCACCCAAAGAAATCCCTGATTTGCCACCTGTCGATGAAGTGGTGAACAAACTATTCCCACGCAAAGATGGCATCCAGACCATGTGTCCAAGATCAACACTTCTTTTTCCAACATTTGCCCAACATTTGATTGATTCGTTTATCAATACGAAAATTAATGTGGAGACTGGTGAGTTTGAGTGGGATAGAACCGATAGCAAGCACGAGATTGGGATAGGTCCTCTGTATGGGGATGAGGTAGAACAGACGAGTCAACTTCGTGAAAAGAGTGAAAAACCTGGGCGGAGGGGACGGCTTAAGACTCAAGTGTTGGAAGGTGGGGAAGAATGGGCACCTTTTCTCTATAATGAAGATGGCACCAAGAAGAAGGAGTTCAGTGCAATCCATGATCCTGATGGGATGAAAACTATCCTTGGGTTGGTCTACAGTTCTGATCCTACCACCAAAAGTTCGATTGAGCAGTCCATCTTTGCATTTGGAGGAAGACGTGCTAATTTGAATCCTAATATCGTTGCATGGAACACTCTCTTGTTGAGGGAGCATAATCGTCTTGCGGGGGAGATTGAGAAGAGTGAACCGGGGTGGGATGATGAACGTGTGTTTCAGACTGCTCGTAATGTCCTTATTGTTATGTATTGCAAGATTGTCATTGAAGAATACATCAAGCACATCAGTGGTGTAAACTTCAAAGTCGAGCCAGGTCCTTGGATGTGGAATGCACCCTGGTACAAAACCAATTGGATGTCTACCGAGTTTGCAATCTTGTATCGCTGGCACGCTCTCATTCCCAATGAGGCTGGTCTAGGTCCAAGCAAGGATGCAGGAGTAATGGAGGCACTATTCAATAATCCTATGTTGCTTGATGACGAGACTGGCCTGGGTGGCAACCTACGTGATATCTTTGTGGACATTAGTCAGACTCGGGTTACATCCTTACAATTGTTCAACACCGAGAAGTGGATGGTTGAACGTGAATCAGCAGCTATCAATCAGGGCCGTGCGAACAATGTTCAGAGTTATGCCGCATACTGCGAATACCTTGACATTGAACCACCCAAGACGTTTGAGGAYATTTCAATGGTGCCTGAAAGGCAGCAAGCCTTGAAGGAGCTTTATGGTACTCCAGATAGGGTAGAATTCTATGTTGGTTTGATTGCGGCAGATCATCCTGCAGGTGGGAAAATCTTTTCAGAGGCCATGACCAAGTTTGTAGCCAACGATGCCTTTAATCAAGCGTTAACCAACCCTCTCCTCTCCCAAAATGTTTGGGAAAATGGTGAGGAGACGTTTGGAAAATATGGTTGGGAGGTGGTAAAGCAAGATCATACCGTCAGGGATCTTCTGGAAAGGAACATTCCAAATGGTCTTGGTGATGCTTTTATTGGGATGACCATTCCAACTCTTTGA

>PTGEs_Skel_mar_MMETSP1039-20121108|5744_1 len=1008

ATGCTCAAAACAACAACAGCAATGAATACTACTCTAAGGTGTTTCGTAGGGCGAGCAGCAGCCCGCACATCTGTGCATAGTCATCATCTAACATCAAAATCACAGCAGCAACTTGCTTTCGTTTCATCATTGCATTCACCATCAACATCGGCATGGCTTTCCAGTAATTATTCAGCAAGATCGTTCTCGGCGGCTKCAACTGGACAAGCACCAGAAATTAAACTCTACCAGTACCACATTTGTCCCTTTTGCAATATTACAAAATCAGTGCTRTCCTATGCCAAATTAGATTATGAGAAGGTGGAGGTAAATCCACTRACAAAACAAGAGTTGAAACCATGGTCAGGAGATTATAAAAAGGTTCCAATAGCTGTGATTGACGAAGAACAAGTGAATGGATCGGAACAAATCATCGATGCGCTACTTGATACACAATATGTGCAGCAATTTTTGCAAAGTCGATGGAATGATGAAGCTAGTGACAAAGATGYCATGACGATGCAACAATTTCAAAACTCGGAAAACTCCCAAAAGTGGATGAGATACGCTGTAGATGATCTTGCAGCSTTGTTGTATCCGAAYAT

42

367368

369

370371372373374375376377378379380381382383384385386387388389390391392393394395396397398399400

401

402403404405406407408409410411

8384

TTGYGGTACWCTTGGKGAYTCTTACGATGCGTTTTCWTATGTCAAGAATGTAGATTCGTTTACTGGTGGTCAAAAGATATCAATTCAGATGCTGGGCTCTTTGGCCATGTACTTTGCAGCATCAAGAGTTAAGTCAAAACGAAACATTACGGATGAAAAGAAAGCATTGAAAGAAGCTCTTGACAAGTTTGAGCACGAGGGACTTGATAATGGAAAGTTATCCTTTTGTTCTGGTGTTTCAACACCTTATTTGGGTGATCTTGCCGTGTTTGGAGTCCTCTACTCTGTAAATGGTTTACAGGCTCATACAGATTCTATCCAGCTTCGAGGAGGTGCTGTGAAAGAATGGTATGATAGAATGCATCTCTCTGTGATGGGAGAAAAAGCGCAGCATCAGATCAAGCTTGGTGTCACTTGTGATTAA

>PTGDs or PtHi_Skel_mar_MMETSP1039-20121108|1431_1 len=648

XXCAACAATACAGACAATACAAACAACAATATGACTTCTTCTTCATCGCGACAGCACGAGCTCCTCTACTTCAACGGCGCCGGAAGAGCAGAAACAATTCGCATCTCACTCCACGCCGCTGGCATCAACTTCACCGACACGAGATTCACCGGCAAAGATTGGCCGGCGATCAAACCAACTACACCACTGGGCGGCGTACCCGTTCTCAAGGTAGATGGTGATTCACACTGTCAGTCTGTTTCGTTAGCCCGTTATGCTGGTAAGCTCGGTGGGTTTTATCCAACTGATCCCCTTCAGGCGCTTATTGTAGACGAGGCGATTGAAACTTTGAATGAGTTGATGGCAGTAGCCCCTAAACACTCTGATCCAGAAGAATTACTCAAGCTTCGTAAAGAGTTCCAAGCTGGCACAATGACAAAGTATATTGACTTCCTCGAGAGTTTGATTCAAAAGAATGGCGGAGTGGGATTTGCTTCTACCCCATCTATCGCCGATCTGACTCTTTGGGGTACAGTCAAGATGATTCGTACTGGGGATTGGACTGGAATTGATGCCACCTTCTTTGAATCTTATGCGGGTATTGTGGCGACTGTGGAGTCGATTGCCAAGGATGATAAGGTTGTTGCCTACTATGCATCGAGGGAGTAA

>PTGT_Skel_mar_MMETSP1039-20121108|17278_1 len=741

GGTTCATTGCCAGCGTTACCTGAAAGTGATGCTGATCTGTTGCAATCGGGTTGGCCATGGAGAGGGCATGTTGACAATCCTTCTTTGCCTCTTGTTCTGAAGAATGTATCAGTTGATATTCCCGCTGGTTCTACGCTAGGAGTTGTCGGCCGAACAGGAAGTGGAAAGAGTTCCCTCTTGCTAACTCTATTCCGTCTTGCTGAGATTGAGGGCAACGCAGGAAGTAGCATCACAATTGATGGGGTTGATATCAGATCACTCAGCCTTCAAGGTCTACGAGATTCAATATCTATCATCCCCCAAACTCCGATTCTTTTTGCTGGAACATTAATGCACAATTTAGATGCATCAGGGAAGGCCACGCCAGAGGATGCTTGGAGAGCATTGGAGTCAGCAAGTCCACAGCTCGCCAAACAGTTTAGAGACTCCAGCGAAGGCTTGAACACAAACATTTCAGAGGGTGGCGAAAACCTTTCTCTTGGTCAAAGACAGCTGATTTGTCTGGCAAGGGCATTAATGAAAAGGAGCAAAATTTTAGTTCTTGACGAGGCTACAAGTAGTATCGACATGAAAACTGATGCCCAAGTTCAAGAGACAATCAGGCGAGAGTTCGTTCAAAAAGGAGTGACAGTAATAACTGTTGCTCATAGATTAGAGACAGTGCTTGGGTACGATAGAATCCTTGTGCTTGATGCTGGCGTTCCAGTTGAGTTTGGCCCACCGGATGAACTCTTGAGGCGX

43

412413414415416417418419

420

421422423424425426427428429430431432

433

434435436437438439440441442443444445446

447

448

449

450

451

452

8586

Documents

images.nature.com€¦ · Web viewSupplementary Information. Supplementary information is provided in three files: a word file containing materials and methods, phylogenetic analysis