1

Supplementary Material

Tissue-Specific Venom Composition and Differential Gene Expression in Sea Anemones.

Research Article

Jason Macrander1, Michael Broe1, and Marymegan Daly1

The supplementary PDF files include:

Supplementary Figures 1 – 8

Supplementary Tables I - II

All alignments for figures were made in the program Geneious (biomatters).

2

Supplementary Figure 1

Protein sequence alignments for candidate Type I KTx with tandem ShK domain noted for M. griffithsi. Redundant fragments were

removed. Sequence prefixes are as follows: As_= Anemonia sulcata, Hc_=Heteractis crispa, Mg_= Megalastics griffithsi. Previously

characterized toxins begin with Genbank ID.

3

4

Supplementary Figure 2

Protein sequence alignments for candidate Type III KTxs. Redundant fragments were removed. Sequence prefixes are as follows:

As_= Anemonia sulcata, Hc_=Heteractis crispa, Mg_= Megalastics griffithsi, Av_= Anemonia viridis, Ae_ =Anthopleura

elegantissima, Am_= Antheopsis maculate, Bg_= Bunodosoma granulifera. Previously characterized toxins are followed by Genbank

ID.

5

6

Supplementary Figure 3

Protein sequence alignments for candidate NaTx. Redundant fragments were removed. Sequence prefixes are as follows: As_=

Anemonia sulcata, Hc_=Heteractis crispa, Mg_= Megalastics griffithsi. Previously characterized toxins begin with Genbank ID.

7

8

Supplementary Figure 4

Protein sequence alignments for candidate type II cytolysins. The signaling region was trimmed for visualization purposes to remove

large gaps. Sequence prefixes are as follows: As_= Anemonia sulcata, Hc_=Heteractis crispa, Mg_= Megalastics griffithsi. Previously

characterized toxins begin with Genbank ID.

9

A) ASIC (2 types)

Type 1

Type 2(incomplete)

B) Acrorhagins 1

C) Acrorhagins 2

D) AETX

E) Class 9a KTx Toxins

F) EGF-like domains

10

G) Type V Potassium Channel Toxins

Supplementary Figure 5

Protein sequence alignments for candidate toxin genes for the A) Acid Sensing Ion Channel

(ASIC) Toxins, B) Acrorhagins1, C) Acrorhagins 2, D) AETX-like toxins, E) class 9a KTx, F)

EGF –like toxins, G) Type V KTx. Redundant fragments were removed. Sequence prefixes are

as follows: As_= Anemonia sulcata, Hc_=Heteractis crispa, Mg_= Megalastics griffithsi.

Previously characterized toxins begin with Genbank ID.

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A)

B)

C)

12

Supplementary Figure 6

Protein alignment of candidate zinc-dependent metalloprotease, astacin-like subfamily (peptidase

family M12A). The top sequence is the nematocyst expressed protein (K7Z9Q9.1) with the

remainder are candidate sequences identified in our bioinformatic pipeline for (A) Anemonia

sulcata, (B) Heteractis crispa, and (C) Megalactis griffithsi. The zinc-binding site/active site in

each alignment is denoted with arrows. To aide in visualization flanking regions were removed,

many transcripts had C-terminal protein domains, including cysteine arrangements that

resembled ShK-like proteins. Please note, in order to focus on this region of the alignment not all

candidate transcripts were included in this figure, please see supplemental File X for entire

alignment.

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14

Supplementary Figure 7

Alignment of BPTI/Kunitz family serine protease inhibitors. Redundant fragments were

removed. Sequence prefixes are as follows: As_= Anemonia sulcata, Hc_=Heteractis crispa,

Mg_= Megalastics griffithsi. Previously characterized toxins begin with Genbank ID.

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Supplementary Figure 8

Protein alignment of the membrane-attack complex/perforin (MACPF) toxins to Trinity sequences of toxin candidates, likely

assembled toxin fragments. Sequence prefixes are as follows: As_= Anemonia sulcata, Hc_=Heteractis crispa, Mg_= Megalastics

griffithsi. Previously characterized toxins begin with Genbank ID.

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Supplementary Figure 9

Protein alignment for the phospholipase A2 (PLA2) candidates. Sequence prefixes are as follows: As_= Anemonia sulcata,

Hc_=Heteractis crispa, Mg_= Megalastics griffithsi. Previously characterized toxins begin with Genbank ID.

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Supplemental Table I. Cnidarian Toxins Used to Identify Toxin Candidates

Entry Protein names Length Organism

P0DMX3 Cytolysin EnT 40 Entacmaea quadricolor P0DMX4 Cytolysin SmT-1 40 Stichodactyla mertensii P30833 Cytolysin tenebrosin-A 20 Actinia tenebrosa P30834 Cytolysin tenebrosin-B 20 Actinia tenebrosa P0CG44 Cytolysin Uc-1 29 Urticina crassicornis C5NSL2 DELTA-actitoxin-Aas1a 213 Anthopleura asiatica P01532 Delta-actitoxin-Ael1a 47 Anthopleura elegantissima P0C1F0 Delta-actitoxin-Ael1b 47 Anthopleura elegantissima P0C1F1 Delta-actitoxin-Ael1c 47 Anthopleura elegantissima P0C1F3 Delta-actitoxin-Ael1d 47 Anthopleura elegantissima B3EWF9 Delta-actitoxin-Ael2d 42 Anthopleura elegantissima P61914 DELTA-actitoxin-Aeq1a 214 Actinia equina Q93109 DELTA-actitoxin-Aeq1b 214 Actinia equina Q9Y1U9 DELTA-actitoxin-Aeq1c 214 Actinia equina Q9NJQ2 Delta-actitoxin-Aeq2a 82 Actinia equina B1NWU2 Delta-actitoxin-Aeq2b 1 82 Actinia equina B1NWU3 Delta-actitoxin-Aeq2b 2 82 Actinia equina B1NWU4 Delta-actitoxin-Aeq2b 3 82 Actinia equina B1NWU5 Delta-actitoxin-Aeq2c 82 Actinia equina B1NWU6 Delta-actitoxin-Aeq2d 81 Actinia equina P69943 Delta-actitoxin-Aer1a 47 Anemonia erythraea B9W5G6 DELTA-actitoxin-Afr1a 179 Actinia fragacea P10453 Delta-actitoxin-Afv1a 47 Anthopleura fuscoviridis P10454 Delta-actitoxin-Afv1b 48 Anthopleura fuscoviridis P69929 Delta-actitoxin-Amc1a 233 Antheopsis maculata P69930 Delta-actitoxin-Amc2a 80 Antheopsis maculata P69928 Delta-actitoxin-Amc3a 74 Antheopsis maculata P0C5F4 Delta-actitoxin-Aspp1a 47 Anthopleura sp. P0C5F5 Delta-actitoxin-Aspp1b 47 Anthopleura sp. P0C5F6 Delta-actitoxin-Aspp1c 47 Anthopleura sp. P0C5F7 Delta-actitoxin-Aspp1d 47 Anthopleura sp. P61915 DELTA-actitoxin-Ate1a 179 Actinia tenebrosa P01533 Delta-actitoxin-Avd1a 46 Anemonia sulcata P01528 Delta-actitoxin-Avd1c 47 Anemonia sulcata P0DL49 Delta-actitoxin-Avd1c 1 80 Anemonia viridis P0DL50 Delta-actitoxin-Avd1c 2 80 Anemonia viridis P0DL51 Delta-actitoxin-Avd1c 3 80 Anemonia viridis B1NWT7 Delta-actitoxin-Avd1c 4 79 Anemonia viridis P01529 Delta-actitoxin-Avd1d 46 Anemonia sulcata P0DL52 Delta-actitoxin-Avd1e 1 80 Anemonia viridis P0DL53 Delta-actitoxin-Avd1e 2 80 Anemonia viridis P0DL54 Delta-actitoxin-Avd1e 3 80 Anemonia viridis P0DMZ1 Delta-actitoxin-Avd1h 81 Anemonia viridis P01535 Delta-actitoxin-Avd2a 27 Anemonia sulcata C3TS08 Delta-actitoxin-Avd2b 1 72 Anemonia viridis C3TS04 Delta-actitoxin-Avd2b 2 71 Anemonia viridis C3TS10 Delta-actitoxin-Avd2b 3 72 Anemonia viridis C3TS06 Delta-actitoxin-Avd2b 4 71 Anemonia viridis C3TS07 Delta-actitoxin-Avd2c 61 Anemonia viridis P01530 Delta-actitoxin-Axm1a 49 Anthopleura xanthogrammica P01531 Delta-actitoxin-Axm1b 49 Anthopleura xanthogrammica P0C5F8 Delta-actitoxin-Axm1c 47 Anthopleura xanthogrammica P0C5G0 Delta-actitoxin-Axm1d 47 Anthopleura xanthogrammica P0C5F9 Delta-actitoxin-Axm1e 47 Anthopleura xanthogrammica P0C5G1 Delta-actitoxin-Axm1f 47 Anthopleura xanthogrammica P0C5G2 Delta-actitoxin-Axm1g 47 Anthopleura xanthogrammica P0C5G3 Delta-actitoxin-Axm1h 49 Anthopleura xanthogrammica P82803 Delta-actitoxin-Bcg1a 48 Bunodosoma cangicum P0C7P9 Delta-actitoxin-Bcg1b 48 Bunodosoma cangicum P0C7Q0 Delta-actitoxin-Bcg1c 48 Bunodosoma cangicum P86459 Delta-actitoxin-Bcg1d 47 Bunodosoma cangicum Q7M425 Delta-actitoxin-Bcs1a 48 Bunodosoma caissarum P0C1F4 Delta-actitoxin-Bgr2a 48 Bunodosoma granuliferum P0C1F5 Delta-actitoxin-Bgr2b 48 Bunodosoma granuliferum

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Supplemental Table I. (Continued)

P0C280 Delta-actitoxin-Cgg1a 47 Condylactis gigantea P0CH42 Delta-actitoxin-Cps1a 47 Condylactis passiflora P0DMX1 Delta-actitoxin-Cps1b 47 Condylactis passiflora P0DMZ2 Delta-actitoxin-Dar1b 31 Dofleinia armata P09949 Delta-actitoxin-Eqd1a 32 Entacmaea quadricolor Q5I4B8 DELTA-actitoxin-Oor1a 165 Oulactis orientalis Q5I2B1 DELTA-actitoxin-Oor1b 173 Oulactis orientalis C9EIC7 DELTA-actitoxin-Ucs1a 221 Urticina crassicornis E3P6S4 Delta-aiptatoxin-Adi1a 90 Aiptasia diaphana P0DL55 DELTA-alicitoxin-Pse1a 226 Phyllodiscus semoni P0DL56 DELTA-alicitoxin-Pse1b 226 Phyllodiscus semoni P58911 DELTA-alicitoxin-Pse2a 501 Phyllodiscus semoni P58912 DELTA-alicitoxin-Pse2b 488 Phyllodiscus semoni B1NWS4 Delta-edwarditoxin-Nvc1b 85 Nematostella vectensis B1NWR6 Delta-edwarditoxin-Nvc1c 85 Nematostella vectensis P0C5G6 Delta-halcutoxin-Hcg1a 47 Halcurias carlgreni P49127 Delta-hormotoxin-Cpt1a 79 Calliactis parasitica P14531 Delta-hormotoxin-Cpt1b 79 Calliactis parasitica P11494 DeltaKappa-actitoxin-Avd4a 43 Anemonia sulcata P59084 DeltaKappa-actitoxin-Avd4b 43 Anemonia sulcata Q86FQ0 DELTA-sagatoxin-Srs1a 216 Sagartia rosea P30832 Delta-stichotoxin-Hcr1a 48 Heteractis crispa P30783 Delta-stichotoxin-Hcr1b 48 Heteractis crispa P30784 Delta-stichotoxin-Hcr1c 48 Heteractis crispa P30785 Delta-stichotoxin-Hcr1d 47 Heteractis crispa P30831 Delta-stichotoxin-Hcr1e 48 Heteractis crispa P0C5G5 Delta-stichotoxin-Hcr3a 47 Heteractis crispa P58691 DELTA-stichotoxin-Hcr4a 175 Heteractis crispa P0C1F8 DELTA-stichotoxin-Hcr4b 177 Heteractis crispa Q9U6X1 DELTA-stichotoxin-Hmg2a 211 Heteractis magnifica P0DMX2 DELTA-stichotoxin-Hmg2b 177 Heteractis magnifica P01534 Delta-stichotoxin-Rpa1a 48 Radianthus paumotensis P08380 Delta-stichotoxin-Rpa1b 48 Radianthus paumotensis Q76CA3 Delta-stichotoxin-Sgt2a 74 Stichodactyla gigantea Q76CA0 Delta-stichotoxin-Sgt3a 84 Stichodactyla gigantea B1B5I9 Delta-stichotoxin-Shd3a 84 Stichodactyla haddoni P19651 Delta-stichotoxin-She1a 48 Stichodactyla helianthus P81662 DELTA-stichotoxin-She4a 176 Stichodactyla helianthus P07845 DELTA-stichotoxin-She4b 175 Stichodactyla helianthus Q5R231 DELTA-thalatoxin-Avl1a 226 Actineria villosa D2YZQ3 DELTA-thalatoxin-Avl1b 226 Actineria villosa Q76DT2 DELTA-thalatoxin-Avl2a 498 Actineria villosa D2KX90 Delta-thalatoxin-Cad1a 84 Cryptodendrum adhaesivum D2KX91 Delta-thalatoxin-Hhe1a 84 Heterodactyla hemprichii D2KX92 Delta-thalatoxin-Tas1a 84 Thalassianthus aster P0C1H0 Equinatoxin-1 20 Actinia equina P0C1H1 Equinatoxin-1'' 20 Actinia equina P0C1H2 Equinatoxin-3 20 Actinia equina P58305 Granulitoxin 21 Bunodosoma cangicum P39088 Hemolytic toxin 54 Heteractis magnifica Q52SK7 Hydralysin 244 Hydra vulgaris Q86LR2 Hydralysin-1 244 Hydra viridissima Q564A5 Hydralysin-2 244 Hydra viridissima Q52SK6 Hydralysin-3 218 Hydra viridissima P61541 Kappa-actitoxin-Ael2a 42 Anthopleura elegantissima P81897 Kappa-actitoxin-Aeq4a 36 Actinia equina Q0EAE5 Kappa-actitoxin-Aer3a 83 Anemonia erythraea P0DMX6 Kappa-actitoxin-Avd4a 76 Anemonia viridis P0DMX7 Kappa-actitoxin-Avd4c 76 Anemonia viridis P0DMX8 Kappa-actitoxin-Avd4d 76 Anemonia viridis P0DMX9 Kappa-actitoxin-Avd4e 76 Anemonia viridis P0DMY0 Kappa-actitoxin-Avd4f 80 Anemonia viridis P0DMY1 Kappa-actitoxin-Avd4g 76 Anemonia viridis P0DMY2 Kappa-actitoxin-Avd4h 77 Anemonia viridis P0DMY3 Kappa-actitoxin-Avd4i 80 Anemonia viridis

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Supplemental Table I. (Continued)

P0DMY4 Kappa-actitoxin-Avd4j 82 Anemonia viridis P0DMY5 Kappa-actitoxin-Avd4k 80 Anemonia viridis P0DMY6 Kappa-actitoxin-Avd4l 76 Anemonia viridis P0DMY7 Kappa-actitoxin-Avd4m 76 Anemonia viridis P0DMY8 Kappa-actitoxin-Avd4n 76 Anemonia viridis Q9TWG1 Kappa-actitoxin-Avd6a 36 Anemonia sulcata C0HJC2 Kappa-actitoxin-Bcs3a 38 Bunodosoma caissarum C0HJC3 Kappa-actitoxin-Bcs3b 37 Bunodosoma caissarum C0HJC4 Kappa-actitoxin-Bcs4a 50 Bunodosoma caissarum P29186 Kappa-actitoxin-Bgr1a 37 Bunodosoma granuliferum P86862 KappaPI-actitoxin-Ael3a 65 Anthopleura elegantissima Q9TWG0 KappaPI-actitoxin-Avd3b 58 Anemonia sulcata Q9TWF9 KappaPI-actitoxin-Avd3c 58 Anemonia sulcata Q9TWF8 KappaPI-actitoxin-Avd3d 59 Anemonia sulcata B1B5I8 KappaPI-stichotoxin-Shd2a 81 Stichodactyla haddoni O16846 Kappa-stichotoxin-Hmg1a 74 Heteractis magnifica E2S061 Kappa-stichotoxin-Sgt4a 74 Stichodactyla gigantea P0C7W7 Kappa-stichotoxin-Shd1a 28 Stichodactyla haddoni E2S062 Kappa-stichotoxin-Shd5a 74 Stichodactyla haddoni P29187 Kappa-stichotoxin-She3a 35 Stichodactyla helianthus E2S063 Kappa-stichotoxin-Smt1a 74 Stichodactyla mertensii E2S064 Kappa-thalatoxin-Cad2a 75 Cryptodendrum adhaesivum E2S065 Kappa-thalatoxin-Hhe2a 75 Heterodactyla hemprichii E2S066 Kappa-thalatoxin-Tas2a 75 Thalassianthus aster P0DMW8 Kunitz-type proteinase inhibitor AEPI-III 35 Actinia equina P0DMW9 Kunitz-type proteinase inhibitor AEPI-IV 37 Actinia equina P58689 Magnificalysin I 20 Heteractis magnifica P58690 Magnificalysin II 22 Heteractis magnifica K7Z9Q9 Nematocyst expressed protein 6 287 Nematostella vectensis P84851 Nephrotoxin PsTX-115 23 Phyllodiscus semoni B1NWS1 Neurotoxin Nv1-116.27.1 85 Nematostella vectensis P0CH90 Neurotoxin Nv1-116.37.1 85 Nematostella vectensis P0CH46 Neurotoxin Nv1-116.39.1 85 Nematostella vectensis B1NWS8 Neurotoxin Nv1-116.40.1 85 Nematostella vectensis A7SCE5 Neurotoxin Nv1-116.41.1 85 Nematostella vectensis B1NWR7 Neurotoxin Nv1-116.45.1 85 Nematostella vectensis Q76CA1 OMEGA-stichotoxin-Sgt1a 86 Stichodactyla gigantea B1B5J0 OMEGA-stichotoxin-Shd4a 86 Stichodactyla haddoni P86780 Phospholipase A2 39 Bunodosoma caissarum D2X8K2 Phospholipase A2 A2-actitoxin-Cgg2a 119 Condylactis gigantea A7LCJ2 Phospholipase A2 A2-actitoxin-Ucs2a 155 Urticina crassicornis Q8WS88 Phospholipase A2 A2-hormotoxin-Apt1a 156 Adamsia palliata P61542 Pi-actitoxin-Ael2b 42 Anthopleura elegantissima P0DMW6 PI-actitoxin-Aeq3a 59 Actinia equina P0DMW7 PI-actitoxin-Aeq3b 59 Actinia equina C0HJB1 Pi-phymatoxin-Pcf1a 32 Phymanthus crucifer R4ZCU1 Precursor protein UG 162 Urticina grebelnyi A7RMN1 Predicted protein NvePTx1 84 Nematostella vectensis P0CH45 Putative delta-edwarditoxin-Nvc1a 85 Nematostella vectensis P0DL61 Small cysteine-rich protein 64 Anemonia viridis P0DL60 Small cysteine-rich protein 69 Metridium senile C0H690 Small cysteine-rich protein 1 89 Acropora millepora C1KIY9 Small cysteine-rich protein 1 79 Montastraea faveolata C0H693 Small cysteine-rich protein 1 1 81 Montipora capitata C0H694 Small cysteine-rich protein 1 2 81 Montipora capitata C0H691 Small cysteine-rich protein 2 79 Acropora millepora C1KIZ0 Small cysteine-rich protein 2 68 Montastraea faveolata C0H692 Small cysteine-rich protein 3 83 Acropora millepora C1KIZ3 Small cysteine-rich protein 4 81 Montastraea faveolata C1KIZ5 Small cysteine-rich protein 6 81 Montastraea faveolata B2ZG38 Small cysteine-rich protein 8 74 Montastraea faveolata C1KIZ4 Small cystein-rich protein 5 68 Montastraea faveolata B2G331 Tau-stichotoxin-Hcr2b 78 Heteractis crispa C0HJF4 Tau-stichotoxin-Hcr2c 56 Heteractis crispa C0HJF3 Tau-stichotoxin-Hcr2d 56 Heteractis crispa

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Supplemental Table I. (Continued)

P43318 Toxic phospholipase A2 32 Rhopilema nomadica P86469 Toxin Bcg III 15.67 29 Bunodosoma cangicum P86466 Toxin Bcg III 21.00 27 Bunodosoma cangicum P86467 Toxin Bcg III 23.41 28 Bunodosoma cangicum P86463 Toxin Bcg III 25.52 33 Bunodosoma cangicum P86462 Toxin Bcg III 28.78 37 Bunodosoma cangicum P86464 Toxin Bcg III 29.21 36 Bunodosoma cangicum P86461 Toxin Bcg III 31.16 38 Bunodosoma cangicum P86468 Toxin Bcs III 15.09 45 Bunodosoma caissarum P86470 Toxin BcV 33 Bunodosoma caissarum Q9GNN8 Toxin CaTX-A 463 Carybdea alata A7L035 Toxin CfTX-1 456 Chironex fleckeri A7L036 Toxin CfTX-2 462 Chironex fleckeri P58762 Toxin CqTX-A 462 Chiropsalmus quadrigatus Q9GV72 Toxin CrTX-A 450 Carybdea rastonii P0C1G1 Toxin Up-1 46 Urticina piscivora P0DMX5 U-actitoxin-Ael2c 70 Anthopleura elegantissima Q3C258 U-actitoxin-Aeq5a 70 Actinia equina Q3C257 U-actitoxin-Aeq5b 72 Actinia equina Q3C256 U-actitoxin-Aeq6a 83 Actinia equina Q3C255 U-actitoxin-Aeq6b 83 Actinia equina P69944 U-actitoxin-Aer2a 59 Anemonia erythraea P69945 U-actitoxin-Aer2b 59 Anemonia erythraea P0DN04 U-actitoxin-Avd10a 86 Anemonia viridis P0DN05 U-actitoxin-Avd11a 70 Anemonia viridis B1NWT3 U-actitoxin-Avd1f 72 Anemonia viridis B1NWT5 U-actitoxin-Avd1g 83 Anemonia viridis P0DN06 U-actitoxin-Avd3f 82 Anemonia viridis P0DN07 U-actitoxin-Avd3g 82 Anemonia viridis P0DN09 U-actitoxin-Avd3h 82 Anemonia viridis P0DN10 U-actitoxin-Avd3i 82 Anemonia viridis P0DN11 U-actitoxin-Avd3j 82 Anemonia viridis P0DN12 U-actitoxin-Avd3k 82 Anemonia viridis P0DN13 U-actitoxin-Avd3l 82 Anemonia viridis P0DN14 U-actitoxin-Avd3m 82 Anemonia viridis P0DN15 U-actitoxin-Avd3n 87 Anemonia viridis P0DN16 U-actitoxin-Avd3o 87 Anemonia viridis P0DN17 U-actitoxin-Avd3p 82 Anemonia viridis P0DN18 U-actitoxin-Avd3q 82 Anemonia viridis P0DN19 U-actitoxin-Avd3r 72 Anemonia viridis P0DN20 U-actitoxin-Avd3s 80 Anemonia viridis B1NWT4 U-actitoxin-Avd7a 76 Anemonia viridis P0DMZ3 U-actitoxin-Avd8a 79 Anemonia viridis P0DMZ4 U-actitoxin-Avd8b 79 Anemonia viridis P0DMZ5 U-actitoxin-Avd8c 76 Anemonia viridis P0DMZ6 U-actitoxin-Avd8d 83 Anemonia viridis P0DMZ7 U-actitoxin-Avd8e 84 Anemonia viridis P0DN00 U-actitoxin-Avd9a 80 Anemonia viridis P0DN01 U-actitoxin-Avd9b 80 Anemonia viridis P0DN02 U-actitoxin-Avd9c 80 Anemonia viridis P0DN03 U-actitoxin-Avd9d 79 Anemonia viridis P86465 U-actitoxin-Bcg2a 30 Bunodosoma cangicum P84919 U-actitoxin-Bcs2a 41 Bunodosoma caissarum G0W2H8 U-actitoxin-Bgr3a 74 Bunodosoma granuliferum G0W2H9 U-actitoxin-Bgr3b 74 Bunodosoma granuliferum G0W2I0 U-actitoxin-Bgr3c 74 Bunodosoma granuliferum G0W2I1 U-actitoxin-Bgr3d 79 Bunodosoma granuliferum G0W2H7 U-AITX-Bg1a 41 Bunodosoma granuliferum C0HJB4 U-homostoxin-Hdu1a 29 Homostichanthus duerdeni P11495 U-metritoxin-Msn1a 36 Metridium senile P0DMD7 U-metritoxin-Msn2a 90 Metridium senile

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Supplemental Table II. Nematocysts measurements and counts for focal taxa and congeners. A. sulcata A. viridis (smargdina) A. viridis (rustica) H. crispa M. comatus M. hemprichii

Length Width Length N=10 Length N = 10 Length Length Length

Type M R W R M SD R M SD R R N n R N R N

C

basitrichous isorhiza 17.1 13.8 - 20.6 2.5 2.0 - 2.6 16.9 0.9 15.3 - 18.6 17.1 0.9 15.6-19.5 14.8 - 22.5 14 132 15.23 - 48.16 765 17.0 - 30.3 163

basitrichs 24.5 - 33.6 4 21 29.91 - 40.34 10 - -

Haplonemes isorhizas 25.5 21.2 - 30.0 5.9 4.6 - 7.8 15.0 - 31.2 16 121

atrichs 34.1 1.5 31.6 - 36 33.9 2.9 30.1 - 39.9

microbasic p-

mastigophores 25.6 - 35.9 4 21

T Spirocysts 34.6 23.4 - 42.9 3.3 2.6 - 3.9 15.0 - 31.2 16 121

basitrichous isorhiza 33.3 22.5 - 41.6 3.4 2.5 - 3.9 32.3 2.9 26.2 - 35.3 34.6 1.6 32.2 - 37.4 12.5 - 25.7 16 131

F

p-mastigophores 22.3 18.7 - 26.2 5.4 3.9 - 6.5 24.6 3.8 20.5 - 321 23.2 2.6 19.5 - 28.4 20.0 - 37.3 13 93

basitrichous isorhiza 38.9 32.5 - 45.5 4.4 3.3 - 5.8 34.8 3.2 29.9 - 40.6 34.1 3.3 26.9 - 40.7 22.4 - 36.3 11 76 32.31 - 63.63 300 33.8 - 72.8 116

basitrichous isorhiza 12.1 7.8 - 14.3 2.3 1.6 - 2.8 13.1 0.5 12.3 - 14 13.4 1.2 11.5 - 15.4 12.3 - 21.4 12 80 17.91 - 31.97 63 18.8 - 26.1 54

Basitrich 3 37.67 - 59.22 12 54 1

Baitrich 4 - - 150+ 1

Spirocysts 17.2 - 27.1 8 29 23.24 - 39.54 41 18.4 - 27.0 10

A

basitrich1 34.56 - 78.83 2973 53.5 - 76.8 93

basitrich 3 42.44 - 70.69 36 76.3 - 81.7 2

Spirocysts 18 - 31.22 60 18.8 - 25 20

a

Haplonemes isorhizas 43.4 35.0 - 52.0 4.5 3.7 - 5.0

atrichs 41.9 1.8 38.6 - 44.9 43.2 3.4 38.1 - 49.2

basitrichous isorhiza 18.3 1 15.3 - 18.6 19.2 2.5 16 - 25.5

P

basitrichous isorhiza 32.7 25.0 - 37.7 3.5 2.6 - 4.0 28.5 3.2 22.9 - 31.7 30.6 2.5 24 - 34.8 15.2 - 27.5 12 97

Spirocysts 17.2 - 27.1 29 8

microbasic p-

mastigophores 20.0 - 37.3 13 93

Citation Schmidt 1972 Chintiroglou and Karalis (2000) Dunn (1981) Ardelean and Fautin (2004)

C: Column, T: Tentacles, F: Filament, A: Acrospheres, a: acrorhagi, P: Pharynx, M: measured nematocyst length, W: measured nematocyst width,

R: nematocyst measreument range (width or length); SD: Standard Deviation, N: number of nematocysts measured, n: number of nematocysts

observed.