657 Supporting Information. Arboleda-Baena, C.M; Freilich, M. A; Pareja, C.B; Logares, R; De la 658 Iglesia, R; Navarrete, S.A. Microbial communities network structure across strong environmental 659 gradients: How do they compare to macroorganisms?. Ecology

660 Appendix S1

661 Table S1. Description of intertidal zone surveys of macroorganisms along sites in central Chile.

662 The majority of surveys took place in 2003 (321 surveys). Additional sampling took place in 2004

663 (124 surveys), 1999 (43 surveys), and 1998 (85 surveys). The surveys included 102 high intertidal,

664 267 mid intertidal, and 247 low intertidal quadrats. The study site for microbial zonation was

665 ECIMN, 33º 50’ S, 71º 63’ W.

TOTAL SOUTH WEST SURVEYS YEAR LAT LONG SITE BY YEAR 1998 33.50 71.63 ECIMN 1998 33.50 71.63 ECIMN

1998 33.50 71.63 ECIMN 85 1998 33.51 71.63 LCRUC 1998 33.51 71.63 LCRUC 1998 33.51 71.63 LCRUC 1999 33.50 71.63 ECIMN 1999 33.51 71.63 LCRUC 43 2003 33.42 71.70 PTRLN 2003 33.43 71.71 PTRLS

2003 33.45 71.68 TABO

2003 33.49 71.64 SAL 321 2003 33.50 71.63 ECIMN 2003 33.51 71.63 LCRUC 2003 33.55 71.61 CART 2003 33.56 71.62 PELA 2003 33.59 71.62 SAN 2004 33.45 71.68 TABO 2004 33.50 71.63 ECIMN

2004 33.50 71.63 ECIMN 124 2004 33.51 71.63 LCRUC 2004 33.55 71.61 CART 2004 33.55 71.61 CART

666

34

667 Appendix S1

668 Table S2. Description and main characteristics of the intertidal zones and control samples of the

669 microbial community (16S rRNA sequencing).

Sample ng Number of Number of Number Number # ID Treatment Extracted dDNA/ reads after OTUs after of reads of OTUs (cm2) uL rarefaction rarefaction

1 560_HighZone High Zone 22826 63.6 56862 907 16888 848 2 561_HighZone High Zone 22139 27.6 49140 980 16888 937 3 568_HighZone High Zone 26536 30.4 38831 746 16888 722 4 569_HighZone High Zone 25057 48.2 59919 958 16888 926 5 570_HighZone High Zone 35562 24.0 53583 666 16888 644 6 577_HighZone High Zone 32009 41.0 72215 792 16888 713 7 578_HighZone High Zone 29780 47.2 60989 556 16888 499 8 579_HighZone High Zone 26944 33.4 88915 665 16888 571 9 554_MiddleZone Middle Zone 18428 98.6 92795 1188 16888 989 10 562_MiddleZone Middle Zone 20842 41.8 80676 877 16888 759 11 563_MiddleZone Middle Zone 22313 32.2 57616 749 16888 694 12 564_MiddleZone Middle Zone 24636 45.6 59340 884 16888 816 13 571_MiddleZone Middle Zone 15625 94.8 31309 893 16888 872 14 572_MiddleZone Middle Zone 21277 110.0 29862 881 16888 864 15 580_MiddleZone Middle Zone 25540 97.2 34865 895 16888 872 16 582_MiddleZone Middle Zone 24194 30.8 71677 1316 16888 1190 17 556_LowZone Low Zone 19953 52.8 191992 2275 16888 1775 18 557_LowZone Low Zone 21375 23.8 71386 1032 16888 989 19 566_LowZone Low Zone 17688 19.9 16888 392 16888 392 20 574_LowZone Low Zone 17302 35.2 62217 1481 16888 1359 21 583_LowZone Low Zone 18560 68.0 52117 1367 16888 1299 22 584_LowZone Low Zone 22465 13.5 60638 1108 16888 1040 23 585_LowZone Low Zone 26207 26.8 45649 1275 16888 1198 Control Rock 16888 24 IROCA Surface 21657 3.7 41564 304 299 Control Rock 16888 25 IIROCA Surface 22202 3.4 42828 330 326 Control Rock 16888 26 IIIROCA Surface 18988 2.1 32654 289 287 Control Rock 16888 27 IVROCA Surface 17624 4.5 18342 261 261 Control Rock 16888 28 VROCA Surface 21407 2.0 22424 257 256 670

671

672

673

35

674 Appendix S1

675 Figure S1. Graphic representation of zonation of the principal Orders of A) Macroorganisms and

676 B) Microorganisms across the High, Middle and Low intertidal zones. The coefficients of variation

677 (CV%) across tides for each Order of C) macro- and D) micro-organisms are shown below. The

678 line across the bars represents the mean CV%.

A. 1. Annelida:Canalipalpata B. 1. Actinobacteria:Microtrichales 2. Arthropoda:Decapoda 3. Arthropoda:Sessilia 2. Bacteroidetes:Chitinophagales 4. Chlorophyta :Bryopsidales 3. Bacteroidetes:Flavobacteriales 5. Chlorophyta :Cladophorales 6. Chlorophyta :Ulvales 4. Bacteroidetes:Rhodothermales 7. Chlorophyta:Cladophorales 8. Chordata:Stolidobranchia 5. Cyanobacteria:Nostocales 9. Cnidaria:Actiniaria 6. Cyanobacteria:Phormidesmiales 10. Echinodermata:Forcipulatida 11. :Chitonida 7. Planctomycetes:Pirellulales 12. Mollusca:Ellobiida 13. Mollusca:Lepetellida 8. Planctomycetes:Planctomycetales 14. Mollusca: 9. Proteobacteria:Alteromonadales 15. Mollusca:Mytilida 16. Mollusca:Neogastropoda 10. Proteobacteria:Caulobacterales 17. Mollusca: 18. Mollusca:Siphonariida 11. Proteobacteria:Cellvibrionales 19. Mollusca: 20. Ochrophyta :Ectocarpales 12. Proteobacteria:Incertae_Sedis 21. Ochrophyta:Fucales 13. Proteobacteria:NA 22. Ochrophyta:Laminariales 23. Ochrophyta:NA 14. Proteobacteria:Oceanospirillales 24. Ochrophyta:Ralfsiales 25. Rhodophyta:Bangiales 15. Proteobacteria:Rhizobiales 26. Rhodophyta:Ceramiales 16. Proteobacteria:Rhodobacterales 27. Rhodophyta:Corallinales 28. Rhodophyta:Gelidiales 17. Proteobacteria:Rickettsiales 29. Rhodophyta:Gigartinales 30. Rhodophyta:Halymeniales 18. Proteobacteria:Sphingomonadales 31. Rhodophyta:Hildenbrandiales 19. Proteobacteria:Thiohalorhabdales 32. Rhodophyta:Nemaliales 33. Rhodophyta:Peyssonneliales 20. Proteobacteria:Thiotrichales 34. Rhodophyta:Plocamiales 35. Rhodophyta:Rhodymeniales 21. Verrucomicrobia:Verrucomicrobiales

HIGH MIDDLE LOW HIGH MIDDLE LOW C INTERTIDAL ZONE D INTERTIDAL ZONE 200 200

150 150 125.2% 100 100 CV % CV % 62.7% 50 50

0 0 679 1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21

680

681

682

683

684

36

685 Appendix S1

686 Table S3. Cover of sessile or density of mobile macroalgal and invertebrate species at

687 the a) high, b) middle, and c) low intertidal zone. Data are averages per tidal zone from all

688 surveys described in Table S1.

689 A) Mobile species (Density)

Total High Mid Low % All # Species Organism type Motility All Average Average Average surveys surveys 1 Acanthopleura echinata Chiton Mobile 0.00 0.06 2.74 692 0.20 2 Chaetopleura peruviana Chiton Mobile 0.00 0.00 0.84 208 0.06 3 Chiton cummingii Chiton Mobile 0.00 0.04 0.00 12 <0.001 4 Chiton granosus Chiton Mobile 2.55 14.20 4.23 5096 1.44 5 Chiton latus Chiton Mobile 0.00 0.03 0.13 40 0.01 6 Tonicia chilensis Chiton Mobile 0.00 0.00 1.13 280 0.08 7 Tonicia elegans Chiton Mobile 0.00 0.01 0.19 52 0.01 8 Tonicia spp Chiton Mobile 0.00 0.01 0.18 48 0.01 9 Lottia orbigny Limpet Mobile 0.00 0.03 0.02 12 <0.001 10 Scurria spp. Limpet Mobile 0.00 0.16 0.00 44 0.01 11 Fissurella costata Limpet Mobile 0.00 0.00 0.32 80 0.02 12 Fissurella crassa Limpet Mobile 0.00 2.55 0.57 820 0.23 13 Fissurella cummingii Limpet Mobile 0.00 0.13 0.13 68 0.02 14 Fissurella limbata Limpet Mobile 0.00 0.67 1.91 652 0.18 15 Fissurella maxima Limpet Mobile 0.00 0.00 0.10 24 0.01 16 Fissurella picta Limpet Mobile 0.00 0.23 0.05 74 0.02 17 Fissurella pulchra Limpet Mobile 0.00 0.25 0.00 68 0.02 18 araucana Limpet Mobile 3.45 18.87 1.47 5753 1.62 19 Scurria ceciliana Limpet Mobile 3.96 15.30 0.65 4649 1.31 20 Scurria variabilis Limpet Mobile 19.45 287.58 1.25 79076 22.30 21 Scurria plana Limpet Mobile 0.00 1.73 0.02 466 0.13 22 Scurria scurra Limpet Mobile 0.00 0.52 4.32 1208 0.34 23 Scurria zebrina Limpet Mobile 0.71 2.04 0.71 793 0.22 24 atra Snail Mobile 0.00 0.00 8.39 2072 0.58 25 araucana Snail Mobile 2269.29 0.87 0.02 231704 65.33 26 Echinolittorina peruviana Snail Mobile 66.47 0.34 0.00 6872 1.94 27 Trimusculus peruvianus pulmonate Limpet Mobile 0.00 0.00 0.03 8 <0.001 28 Concholepas concholepas Snail Mobile 0.00 0.57 1.15 436 0.12 29 Siphonaria lessoni pulmonate snail Mobile 40.82 29.76 0.11 12139 3.42 30 Taliepus dentatus Crab Mobile 0.00 0.00 0.05 12 <0.001 31 Acanthocyclus gayi Crab Mobile 0.00 0.45 0.60 268 0.08 32 Acanthocyclus hassleri Crab Mobile 0.00 0.09 0.31 100 0.03 33 Heliaster helianthus Sea star Mobile 0.16 0.10 0.70 216 0.06 34 Stichaster striatus Sea star Mobile 0.00 0.03 2.35 588 0.17 35 Phragmatopoma sp polychaete worm Mobile 0.00 0.00 0.06 15 <0.001 690

691

37

692 B) Sessile species (Cover)

Total Organism High Mid Low % All # Species Motility All type Average Average Average surveys surveys 1 Brachidontes granulata Mussel Sessile 0.00 0.07 0.18 63 0.10 2 Perumytilus purpuratus Mussel Sessile 3.94 27.91 1.40 8199 12.39 3 Semimytilus algosus Mussel Sessile 0.00 0.03 0.09 30 0.05 4 Jhelius cirratus Barnacle Sessile 40.24 8.71 0.01 6432 9.72 5 Nothochthamalus scabrosus Barnacle Sessile 0.34 5.08 0.03 1399 2.11 6 Austromegabalanus psittacus Barnacle Sessile 0.00 0.00 0.02 4 0.01 7 Balanus spp Barnacle Sessile 0.00 0.00 0.10 24 0.04 8 Nothobalanus flosculus Barnacle Sessile 0.01 0.55 8.14 2159 3.26 9 Balanus laevis Barnacle Sessile 0.00 0.13 0.05 48 0.07 10 Anthotoe spp. Anemone Sessile 0.00 0.00 0.02 6 0.01 11 Isolauctis spp. Anemone Sessile 0.00 0.00 0.00 1 <0.001 12 Phymactis spp. Anemone Sessile 0.00 0.31 1.30 403 0.61 13 Pyura chilensis sea squirt Sessile 0.00 0.01 0.04 11 0.02 14 Centroceras spp. Macroalgae Sessile 0.00 0.08 0.07 38 0.06 15 Ceramium spp. Macroalgae Sessile 0.02 0.83 0.10 248 0.37 16 Chaetomorpha_spp Macroalgae Sessile 0.00 0.01 0.00 4 0.01 17 Cladophora spp. Macroalgae Sessile 0.01 0.01 0.07 20 0.03 18 Ectocarpus silicosus Macroalgae Sessile 0.01 0.00 0.00 1 <0.001 19 Polysiphonia_spp Macroalgae Sessile 0.03 3.92 0.36 1140 1.72 20 Rama novae-zelandiae Macroalgae Sessile 0.00 0.03 0.45 117 0.18 21 Rhizoclonium cilindricum Macroalgae Sessile 0.00 0.00 0.02 6 0.01 22 Enteromorpha compressa Macroalgae Sessile 0.00 0.02 0.00 6 0.01 23 Pyropia _spp Macroalgae Sessile 0.84 0.59 0.03 251 0.38 24 Scythosiphon lomentaria Macroalgae Sessile 0.00 0.05 0.00 14 0.02 25 Ulva rigida Macroalgae Sessile 0.30 1.13 0.17 377 0.57 26 Petalonia fascia Macroalgae Sessile 0.01 0.14 0.03 45 0.07 27 Sacorthalia sp Macroalgae Sessile 0.00 0.05 0.01 17 0.03 28 Adenocystis utricularis Macroalgae Sessile 0.00 0.00 0.00 1 <0.001 29 Ahnfeltiopsis spp. Macroalgae Sessile 0.00 0.00 0.01 3 <0.001 30 Chondrus canaliculatus Macroalgae Sessile 0.00 0.00 0.02 5 0.01 31 Codium dimorphum Macroalgae Sessile 0.00 0.90 0.94 471 0.71 32 Colpomenia sinuosa Macroalgae Sessile 0.00 0.04 0.00 10 0.02 33 Gelidium rex Macroalgae Sessile 0.00 0.00 0.40 100 0.15 34 Gelidium spp. Macroalgae Sessile 0.02 2.86 7.21 2545 3.85 35 Grateloupia Macroalgae Sessile 0.00 0.00 0.02 5 0.01 36 Gymnogongrus furcellatus Macroalgae Sessile 0.00 0.00 0.52 129 0.19 37 Laurencia chilensis Macroalgae Sessile 0.00 0.05 0.00 14 0.02 38 Mazzaella laminarioides Macroalgae Sessile 0.66 10.78 0.07 2961 4.48 39 Nothogenia Macroalgae Sessile 0.00 0.04 0.00 11 0.02 40 Petroglossum spp. Macroalgae Sessile 0.00 0.00 0.00 1 <0.001 41 Plocamium cartilageneum Macroalgae Sessile 0.00 0.00 0.02 4 0.01 42 Prionitis spp Macroalgae Sessile 0.00 0.01 0.00 4 0.01 43 Rhodymenia sp Macroalgae Sessile 0.00 0.00 0.09 23 0.03 44 Schottera nicaensis Macroalgae Sessile 0.00 0.01 0.08 22 0.03 45 Durvillaea antarctica Macroalgae Sessile 0.00 0.36 6.22 1632 2.47 46 Lessonia nigrescens Macroalgae Sessile 0.00 0.01 84.91 20976 31.70 47 Macrocystis pyrifera Macroalgae Sessile 0.00 0.00 0.05 12 0.02 48 Corallina offcinalis var. chilensis Macroalgae Sessile 0.01 0.63 8.28 2216 3.35 49 Hildenbrandia lecanelieri Macroalgae Sessile 6.16 4.41 3.40 2646 4.00 50 Lithothamnion spp. Macroalgae Sessile 0.00 0.52 41.01 10269 15.52 51 Ralfsia californica Macroalgae Sessile 1.02 0.44 0.02 227 0.34 52 Ulvella spp. Macroalgae Sessile 0.09 1.20 0.05 341 0.52 53 Peysonella spp. Macroalgae Sessile 3.78 0.26 0.00 456 0.69

38

693 Appendix S1

694 Figure S2. Relative abundance of the microbial community (Bacteria and Archaea) at the levels

695 of Phylum and Class in samples from the high to low the intertidal zones from left to right. The

696 composition in the Control for the rock surface is also shown.

High Intertidal Zone Middle Intertidal Zone Low Intertidal Zone Control

100

Acidobacteria Kiritimatiellaeota Actinobacteria Lentisphaerae 75 Alphaproteobacteria Marinimicrobia_(SAR406_clade) Bacteroidetes Nanoarchaeaeota Chloroflexi Other Cyanobacteria Patescibacteria 50 Deinococcus−Thermus Planctomycetes Deltaproteobacteria Spirochaetes Euryarchaeota Thaumarchaeota Fibrobacteres Unclassified 25 Firmicutes Unresolved_Proteobacteria

% of 16S rRNA gene copies recovered Fusobacteria Verrucomicrobia Gammaproteobacteria WPS−2

0 Control_I Control_II Control_V Control_III Control_IV LowZone_556 LowZone_557 LowZone_566 LowZone_574 LowZone_583 LowZone_584 LowZone_585 HighZone_560 HighZone_561 HighZone_568 HighZone_569 HighZone_570 HighZone_577 HighZone_578 HighZone_579 MiddleZone_554 MiddleZone_562 MiddleZone_563 MiddleZone_564 MiddleZone_571 MiddleZone_572 MiddleZone_580 MiddleZone_582 697 Sample

698 Widely distributed groups in the Chilean rocky intertidal shore belong to Alphaproteobacteria,

699 Gammaproteobacteria, Bacteroidetes, Cyanobacteria, Planctomycetes, Actinobacteria,

700 Verrucomicrobia; these groups were previously described in biofilms of similar systems (Lee et

701 al. 2014, Taylor et al. 2014, Tan et al. 2015, Kerfahi et al. 2020), but this studies also described

702 groups that we do not find abundant, like Chloroflexi, Gemmatinomadetes, Chlorobi (Kerfahi et

703 al. 2020), Deltaproteobacteria and Firmicutes (Tan et al. 2015). All this groups have been widely

704 described on marine biofilms (Dang and Lovell 2016).

705 References

706 Dang, H., and C. R. Lovell. 2016. Microbial Surface Colonization and Biofilm Development in

39

707 Marine Environments. Microbiology and Molecular Biology Reviews 80:91–138.

708 Kerfahi, D., B. P. Harvey, S. Agostini, K. Kon, R. Huang, J. M. Adams, and J. M. Hall-Spencer.

709 2020. Responses of Intertidal Bacterial Biofilm Communities to Increasing pCO2.

710 Marine Biotechnology 22:727–738.

711 Lee, O. O., H. C. Chung, J. Yang, Y. Wang, S. Dash, H. Wang, and P.-Y. Qian. 2014. Molecular

712 Techniques Revealed Highly Diverse Microbial Communities in Natural Marine Biofilms

713 on Polystyrene Dishes for Invertebrate Larval Settlement. Microbial Ecology 68:81–93.

714 Tan, E. L.-Y., M. Mayer-Pinto, E. L. Johnston, and K. A. Dafforn. 2015. Differences in Intertidal

715 Microbial Assemblages on Urban Structures and Natural Rocky Reef. Frontiers in

716 Microbiology 6.

717 Taylor, J. D., R. Ellis, M. Milazzo, J. M. Hall-Spencer, and M. Cunliffe. 2014. Intertidal epilithic

718 bacteria diversity changes along a naturally occurring carbon dioxide and pH gradient.

719 FEMS Microbiology Ecology 89:670–678.

720

721

722

723

724

725

726

727

728

729

40

730 Appendix S1

731 Table S3. Main taxa of microorganisms and their abundance expressed as percentage of total

732 found in each tidal zone and their contribution to the total number of reads (% Total) The

733 representation of these taxa in the control rock surface is also shown. Figures in red correspond to

734 the most abundant taxa of major microbial phyla, i.e., relative abundance > 1% (Pedrós-Alió,

735 2012).

Rock High Middle Low surface TAXA Zone Zone Zone control % % % % Acidobacteria 0.03 0.10 0.29 0.02 Actinobacteria 0.55 0.46 2.56 1.27 Alphaproteobacteria 47.16 43.56 39.99 18.62 Bacteroidetes 31.44 40.09 29.65 8.49 Chloroflexi 0.05 0.21 0.43 0.03 Cyanobacteria 5.61 4.76 2.95 2.02 Deinococcus-Thermus 0.01 0.00 0.13 0.06 Deltaproteobacteria 0.25 0.50 0.70 0.83 Euryarchaeota 0.00 0.00 0.00 0.28 Fibrobacteres 0.00 0.00 0.00 0.03 Firmicutes 0.04 0.04 0.17 6.06 Fusobacteria 0.02 0.01 0.16 0.00 Gammaproteobacteria 11.21 7.01 10.63 60.86 Kiritimatiellaeota 0.00 0.00 0.04 0.01 Lentisphaerae 0.01 0.00 0.01 0.06 Marinimicrobia_(SAR406_clade) 0.00 0.01 0.00 0.24 Nanoarchaeaeota 0.00 0.00 0.00 0.12 Other 0.01 0.01 0.04 0.14 Patescibacteria 0.02 0.01 0.06 0.04 Planctomycetes 2.17 1.96 9.52 0.34 Spirochaetes 0.00 0.00 0.01 0.04 Thaumarchaeota 0.01 0.00 0.01 0.14 Unclassified 0.34 0.20 0.34 0.10 Unresolved_Proteobacteria 0.79 0.60 0.23 0.10 Verrucomicrobia 0.25 0.44 1.97 0.09 WPS-2 0.03 0.03 0.12 0.00 TOTAL 100 100 100 100 736 References

737 Pedrós-Alió, C., 2012. The rare bacterial biosphere. Annual review of marine science, 4, pp.449- 738 466.

41