DETAILED RESULTS ON FUNGAL AND BACTERIAL COMMUNITIES COLONIZING THE COMPOST-INCUBATED POLYMER CARD SURFACE

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1 DETAILED RESULTS ON FUNGAL AND BACTERIAL COMMUNITIES COLONIZING THE COMPOST-INCUBATED POLYMER CARD SURFACE PART 1: FUNGAL COMMUNITIES COLONIZING THE COMPOST-INCUBATED POLYMER CARD SURFACE Fungal OTUs richness, Shannon and Evenness diversity indices. Regarding fungal diversity metrics, compost-incubated, and PBAT polymer cards displayed the lower fungal richness, with less than 47 fungal OTUs assigned, while a high number of fungal OTUs was assigned on the PA66 card surface with a mean of 114 (± 15) OTUs (Table S1). The Shannon index ranged from 1.89 to 3.83, suggesting that the fungal communities adhered on polymer surfaces were less diversified than the bacterial communities. As previously observed with the bacterial communities, the, and PBAT polymer cards displayed the lowest Shannon index. The Evenness index values suggested the enrichment of a subset of fungal OTUs on and PBAT card surfaces. Table S1: Fungal OTU richness and diversity indices from the compost and the different polymers. Each value represents the mean ± standard error of the mean). Number of OTU Shannon Evenness Compost 67 (± 5) 3.52 (± 0.07) 0.84 (± 0.01) Polyolefin EVOH 81 (± 5) 3.23 (± 0.08) 0.74 (± 0.02) PP 50 (± 3) 3.11 (± 0.10) 0.80 (± 0.03) Polyester PBAT 43 (± 5) 1.89 (± 0.18) 0.50 (± 0.03) 96 (± 8) 3.71 (± 0.16) 0.81 (± 0.02) 46 (± 8) 2.77 (± 0.33) 0.72 (± 0.06) 36 (± 3) 1.92 (± 0.09) 0.53 (± 0.03) PLLA 85 (± 6) 3.24 (± 0.17) 0.73 (± 0.03) Polyamide PA (± 15) 3.83 (± 0.10) 0.81 (± 0.02) S1

2 Fungal community structure. PCoA analyses revealed that the fungal communities colonizing the, and PBAT cards were discriminated along the first principal coordinates that account for 27.43% of the total variability. These communities also appeared clearly different from the five other polymers and from the autochthonous bacterial community of the surrounding compost, which clustered together along the second principal coordinates (25.62% of the variability; see below Fig. S1). Sample Compost EVOH PA66 PP PBAT PLLA Fig. S1: Principal coordinate analysis (PCoA) of the fungal communities associated with the different polymer cards and the compost. Two-dimensional PCoA plot was based on the Bray-Curtis distance matrix. Percentage of the diversity distribution explained by each axis is indicated on the figure. S2

3 Fungal community composition associated to polymer card. With the exception of the sequences assigned to environmental and unclassified at the family level, Shiraiaceae was a dominant fungal family detected at the surface of PBAT and (see below Fig. S2), with (± 17.60)% and 9.49 (± 6.32)% of the total sequences assigned at the taxonomic level, respectively, values which appeared higher than the one obtained with the compost samples (2.28 ± 0.93%). Herpotrichiellaceae was dominant among the fungal communities attached on the surfaces of the cards made of (13.21 ± 2.13% of the sequences) and PA66 (8.97 ± 4.40%) and Mortierellaceae was significantly present on the two polyolefin cards studied (EVOH (6.37 ± 3.06%) and PP (11.17 ± 0.48%), but also on two of the five polyester cards ( (7.17 ± 1.08%) and PLLA (16.29 ± 9.67%). Members of Nectriaceae were predominant in -attached fungal communities, with (± 4.85)% of the sequence assigned to this family (see below Fig. S2 and SI4 Table S6). Phylum Ascomycota Basidiomycota Blastocladiomycota Chytridiomycota Early diverging fungal lineages Glomeromycota Proportion (%) Class Dothideomycetes Eurotiomycetes Leotiomycetes Pezizomycetes Sordariomycetes Agaricomycetes Blastocladiomycetes Chitriomycetes Monoblepharidomycetes Unclassified Glomeromycetes Others Compost EVOH Family Davidiellaceae Leptosphaeriaceae Phaeosphaeriaceae Shiraiaceae Testudinaceae Herpotrichiellaceae Helotiaceae Pezizaceae Pyronemataceae Bionectriaceae Chaetomiaceae Clavicipitaceae Coniochaetaceae Cordycipitaceae Nectriaceae Plectosphaerellaceae Auriculariaceae Entolomataceae Sebacinaceae Physodermataceae Megachytriaceae Rhizophydiaceae Harpochytriaceae Unknown Unclassified Environmental Other (<1%) Fig. S2: Heatmap representing relative abundance of fungal communities inhabiting the surface of the compost-incubated polymer cards. Only family representing at least 1% of relative abundance from the total sequences list in at least one condition were represented (see SI4 Table S6 for detailed fungal families) S3 PP PBAT PLLA PA66 Monoblepharidaceae Mortierellaceae Endogonaceae Legeriomycetaceae Glomeraceae

4 Comparison of fungal communities colonizing polymers: unique and shared OTUs. A total of 21 fungal OTUs were shared by the three community clusters and accounted for 66.50%, 82.21% and 73.82% of total classified sequences for, and, respectively (see below Fig. S3). 31 OTUs 4490 sequences 2 OTUs = 15.99% 4 OTUs = 21.60% 6 OTUs 2.56% 3.28% 21 OTUs = 66.50% = 73.82% 4 OTUs = 82.21% 4 OTUs = 14.94% = 2.37% = 1.29% = 5.79% 33 OTUs 4477 sequences 4 OTUs 9.63% 33 OTUs 4476 sequences Fig. S3: Three-set Venn diagram representing the unique and shared fungal OTUs (in bold) among polymers. Percentages indicate the corresponding proportions of associated sequences. S4

5 Drivers of fungal colonization: relationships between material surface properties and fungal taxa of biofilms. The best correlations (Fig. S4) were obtained for the fungal composition dissimilarity matrices from and the polar component (r 2 = 0.552) and with the energy of surface (r 2 = 0.533). The and PBAT cards were associated with the dispersive component (r 2 = 0.430) for the fungal community patterns. The cristallinity did not appear to be an important determinant of fungal community structure on polymer surface (r 2 = ) since the correlation did not reach a statistical significance (p > 0.01). Fig. S4: Polymer card properties (see Table 1) fitted onto fungal community PCoA (see above Fig. S1) after pyrosequencing and taxonomic assignment by GnS-PIPE. Arrows represent the contribution of each property to the discrimination of the bacterial diversity in contact with polymer card surface. A long arrow shows a strong correlation. 95% confidence ellipses are drawn for each plastic. Explained variations for the first five dimensions are indicated on the figure (labels and bar plot). S5

6 PART 2: BACTERIAL FAMILIES COLONIZING THE COMPOST-INCUBATED POLYMER CARD SURFACE Bacterial community composition associated to polymer card. The most represented bacterial families observed at the surface of the eight polymer types (see below Fig. S5 and SI4 Table S3) were Chitinophagaceae ( % of the total pyrosequencing sequences at the family level), Flavobacteriaceae ( %), Cytophagaceae ( %) and Bacteroidaceae ( %). Relevantly, Pseudomonadaceae was a dominant family detected on the card surfaces, with 8.09 (± 2.60)% of the total sequences assigned at this taxonomic level. This family was also abundant on EVOH, PBAT and PLLA cards with 3.87 (± 0.50)%, 3.24 (± 0.71)%, 3.75 (± 0.98)% of the total sequences, respectively. Sphingomonadaceae was dominant among the bacterial communities attached on the surfaces of the cards made of (5.59 ± 0.38% of the sequences) and (7.27 ± 2.22%), and Comamonadaceae was enriched on PBAT (23.57 ± 0.99%), (23.00 ± 6.20%) and (7.02 ± 1.39%) cards, whereas less than 3.44% of the total sequences were assigned to this family for the 5 other polymers. Interestingly, relative abundance of Bradyrhizobiaceae and Caulobacteraceae was significantly higher on the surfaces of, and PBAT than in the surrounding compost. S6

7 Phylum Acidobacteria Bacteroidetes Chloroflexi Firmicutes Planctomycetes Proteobacteria Class Holophagae Bacteroidia Cytophagia Flavobacteria Sphingobacteria Anaerolineae Clostridia Phycisphaerae Planctomycetia Alphaproteobacteria Betaproteobacteria Deltaproteobacteria Gammaproteobacteria Others Family Holophagaceae Bacteroidaceae Cytophagaceae Flavobacteriaceae Chitinophagaceae Sphingobacteriaceae Anaerolineaceae Clostridiaceae Lachnospiraceae Phycisphaeraceae Planctomycetaceae Bradyrhizobiaceae Caulobacteraceae Hyphomicrobiaceae Methylocystaceae Phyllobacteriaceae Rhizobiaceae Rhodospirillaceae Sphingomonadaceae Burkholderiaceae Comamonadaceae Oxalobacteraceae Rhodocyclaceae Bdellovibrionaceae Desulfuromonadaceae Geobacteraceae Polyangiaceae Sandaracinaceae Coxiellaceae Legionellaceae Methylococcaceae Moraxellaceae Pseudomonadaceae Sinobacteraceae Xanthomonadaceae Unknown Unclassified Other (<1%) Proportion (%) Compost EVOH PP PBAT PLLA PA Fig. S5: Heatmap representing relative abundance of bacterial communities inhabiting the surface of the compost-incubated polymer cards. Only family representing at least 1% of relative abundance from the total sequences list in at least one condition were represented (see SI4 Table S3 for detailed bacterial families) S7