Supplementary Material Microbial communities ...

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SSP: sieved agriculture soil mixed with perlite; T6: cellulose mat; T8: (PLA/PHB_w) original PLA/PHB transparent foil; T12: (PLA/PHB_b) original carbon.
Supplementary Material Microbial communities responsible for the degradation of poly(lactic acid)/poly(3-hydroxybutyrate) blend mulches in soil burial respirometric tests.

Lenka Jeszeová1, Andrea Puškárová1, Mária Bučková1, Lucia Kraková1, Tomáš Grivalský1, Martin Danko2, Katarína Mosnáčková2, Štefan Chmela2, Domenico Pangallo1, 3, *

(1) Institute of Molecular Biology, Slovak Academy of Sciences, Dúbravská cesta 21, 845 51 Bratislava, Slovakia. (2) Polymer Institute, Slovak Academy of Sciences, Dúbravská cesta 9, 845 41 Bratislava, Slovakia. (3) Caravella s.r.o., Tupolevova 2, 851 01 Bratislava, Slovakia

*Corresponding author: [email protected]

1

Table S1. Clustering (based on ITS-PCR and Qiaxcel electrophoresis), identification of bacterial isolates and their polymers degradation abilities. Qiaxcel profile bp 252, 448

263, 410, 422 281, 450

Isolates

SSP SSP SSP SSP SSP SSP

BA 1 BA5 BA6 BA7 BA8 BA9

SSP BA10

Species identification on the basis of the highest 16S rRNA similarity score

Order

Phylum

MF319408 Bacillus sp. 100% / 100%

Bacillales

Firmicutes

Micrococcales

Actinobacteria

KY127420 Brevibacterium frigoritolerans 100% / 100% KX507092 Bacillus subtilis 100% / 100%

SSP BA11 SSP BA12 SSP BA13 SSP SSP SSP SSP SSP SSP

KX507092 Bacillus subtilis 100% / 100%

BA14 BA15 BA16 BA17 BA18 AIA2

SSP AIA3 SSP FSS5 SSP FSS6 SSP FSS7 SSP FSS8

Bacillales KX507088 Bacillus subtilis 100% / 100%

KX507088 Bacillus subtilis 100% / 100%

Firmicutes

Polymer degradation assays PHB PLA/PHB PLA blend + +++ + + + +++ + + + ++++ +++ +++ +++ +++ -

-

+++

-

-

+++ +++

++++

+++

+++

++++ -

+++ +++ +++ +

++ + -

+++

+++

+

+++ +++

+++ +++

+++ +++

+++

+++

+++

+++

+++

+++ 2

SSP FSS9 SSP LB10-1 SSP LB10-4 SSP LB10-5 LB10-7 LB10-

240, 455

SSP SSP 10 SSP 11 SSP 12 SSP

200, 607

SSP SSP SSP SSP

AIA5 AIA7 AIA8 BA2

312

SSP BA3

588

SSP BA4

358, 600

SSP AIA1

552

SSP SSP SSP SSP

KX507088 Bacillus subtilis 100% / 100% KX507088 Bacillus subtilis 100% / 100% KX507092 Bacillus subtilis 100% / 100%

LB10LB10AIA4

AIA10 AIA12 AIA9 AIA11

MF062987 Bacillus pseudomycoides 100% / 100%

MF377573 Bacillus sp. 100% / 99% MF319407 Bacillus sp. 100% / 100% KX859155 Pseudomonas sp. 100% / 100% KR673340 Pseudomonas sp. 100% / 100%

KX504218 Arthrobacter globiformis 100% / 99%

Pseudomonadales

Micrococcales

Proteobacteria

Actinobacteria

+++

+++

+++

++++

-

-

-

-

++++

++++

++++

+++

+++

-

-

+++

-

+++

-

-

+++

+++

-

+++

++

+++

+

+ -

+ ++ -

+ ++ -

-

+++

-

+++

+++

+++

-

-

+

-

+++

-

-

++

+

++++

-

+ 3

496

SSP SM1 SSP SM2 SSP SM3

611

SSP SSP SSP SSP SSP

522

SSP SM9

402, 603

403, 626

SM4 SM5 SM6 SM7 SM8

SSP FSS1 SSP FSS2 SSP FSS3 SSP FSS4 SSP FSS10 SSP LB10-2 SSP LB10-3 SSP SSP SSP SSP 13 SSP 14 SSP 15

LB10-6 LB10-8 LB10-9 LB10LB10LB10-

KR265734 Microbacterium sp. 100% / 99% KR265734 Microbacterium sp. 100% / 100%

JQ890541 Arthrobacter sp. 100% / 100% MF796783 Paenarthrobacter nitroguajacolicus 100% / 100%

Micrococcales

Actinobacteria

Micrococcales

Actinobacteria

Micrococcales

Actinobacteria

MF872589 Acinetobacter sp. 100% / 97%

KC178577 Acinetobacter ursingii 99% / 99% KC178577 Acinetobacter ursingii 100% / 99%

Pseudomonadales

Proteobacteria

-

-

-

-

-

+++

-

-

+++

-

+ -

-

-

+

-

-

+

-

-

-

-

-

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

+++

4

SSP LB1017 SSP LB1018

KC178577 Acinetobacter ursingii 100% / 99%

477

T6 FSS32

595

T6 LB10-26

632

T6 LB10-27

JF508423 Saccharothrix sp. 100% / 100% KX817875 Pseudoxanthomonas sp. 100% / 99% MG266274 Pseudarthrobacter sp. 100% / 100%

352, 510, 600

T6 T6 T6 T6

405

T8 FSS76

477

T8 LB10-49 T8 FSS75 T8 FSS62

451, 558

T8 FSS38

425

T8 T8 T8 T8 T8

355, 542

T8 LB10-50

FSS30 LB10-25 FSS29 LB10-28

FSS39 LB10-61 FSS63 FSS74 FSS40

KX767103 Bacillus idriensis 99% / 99% KY419849 Streptomyces sp. 99% / 100%

JF508423 Saccharothrix sp. 100% / 100% FJ532408 Streptomyces albovinaceus 99% / 100%

Pseudonocardiales

Actinobacteria

Xanthomonadales

Proteobacteria

Micrococcales

Actinobacteria

Bacillales

Streptomycetales

Pseudonocardiales

Streptomycetales

FM998727 Streptomyces cyaneogriseus 100% / 99% JN863488 Paenibacillus sp. 99% / 99%

Firmicutes

-

+++

+++

-

-

-

+++

++

+++

-

-

-

-

++

++

+++ +

++++ +

+ + -

++

++

+

+ +++

+++ ++ ++

+ + +++

++

+++

-

+ +++

++ ++++

+++ +++ ++++

++++

++++

++++

Actinobacteria

Actinobacteria

Actinobacteria

5

355, 613 300, 507, 600

T8 LB10-48 T8 LB10-47 T8 FSS77

KX959966 Paenibacillus lautus 99% / 100% JF912451 Bacillus sp. 100% / 99%

Bacillales

Firmicutes

T8 FSS37 T8 LB10-59 410

T12 AIA45

425

T12 FSS34 LB10-

444

T12 18 T12 T12 20 T12

LB10-

448

T12 19 T12 21 T12 T12

550

T12 AIA16

433

T12 FSS41 T12 FSS44

450

401, 450

KX035034 Streptomyces sp. 100% / 100% FM998727 Streptomyces cyaneogriseus 100% / 99%

Streptomycetales

Actinobacteria

FSS33 LB10AIA15

KF595308 Streptomyces sp. 100% / 100%250, MG205632 Rhodococcus jialingiae 100% / 100%

LB10FSS43 AIA46

T12 LB1067

Corynebacteriales

Actinobacteria

HE862284 Rhodococcus jostii 100% / 100% MF359740 Rhodococcus sp. 100% / 100% KU382287 Streptomyces sp. 100% / 99% KY753363 Streptomyces ciscaucasicus 100% / 100%

Streptomycetales

Actinobacteria

++++ ++++ ++

++ ++ ++++

++++ ++++ +

+ +

+ +

+ +

++

+

++++

++

++

+++

-

+

-

+ +

+ -

+

+

++

+++

-

-

-

-

-

-

+

+ -

+ -

+

+

+

+

++ +

+++

++

+++

+++

6

437 476 250, 476 250, 500

300, 500 352, 510, 600

618

T12 LB1066 T12 LB1065 T12 LB1055 T12 LB1058 T12 LB1047 T2 LB10-17 T2 AIA13 T2 FSS48 T12 LB1060 T12 AIA14 T12 FSS42 T12 LB1064 T12 FSS36

KY945877 Streptomyces sp. 100% / 100% GQ340513 Bacillus licheniformis 99% / 100% KY082733 Bacillus licheniformis 100% / 100% MF464018 Bacillus sp. 100% / 100% KY312760 Bacillus sp. 100% / 100%

Bacillales

Firmicutes

JQ357579 Bacillus megaterium 100% / 100% MF101078 Bacillus idriensis 100% / 100%

JN656234 Arthrobacter sp. 99% / 99%

Micrococcales

Actinobacteria

T12 FSS35 T12 FSS69 458

T21 AIA11

550

T21 AIA10 T21 AIA12 T21 LB1054

MF525917 Rhodococcus sp. 100% / 100% MF359740 Rhodococcus sp. 100% / 100% MF359740 Rhodococcus sp. 100% / 100%

Corynebacteriales

Actinobacteria

++

++

++

++

+++

++++

+

++

+

-

-

-

+

+

+

+ +++

+ + +++

++ ++++

+

+

-

-

++ -

-

+++

+

+

-

-

+ -

++++

++++

++++

++++ ++++

++++ +++

++++ ++++

++++

+++

++++

7

T21 21 T21 T21 60 T21 52 T21 53 T21

LB10-

-

-

+

AIA57 LB10-

+++ -

++ ++++

++++ +

++

-

-

-

+++

-

++++

++++

+++

AIA57 LB10-

+++ -

++ ++++

++++ +

++

++

-

250, 450

T21 T21 70 T21 73 T21

++++

++

+++

558

T21 AIA58

-

-

++

450, 611

T21 AIA59

+++

++++

++

+++

++++

++

++

++

+++

477

462

400, 478

LB10LB10AIA56

LB10AIA9

T21 LB1071 T21 LB1072

JF508423 Saccharothrix sp. 99% / 100% JQ419609 Saccharothrix sp. 100% / 99% KF730780 Streptomyces lateritius 99% / 100%

Pseudonocardiales

Streptomycetales CP021748 Streptomyces alboflavus 99% / 99% DQ286334 Bacillus thuringiensis 100% / 100% KU060121 Micrococcus sp. 99% / 99% KY514151 Arthrobacter oryzae 100% / 100% KY514151 Arthrobacter oryzae 100% / 100%

Actinobacteria

Actinobacteria

Bacillales

Firmicutes

Micrococcales

Actinobacteria

SSP: sieved agriculture soil mixed with perlite; T6: cellulose mat; T8: (PLA/PHB_w) original PLA/PHB transparent foil; T12: (PLA/PHB_b) original carbon black PLA/PHB filled foil; T21: (PLA/PHB_b 90D sun) carbon black PLA/PHB foil exposed 90 days to sun light. ++++: very rapid and extensive positive reaction; +++: quick and extensive positive reaction; ++: good positive reaction; +: weak positive reaction; -: negative reaction.

8

Table S2. Identification of fungal isolates and their polymers degradation abilities. Isolate

Species identification on the basis of the highest ITS similarity score

SSP FSS15-f

NR_148193 Cladosporium subcinereum 100% / 100% KX079893 Cladosporium sp. 98% / 100% KT269200 Cladosporium sp. 99% / 99% KY398831 Cladosporium sp. 99% / 100%

SSP PDA16-f SSP FSS14-f SSP PDA16E-f SSP SAB20-f SSP SAB18-f SSP SAB23-f SSP SAB25-f SSP FSS30-f SSP SAB3-f SSP FSS11A-f SSP FSS11B-f SSP SAB10-f SSP PDA24-f

AB190399 Phialophora intermedia 99% / 99% AB190399 Phialophora intermedia 100% / 99% AB190399 Phialophora intermedia 99% / 99% AB190399 Phialophora intermedia 99% / 99% AB190399 Phialophora intermedia 100% / 99% AB190399 Phialophora intermedia 100% / 99% AF443929 Trichoderma harzianum 98% / 98% AF443929 Trichoderma harzianum 100% / 97% KR364588 Fusarium oxysporum 99% / 99% KX237567 Geotrichum candidum 100% / 1008%

Order

Capnodiales

Chaetothyriales

Hypocreales

Saccharomycetales

Phylum

Ascomycota

Ascomycota

Ascomycota

Ascomycota

Polymer degradation assays PHB PLA/PHB PLA blend +++

+++

+++

++ ++

++ +

++ ++

++

+

++

++

+

+

++

++

++

++

++

++

++

++

++

++

++

++

++

+

++

++

++

++

+++

+++

+++

++

+

++

++

++

++

9

SSP PDA17B-f SSP FSS7-f SSP SAB12-f SSP FSS8-f SSP FSS22-f SSP PDA17A-f SSP SAB13-f

T6 PDA1-f T6 PDA2-f T6 PDA3-f T6 PDA4-f T6 SAB1-f T6 SAB3-f T8 PDA1-f T8 PDA2-f T8 PDA4-f T8 PDA5-f T8 PDA6-f T8 SAB1-f

KF298066 Galactomyces candidum 100% / 99% JN636809 Candida sp. 99% / 99% KM877470 Aureobasidium pullulans 100% / 99% EU484210 Umbelopsis isabellina 100% / 99% KX008633 Penicillium sp. 100% / 99% NR_077155 Penicillium melinii 99% / 99% DQ888734 Penicillium melinii 99% / 100% KP900325 Penicillium skrjabinii 100% / 99% KY425720 Penicillium sp. 100% / 100% KP267009 Fusarium oxysporum 100% / 100% KJ702597 Fusarium sp. 100% / 100% LT746274 Fusarium solani 100% / 100% MG519516 Fusarium sp. 100% / 100% KX359599 Paraphaeosphaeria sporulosa 100% / 99% MG519516 Fusarium sp. 100% / 99% KC329616 Fusarium oxysporum 100% / 97% MG543779 Fusarium oxysporum 100% / 100% KJ847746 Fusarium oxysporum 98% / 99% KY225678 Trichoderma harzianum 100% / 100%

Dothideales

Ascomycota

Umbelopsidales

Mucoromycota

Eurotiales

Ascomycota

Eurotiales

Hypocreales

Pleosporales

Hypocreales

Ascomycota

Ascomycota

Ascomycota

Ascomycota

+

+

++

+

+

+

+++

+++

+++

++

++

++

+

+

+

+

+

+

+

+

+

++

++

++

++ ++

+++ ++++

+ ++

+ ++ ++++

++++ ++++ ++++

+ +++ +

++++

++

+

++ ++

+++ +++

+ ++

+++

+++

++

++

+++

+

-

++++

++++ 10

T8 PDA8-f T8 FSS2-f

KJ191439 Absidia sp. 100% / 99% KY788057 Cladosporium cladosporioides 100% / 100%

T12 PDA2-f T12 PDA3-f

JF745093 Absidia sp. 100% / 100% JX975889 Mortierella lignicola 100% / 100% KY419559 Fusarium equiseti 100% / 100%

T12 SAB1-f

T21 SAB1-f T21 SAB2-f T21 PDA1-f T21 SAB3-f T21 PDA4-f T21 SAB5-f T21 PDA5-f

KU509526 Bjerkandera adusta 100% / 100% KX343172 Mortierella sp. 99% / 99% KM404167 Actinomucor elegans 100% / 100% KY750722 Aspergillus fumigatus 100% / 99% KX788164 Penicillium sp. 100% / 100% KX347463 Fusarium solani 99% / 99% MF166765 Fusarium equiseti 100% / 100%

Mucorales Capnodiales

Mucoromycota Ascomycota

+++ ++

++++ ++

++ -

Mucorales Mortierellales

Mucoromycota

+ ++++

++ +++

+ -

Hypocreales

Ascomycota

++++

++++

+++

Polyporales

Basidiomycota

+++

++++

+++

Mortierellales Mucorales

Mucoromycota

++ +++

++++ +++

+++ ++++

Eurotiales

Ascomycota

+

+++

-

Ascomycota

+ ++ ++++

++ ++++ +++

+ ++ +++

Hypocreales

SSP: sieved agriculture soil mixed with perlite; T6: cellulose mat; T8: (PLA/PHB_w) original PLA/PHB transparent foil; T12: (PLA/PHB_b) original carbon black PLA/PHB filled foil; T21: (PLA/PHB_b 90D sun) carbon black PLA/PHB foil exposed 90 days to sun light. ++++: very rapid and extensive positive reaction; +++: quick and extensive positive reaction; ++: good positive reaction; +: weak positive reaction; -: negative reaction.

11

Table S3. Bacterial microbiome detected in soil and on polymeric samples by DGGE-cloning approach. Sample SSP

Sequenced Clone / No. of clones K-253-14-36 / 3 K-253-14-30 / 14 K-253-14-15 / 14 K-253-14-32 / 4 K-253-14-33 / 4 K-253-14-5 / 3 K-253-14-40 / 4 K-253-14-39 / 4 K-253-14-42 / 3 K-253-14-21 / 3 K-253-14-35 / 14 K-253-14-7 / 14

Identification on the basis of the highest BLAST hit 16S rRNA similarity score JF417817 Uncultured bacterium 100% / 96% LN567865 Uncultured bacterium 100% / 99% JQ426278 Uncultured bacterium 100% / 100% AY493983 Uncultured bacterium 99% / 99% MF116285 Uncultured Agromyces sp. 100% / 100% JQ356905 Uncultured bacterium 100% / 99% EU133872 Uncultured bacterium 100% / 100% AJ275891 Uncultured Nitrosomonas sp. 100% / 100% JF829432 Uncultured bacterium 100% / 99% JQ379484 Uncultured bacterium 100% / 96% HQ190351 Uncultured bacterium 100% / 99% EU122711 Uncultured Acidobacteria bacterium 100% / 99%

Best BLAST hit using the bacterial and archaeal 16S rRNA database NR_149251 Longimicrobium terrae 91% / 91% NR_074708 Gemmatimonas aurantiaca 100% / 83% NR_132675 Gemmatirosa kalamazoonesis 98% / 86% NR_112972 Aciditerrimonas ferrireducens 99% / 89% np

Order

Phylum

Longimicrobiales Gemmatimonadal es Gemmatimonadal es Acidimicrobiales

Gemmatimonade tes Gemmatimonade tes Gemmatimonade tes Actinobacteria

Micrococcales

Actinobacteria

NR_137235 Burkholderia rhizosphaerae 99% / 88% NR_114127 Derxia gummosa 100% / 91% np

Burkholderiales

Proteobacteria

Burkholderiales

Proteobacteria

Nitrosomonadales

Proteobacteria

NR_102479 Thioflavicoccus mobilis 99% / 83% NR_109657 Dehalogenimonas alkenigignens 100% / 82% NR_151905 Vicinamibacter silvestris 99% / 81% NR_151905 Vicinamibacter silvestris 100% / 90%

Chromatiales

Proteobacteria

Dehalococcoidia

Chloroflexi

Acidobacteria subdivision 6 Acidobacteria subdivision 6

Acidobacteria Acidobacteria

12

K-253-14-41 / 4 K-253-14-13 / 4 K-253-14-47 / 4 K-253-14-6 / 4

T6

K-9-16-22 / 15 K-9-16-16 / 2 K-9-16-19 / 4 K-9-16-8 / 3 K-9-16-42 / 8 K-9-16-2 / 5 K-9-16-14 / 2 K-9-16-35 / 8 K-9-16-21 / 3 K-9-16-13 / 2 K-9-16-9 / 11

KF836153 Uncultured bacterium 100% / 98% JN697135 Uncultured Nitrospira sp. 100% / 99% HG327388 Uncultured bacterium 100% / 99% NR_146376 Lacunisphaera limnophila 100% / 97%

NR_147754 Rufibacter glacialis 100% / 83% np

Cytophagales

Bacteroidetes

Nitrospirales

Nitrospirae

NR_114396 Nitrospira japonica 98% / 98% np

Nitrospirales

Nitrospirae

Opitutales

Verrucomicrobia

GQ202639 Uncultured bacterium 100% / 100% HQ597149 Uncultured Acidobacteria 100% / 100% GQ214128 Uncultured bacterium 100% / 96% GQ214128 Uncultured bacterium 100% / 96% HQ597726 Uncultured Acidobacteria 100% / 99% HQ598467 Uncultured Acidobacteria bacterium 100% / 99% KY302209 Uncultured Massilia sp. 100% / 100% EF540374 Uncultured soil bacterium 99% / 99% KJ664622 Uncultured bacterium 100% / 100% HQ264893 Uncultured bacterium 100% / 98%

NR_151905 Vicinamibacter silvestris 100% / 92% NR_151905 Vicinamibacter silvestris 100% / 90% NR_151905 Vicinamibacter silvestris 100% / 90% NR_151905 Vicinamibacter silvestris 100% / 90% NR_151905 Vicinamibacter silvestris 100% / 90% NR_137410 Terriglobus albidus 99% / 85%

Acidobacteria subdivision 6 Acidobacteria subdivision 6 Acidobacteria subdivision 6 Acidobacteria subdivision 6 Acidobacteria subdivision 6 Acidobacteriales

Acidobacteria

np

Burkholderiales

Proteobacteria

NR_109152 Cupriavidus alkaliphilus 100% / 89% NR_044309 Steroidobacter denitrificans 97% / 87% NR_126260 Acidibacter ferrireducens 90% / 95%

Burkholderiales

Proteobacteria

Nevskiales

Proteobacteria

JF319219 Uncultured Acidobacteria 100% / 100%

NR_041981 Kofleria flava 99% / 82%

Acidobacteria Acidobacteria Acidobacteria Acidobacteria Acidobacteria

unclassified Proteobacteria Gammaproteobact eria Myxococcales Proteobacteria

13

K-9-16-47 / 9 K-9-16-29 / 2 K-9-16-27 / 11 K-9-16-1 / 8 K-9-16-25 / 2 K-9-16-45 / 5

T8

K-10-16-4 / 8 K-10-16-41 / 5 K-10-16-26 / 3 K-10-16-29 / 2 K-10-16-7 / 13 K-10-16-8 / 16

KY041865 Uncultured Desulfonatronum sp. 100% / 100% DQ833488 Uncultured bacterium 100% / 96% FM253679 Uncultured Chloroflexi bacterium 100% / 100% JQ383030 Uncultured bacterium 100% / 96% EF540397 Uncultured bacterium 100%/ 100% JQ374231 Uncultured bacterium 100% / 96% MG557797 Uncultured Pseudomonas sp. 100% / 100% KF182949 Uncultured gamma proteobacterium 100% / 96% AB769234 Uncultured Geobacter sp. 100% / 100% KC215462 Uncultured Desulfomicrobium sp. 100% / 100% HQ729826 Uncultured Acidobacteria bacterium 100% / 98% FJ444739 Uncultured bacterium 100% / 95%

np

Desulfovibrionales

Proteobacteria

NR_132293 Thermomarinilinea lacunifontana 100% / 82% NR_109544 Ornatilinea apprima 98% / 82%

Anaerolineales

Chloroflexi

Anaerolineales

Chloroflexi

NR_041354 Bellilinea caldifistulae 50% / 79% NR_074708 Gemmatimonas aurantiaca 100% / 90% NR_108511 Chryseolinea serpens 100% / 87%

Anaerolineales

Chloroflexi

Gemmatimonadal es Cytophagales

Gemmatimonade tes Bacteroidetes

np

Pseudomonadales

Proteobacteria

NR_112675 Methylomarinum vadi 100% / 87%

Methylococcales

Proteobacteria

np

Proteobacteria

np

Desulfuromonadal es Desulfovibrionales

NR_151905 Vicinamibacter silvestris 100% / 89%

Acidobacteria subdivision 6

Acidobacteria

NR_151905 Vicinamibacter silvestris 100% / 89%

Acidobacteria subdivision 6

Acidobacteria

Proteobacteria

14

K-10-16-25 / 8

HQ729884 Uncultured Acidobacteria bacterium 99% / 97% AB733567 Uncultured Bacillus sp. 100% / 100% KY515440 Uncultured Paenibacillus sp. 100% / 100% LC269949 Uncultured Pseudonocardia sp. 100% / 100% HE647633 Uncultured bacterium 100% / 97%

NR_151905 Vicinamibacter silvestris 100% / 90%

Acidobacteria subdivision 6

Acidobacteria

np

Bacillales

Firmicutes

np

Bacillales

Firmicutes

np

Pseudonocardiales

Actinobacteria

NR_109544 Ornatilinea apprima 98% / 82%

Anaerolineales

Chloroflexi

K-19-16-10 / 10

KF145634 Uncultured bacterium 100% / 95%

Anaerolineales

Chloroflexi

K-19-16-8 / 7

GU941080 Uncultured bacterium 100% / 100% KC432420 Uncultured bacterium 100% / 98%

NR_132293 Thermomarinilinea lacunifontana 100% / 80% NR_074383 Anaerolinea thermophila 89% / 82% NR_132293 Thermomarinilinea lacunifontana 91% / 80% np

Anaerolineales

Chloroflexi

Anaerolineales

Chloroflexi

Burkholderiales

Proteobacteria

NR_114629 Acidiferrobacter thiooxydans 99% / 89% NR_108192 Solirubrobacter ginsenosidimutans 100% / 91% NR_132675 Gemmatirosa kalamazoonesis 100% / 84%

Acidiferrobacteral es Solirubrobacterale s

Proteobacteria

Gemmatimonadal es

Gemmatimonade tes

K-10-16-5 / 10 K-10-16-1 / 9 K-10-16-34 / 21 K-10-16-42 / 5

T12

K-19-16-36 / 7 K-19-16-1 / 17 K-19-16-29 / 2 K-19-16-16 / 5 K-19-16-37 / 3

MG459259 Uncultured Burkholderia sp. 100% / 100% KC331240 Uncultured bacterium 99% / 99% HQ318613 Uncultured soil bacterium 100% / 100% FQ658847 Uncultured bacterium 100% / 99%

Actinobacteria

15

K-19-16-46 / 7 K-19-16-40 / 6 K-19-16-33 / 5 K-19-16-39 / 2 K-19-16-42 / 5 K-19-16-30 / 14 K-19-16-28 / 10

T21

K-11-16-24 / 10 K-11-16-30 / 5 K-11-16-50 / 7 K-11-16-1 / 10 K-11-16-13 / 5 K-11-16-25 / 2 K-11-16-9 / 7

HQ598260 Uncultured Acidobacteria 100% / 97% JN038787 Uncultured Acidobacteria 99% / 95% KC540997 Uncultured bacterium 100% / 96% HQ729833 Uncultured Acidobacteria bacterium 100% / 99% FJ444642 Uncultured bacterium 100% / 99% JX489863 Uncultured soil bacterium 100% / 95% FJ175009 Uncultured bacterium 100% / 98%

NR_151905 Vicinamibacter silvestris 100% / 90% NR_151905 Vicinamibacter silvestris 99% / 88% NR_151905 Vicinamibacter silvestris 99% / 82% NR_151905 Vicinamibacter silvestris 100% / 89%

Acidobacteria subdivision 6 Acidobacteria subdivision 6 Acidobacteria subdivision 6 Acidobacteria subdivision 6

Acidobacteria

NR_151905 Vicinamibacter silvestris 100% / 90% NR_117436 Ohtaekwangia kribbensis 100% / 91% NR_117435 Ohtaekwangia koreensis 100% / 94%

Acidobacteria subdivision 6 Cytophagales

Acidobacteria

Cytophagales

Bacteroidetes

HQ597745 Uncultured Acidobacteria bacterium 100% / 100% HQ121155 Uncultured bacterium 100% / 99% KF712822 Uncultured bacterium 100% / 100% DQ125910 Uncultured bacterium 100% / 100% KJ766171 Uncultured bacterium 100% / 99% JQ373103 Uncultured bacterium 100% / 99% KF183039 Uncultured gamma proteobacterium 100% / 99%

NR_151905 Vicinamibacter silvestris 100% / 90%

Acidobacteria subdivision 6

Acidobacteria

NR_151905 Vicinamibacter silvestris 100% / 91% NR_134120 Paludibaculum fermentans 100% / 85% NR_028927 Sporomusa ovata 100% / 86% NR_126172 Povalibacter uvarum 100% / 90% NR_126172 Povalibacter uvarum 100% / 95% NR_114629 Acidiferrobacter thiooxydans 99% / 90%

Acidobacteria subdivision 6 Solibacterales

Acidobacteria

Selenomonadales

Firmicutes

Nevskiales

Proteobacteria

Nevskiales

Proteobacteria

Acidiferrobacteral es

Proteobacteria

Acidobacteria Acidobacteria Acidobacteria

Bacteroidetes

Acidobacteria

16

K-11-16-15 / 2 K-11-16-21 / 5 K-11-16-22 / 7

LN555100 Uncultured Nitrospira sp. 100% / 100% KC432418 Uncultured bacterium 100% / 98% KF145634 Uncultured bacterium 100% / 95%

K-11-16-29 / 8

KF145634 Uncultured bacterium 100% / 95%

K-11-16-39 / 12

JQ825200 Uncultured bacterium 100% / 98% HQ697782 Uncultured bacterium 100% / 97% HQ697782 Uncultured bacterium 100% / 96%

K-11-16-7 / 13 K-11-16-5 / 7

np

Nitrospirales

Nitrospirae

NR_041354 Bellilinea caldifistulae 100% / 88% NR_132293 Thermomarinilinea lacunifontana 100% / 80% NR_132293 Thermomarinilinea lacunifontana 100% / 80% NR_074383 Anaerolinea thermophila 99% / 81% NR_117435 Ohtaekwangia koreensis 100% / 88% NR_117435 Ohtaekwangia koreensis 100% / 89%

Anaerolineales

Chloroflexi

Anaerolineales

Chloroflexi

Anaerolineales

Chloroflexi

Anaerolineales

Chloroflexi

Cytophagales

Bacteroidetes

Cytophagales

Bacteroidetes

SSP: sieved agriculture soil mixed with perlite; T6: cellulose mat; T8: (PLA/PHB_w) original PLA/PHB transparent foil; T12: (PLA/PHB_b) original carbon black PLA/PHB filled foil; T21: (PLA/PHB_b 90D sun) carbon black PLA/PHB foil exposed 90 days to sun light.

17

Table S4. Eukaryotic microbiome detected in soil and on polymeric samples by DGGE-cloning approach. Sample

Sequenced Clone / No. of clones

SSP

K-254-14-2 / 3 K-254-14-15 / 2 K-254-14-5 / 11 K-254-14-38 / 4 K-254-14-6 / 2 K-254-14-12 / 6 K-254-14-10 / 2 K-254-14-27 / 12 K-254-14-4 / 15 K-254-14-3 / 22

Identification on the basis of the highest BLAST hit ITS similarity score HE610118 Desmochloris halophila 100% / 100% FR865573 Chlamydomonas sphagnophila 100% / 100% FJ554389 Oxytrichidae environmental clone 100% / 98% DQ241756 Halteria grandinella 100% / 99% FJ553679 Oxytrichidae environmental clone 100% / 97% GU055550 Uncultured Hypocreales clone 100% / 100% KF800342 Uncultured fungus 100% / 100% KT215208 Arthrobotrys amerospora 100% / 100% KY270893 Pseudogymnoascus sp. 100% / 99% HG935918 Uncultured Pseudeurotium sp. 100% / 99%

BLAST search excluding the Order uncultured/environmental sample sequences

Phylum

np

Ulvales

Chlorophyta

np

Chlamydomonadales Chlorophyta

AJ286799 Oxytricha longigranulosa 100% / 96%

Sporadotrichida

Ciliophora

np

Sporadotrichida

Ciliophora

KM222032 Hemiamphisiella terricola 100% / 97%

Stichotrichida

Ciliophora

EU873529 Dendroclathra lignicola 100% / 79%

Hypocreomycetidae incertae sedis

Ascomycota

AB916509 Venturia sp. 100% / 97% np

Venturiales

Ascomycota

Orbiliales

Ascomycota

np

Leotiomycetes incertae sedis

Ascomycota

np

Leotiomycetes incertae sedis

Ascomycota

18

K-254-14-17 / 2 K-254-14-22 / 2 K-254-14-45 / 5 K-254-14-18 / 8 K-254-14-43 / 4

T6

K-5-16-22 / 2 K-5-16-5 / 5 K-5-16-10 / 14 K-5-16-13 / 5 K-5-16-17 / 18 K-5-16-14 / 12 K-5-16-41 / 44

T8

K-7-16-9 / 3

AJ972828 Uncultured Ascomycota fungus 100% / 100% JN392112 Uncultured Peziza clone 100% / 99% HG937009 Chrysosporium sp. 100% / 100% KX359606 Uncultured Solicoccozyma terricola 100% / 99% KX359606 Uncultured Solicoccozyma terricola 100% / 100%

MG076777 Pleosporales sp. 100% / 97%

Pleosporales

Ascomycota

np

Pezizales

Ascomycota

np

Onygenales

Ascomycota

np

Filobasidiales

Basidiomycota

np

Filobasidiales

Basidiomycota

FJ553398 Oxytrichidae environmental clone 99% / 99% KX710251 Preussia sp. 99% / 100% JX270604 Uncultured Geomyces sp. 100% / 99% JX270387 Uncultured Geomyces sp. 100% / 98% KY977542 Uncultured Pseudeurotium bakeri 100% / 100% JN392112 Uncultured Peziza clone 100% / 99% JN392106 Uncultured Coprinus sp. 100% / 99%

DQ241756 Halteria grandinella 97% / 93%

Sporadotrichida

Ciliophora

np

Pleosporales

Ascomycota

np

Leotiomycetes incertae sedis Leotiomycetes incertae sedis Leotiomycetes incertae sedis

Ascomycota

np

Pezizales

Ascomycota

np

Agaricales

Basidiomycota

FR865573 Chlamydomonas sphagnophila 100% / 100%

np

Chlamydomonadales Chlorophyta

np np

Ascomycota Ascomycota

19

K-7-16-10 / 3 K-7-16-32 / 13 K-7-16-41 / 9 K-7-16-44 / 24 K-7-16-22 / 15 K-7-16-33 / 15 K-7-16-45 / 18 T12

K-6-16-8 / 2 K-6-16-10 / 4 K-6-16-40 / 2 K-6-16-42 / 2 K-6-16-5 / 2 K-6-16-22 / 30 K-6-16-2 / 2

FJ552794 Uncultured stichotrichid 99% / 97% JX270604 Uncultured Geomyces sp. 100% / 100% KY977601 Uncultured Pseudogymnoascus sp. 100% / 99% HG935918 Pseudeurotium sp. 100% / 99% KT215208 Uncultured Arthrobotrys amerospora 100% / 100% MG515307 Fusarium solani 100% / 99% KF296737 Uncultured fungus 100% / 97%

DQ241756 Halteria grandinella 97% / 96% np

Sporadotrichida

Ciliophora

Leotiomycetes incertae sedis

Ascomycota

np

Leotiomycetes incertae sedis

Ascomycota

np

Ascomycota

np

Leotiomycetes incertae sedis Orbiliales

np

Hypocreales

Ascomycota

AY461837 Coprinus cordisporus 100% / 97%

Agaricales

Basidiomycota

DQ241756 Halteria grandinella 100% / 99% FJ553398 Oxytrichidae environmental clone 100% / 99% FJ553795 Uncultured stichotrichid 100% / 96% FR865573 Chlamydomonas sphagnophila 100% / 100% JQ388941 Pleosporales sp. 100% / 100% JX270604 Geomyces sp. 100% / 99% JX270343 Geomyces sp. 100% / 100%

np

Sporadotrichida

Ciliophora

DQ241756 Halteria grandinella 97% / 93%

Sporadotrichida

Ciliophora

DQ241756 Halteria grandinella 97% / 96% np

Sporadotrichida

Ciliophora

np

Pleosporales

Ascomycota

np

Leotiomycetes incertae sedis Leotiomycetes incertae sedis

Ascomycota

np

Ascomycota

Chlamydomonadales Chlorophyta

Ascomycota

20

K-6-16-28 / 8 K-6-16-12 / 16 K-6-16-37 / 9 K-6-16-29 / 7 K-6-16-38 / 2 K-6-16-31 / 12 K-6-16-32 / 2

T21

K-8-16-4 / 4 K-8-16-31 / 4 K-8-16-13 / 4 K-8-16-20 / 19 K-8-16-1 / 4 K-8-16-30 / 7 K-8-16-19 / 37

JX270604 Geomyces sp. 100% / 100% KY977542 Uncultured Pseudeurotium bakeri 100% / 99% KY977542 Uncultured Pseudeurotium bakeri 100% / 99% KY977601 Pseudogymnoascus sp. 100% / 99% JN392112 Uncultured Peziza clone 100% / 98% MF939662 Mortierella alpina 100% / 100% FN397311 Uncultured fungus 100% / 98%

np

Leotiomycetes incertae sedis Leotiomycetes incertae sedis

Ascomycota

np

Leotiomycetes incertae sedis

Ascomycota

np

Leotiomycetes incertae sedis

Ascomycota

np

Pezizales

Ascomycota

np

Mortierellales

Mucoromycota

KX343169 Mortierella alpina 100% / 97%

Mortierellales

Mucoromycota

FR865573 Chlamydomonas sphagnophila 100% / 100% AF508755 Cyrtohymena citrina 100% / 100% JX270604 Geomyces sp. 100% / 99% JX270476 Pseudeurotium sp. 100% / 100% KC966070 Uncultured fungus 100% / 99% KC966070 Uncultured fungus 100% / 99% EU292388 Uncultured fungus clone 100% / 100%

np

Chlamydomonadales Chlorophyta

np

Sporadotrichida

Ciliophora

np

Leotiomycetes incertae sedis Leotiomycetes incertae sedis Pleosporales

Ascomycota

Pleosporales

Ascomycota

Thelephorales

Basidiomycota

np

np KT268587 Herpotrichia sp. 100% / 99% KT268587 Herpotrichia sp. 100% / 99% AF184743 Thelephoraceae sp. 100% / 98%

Ascomycota

Ascomycota Ascomycota

21

K-8-16-33 / 21

KX08068 Mortierella alpina 100% / 100%

np

Mortierellales

Mucoromycota

SSP: sieved agriculture soil mixed with perlite; T6: cellulose mat; T8: (PLA/PHB_w) original PLA/PHB transparent foil; T12: (PLA/PHB_b) original carbon black PLA/PHB filled foil; T21: (PLA/PHB_b 90D sun) carbon black PLA/PHB foil exposed 90 days to sun light.

22