Supplementary Materials

Table S1. Origin data of Antarctic bacterial strains included in this study.

Strain ID* Sampling Location Sampling Date Strain in Coculture So1b Fildes Bay (a) 2013/2014 So13.3 So13.3 Fildes Bay (a) 2013/2014 So1b So64.6b Ardley Island (ASPA 150) 2013/2014 So32D So5b Collins Glacier beach area (ASPA 125) 2013/2014 So27D Se14.01b Armonía point (b) (ASPA 133) 2013/2014 Se41.02b Se5.02b Hannah point, (c) (ASPA 126) 2013/2014 Se5.01b Se18.01b Fildes Bay (a) 2013/2014 Se63.02b Se28.01b Dee- (a) 2013/2014 Se32.01b Se32.01b Dee-Greenwich Island (a) 2013/2014 Se28.01b Se41.02b Armonía point (ASPA 133) 2013/2014 Se14.01b Wa41.01b Armonía point (ASPA 133) 2013/2014 Dwa41.01b So64.3b Ardley Island (ASPA 150) 2013/2014 So64.6b So27D Collins Glacier beach area (ASPA 125) 2013/2014 So5b So10b Collins Glacier beach area (ASPA 125) 2013/2014 So2b So2b Collins Glacier beach area (ASPA 125) 2013/2014 So10b So9b Collins Glacier beach area (ASPA 125) 2013/2014 So6b Dwa41.01b Fildes Bay (a) 2013/2014 Wa41.01 Se4.02b Hannah point, Livingston Island (ASPA 126) 2013/2014 Se5.02b So11b Collins Glacier beach area (ASPA 125) 2013/2014 SoD9b Se63.02b Fildes Bay (a) 2013/2014 Se18.01 So1d Collins Glacier beach area (ASPA 125) 2013/2014 So5b So13b Fildes Bay (a) 2016 So02b So3.2b Island (ASPA 112) 2016 Se32.02b Se32.02b Robert Island (ASPA 112) 2013/2014 So3.2b Se5.01b Hannah point, Livingston Island (ASPA 126) 2013/2014 Se5.02b So6b Collins Glacier beach area (ASPA 125) 2013/2014 So9b SoD9b Collins Glacier beach area (ASPA 125) 2013/2014 So11b So02b Fildes Bay (a) 2016 So13b Se16.2.2 Collins Glacier beach area (ASPA 125) 2016 Se16.2.3 Se16.2.3 Collins Glacier beach area (ASPA 125) 2016 Se16.2.2 Se16.2.5 Collins Glacier beach area (ASPA 125) 2016 Se16.2.3 So16.8 Armonía point (ASPA 133) 2016 So16.17 So16.17 Armonía point (ASPA 133) 2016 So16.8 So16.14.3 Robert Island (ASPA 112) 2016 So16.14.1 *Name codes So: soil, Se: sediment, Wa: water, Dwa: 10 m deep water, .ASPA: Antarctic Specially Protected Area number. (a) Area with human presence. (b) Lake sediment 0.3 m deep. (c) Marine sediment 0.3 m deep.

Table S2. Composition of culture media used in this study.

Media ID Composition Per Liter Peptone (2.0 g) M1 Yeast extract (4.0 g) Starch (1 0.0 g) Yeast extract (4.0 g) ISP2 Malt extract (10.0 g) Glucose (4.0 g) Mannitol (40.0 g) Maltose (40.0 g) Yeast extract (10.0 g) M2 K2HPO4 (2.0 g) MgSO4.7H2O (0.5 g) FeSO4.7H2O (0.01 g) Starch (10.0 g) Casein (0.3 g) KNO3 (2.0 g) SCA NaCl (2.0 g) K2HPO4 (2.0 g) MgSO4.7H2O (0.05 g) CaCO3 (0.02 g) 2 of 9

FeSO4.7H2O (0.01 g) Yeast extract (4g) Malt extract (10g) IMA Glucose (4g) Mannitol (40g) Peptidic digest of animal tissue (5.0 g), SDB Casein pancreatin disgest (5.0 g), Dextrose (40.0 g) Glicerol (30g) Peptone (2g) K2HPO4 (1g) NaCl (1g) MgSO4.7H2O (0.5g) Trace solution * (5mL) *Trace solution CCA CaCl2x2H2O (3g) MnSO4 (0.2g) ZnCl2 (0.1g) CuSO4.5H2O (0.025g) Na2B4O7x10H2O (0.02g) CoCl2 (0.004g) (NH4)6Mo7O24 4H2O (0.01g) Yeast extract (3g) Peptone (5g) YEME Malt extract (3g) Glucose (10g) Sucrose (170g) Yeast extract (4g) Malt extract (10g) GYA Glucose (4g) CaCO3 (2g) Starch (20g) Sucrose (150 g) Yeast extract (20 g) YES MgSO4x7H2O (0.5 g) ZnSO4x7H2O (0.01 g) CuSO4x5H2O (0.005 g) Starch (10.0 g) CaCO3 (2.0 g) (NH4)2SO4 (2.0 g) K2HPO4 (1.0 g) ISP4 MgSO4.7H2O (1.0 g) NaCl (1.0 g) FeSO4.7H2O (1 mg) MnCl2.7H2O (1.0 mg) ZnSO4.7H2O (1.0 mg) ES 100g of Antarctic soil (1 hour agitation prior filtering) R2YE [1] [1] Ghimire, G. P.; Koirala, N.; Sohng, J. K. Activation of cryptic hop genes from streptomyces peucetius ATCC 27952 involved in hopanoid biosynthesis. J. Microbiol. Biotechnol. 2015, doi:10.4014/jmb.1408.08058. 3 of 9

Table S3. Biosynthetic gene clusters (BGCs) identified on the genomes of selected Antarctic bacteria strains as potential source of novel antimicrobial compounds.

Dwa41.01b Region Type From To Most similar known cluster Similarity Region 1 ladderane,fatty_acid 75,852 118,134 p seudomonas quinolone signal 27% Region 2 saccharide 271,126 294,546 Region 3 saccharide 364,464 380,12 Region 4 saccharide 1,126,731 1,151,389 Region 5 saccharide 1,404,293 1,426,859 Region 6 halogenated 1,466,276 1,491,000 WAP-8294A2 30% Region 7 saccharide 1,500,477 1,535,607 Region 8 saccharide 1,545,784 1,566,616 Region 9 saccharide 1,706,299 1,752,277 exopolysaccharide 20% Region 10 saccharide 2,181,871 2,200,672 Region 11 saccharide 2,498,222 2,537,719 Region 12 terpene 2,585,329 2,604,912 Region 13 acyl_amino_acids 2,666,701 2,726,921 Region 14 arylpolyene 3,204,781 3,248,883 APE Vf 35% Region 15 hserlactone 3,275,936 3,296,523 Region 16 fatty_acid 3,375,525 3,398,099 Region 17 saccharide 3,584,363 3,622,907 K53 capsular polysaccharide 15% Region 18 saccharide 3,827,884 3,872,531 Region 19 saccharide 4,186,168 4,207,395 lipopolysaccharide 5% Region 20 terpene 4,301,655 4,325,256 carotenoid 100% Region 21 saccharide 4,346,330 4,403,382 Region 22 saccharide 4,479,885 4,530,756 lipopolysaccharide 14% Region 23 saccharide 4,615,542 4,639,123 O-antigen 14% Region 24 saccharide 4,812,304 4,836,129 lipopolysaccharide 5% Region 25 saccharide 4,840,049 4,860,600 Region 26 acyl_amino_acids,sacc 5,179,912 5,267,121 K53 capsular polysaccharide 10% Region 27 fatty_acid 5,268,349 5,289,470 polyhydroxyalkanoate 50% So1b Region Type From To Most similar known cluster Similarity Region 1 saccharide,T3PKS 333,107 430,53 dipeptide aldehydes 11% Region 2 saccharide,terpene 435,276 473,443 Region 3 NRPS,betalactone 536,424 618,887 fengycin 100% Region 4 saccharide 665,202 685,619 Region 5 transAT-PKS- 742,139 842,379 bacillaene Similarity Region 6 saccharide 1,006,994 1,027,472 Region 7 saccharide 1,334,496 1,356,064 Region 8 fatty_acid 1,385,610 1,406,452 chejuenolide A / chejuenolide B 11% Region 9 terpene 1,433,936 1,454,715 Region 10 saccharide 1,468,497 1,485,149 zwittermicin A 18% Region 11 fatty_acid 1,501,606 1,522,487 Region 12 halogenated 1,705,352 1,725,585 Region 13 saccharide 1,785,201 1,807,273 Region 14 NRPS 2,137,424 2,201,286 surfactin 78% Region 15 saccharide 2,219,940 2,262,274 Region 16 lanthipeptide 2,332,643 2,357,099 subtilomycin 100% Region 17 saccharide 2,485,752 2,506,785 Region 18 saccharide 2,758,084 2,778,117 bacillomycin D 20% Region 19 saccharide 2,806,497 2,826,120 Region 20 saccharide,other 2,852,963 2,910,931 bacilysin 100% Region 21 sactipeptide,head_to_t 2,917,688 2,939,298 subtilosin A 100% Region 22 saccharide 3,072,095 3,126,189 teichuronic acid 100% Region 23 saccharide 3,194,906 3,256,035 K53 capsular polysaccharide 15% Region 24 saccharide 3,277,735 3,303,013 Region 25 NRPS 3,458,208 3,507,248 bacillibactin 100% Region 26 saccharide 3,532,514 3,563,347 Region 27 saccharide 3,587,706 3,621,555 scytophycin 11% Region 28 saccharide 3,666,012 3,716,386 4 of 9

Region 29 saccharide 3,915,985 3,956,517 Se16.2.3 Region Type From To Most similar known cluster Similarity Region 1 saccharide 61,393 85,833 O-antigen 9% Region 2 saccharide 258,489 282,201 lipopolysaccharide 5% Region 3 saccharide 284,796 306,672 pyoverdin 1% Region 4 acyl_amino_acids, 626,909 711,754 K53 capsular polysaccharide 10% Region 5 fatty_acid 716,652 734,902 polyhydroxyalkanoate 50% Region 6 fatty_acid 882,123 909,289 pseudomonas quinolone signal 27% Region 7 saccharide 1,066,648 1,089,120 Region 8 saccharide 1,159,045 1,174,700 Region 9 saccharide 1,922,274 1,946,138 Region 10 halogenated 2,261,080 2,285,942 WAP-8294A2 30% Region 11 saccharide 2,501,139 2,547,133 exopolysaccharide 20% Region 12 saccharide 2,674,663 2,695,176 Region 13 saccharide 2,973,950 2,996,371 Region 14 saccharide 3,293,374 3,332,872 Region 15 terpene 3,380,530 3,400,071 Region 16 acyl_amino_acids 3,461,782 3,522,558 Region 17 arylpolyene 4,000,001 4,043,292 APE Vf 40% Region 18 hserlactone 4,071,169 4,091,756 Region 19 fatty_acid 4,170,318 4,194,832 Region 20 saccharide 4,379,674 4,419,034 K53 capsular polysaccharide 15% Region 21 saccharide 4,623,223 4,667,880 Region 22 saccharide 4,981,635 5,002,858 lipopolysaccharide 5% Region 23 terpene 5,097,337 5,120,843 carotenoid 83% Region 24 saccharide 5,142,147 5,198,880 Region 25 saccharide 5,274,999 5,326,287 lipopolysaccharide 17% SoD9b Region Type From To Most similar known cluster Similarity Region 1 fatty_acid,saccharide 202,78 235,29 eicoseicosapentaenoic acid 15% Region 2 fatty_acid 525,312 546,261 Region 3 saccharide 569,733 626,198 lipopolysaccharide 54% Region 4 saccharide 661,028 703,576 Region 5 saccharide 727,413 749,236 Region 6 fatty_acid 1,011,227 1,036,777 Region 7 saccharide 1,138,471 1,168,349 Region 8 bacteriocin 1,190,247 1,198,754 Region 9 bacteriocin 1,497,254 1,508,128 Region 10 saccharide 1,876,199 1,898,314 O-antigen 14% Region 11 halogenated 2,292,133 2,311,229 Region 12 saccharide 2,459,986 2,490,582 pseudopyronine A / 18% Region 13 NRPS 2,503,908 2,550,336 streptobactin 23% Region 14 saccharide 3,644,796 3,664,219 Region 15 saccharide 3,853,356 3,881,455 Region 16 saccharide 3,885,147 3,905,260 Region 17 arylpolyene,halogenat 4,222,695 4,266,297 APE Vf 35% Region 18 saccharide 4,286,303 4,308,080 So63.02b Region Type From To Most similar known cluster Similarity Region 1 saccharide 521,771 553,102 Region 2 saccharide 644,353 690,098 kosinostatin 3% Region 3 saccharide 1,026,342 1,063,007 Region 4 saccharide 1,114,870 1,154,348 Region 5 saccharide 1,488,720 1,563,242 phosphonoglycans 9% Region 6 saccharide 1,926,695 1,951,760 Region 7 fatty_acid 2,209,409 2,232,061 Region 8 saccharide,betalactone 2,448,883 2,512,420 microansamycin 7% Region 9 saccharide 2,527,091 2,550,083 Region 10 saccharide 2,699,537 2,732,407 Region 11 terpene 2,914,074 2,934,982 carotenoid 28% Region 12 saccharide 3,222,694 3,245,310 5 of 9

Region 13 saccharide 3,426,715 3,445,479 Region 14 saccharide 3,473,784 3,494,764 So64.6b Region Type From To Most similar known cluster Similarity Region 1 saccharide 402,957 428,867 Region 2 saccharide 666,331 689,678 Region 3 saccharide 717,381 736,371 Region 4 saccharide 833,072 877,204 glycopeptidolipid 5% Region 5 saccharide 883,546 910,762 Region 6 saccharide 983,304 1,024,979 nogabecin 8% Region 7 lassopeptide,hserlacto 1,448,221 1,470,598 Region 8 fatty_acid 1,532,944 1,561,119 Region 9 saccharide 1,842,988 1,871,436 Region 10 saccharide 2,255,557 2,285,856 O&K-antigen 4% Region 11 halogenated 2,349,570 2,368,140 Region 12 saccharide 2,549,397 2,570,096 lipopolysaccharide 5% Region 13 saccharide 2,722,855 2,745,868 gellan polysaccharide 7% Region 14 saccharide,lassopeptid 3,440,244 3,499,720 Region 15 T3PKS 3,889,391 3,930,452 Region 16 terpene 4,072,179 4,096,869 carotenoid 30% Region 17 fatty_acid 4,115,483 4,136,445 Region 18 saccharide 4,154,895 4,177,593 Region 19 halogenated 4,231,676 4,252,788 Region 20 saccharide 4,417,107 4,453,510 So13.3 Region Type From To Most similar known cluster Similarity Region 1 lanthipeptide 68,939 91,332 SapB 100% Region 2 fatty_acid 181,707 201,626 actinomycin D 10% Region 3 T3PKS 366,627 406,159 A-47934 23% Region 4 NRPS,other 460,304 527,487 actinomycin D 89% Region 5 CDPS,T3PKS 913,307 960,433 alkylresorcinol 100% Region 6 lassopeptide 1,067,644 1,090,272 clavulanic acid 8% Region 7 NRPS 1,403,665 1,453,533 deimino-antipain 66% Region 8 terpene 1,517,889 1,534,894 ebelactone 5% Region 9 fatty_acid,T2PKS,NRP 1,561,713 1,657,640 spore pigment 83% Region 10 saccharide,terpene 1,799,022 1,834,067 pristinol 100% Region 11 NRPS- 1,855,463 1,902,275 marineosin A / marineosin B 86% Region 12 saccharide 2,110,127 2,134,781 Region 13 saccharide 2,279,201 2,299,855 daptomycin 10% Region 14 saccharide 2,382,299 2,405,253 steffimycin D 5% Region 15 ectoine,saccharide 2,924,964 2,969,079 ectoine 100% Region 16 saccharide 3,110,074 3,133,370 Region 17 saccharide 3,256,328 3,306,690 acarviostatin I03 / acarviostatin 29% Region 18 saccharide 3,318,888 3,351,341 glycopeptidolipid 5% Region 19 fatty_acid 3,574,310 3,595,405 Region 20 siderophore 3,643,627 3,657,431 Region 21 saccharide 3,753,455 3,778,710 Region 22 saccharide 4,115,700 4,140,963 Region 23 saccharide 4,296,616 4,316,343 Region 24 saccharide 4,469,278 4,496,601 cathomycin 20% Region 25 saccharide 4,690,571 4,715,758 neocarzinostatin 6% Region 26 saccharide 4,917,327 4,950,799 Region 27 melanin,saccharide 5,141,629 5,170,222 melanin 100% Region 28 halogenated 5,542,357 5,562,337 Region 29 saccharide 5,618,194 5,657,873 TP-1161 41% Region 30 saccharide 5,886,722 5,928,571 napyradiomycin A80915C 9% Region 31 saccharide 6,092,627 6,116,746 Region 32 T1PKS,fatty_acid,NRP 6,242,873 6,377,035 macrotermycins 46% Region 33 saccharide 6,508,813 6,527,699 Region 34 saccharide,other,halog 6,531,244 6,581,567 A-503083 A / A-503083 B / A- 3% Region 35 siderophore 6,943,207 6,957,918 ficellomycin 3% Region 36 NRPS 7,092,437 7,137,408 enduracidin 10% 6 of 9

Region 37 bacteriocin 7,142,716 7,153,177 Region 38 saccharide 7,155,120 7,188,902 ashimides 12% Region 39 saccharide 7,368,836 7,390,778 belactosin A / belactosin C 20% Region 40 terpene 7,486,846 7,510,747 hopene 76% Region 41 NRPS,fatty_acid 7,755,098 7,815,348 myxothiazol 28% Region 42 terpene 7,922,434 7,942,364 nenestatin 3% Region 43 lanthipeptide,terpene 7,942,457 7,984,662 geosmin 100% Region 44 saccharide 8,017,247 8,057,061 Region 45 T1PKS,LAP 8,059,656 8,109,323 maduropeptin 3% Region 46 NRPS,fatty_acid 8,118,137 8,162,078 diisonitrile antibiotic SF2768 61% Region 47 T1PKS,T2PKS,fatty_ac 8,241,505 8,323,840 lysolipin I 65% Region 48 NRPS-like 8,454,616 8,497,189 borrelidin 5% Region 49 bacteriocin 8,508,335 8,520,242 Region 50 melanin 8,793,470 8,803,892 melanin 28% Region 51 NRPS 9,104,379 9,167,226 qinichelins 44% Region 52 saccharide 9,250,594 9,271,013 cosmomycin C 3% Region 53 lanthipeptide 9,293,335 9,315,935 polyoxin A / polyoxin H 5% Region 54 saccharide 9,436,178 9,469,306 acarviostatin I03 / acarviostatin 18%

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Figure S1. HPLC-MS/MS spectra of organic crude extracts from the Antarctic strain Streptomyces fildesensis So13.3 under different culture conditions: (a) IMA medium with no elicitation treatment (basal condition) and (b) IMA medium with coculture elicitation strains So13.3 + So1b. Peak correspondent to putative Actinomycin D is showed and labeled with the intensity of this compound detected for each culture condition.

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Figure S2. Maximum-likelihood phylogenetic tree based on complete 16S rRNA gene sequences showing the genetic distances between Antarctic bacterial strains (highlighted in color and bold) and closely related species based on 16S rRNA reference sequences from DDBJ/ENA/GenBank database. Numbers at nodes represent the bootstrap support (%). Accession number of 16S rRNA gene sequences are shown after the species name.

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Figure S3. Chemical structures predicted for biosynthetic gene cluster similar to Actinomycin D from the Antarctic strain So13.3 genome (Scaffolds 1-3) compared to the structure of the known drug. The predicted structures were based on the core biosynthetic genes using PRISM 4 (http://grid.adapsyn.com/prism/#!/prism).