Supplementary Airborne Bacterial and Eukaryotic Community Structure Across the United Kingdom Revealed by High-Throughput Sequencing

Hokyung Song, Ian Crawford, Jonathan R. Lloyd *, Clare H. Robinson, Christopher Boothman, Keith Bower, Martin Gallagher *, Grant Allen and David Topping

Department of Earth and Environmental Sciences, The University of Manchester, Manchester M13 9PL, UK; [email protected] (H.S.); [email protected] (I.C.); [email protected] (C.H.R.); [email protected] (C.B.); [email protected] (K.B.); [email protected] (G.A.); [email protected] (D.T.) * Correspondence: [email protected] (J.R.L.); [email protected] (M.G.)

Supplementary Figures

Figure S1. 96 hour HYSPLIT Lagrangian back trajectories of air sampled at: (a) Chilbolton at 950 m, (b) Chilbolton at 380 m, and (c) Weybourne at 724 m.

Figure S2. Canonical correspondence analysis result plot. Each of CH10, CH20, CH30 indicates samples collected in Chilbolton at high elevation (range of 950-1000 m asl) in the time duration of 10 min, 20 min, and 30 min. Each of W10, W20, W30 indicates samples collected in Weybourne (height range of 650-1000 m) in the time duration of 10 min, 20 min, and 30 min.

Figure S3. Number of bacterial OTUs. Each of CH10, CH20, CH30 indicates samples collected in Chilbolton at high elevation (range of 950-1000 m asl) in the time duration of 10 min, 20 min, and 30 min. Each of W10, W20, W30 indicates samples collected in Weybourne (height range of 650-1000 m) in the time duration of 10 min, 20 min, and 30 min.

Figure S4. Redundancy analysis result plot. Each of CH10, CH30 indicates samples collected in Chilbolton at high elevation (range of 950-1000 m asl) in the Table 10. min and 30 min. Each of CL10, CL20, CL30 indicates samples collected in Chilbolton at low elevation (range of 300-450 m asl) in the time duration of 10 min, 20 min, and 30 min. Each of W10, W20, W30 indicates samples collected in Weybourne (height range of 650-1000 m) in the time duration of 10 min, 20 min, and 30 min.

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Figure S5. Number of Eukaryotic OTUs. Each of CH10, CH30 indicates samples collected in Chilbolton at high elevation (range of 950–1000 m asl) in the time duration of 10 min and 30 min. Each of CL10, CL20, CL30 indicates samples collected in Chilbolton at low elevation (range of 300–450 m asl) in the time duration of 10 min, 20 min, and 30 min. Each of W10, W20, W30 indicates samples collected in Weybourne (height range of 650–1000 m) in the time duration of 10 min, 20 min, and 30 min. . Supplementary tables.

Table S1. Detailed information of collected filter samples. “lat” stands for latitude, “lon” stands for longitude, and “alt” stands for altitude. Relative humidity was calculated using the Arden Buck equation (Buck 1981).

Average Start End Start End Average Start End Relative Filter ID Run Location Latitude/Longitude Latitude/Longitude Altitude Altitude Temperature Time Time Humidity (°) (°) (km) (km) (℃) (%) leg 1 Chilbolton 11:26:30 11:37:40 50.7962/−2.6726 51.0184/−1.7314 0.436 0.354 12.995 48.508 Chilbolton low 30 min leg 2 Chilbolton 11:44:38 11:54:37 51.1384/−1.4495 50.8815/−2.3144 0.378 0.406 13.817 46.741 leg 3 Chilbolton 11:57:11 12:08:01 50.8672/−2.3676 51.1309/−1.4658 0.385 0.320 14.089 45.837 Chilbolton low 10 min leg 1 Chilbolton 11:26:30 11:37:40 50.7962/−2.6726 51.0184/−1.7314 0.436 0.354 12.995 48.508 leg 1 Chilbolton 11:44:38 11:54:37 51.1384/−1.4495 50.8815/−2.3144 0.378 0.406 13.817 46.741 Chilbolton low 20 min leg 2 Chilbolton 11:57:11 12:08:01 50.8672/−2.3676 51.1309/−1.4658 0.385 0.320 14.089 45.837 leg 1 Chilbolton 12:14:15 12:24:19 51.0898/−1.5581 50.8384/−2.4859 0.975 0.974 8.196 62.864 Chilbolton high 30 min leg 2 Chilbolton 12:28:47 12:39:36 50.7926/−2.3218 51.1414/−1.4436 0.979 0.976 8.374 61.906 leg 3 Chilbolton 12:44:10 12:54:06 51.1324/−1.4567 50.9039/−2.3721 0.981 0.963 8.620 60.722 Chilbolton high 10 min leg 1 Chilbolton 12:14:15 12:24:19 51.0898/−1.5581 50.8384/−2.4859 0.975 0.974 8.196 62.864 leg 1 Chilbolton 12:28:47 12:39:36 50.7926/−2.3218 51.1414/−1.4436 0.979 0.976 8.374 61.906 Chilbolton high 20 min leg 2 Chilbolton 12:44:10 12:54:06 51.1324/−1.4567 50.9039/−2.3721 0.981 0.963 8.620 60.722 leg 1 Weybourne 13:36:05 13:46:18 52.9406/0.55184 52.8147/1.5293 0.973 0.677 7.519 37.590 Weybourne 30 min leg 2 Weybourne 13:50:10 13:59:16 52.8763/1.3028 52.9563/0.36313 0.670 0.673 8.515 56.670 leg 3 Weybourne 14:02:10 14:13:10 52.9452/0.4676 52.8768/1.5355 0.679 0.670 8.181 55.923 Weybourne 10 min leg 1 Weybourne 13:36:05 13:46:18 52.9406/0.55184 52.8147/1.5293 0.973 0.677 7.519 37.590 leg 1 Weybourne 13:50:10 13:59:16 52.8763/1.3028 52.9563/0.36313 0.670 0.673 8.515 56.670 Weybourne 20 min leg 2 Weybourne 14:02:10 14:30:10 52.9452/0.4676 53.1219/0.8797 0.679 2.593 7.596 56.309

Table S2. Blast result of the 20 most abundant bacterial OTUs. Among 100 best hits, only the hit results with highest similarity, highest score, and lowest e-value are shown. If there are multiple best hit sequences with the same similarity, e-value, and hit score, they are shown together.

OTU ID Genus (Silva Database) Blast (NCBI NT Database) Similarity E−value Hit Score OTU008 Gordonia Gordonia iterans 100 1.96E-127 466 OTU009 Psychroglaciecola uncultured bacterium OTU010 Burkholderiaceae_unclassified Hydrogenophaga soli 99.206 4.24E-124 455 OTU011 Lautropia uncultured bacterium OTU013 Morganella Morganella morganii, Providencia rustigianii, Salmonella enterica, Escherichia coli 100 1.96E-127 466 OTU014 Candidatus_Udaeobacter uncultured bacterium OTU015 Fusobacterium Fusobacterium nucleatum 100 7.01E-127 464 Janibacter indicus, , Janibacter terrae, Knoellia locipacati, Janibacter anophelis, OTU016 Intrasporangiaceae_unclassified Janibacter melonis, Janibacter limosus, Knoellia sinensis, Janibacter massiliensis, Knoellia 100 1.96E-127 466 subterranea OTU017 Chryseobacterium Chryseobacterium hominis 100 1.96E-127 466 OTU018 Sphingomonas uncultured bacterium OTU019 Pajaroellobacter uncultured bacterium Streptococcus oralis, Streptococcus mitis, Streptococcus infantis, Streptococcus pyogenes, OTU020 Streptococcus 100 7.01E-127 464 Streptococcus australis, Streptococcus sanguinis, Streptococcus pneumoniae OTU021 Nakamurella uncultured bacterium Arthrobacter davidanieli, Arthrobacter russicus, Psychromicrobium silvestre, Psychromicrobium OTU022 Micrococcaceae_unclassified 100 1.96E-127 466 lacuslunae OTU023 Actinomyces Actinomyces oris, Actinomyces naeslundii, Actinomyces viscosus 100 1.96E-127 466 OTU024 Rubellimicrobium uncultured bacterium Chryseobacterium bernardetii, Bergeyella porcorum, Chryseobacterium taichungense, OTU026 uncultured 95.635 4.33E-109 405 Chryseobacterium moechotypicola OTU028 Burkholderiaceae_unclassified Tepidimonas taiwanensis 99.6 1.18E-124 457 OTU029 Gemella Gemella haemolysans, Gemella morbillorum 100 1.96E-127 466 OTU030 Devosia uncultured bacterium

Table S3. Blast result of the 20 most abundant eukaryotic OTUs. Among 100 best hits, only the hit results with highest similarity, highest score, and lowest e-value are shown. If there are multiple best hit sequences with the same similarity, e-value, and hit score, they are shown together.

OTU ID Genus Blast Result Similarity E-value Hit Score OTU0002 Embryophyta unclassified Pinus luchuensis 100 4.81E-50 207 Cladosporium tenuissimum, Cladosporium halotolerans, Cladosporium xylophilum, Cladosporium oxysporum, Cladosporium exasperatum, Cladosporium herbarum, Panicum hallii, Cladosporium anthropophilum, Selaginella moellendorffii, Cladosporium xanthochromaticum, Cladosporium velox, Cladosporium tenellum, Cladosporium subinflatum, Cladosporium sphaerospermum, OTU0001 Cladosporium Cladosporium ramotenellum, Cladosporium pulvericola, Cladosporium pseudocladosporioides, 100 6.07E-49 204 Cladosporium perangustum, Cladosporium parasubtilissimum, Cladosporium parahalotolerans, Cladosporium neolangeronii, Cladosporium needhamense, Cladosporium lycoperdinum, Cladosporium limoniforme, Cladosporium langeronii, Cladosporium inversicolor, Cladosporium funiculosum, Cladosporium floccosum, Cladosporium dominicanum OTU0010 Phoma Epicoccum sorghinum (also known as Phoma sorghina), Epicoccum nigrum, Curvularia kusanoi 100 6.07E-49 204 OTU0012 Chlamydomonadal unclassified Chloromonas subdivisa 100 7.65E-48 200 Vigna unguiculata, Brassica oleracea, Brassica rapa, Eutrema salsugineum, Brassica carinata, Brassica napus, Brassica juncea, Raphanus sativus, Raphanus raphanistrum, Brassica oleracea, OTU0008 Brassicales unclassified 100 4.81E-50 207 Brassica nigra, Arabis alpina, Thlaspi arvense, Nasturtium officinale, Matthiola longipetala, Sisymbrium orientale, Isatis tinctoria, Draba nemorosa, Brassica tournefortii Moesziomyces antarcticus, Ustilago maydis, Sporisorium reilianum, Pseudozyma hubeiensis, OTU0009 Ustilaginaceae unclassified Melanopsichium pennsylvanicum, Ustilago williamsii, Ustilago striiformis, Ustilago cynodontis, 100 1.71E-49 206 Tranzscheliella hypodytes, Macalpinomyces bursus, Melanopsichium pennsylvanicum OTU0016 Pleosporales unclassified Torula herbarum 100 6.07E-49 204 OTU0027 Blumeria Blumeria graminis 99.091 2.82E-47 198 OTU0032 Pleosporales unclassified Phaeosphaeria graminis, Neocucurbitaria acanthocladae, Alternaria alternariae 100 6.07E-49 204 OTU0015 Hemiptera Aulacorthum solani 99.194 5.43E-55 224 OTU0014 Ranunculales unclassified Ranunculus japonicus 100 4.81E-50 207 OTU0029 Blumeria Blumeria graminis 100 6.07E-49 204 OTU0025 Sclerotinia Sclerotinia sclerotiorum, Botrytis cinerea, Sclerotinia trifoliorum 100 6.07E-49 204 Dothidea berberidis, Dothidea muelleri, Dothidea sambuci, Sydowia polyspora, Delphinella OTU0017 Delphinella 100 6.07E-49 204 strobiligena, Dothidea ribesia, Dothidea insculpta, Dothidea hippophaeos OTU0020 Chlorophyta unclassified Apatococcus lobatus 100 6.07E-49 204 Gossypium turneri, Klebsiella pneumoniae, Gossypium raimondii, Gossypioides kirkii, Passiflora cincinnata, Gossypium arboreum, Gossypium hirsutum, Bacopa monnieri, Polygala alba, Ceanothus herbaceus, Plantago patagonica, Veronica americana, Cucumis melo, Lagenaria siceraria, Hibiscus sabdariffa, Calceolaria integrifolia, Plocosperma buxifolium, Morina longifolia, Desfontainia spinosa, Columellia oblonga, Veronica undulata, Elaeagnus oldhamii, Plantago asiatica, Hibiscus OTU0018 Embryophyta unclassified acetosella, Hibiscus surattensis, Hibiscus cannabinus, Hibiscus radiatus, Ileostylus micranthus, 100 4.81E-50 207 Corallocarpus bainesii, Octomeles sumatrana, Ecballium elaterium, Lagenaria breviflora, Schizopepon bryoniifolius, Seyrigia humbertii, Marah macrocarpa, Gurania tubulosa, Ochroma pyramidale, Grewia occidentalis, Proboscidea louisiana, Phyla lanceolata, Euthystachys abbreviata, Nematanthus hirsutus, Buddleja davidii, Datisca glomerata, Tetrameles nudiflora, Abobra tenuifolia Piofontia schultzii, Piofontia heterophylla, Diplostephium foliosissimum, Piofontia jenesana, Piofontia obtusa, Diplostephium oblanceolatum, Piofontia tenuifolia, Piofontia antioquensis, Piofontia phylicoidea, Diplostephium gynoxyoides, Diplostephium barclayanum, Diplostephium goodspeedii, Piofontia mutiscuana, Diplostephium sagasteguii, Piofontia camargoana, Exostigma notobellidiastrum, Diplostephium jelskii, Laennecia sophiifolia, Piofontia floribunda, Diplostephium hartwegii, Piofontia tachirensis, Piofontia revoluta, Piofontia violacea, Diplostephium cinereum, Piofontia colombiana, Piofontia rosmarinifolia, Westoniella kohkemperi, Diplostephium glandulosum, Diplostephium espinosae,Baccharis genistelloides, Blakiella bartsiifolia, Diplostephium juniperinum, Piofontia costaricensis, Piofontia apiculata, Piofontia eriophora, OTU0022 Diplostephium 100 4.81E-50 207 Piofontia oblongifolia, Diplostephium callilepis, Piofontia rhododendroides, Diplostephium pulchrum, Piofontia alveolata, Aztecaster matudae, Piofontia rupestris, Diplostephium oxapampanum, Piofontia cinerascens, Hinterhubera ericoides, Piofontia rhomboidalis, Diplostephium empetrifolium, Piofontia coriacea, Piofontia juajibioyi, Piofontia huertasii, Diplostephium crypteriophyllum, Piofontia lacunosa, Diplostephium cajamarquillense, Piofontia inesiana, Diplostephium gnidioides, Floscaldasia hypsophila, Baccharis tricuneata, Diplostephium lechleri, Piofontia venezuelensis, Archibaccharis asperifolia, Piofontia romeroi, Diplostephium azureum, Olearia lyrata, Heterotheca villosa, Heterotheca stenophylla, Gutierrezia sarothrae, Xanthisma spinulosum, Grindelia squarrosa OTU0044 Itersonilia Itersonilia perplexans, Tausonia pullulans 100 6.07E-49 204 OTU0033 Embryophyta unclassified Cucurbita pepo, Cucurbita maxima, Cucurbita moschata 100 4.81E-50 207 OTU0021 Tetracladium Articulospora proliferata 99.091 2.82E-47 198