DNA metabarcoding reveals changes in the contents of carnivorous plants along an elevation gradient

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Supplemental Information for:

DNA metabarcoding reveals changes in the contents of carnivorous plants along an elevational gradient

Littlefair, J.E., Zander, A., de Sena Costa, C., Clare, E.L.

Table S1 Climate variables downloaded from www.worldclim.org/bioclim in March 2018 (Version 2.0)1

1_{Fick, S.E. and R.J. Hijmans, 2017. Worldclim 2: New 1-km spatial resolution climate surfaces for global land areas. International Journal of Climatology.}

Elevation Annual mean temp ᵒC

Max temp of warmest month ᵒC

Min temp of coldest month ᵒC Annual Precipitation (mm) 600m 8.5 23.3 -3.6 1021 800m 7.4 21.6 -4.3 1247 1000m 6 19.3 -5 1296 1200m 5.3 19.1 -6.6 1319 1450m 4.4 17.8 -7.3 1402

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Table S2 Full table of statistical test results

LMM

Model 1 formulation: square-root(plant MOTU richness) ~ elevation + (1|site) AIC: 681.5

Estimate Standard error T value

Intercept 4.47 0.394 11.3

Elevation -0.0006 0.0004 -1.63

Model 2 formulation: square-root(plant MOTU richness) ~ (1|site) AIC: 682.1

Estimate Standard error T value

Intercept 3.85 0.127 30.23

ANOVA

Model formulation: square-root(plant MOTU richness) ~ site

Degrees of freedom Sum of squares Mean squares F value P value

Site 4 13.41 3.35 1.34 0.257

Residuals 175 438 2.50

Partial Mantel test

Model formulation: mantel.partial (xdist= community, ydist= elevation, zdist=geographic, method=”pearson”, permutations=999) Mantel statistic r: 0.0589

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Table S3 Full list of taxa recovered by BLAST search against a custom curated reference database

Species match at 99%

and 100% similarity Order

Known

distribution Pitcher presence

Acanthiophilus helianthi Diptera Europe FX8_B128

Adineta gracilis Adinetida

Europe and

further FX22_E142 FX24_E144 FX6_B126 LM1

Adineta vaga Adinetida

Europe and

further CB1_G182 CB1_I194 CB2_G183 CB2_H189 CB3_G184 CB3_I196 CB4_F179

CB4_G185 CB4_I197 CB5_F180 CB6_F181 CB6_G187 CB6_I199 CBF177

CBI195 LE1090 LM20 LM31

Aedes cinereus Diptera

Europe and

further CB1_B152 CB1_H188 CB2_B153 CB3_B154 CB3_L172 CB4_B155 CB4_C161

CB4_I197 LE24_104 LM19 LM33

Aedes sp. Diptera Global CB1_B152 CB1_H188 CB2_B153 CB3_B154 CB3_L172 CB4_B155 CB4_C161

CB4_I197 LE24_104 LM19 LM33

Agonum muelleri Coleoptera Europe CB5_CA150

Alcis repandata Lepidoptera Europe LTC54

Aphis fabae Hemiptera

Europe and

further LM17 LTC51

Aphis hederae Hemiptera

Europe and

further LM17 LTC51

Athalia sp. Hymenoptera

Europe and

further CB4_F179

Atolmis rubricollis Lepidoptera Europe CBI198

Aulacorthum solani Hemiptera

Europe and

further LTE62 LTE64

Biston strataria Lepidoptera Europe CB2_E171

Biston stratarius Lepidoptera Europe CB2_E171

Calliphora sp. Diptera Global CBI195

(4)

4

LE21101 LE484 LM11 LM2 LM28 LM32 LTE61

Calliphora vomitoria Diptera

Europe and

further LM32 LM39 LTI70

Calyptratae sp. Diptera Global CB4_H191 LM2

Cantharis livida Coleoptera Europe CB5_H192

Carychium minimum Eupolmonata Europe CB5_CA150

Catops coracinus Lepidoptera Europe LE38118

Catops tristis Lepidoptera Europe LE28_109 LE38118

Ceratopogonidae sp. Ceratopogonidae Global CB4_C161

Craspedosoma rawlinsii

Craspedosomatida

e Europe CB2_E171 CB2_G183 LM33

Cryphia algae Lepidoptera Europe CB3_CA148 FX23_E143

Culex apicalis Diptera

Europe and

further CB1_B152

Culex pipiens Diptera

Europe and

further CB1_H188 CB2_E171 CB5_F180 CB6_G187 LE23_103 LE31_111

Culex territans Diptera

Europe and

further CB1_B152

Culex torrentium Diptera Europe CB6_G187 LT671

Cyclophora linearia Lepidoptera Europe CB4_CA149

Cynomya mortuorum Diptera

Europe and

further CB2_C159 LE1595 LE28_109 LE32_112

Delia sp. Diptera Europe CB1_A146 CB1_G182 CB2_C159 CB2_E171 CB2_G183 CB2_H189 CB3_H190

CB3_L172 CB4_G185 CB4_H191 CB5_CA150 CB5_H192 FX24_E144 FX4_A124

FX5_A125 FX6_B126 FX7_B127 LE21101 LE27_107 LE40120 LM1

LM19 LTC54 LTD57 LTE61

Diptera sp. Diptera Europe CB1_A146 CB1_E170 CB2_E171 CB3_L172 CB4_C161 CB4_I197 CBC162

CBI198 FX22_E142 FX4_A124 FX6_B126 FX7_B127 LE1191 LE1292

LE1393 LE21101 LE22_108 LE23_103 LE26_106 LE28_109 LE31_111

(5)

5

LM24 LM34 LM35 LM36 LM39 LT671 LTA43

LTB47 LTD56 LTE61 LTE62 LTE64 LTG74 LTH77

Drepanosiphum

platanoidis Hemiptera Europe CB1_H188

Drosophila immigrans Diptera Europe LE787

Drosophila suzukii Diptera Europe CB1_C158 CB2_C159 CB2_E171 CB4_H191 CBC162 CBD168 Fr3a123

FX20_D140 LE1292 LE21101 LE23_103 LE25_105 LE28_109 LE30_110

LE31_111 LE33113 LE787 LE989 LM10 LM28 LM3

LM36 LM37 LM8 LT671 LTB48 LTC54 LTD57

LTH77 LTI67 LTI70

Drosophilidae sp. Diptera Europe CB2_E171 CB4_I197 FX7_B127 LE1292 LE22_108 LE23_103 LE28_109

LE31_111 LE38118 LM23 LM24 LM36 LTB47 LTE62

LTE64

Drymeia hamata Diptera Europe CBD165

Eilema depressa Lepidoptera Europe LE24_104

Eilema griseolum Lepidoptera

Europe and

further CB4_CA149

Eilema lurideola Lepidoptera Europe CB2_B153

Eilema lutarellum Lepidoptera

Europe and

further CB2_B153

Elachistidae gen. Lepidoptera Global CB2_H189

Ematurga atomaria Lepidoptera

Europe and

further FX24_E144 LTB46

Euclidia glyphica Lepidoptera

Europe and

further CB1_A146

Eutrichota sp. Diptera

Europe and

further LM33

Fannia canicularis Diptera

Europe and

further CB4_H191 LE21101 LM2

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6

Helina impuncta Diptera

Europe and

further CB2_G183 LE1494 LE21101 LE484 LM11 LM28 LM32

LM33 LTA42 LTB48 LTC55 LTD56 LTE61

Hyalopterus pruni Hemiptera

Europe and

further CBF177

Ichneumonidae sp. Hymenoptera Global LE23_103 LM1

Idaea biselata Lepidoptera Europe CB1_C158

Lasiocampa quercus Lepidoptera Europe LTB46

Lepidoptera sp. Lepidoptera Global CB2_B153 CB2_H189 CB4_C161

Leskia sp. Diptera Global FX6_B126 LE38118 LM16 LTE61

Leucania sp. Lepidoptera Global CB2_B153

Limax cinereoniger Stylommatophora Europe LTA43

Limnophyes edwardsi Diptera Europe LTH77

Limonia sp. Diptera Europe and further LE21101 Limoniidae sp. Diptera Europe and further LE26_106

Lucilia ampullacea Diptera Europe CB1_E170 CB3_I196 CB4_I197 CB6_I199 CBI198 LE1191 LE1292

Lucilia bufonivora Diptera Europe FX25_E145 LE23_103

Lucilia caesar Diptera Europe CB1_E170 CB2_CA147 CB3_L172 CB6_I199 LE1191 LE25_105 LM11

LTA41 LTA43 LTD56 LTE61

Lucilia cf. Diptera Europe CB1_E170 CB2_CA147 CB3_L172 CB6_I199 LE25_105 LM11 LTA41

LTA43 LTD56 LTE61

Lucilia cupri Diptera Europe LE888

Lucilia illustris Diptera Europe CB1_E170 CB2_CA147 CB3_L172 CB6_I199 LE1191 LE25_105 LM11

LTA41 LTA43 LTD56 LTE61

Lucilia richardsi Diptera Europe CB4_I197 CBD165 CBD168 CBI195 LE888 LM2 LM32

LTD58

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7

Lucilia silvarum Diptera Europe CB4_I197 CBD165 CBD168 CBI195 LE888 LM2 LM32

LTD58 Megaselia giraudii Diptera

Europe and

further LE1393 LE27_107 LE32_112 LM1 LM23 LM24 LTE62

LTG74 LTH77

Megaselia rufipes Diptera

Europe and

further LE32_112 LTG74

Megaselia subtumida Diptera Europe LE32_112 LTE62

Melanostoma cf. Diptera

Europe and

further CBC162 FX22_E142 FX4_A124 LE1191 LE26_106 LE484 LM1

LM11 LM16 LM35 LM39 LT671 LTD56 LTH77

Melanostoma mellinum Diptera

Europe and

further CBC162 FX22_E142 LE1191 LE484 LM1 LM11 LM34

LT671 LTD56

Mesembri meridiana Diptera Europe CBD167 Fr17d137 Fr3a123 LE1292 LE25_105 LE28_109 LM32

LM6 LTC54 LTD56 LTI70

Mesoligia furuncula Diptera Europe FX23_E143

Metriocnemus

albolineatus Diptera

Europe and

further CB3_I196

Micropsectra recurvata Diptera Europe LE22102 LE484

Morellia aenescens Diptera Europe LE1898

Musca autumlis Diptera Europe CB1_H188 CB4_H191 CBD165 Fr3a123 LE22102 LE28_109 LM11

LM19 LM32 LM35 LT675

Musca domestica Diptera Europe CBC162 CBD165 CBD167 CBI198 Fr3a123 LE1494 LE1898

LE21101 LE484 LM11 LM2 LM28 LM32 LTE61

Muscidae sp. Diptera Global CB1_A146 CB1_E170 CB3_L172 LE21101

Muscina levida Diptera Europe CB4_G185 CB6_E175 CBD165 CBD168 Fr17d137 FX22_E142 FX23_E143

FX25_E145 FX8_B128 LE1292 LE1494 LE1898 LE22_108 LE23_103

LE25_105 LE26_106 LE28_109 LE484 LM10 LM34 LTA41

(8)

8

Myospila meditabunda Diptera Europe CBD167 LE21101 LTD56

Mythimna albipuncta Diptera Europe CB2_C159 CB3_G184 CB6_E175 LM25

Mythimna conigera Diptera Europe LTE61

Mythimna impura Diptera Europe CBF177 LE32_112 LM20

Mythimna pallens Diptera Europe CB2_B153

Noctua comes Lepidoptera Europe CB2_H189 CB5_H192

Noctua pronuba Lepidoptera Europe CB2_H189

Noctua sp. Lepidoptera Europe CB2_H189 CB3_G184

Noctuidae sp. Lepidoptera Europe CB2_B153

Ochlerotatus cataphylla Diptera Europe CB1_C158

Ochropleura plecta Lepidoptera Europe CB3_G184

Pardosa prativaga Araneae Europe LTD58

Pegomya winthemi Diptera

Europe and

further CBI195

Pegoplata infirma Diptera

Europe and

further LM10

Pelosia muscerda Lepidoptera Europe LE24_104 LM34

Phaonia subventa Diptera

Europe and

further LE1595 LE22_108 LM35

Phaonia tuguriorum Diptera

Europe and

further CB1_H188 CB4_C161 CB4_H191 Fr17d137 FX6_B126 FX8_B128 LE21101

LE31_111 LE32_112 LM1 LM39 LTA41 LTB49

Phiaris bipunctana Lepidoptera

Europe and

further LTI67

Phoridae sp. Diptera Global CB2_C159 CBI198 FX1_A121 FX22_E142 FX6_B126 FX7_B127 LM35

LTD58 LTG74

Phytomyza ranunculi Diptera

Europe and

further CB4_C161

Platycheirus sp. Diptera Global CBC162 FX22_E142 LE1191 LE484 LM1 LM11 LM34

(9)

9

Plodia cf. Lepidoptera Global LE34114 LTD60 LTE63 LTG72

Plodia interpunctella Lepidoptera Global LE34114 LTD60 LTE63 LTG72

Polietes lardarius Diptera Europe CB2_B153 CB3_C160 CB4_CA149 CB5_H192 CBC162 CBD165 CBD167

CBF177 CBI195 CBI198 Fr13c133 Fr17d137 Fr3a123 LE1090

LE1191 LE1494 LE1595 LE1898 LE1999 LE21101 LE22_108

LE23_103 LE24_104 LE25_105 LE28_109 LE30_110 LE31_111 LE32_112

LE33113 LE484 LE787 LE989 LM1 LM11 LM16

LM23 LM3 LM32 LM35 LM36 LM37 LM39

LM6 LM8 LTA41 LTA43 LTB47 LTB48 LTB49

LTC52 LTC54 LTC55 LTD56 LTD57 LTE61 LTE64

LTG74 LTH77 LTH79 LTI67 LTI70

Polistes dominula Hymenoptera Europe CB2_G183

Polistes gallicus Hymenoptera

Europe and

further CB2_G183

Pollenia rudis Diptera

Europe and

further CB1_A146 CB4_F179 LE31_111 LM6

Propylea

quatuordecimpunctata Diptera Global CB5_F180 LM34

Pseudococcyx mughiana Lepidoptera Europe LTI67

Psocoptera sp. Psocoptera Global LM35

Rhagonycha fulva Coleoptera Europe CB4_C161 CB4_F179

Sarcophaga africa Diptera Europe CB2_G183 CB6_G187 FX22_E142 FX24_E144 FX4_A124 FX6_B126 LE20100

LM34 LTB46

Sarcophaga aratrix Diptera Europe CB6_G187 FX21_E141 FX22_E142 FX24_E144 FX25_E145 FX4_A124 FX6_B126

FX8_B128 LM1 LM34 LTB46

Sarcophaga caerulescens Diptera Europe LM34 LTB46

Sarcophaga carnaria Diptera Global CB1_A146 CB1_B152 CB1_C158 CB1_G182 CB1_H188 CB2_C159 CB2_CA147

CB2_E171 CB2_G183 CB3_C160 CB3_CA148 CB3_G184 CB3_L172 CB4_B155

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10

CBD165 FX1_A121 FX22_E142 FX24_E144 FX4_A124 FX5_A125 FX6_B126

LE1898 LE20100 LE21101 LE22102 LE27_107 LE32_112 LM1

LM10 LM19 LM33 LM34 LM37 LM39 LM6

LTA41 LTA43 LTB46 LTB47 LTG74

Sarcophaga dissimilis Diptera Europe LM28

Sarcophaga granulata Diptera Europe CB2_C159 CB2_CA147 CB3_G184 CB3_L172 CB4_G185 FX21_E141 FX22_E142

FX4_A124 LE20100 LE22102 LM19 LM34 LTA41 LTA43

LTB46 LTB47 LTG74

Sarcophaga incisilobata Diptera Europe CB2_B153 LE20100 LTB46

Sarcophaga jeanleclercqi Diptera Europe CB1_B152 CB1_C158 CB1_H188 CB2_C159 CB2_E171 CB2_G183 CB3_CA148

CB3_G184 CB4_B155 CB4_H191 CB5_H192 CB6_G187 CBD165 FX1_A121

FX22_E142 FX24_E144 FX4_A124 FX5_A125 FX6_B126 LE1898 LE20100

LE21101 LE22102 LE32_112 LM1 LM10 LM19 LM33

LM34 LM37 LM39 LTA41 LTA43 LTB46 LTB47

LTG74

Sarcophaga lehmanni Diptera Europe CB1_A146 CB1_B152 CB1_C158 CB1_G182 CB1_H188 CB2_C159 CB2_CA147

CB2_E171 CB2_G183 CB3_B154 CB3_C160 CB3_CA148 CB3_G184 CB3_H190

CB3_L172 CB4_B155 CB4_C161 CB4_G185 CB4_H191 CB4_I197 CB5_CA150

CB5_H192 CB6_E175 CB6_G187 CB6_I199 CBD165 FX1_A121 FX22_E142

FX24_E144 FX4_A124 FX5_A125 FX6_B126 LE1090 LE1191 LE1898

LE20100 LE21101 LE22102 LE27_107 LE32_112 LM1 LM10

LM19 LM31 LM32 LM34 LM39 LM6 LTA41

LTA43 LTB46 LTB47 LTG74

Sarcophaga noverca Diptera Europe CBD165 FX21_E141 FX6_B126 LM34

Sarcophaga okaliana Diptera Europe LE20100 LTB46

Sarcophaga pumila Diptera Europe FX21_E141

Sarcophaga pyrenaica Diptera Europe CB1_B152 CB1_C158 CB1_H188 CB2_C159 CB2_E171 CB2_G183 CB3_CA148

(11)

11

FX22_E142 FX24_E144 FX4_A124 FX5_A125 FX6_B126 LE1898 LE20100

LE21101 LE22102 LE32_112 LM1 LM10 LM19 LM33

LM34 LM37 LM39 LTA41 LTA43 LTB46 LTB47

LTG74

Sarcophaga rosellei Diptera Europe CBD165 LE20100 LM34 LTB46

Sarcophaga sexpunctata Diptera Europe CBD165

Sarcophaga similis Diptera Europe CB6_G187 FX21_E141 FX22_E142 FX24_E144 FX25_E145 FX4_A124 FX6_B126

FX8_B128 LM1 LTB46

Sarcophaga subvicina Diptera

Europe and

further CB1_A146 CB1_B152 CB1_C158 CB1_G182 CB1_H188 CB2_C159 CB2_CA147

CB2_E171 CB2_G183 CB3_B154 CB3_C160 CB3_CA148 CB3_G184 CB3_L172

CB4_B155 CB4_C161 CB4_G185 CB4_H191 CB5_CA150 CB5_H192 CB6_G187

CB6_I199 CBD165 FX1_A121 FX21_E141 FX22_E142 FX24_E144 FX4_A124

FX5_A125 FX6_B126 LE1898 LE20100 LE21101 LE22102 LE27_107

LE32_112 LM1 LM10 LM19 LM33 LM34 LM37

LM39 LM6 LTA41 LTA43 LTB46 LTB47 LTG74

Sarcophaga vagans Diptera Europe FX22_E142 FX6_B126 LE787 LM1 LM34 LM39 LM6

LTD56 LTI70

Sarcophaga variegata Diptera Europe CB1_A146 CB1_B152 CB1_C158 CB1_G182 CB1_H188 CB2_C159 CB2_CA147

CB2_E171 CB2_G183 CB3_B154 CB3_C160 CB3_CA148 CB3_G184 CB3_H190

CB3_L172 CB4_B155 CB4_C161 CB4_G185 CB4_H191 CB4_I197 CB5_CA150

CB5_H192 CB6_E175 CB6_G187 CB6_I199 CBD165 FX1_A121 FX22_E142

FX24_E144 FX4_A124 FX5_A125 FX6_B126 LE1090 LE1191 LE1898

LE20100 LE21101 LE22102 LE27_107 LE32_112 LM1 LM10

LM19 LM31 LM32 LM34 LM39 LM6 LTA41

LTA43 LTB46 LTB47 LTG74

Sarcophaga vicina Diptera Europe CB1_C158

Sarcophagidae sp. Diptera Global CB2_G183 CB6_G187 CBD165 FX22_E142 FX24_E144 FX4_A124 FX6_B126

(12)

12

Saturnia pavonia Lepidoptera Europe CBF177 LM20

Scaptomyza flava Diptera Europe LTC55

Scathophaga cf. Diptera

Europe and

further CB2_G183 CB4_H191 FX20_D140 FX23_E143 FX4_A124 FX6_B126 FX7_B127

LE1292 LE23_103 LE25_105 LE30_110 LE32_112

Scathophaga stercoraria Diptera Europe CB2_G183 CB4_H191 FX20_D140 FX23_E143 FX4_A124 FX6_B126 FX7_B127

LE1292 LE23_103 LE25_105 LE30_110 LE32_112

Sepsis cynipsea Diptera Europe CB4_I197 FX20_D140 FX23_E143 FX6_B126 FX7_B127 FX8_B128 LE1595

LE1898 LE1999 LE21101 LE787 LM2 LM28 LM37

LT671 LTD57 LTG74

Sepsis flavimana Diptera Europe LE40120 LM33 LT671 LTD56 LTI66

Sepsis fulgens Diptera Europe CB3_I196 CB4_I197 CBI198 FX7_B127 LM3 LM36

Sepsis neocynipsea Diptera

Europe and

further CB4_I197 FX20_D140 FX23_E143 FX6_B126 FX7_B127 FX8_B128 LE1595

LE1898 LE1999 LE21101 LE787 LM2 LM28 LM37

LT671 LTD57 LTG74

Sepsis orthocnemis Diptera Europe CB3_I196 CB4_I197 CBI198 FX7_B127 LM36

Sepsis violacea Diptera Europe CB4_I197 CBI198 FX7_B127 LM3

Setema cereola Lepidoptera Europe CB2_B153

Simulium crenobium Diptera Europe CBD168 LE484

Simulium vernum Diptera Europe CBD168 LE484

Sphaerophoria scripta Diptera

Europe and

further LE1191 LM39

Sphaerophoria sp. Diptera

Europe and

further LM39

Syrphus vitripennis Diptera

Europe and

further CB3_I196 LE32_112 LM32

Tachinidae gen. Diptera Global CBD168 LM13

Tenuiphantes tenuis Araneae Global CB2_G183

(13)

13

Thricops genarum Diptera Europe CBC162 LE21101 LM28 LM37 LM39

Tortricidae sp. Lepidoptera Global CB1_B152 CBD165 CBD168 LE1898 LE21101 LE22_108 LE27_107

LM11 LM28 LTC55

Triphleba nudipalpis Diptera Europe CB1_H188

Craspedosoma

taurinorum Chordeumatida Europe CB2_E171 CB2_G183 LM33

Promethes sulcator Hymenoptera

Europe and

further CB5_CA150

Sussaba aciculata Hymenoptera Europe LE23_103 LM1

Xestia cnigrum Lepidoptera

Europe and

further CB1_C158 CB1_G182