Posted on Authorea 14 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161065834.45838723/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. htas om h liefrs ie nms ra nti ein(rn&Pthff 08.A h forest- Norway, the In At 1b). 2018). (Fig. Potthoff, vegetation vegetation & low-alpine to as (Bryn shifting to region before referred zone, this commonly transition in a species, areas in plant dominated most ericaceous shrub by in normally dominated is lines ( vegetation forest birch the alpine ecotone, mountain alpine the by 2017). forms covered Tallaksen, temperature also & are local Fennoscandia that Bryn, the increase Stordal, in probably (Rydsaa, areas will Thomas, further lines Large forest expansion & alpine shift Roy, ongoing Wit of Ohlemuller, (de upward the elevation lines Hill, accelerate the an forest in therefore Chen, alpine including increase and 2008; expanding an predicted, from and al., feedbacks are 2014) et climate al., vegetation (Beckage positive et regions documented ecosystems in (K¨orner, have mountain 2012). mountain driven changes studies many into Recent temperature warming, 2011). mainly Although line climate are forest to lines 2009). the forest response of Duncan, alpine elevations a uppermost & higher most in as towards the alpine McGlone, varies moved the use, Hence, treeline, regions to Hulme, land of or many by forests, (Harsch, in one (here) influenced lowland has line decades are regions, and forest boreal last boreal globe, the from the the termed and change across ecotone, during temperate marked composition the This species In is and 1a). areas altitude 2011). (Fig. elevation lati- Barry, forests high High without & elevation-dependent in highlands 2017). and Serreze transitions al., amplification 2015; arctic et ecological Pecl of al., striking 2006; because et (Parmesan, exposed change (Pepin especially climate are warming by ecosystems affected altitude are and world tude the over all Ecosystems processes. sequestration C to linked biota belowground the Introduction in sequestration. shift C the corresponding enhance below a may be fungi line will forest saprotrophic there the lines, and above correlated forest ascomycetes ectomycorrhizal in positively root-associated of rise and while further turnover, predominance strongly With C soil The high and in promote ascomycetes. content likely ectomycorrhizal ericoid line C root-associated forest including Archaeorhizomycetes, percentage of including and mucoromycetes, abundance ecotone: Ergosterol ascomycetes, and line the endophytes. root-associated forest basidiomycetes with septate the more dark of across relatively and biota proportions observed soil fungi we the mycorrhizal higher in Above, were markers, shift ITS2 strong there to fungi. and a forests saprotrophic line, observed 18S We birch the forest mountain biomass. of zone the fungal subalpine metabarcoding transition quantify Below DNA from important to using an stretching used surveyed was ecotone, transects were ergosterol micro-eukaryotes line along while and forest samples fungi the space-for- soil Soil across a collected Using vegetation. biota (C). We low-alpine soil carbon the and ecosystems. in biota different soil changes on between compositional consequences analyse with worldwide, we lines approach, forest time of shift upward causes change Climate Abstract 2021 14, January 3 2 1 Mundra Tonjer Lea-Rebekka communities fungal a belowground indicates in space-for-time shift corresponding regions: alpine in forests Expanding owga nvriyo ieSciences Life of Fakultet University Matematisk-naturvitenskapelige Norwegian Det Oslo i Universitetet Oslo in University 1 ieNybakken Line , 1 laThoen Ella , 3 nesBryn Anders , 1 usMorgado Luis , 1 n H˚avard and Kauserud , 1 1 Synn , euapubescens Betula ø eSey Botnen Smebye ve 1 ssp czerepanovii . 2 Sunil , forest, ) Posted on Authorea 14 Jan 2021 | The copyright holder is the author/funder. All rights reserved. No reuse without permission. | https://doi.org/10.22541/au.161065834.45838723/v1 | This a preprint and has not been peer reviewed. Data may be preliminary. eeain n ii oefna ims sfudaoetefrs ie hc a edet h rsneof presence the to due the be (ii) may ecotone; which line line, forest forest aboveground the in the above changes across found with is groups, correlated biomass strongly factors functional change are fungal and compositional biotic fungi more soil strong taxonomic (iii) and of a and of groups vegetation, is bedrock) terms functional there and in and taxonomic (i) both in climate whether: across changes biota, investigate changes factors, soil to belowground wanted the edaphic and we in above- (soil aimed specifically, between We More abiotic relationships biota. ecotone. the both belowground the to investigate the with biomass) to about fungal vegetation), results for information (all (aboveground proxy these only) 18S (a and (fungi ergosterol couple quantitative only) measured to we we (fungi and and ectone, ITS2 qualitative sites soil, the line both nine from targeted obtain in forest DNA we ecotone ribosomal birch metabarcoding, the of mountain DNA across regions changes eukaryotes) Using the Norway. Phillips compositional across southern belowground 2018; biota across measure al., studied. spread to belowground et well designed (Geisen the not survey web is of a food conducted processes understanding soil C our the belowground invertebrates improve and to To including biota contribution micro-eukaryotes, soil their the Other although of groups 2019), dynamics. functional members al., dominate sequestration C dominating essential et shrubs belowground also C soil dwarf in are soil current ericoid shift protists, in a line, shifting and upwards, role forest both potentially migrates central the forest since expected, Above a the (Hill ErM is 2013). as play plants Thus, and Koide, to heath. melanized & in DSE proposed alpine have Fernandez forming uptake the fungi been 2015; ErM fungi nutrient al., has and between et which promote DSEs (Clemmensen separate decomposition, can Both Michelsen, to Leotiomycetes. to DSE Olsrud, especially hard resistant 2009; that ascomycetes, hyphae be by Read, shown can dominated & been are it Upson, Taxonomically, groups recently (Newsham, 2019). and fungi has vegetation al., ErM arctic It et others as and 2007). in alpine environments sequestration Dickie, Wallander, in same & C common systems & the John, also or some St in are 2008) Kirschbaum, in found (DSE) al., Orwin, endophytes often matter et 2014; fungi, septate Talbot Malcolm, organic dark EcM & 2015; Fernandez, Root-associated of 2008). Tunlid, Koide, 2011). turnover & Read, 2014; Lindahl promote Finzi, & 2019; & (Smith can Frey, Turner, graminoids (Averill, respectively 2014; ecosystems, and and al., (ErM), forest herbs structure et plants mainly northern (Bodeker hosts, ectomycorrhizal ericoid with in (AM), and plant symbiosis mycorrhizal dominate (EcM), their establish arbuscular which shrubs and which 2008). are on and (N) Treseder, fungi, groups trees depending nitrogen (ErM) & main (AM), the mycorrhizal groups Allison, three ericoid as Talbot, The functional and well 2008; 2008). into (EcM), as Read, Read, Saprotrophic divided cycle, & & nutrients. (Smith C Smith be and function global 2015; water can Tunlid, the for fungi & exchange of in (Lindahl Mycorrhizal components roots, host cycles crucial plant plant (P) thus with their organic symbiosis phosphorus are from mutualistic dead fungi C form mycorrhizal recycle fungi fixed and Mycorrhizal and freshly atmosphere. receive decompose strategies the they nutritional fungi to where between C Saprotrophic parasites, transition line releasing into 2018). blurry thus forest grouped Martos, a matter, be the is roughly & can across there Schneider-Maunoury, and although dynamics ways, (Selosse, various saprotrophs, pool in and in processes C symbionts C mutualistic to the soil to similar of contribute fungi fungi, understanding Belowground their including deeper and microorganisms, a communities these belowground for of the by differences important knowledge ecotone. in Improved These regulated are 2019). than 2015). Frey, roles be 2013; vegetation al., al., functional largely et forest et (Parker may birch (Clemmensen decomposition is systems mountain pools to lowland it and resistance C Furthermore, to shrub litters heath. soil compared the the alpine vegetation in of in the shrub independent in faster and heath, plants decomposes alpine forest ericaceous litter the beneath that in vegetation higher documented significantly forest smaller and and pools vegetation shrub C that shrub S under belowground Similarly, shown than and have Parker heath. heaths 2018). faster arctic regions al., alpine was arctic et under turnover and Sorensen and (Bryn higher C 2015; production, alpine change Wookey, significantly primary use from & is increased land Subke, studies content to (Parker, and However, leads warming C shrubs fixation. c ø rensen tal. et Srne ta. 08 hwdta tcswr oetbeneath lowest were stocks C that showed 2018) al., et (Sorensen 2 tal. et Pre ta. 05 hwdta soil that showed 2015) al., et (Parker Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. rvre Soc ta. 00 o 8.Alpieswr agdwt nqemlclrietfir MD.Each (MID). identifiers molecular unique TAReukREV3 with and tagged 1 (Ihrmark (forward) μ were TAReuk454FWD1 (reverse) of primers and ITS4 consisted All region, and 18S. reaction ITS2 for (forward) PCR 2010) the gITS7 al., amplifying primers et for The (Stoeck used PCR (reverse) regions. We were during 18S step. 2012) PCR and introduced al., the ITS2 were et during rDNA introduced controls) the were (negative controls) both (positive DNA negatives amplified communities based mock extraction column while and extraction, a DNA replicates by Technical followed USA). protocol E.Z.N.A. Norcross, extraction the DNA CTAB-Chloroform using a purification using DNA extracted We methods Molecular sdapoiaey20m ffez-re olfrmauigfe n oa olegseo concentrations ergosterol soil total and free measuring We for UK). Southampton, soil determined Ltd, freeze-dried was Analytical concentration of SEAL g (Thermo AutoAnalyse, P mg (mg analyzer HR elemental Soil 200 AA3 flash approximately USA). (SEAL a Waltham, used analyzer by flow Scientific, soil, segmented ThermoFisher freeze-dried a of 1112, C by g Soil EA 0.5 protocol. Flash using manufacturers soil For determined, Finnigan the dH analyses. was following mL different concentration (M.P. Japan) 5 the N Kyoto, beadbeater in for Scientific, and FastPrep-24 subsampled soil (Horiba a and freeze-dried Meter MHz using of 25 pH pulverised g at LAQUA-TWIN-11 0.5 then sec diluted and corporation, 20 we (Labconco h of measurements, freeze-dried 36 rounds pH two was for in sample tubes USA) soil pieces falcon CA, (>2mm) of Biomedicals, in coarse g and USA) 70 plants Approximately MO, where City, removed. hood, were Kansas flow laminar roots a and in wood further of handled and thawed standardized. were samples and Soil transformed skewness zero then a in were analyses provided variables on Soil ranked climatic bedrock were and of environmental bedrock”) types “nutrient-rich All three bedrock”, scale. The 2019): “nutrient-average 1-3 al., slope. bedrock”, et and (“nutrient-poor (Horvath bedrock data dataset temperature, published this mean a AM from precipitation, plants, plant variables annual EcM each environmental plants, aspect, site-specific of ErM following groups: proportion rare the plant levels: the obtained following We calculated three the of we into proportion information, and species overall mosses this each the lichens, as on of well plants, Based abundance as (3). plot, the We per dominant categorized together. species and and them (2) plants analysed common higher -20 therefore (1), at and there we kept diameter mosses often, and were in lichens, Most layers samples cm for layer. 3.8 Soil organic soil a mosses. and mineral -80 using and the litter at radius plants transect, excluding the m orientation green m sample, each between 1.5 plots living in soil 200 with division 11 removed collected representative circle a (i.e. were clear one a above cores along no to within soil distance m was five centre pooled m plot, the 20 and 100 each and corer, every At to West) 1c). soil line located South, other Fig. forest East, each plots together; birch from North, all from mountain km plots (i.e. samples the 99 15 and least below soil transect at distance collected per located m we and 100 site, vegetation Potthoff, from alpine & each stretching to (Bryn At Norway forests for 1a). birch model mountain (Fig. line from forest shift of a distinct set on south- wider based a in was a sites sites on line of Based forest selection high-elevation 2018). Our different 1). nine (Fig. at Norway 2017 central September/October in sampling conducted We sampling and Norway. design southern Study across analysed spread we sites observations, methods replicated our and nine of Materials in generality ecotones the along assess To obtained mycelium. samples recalcitrant soil with fungi ErM/DSE more odMgCl Gold l -1 ° eoefrhrpoesn.Wti ahpo,w eoddteudrtr eeaint pce level species to vegetation understory the recorded we plot, each Within processing. further before C W sn iia rtcla n(asdke,Apud hsn yakn 2019). Nybakken, & Ohlson, Asplund, (Ransedokken, in as protocol similar a using DW) 2 1 , μ 0gm S,0.2 BSA, 20mg/ml l Pyrola μ N epaead24 and template DNA l priori a ® pce,telte aigabti mycorrhiza. arbutoid making latter the species, olDAKtfloigtemnfcuespooo OeaBio-tek, (Omega protocol manufacturers the following Kit DNA Soil addt ie,w eetdnn ie ihacereeaingradient, elevation clear a with sites nine selected we sites, candidate μ NP,0.1 dNTPs, l 3 μ μ atrmx 15.7 mix: master l mlTqGl,1.5 Gold, AmpliTaq l 2 o n or n esrdp sn a using pH measured and hour, one for O ° uigfilwr n ae stored later and fieldwork during C μ dH l μ 10 l 2 ,2.5 O, μ owr rmrand primer forward M μ odBffr 2.5 Buffer, Gold l Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. rtclfr1Swssihl ieet C ecin eernwt nta eauaina 98 at denaturation initial 98 with at run 72 denaturation were of at reactions cycles PCR 32 different: by slightly followed was 18S for protocol 95 at 72 denaturation of at cycles 32 by followed 1.5 ed) h nl1Sdtstcnitdo 3smlsad49 OTUs. 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ITS2 due using variation reads. the default performed 32 intraspecific chimera-free extracting with was for widespread the to were 2013) algorithm a to with al., set Chimeras bimera Due table used et respectively. was We DADA2 reads. (ASV) (Bengtsson-Palme reads, variant ITSx merged. the size 18S used sequence were using we band and amplicon reads data, approach ITS2 an the reverse denovo constructed merging ITS2 and a then when forward for We with nucleotides corrected removed while 8 The and and 18S, and marker. checked 50 for forward this of the corrected used of overlap from independently were rate minimum parameters then INDEL reads correction settings imposed. We estimates higher sequence was default 2.5. that 18S the truncation The to algorithm the no machine-learning set itself. for marker, DADA2 was set ITS2 data the was error the using bp of expected reads variability 200 maximum reverse length of datasets, natural length both to truncate decide In Due A to environment information reads). reads. statistical this called quality used the and (hereafter using low profiles 2016) quality filtering al., sequence of for generated et We processing parameters (Callahan Further 2014). Team, allowed. pipeline using Core DADA2 were removal (R the tags 3.6.0 primers version with MID and performed and tags University was simultaneous primer the data with with at the miss-match separately cluster demultiplexed No computer were 2011). reads high-performance (Martin, CUTADAPT PE Abel raw the The on Oslo. analyses of bioinformatics all performed We the Bioinformatics with amplicons the ligate to chemistry. used V3 was with tag-jumping, (PE) adapters. minimize paired-end by flow-cell bp to sequenced Concentrator-5 250 designed Miseq were a & region) specifically with DNA Clean protocol, MiSeq targeted Illumina ligation DNA using each A with runs for two concentrated (two in 2 equimolar libraries and (Switzerland) to four Qubit SA cleaned Fasteris The pooled the was USA). and with California, pool USA) kit Research, Each measured MA, Concentrator-5 (Zymo Waltham, were pools. Scientific, & four sample Fisher Clean into (Thermo each gel, DNA concentration Kit agarose for ZR-96 Assay 2% concentrations BR with a DNA dsDNA amplicons using fluorometer USA). the electrophoresis California, of gel Research, purification with (Zymo and amplification clean-up positive for individual product before PCR each controlled We μ 10 l ° ° o i.W de nleogto tpa 72 at step elongation final a added We min. 1 for C o 5sc nleogto tpwsicue t72 at included was step elongation final A sec. 45 for C μ ees rmr ernPRratosfrIS ihiiildntrto t95 at denaturation initial with ITS2 for reactions PCR run We primer. reverse M ° ° o 0sc rmranaiga 53 at annealing primer sec, 30 for C 55 at annealing primer sec, 30 for C 4 ° o i,bfr oln ont 4 to down cooling before min, 7 for C ° o 0mn eoecoigdw o4 to down cooling before min, 10 for C ° ° o 0scadelongation and sec 30 for C elongation and sec 30 for C ° ° o min, 5 for C o min, 7 for C < ) Plant 9). ° .The C. ° C. R Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. aeu h ags ru 85OU,1096ras,floe yuasge soyee 66OU,148 OTUs, (626 ascomycetes unassigned by followed reads), 986 saprotrophs 130 that revealed OTUs, which (815 3a), (Fig. group guilds largest functional the to OTUs up assigned made we only reads. dataset, ITS2 Mortierellomycetes/Mucoromycetes ITS2 fungal at the and the For Ascomycetes Archaeorhizomycetes of of proportion data, proportion small 18S highest a the the up up to made along made contrast Eurotiomy- Leotiomycetes decreased In and 2d). Agaricomycetes) Leotiomycetes level. (Fig. (especially (especially class trend Ascomycetes Basidiomycetes opposite while the of an vegetation, to Basi- showed alpine proportion compared and cetes) low abundance, the reads) the relative 072 data, towards in 380 ecotone ITS2 pattern OTUs, different the the a (1522 In showed Ascomycota and data. phyla reads) the 18S 356 by 200 dominated OTUs, was (903 (Basidiomycota). 2d) diomycota 2c). 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Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. odsrmnt gis ..Acaohzmcts(hmr ta. 02 oln ta. 01 eesoe al., et Tedersoo shown 2011; been al., have primers et ITS Rosling employed (Nilsson, 2012; The biases al., 2015). primer et al., by the (Ihrmark et Archaeorhizomycetes explained of Tedersoo e.g. proportion be 2011; against large al., largely discriminate a et can to up Rosling results 2019; made contrasting al., but these et dataset, that Anslan, 18S the suggest the On We dataset. in Lecanoro- dataset. 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We low-alpine birch the the of in part lichen-forming heath EcM and Discussion the more ascomycetes with relatively where associated root-associated above, and was more outlined dataset. vegetation ascomycetes as ITS2 relatively low-alpine root-associated trends the with variation same the in Total the ecotone in 27.12% revealed (ITS2). the and fungi 5c) 4.2% dataset, (Fig. along OTUs 18S and distributed ITS2 the (18S) of OTUs in dataset. 3.3 ordination 25.96% ITS2 GNMDS were was score the level, variation, Species varia- in total plot plot-specific of composition and and fraction community site site- a in Thus, at as variation respectively. dataset explained explained, explaining dataset, ITS2 both in ITS2 explained the effects, important the gradients, in Interaction in equally individual 13.68% 13.44% the about and and within were variability dataset dataset, bles for 18S 18S account varia- the the also (between-site) in in 5b). which 11.27% regional (Fig. variation factors, for diagram compositional plants Plot-specific account ITS2 1). the EcM which the (Table of factors, in understory structured N 4.69% site-specific other were soil explained that in (GNMDS2) to bility, demonstrated addition axes increase analyses in second slight CCA b), the increased a The 5a, datasets, plants (Fig. was both variables forming there environmental In ErM forming site-specific while 3c). EcM of the (Fig. line, the by from amount no vegetation definition, forest increased observed The low-alpine By We content the samples. 3c). the 3b). ergosterol below (Fig. towards soil (Fig. and decreased present the plants decreased soil only in AM content mass while was content P dry line, N and in forest pH total C the while Percent % above the vegetation, b). for as Soil low-alpine and gradient trend communities. the main 5a systematic soil same to Fig. the in forest c, along birch changes and structured the compositional largely 3b forest were (Fig. the birch groups, biota driving mountain plant diagrams, soil the subalpine gradient both with from main In together stretching the 5b). factors, ecotone Fig. edaphic as dataset, the vegetation (ITS2 identified low-alpine alone composi- (GNMDS1) community fungi to axis in and ecotone ordination 5a) the Fig. first across dataset, the gradient (18S micro-eukaryotes clear all a of demonstrated tion datasets (R both ergosterol on analyses and GNMDS samples the composition correlation of community (R strong mass of ascomycetes a Drivers dry vegetation root-associated observed low-alpine the of we were abundance Furthermore, the in read 3a). in other content and abundant (Fig. the C content more forest of percent birch far most the mountain being root- between and subalpine while of vegetation, ascomycetes the trend low-alpine reads). to the root-associated opposite 586 towards compared were an 20 decreased showed saprotrophs OTUs, OTUs and ascomycetes (108 common EcM lichens associated of most OTUs, and abundance (135 40 The reads) pathotrophs fungi. 454 the reads), EcM 932 94 of 43 OTUs, OTUs, half (115 (149 yeasts About ascomycetes reads), root-associated 428 137 reads), OTUs, 7612 (321 fungi EcM reads), 884 6 2 04;Fg )ars h ecotone. the across 4) Fig. =0.45; 2 07) n ecn C percent and =0.73), euapubescens Betula Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. h et,wihmyacutfrtelwrfato fsi bevdhr,det atrdecomposition. faster to due to here, compared observed forests C birch soil alpine of the fraction in lower fungi the saprotrophic for of account proportion may forest which higher the and heath, below a 2015) observed the observed Tunlid, C we & soil general, Lindahl of amount In 2013; lower fast al., the relatively study. et for with our account Clemmensen associated in fungi, 2014; are line saprotrophic al., fungi the et EcM with Many (Bodeker together may, altitude. observations turnover with Our Vaˇsutov´a matter declined line. organic by fungi forest plants soil observed birch EcM host mountain their where what reflected the 2017), with ecotone below Cudlin, the line fungi across EcM in fungi of EcM are abundances not of higher and distribution be content with the must 2, C distributions, data hypothesis percent additional with on dynamics, accordance data stock In have al., C overall only et we about (Clemmensen conclude study composition to community this Hence, fungal collected. in measurements. the that content stock of emphasised C C consequence processes, be overall a cycling community must rather and it the 2017; driver, decomposition However, with a to al., 2015). strongly and as respect and et C correlated regarded with up high Hagenbo content but not the build diagrams, C is 2015; for ordination Percent long-term al., account the vegetation. a also in et low-alpine may to composition Clemmensen the processes leading in 2013; These rate content 2017). al., turnover ergosterol Lindahl, et & mycelial Karltun, (Clemmensen slower Clemmensen, sequestration to Kyaschenko, Clemmensen C due 3. is soil a hypotheses relationship increased had our this content with ergosterol that line and argued in C vegetation, soil low-alpine percent the that al. towards observed we correlation ascomycetes, positive root-associated strong the 2003). (Pinto-Figueroa to Schmidt, ecosystems Other parallel latitude & DSEs. In and Lipson, and (Sterkenburg, altitude Martin, fungi environments high Schadt, in ErM stressful 2019; e.g. alpine with al., in 2015), low along et Lindahl, abundant mutualists the plants, & relatively root-associated Clemmensen, in al., ericaceous are Durling, Archaeorhizomycetes to et be Archaeorhizomycetes Bahr, of linked (Rosling that may abundance are Archaeorhizomycetes confirm Archaeorhizomycetes high they Archaeorhizomycetes the studies The some that of of 2014). indicate that increase may ecology Rosling, hypothesised an vegetation & the observed been James, about root-associated Urbina, also has known between (Menkis, We is is correlation study. Little but positive this longer heath. 2011), strong the in non- alpine and the content with 2015), the al., explain comparison C et towards may in (Clemmensen percent longer, pools mycelia and carbon be melanised soil ascomycetes thus boreal of may of time part soil important necromass retention Fungal the an under 2014). is Koide, in living biomass) & Fernandez fungi dead 2019; fungi (i.e. Kennedy, melanized & DSE Kolka, highly decomposition Heckman, slower (Fernandez, and of induces fungi wall Melanin melanized cell time ErM 2013). hyphal retention Koide, the tundra. & of the (Fernandez arctic polymer desiccation and recalcitrant against Tedersoo in a fungi in is protect abundances especially Melanin may showed hyphae. that plants, relative pattern melanized have ericaceous highest the typically where conditions with had stressful ecotone, line groups in the these is along where data) heath representing (ITS2 ascomycetes, alpine vegetation root-associated the alpine the in of the most clustered plot, nigrum example, fungi, ordination Empetrum For species DSE fungi. to the and similarity root-associated In ErM . sequence plant 2008) represented high Read, showed data & (Smith OTU ITS2 notably common the most line, most in forest lineages. overall Ascomycota the Pezizomycotina of data). above after numerous (ITS2 Ascomycota groups Indeed, in Eurotiomycetes more Several present. and of observed Leotimycetes trend be was general and may primer a data), biases reverse was (18S primer there the Archaeorhizomycota the However, biases, that in provide the dataset. indicated mismatch and of ITS2 data a 2014) Irrespective the 18S al., to inspection, the et compared closer (Hadziavdic in composition a eukaryotes Pezizomycotina community all fungal of amplify the amount to of low 18S expected overview The was general poorly. study, more Cryptomycota this a and in Chytridiomycota used Mucoromycota, we amplify seemingly primers also they and 2015) Cemne ta. 05 hwdasmlrcreainbtenegseo n oa olCsok and stock, C soil total and ergosterol between correlation similar a showed 2015) al., et (Clemmensen oiae.Tegaulices fLoimctsadErtoyee oad h low the towards Eurotiomycetes and Leotiomycetes of increase gradual The dominated. , tal et Vstv,EwrsJnsv,Blra,Cra,& Cermak, Baldrian, Edwards-Jonasova, (Vasutova, . 7 eooaericae Pezoloma tal. et omnEMfungus ErM common a , Tdro ta. 2014), al., et (Tedersoo et Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. ae,S . lmne .C,Foe,G . atr,W . afe,L . ngt . irr N. of shift upward Fierer, rapid A & (2008). R., T. Perkins, Knight, & T., W., Siccama, C., L. Pucko, G., Parfrey, D. Gavin, A., B., W. Osborne, soil. B., Walters, Beckage, in E., communities protistan G. diverse Flores, highly doi:10.1038/ismej.2012.147 of C., biogeography J. Global and Clemente, (2013). plants T., between version. S. package competition Bates, R Mycorrhiza-mediated ‘MuMIn’. Package (2014). (2019). C. K. A. 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(Parker matter. fungal of to cycling this in lead on shifts C may conclude corresponding and ecosystems to through respiration northern 2018), despite soil in al., that higher et line view of Sorensen forest the parallel because abundance in corroborate a loss the raise findings C is with and Our soil there associated shrubification vegetation. strongly shown productivity, low-alpine is aboveground has the which higher study in ecotone, ascomycetes the This across root-associated content respectively. of ergosterol ascomycetes, fungi belowground saprotrophic a root-associated and in represents EcM by shift vegetation, by low-alpine dominated dominated soils treeless soils pattern from to communities, to distribution forest fungal clear belowground birch al., in mountain any et shift subalpine showed corresponding Seppey from protists 2017; stretching the al., ecotone, of et The none (Mahe level, (Arthropodes) distribution phylum their species ecotone. Apicomplexa, and host the of more kingdom of across lifestyle relatively at presence parasitic be However, the the to to on shown 2020). Due dependent been 2013). largely have al., likely commonly Apicomplexa et is is and as (Bates Cercozoa soils pH Ciliophora general, humid soil whereas In in regard Heger types, abundant altitude. others soil and 2014). while communities various al., 2014), (Bates protist in et Michael, between communities observed Shen correlation & protist 2016; a Jorg, Mitchell, high showed influence Cornelia, & also strongly at Stefana, 2016) Theurillat, moisture 2018; Derungs, soils abundance al., soil (Heger, in higher et important that Araujo relatively more abundant shown de their have be 2013; with Studies al., to well et line. corresponds shown & (Biela´nska-Grajner, forest which been Ejsmont-Karabin, lichens annelids the 2006), and where also above study, Ricci, mosses this has & in of Fontaneto present latitudes Rotifera result often 2011; the high Yakovenko, are groups. with at and line dominant altitudes, abundant in and the more is latitudes This are are forest 1990). typically the Procter, nematodes nematodes above 2019; and and abundant al., annelids et more common (Phillips relatively that were altitudes were shown Nematoda, and Rotifera have and and studies Annelida picture Arthropoda Other unbiased groups, an whereas Metazoa line. as been reflect ecotone, abundant have abundant proportions entire most might the equally two the Ascomycota whether The across about fungi unclear biota. remains some was it belowground earlier, that,Metazoa so the pinpointed marker, of found As 18S reads. we the sequence by fungi, amplified as on poorly measured soil focused the explain mainly in may was fungi receiving ecotone. which entire study roles, 2019), the functional our al., different across the Although et root play Mucoromycota in may Hoysted fine of Mucoromycetes litter as 2015; abundance Hence, available Mucoromycotina, relative al., N. that of high soil et observed the abundance of (Field These been exchange higher cryptogams line. also in a C with has forest likely plant symbionts it the However, is important below heath. there are community alpine endophytes, and the fungal saprotrophs, to the as compared dominated known forest Mucoromycetes foremost dataset, are 18S fungi the on Based aue 505 Nature, 78) 4-.doi:10.1038/nature12901 543-+. 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No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. l aata upr h nig fti td r pnyaalbei ra.r thttp://doi.org/[doi], at Dryad.org in available openly are study this of findings the number]. [reference support that data All Gram- the Using accessibility Visualisations treeline Data Data Elegant alpine 2). Create (Version the ‘ggplot2’. Graphics Package at of (2016). W. mar fungi Chang, saprotrophic & H., and Wickham, mycorrhizal of environmental composition Distinct community (2017). ecotone. P. Cudlin, the & drive ordination M., of Cermak, variables P., choice Baldrian, of M., importance Edwards-Jonasova, data. the M., Vasutova, abundance ordinations: parallel species Multiple of (2014). weighting R. and Halvorsen, method & C., (2014). T. K. Son, Abarenkov, van . . . Shotgun R., (2015). Wijesundera, fungi. 10.1126/science.1256688 K. S., soil N. 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Shi, research patterns J., A diversity W. amplification: elevational Liang, Arctic C., of E. C. impacts and Lara, Shen, Processes 77 . (2011). Change, edaphic . G. by Planetary . R. than and Barry, D., Global topo-climatic & by Singer, C., predicted E., M. better Pinto-Figueroa, Serreze, is E., Alps western Yashiro, Swiss A., the Buri, in variables. diversity O., protist Broennimann, Soil W., (2020). V. C. Seppey, > ://WOS:000263083500002 uglEooy 27 Ecology, Fungal ora fBoegah,47 Biogeography, of Journal < ot ISI to Go Mycokeys oeua clg,19 Ecology, Molecular > eoilga 57 Pedobiologia, 1) -3 doi:10.3897/mycokeys.10.4852 1-43. (10), ://WOS:000345326000019 1-2.doi:10.1016/j.funeco.2016.08.010 116-124. , yoria Symbiosis Mycorrhizal 12,8-6 doi:10.1016/j.gloplacha.2011.03.004 85-96. (1-2), 4,8688 doi:10.1111/jbi.13755 866-878. (4), 46,2523 doi:10.1016/j.pedobi.2014.10.001 205-213. (4-6), cec,346 Science, 13.doi:10.1111/j.1365-294X.2009.04480.x 21-31. , omreta 37 Sommerfeltia, 13 cdmcPress. Academic : 61) 08+ o:RN1256688 doi:ARTN 1078-+. (6213), mi,38 Ambio, e htlgs,207 Phytologist, New 1,1-37. (1), ucinlEooy 22 Ecology, Functional 1,21.Rtivdfrom Retrieved 2-10. (1), clg,95 Ecology, csses 21 Ecosystems, 1) 3190-3202. (11), 4,1145-1158. (4), 6,955-963. (6), < (2), Go Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. (40) , (36) (32) 2, sp. albicastaneus Hygrophorus bicolor Piloderma number sp the to proportional alpinus are circles birch of (10) mountain size num- (boreal and are 11 legend), Isolines derma to visualized. (see (1) vegetation) are (low-alpine groups reads. OTUs 01 functional of common from indicate species most ranging on Colors 40 based plot-numbers the forest). dataset the ITS2 Only illustrates to the OTU. arrows according of one green plot bered that represents ordination light Variables GNMDS point while details). (c) Each factors, types. for scores. site-specific plant legend mycorrhizal arrows of (see green distribution green dark the factors, are (p edaphic forest structure represent birch community arrows matri- mountain the red/yellow, OTU colored boreal with rarefied are on the correlated vegetation based low-alpine significantly to the dataset shown to ITS2 belonging belonging values (b) plots Plots and squared sample. while dataset soil R 18S one represents the the point (a) which Each of ces. from plots lines. ordination factor, GNMDS regression random 5. squares a Figure least pro- as represent relationships (b) sites lines The and with content. The content carbon extracted. models carbon soil were mixed and soil reads) and using of ergosterol while modelled (number (a) endophytes forest, were septate between birch dark relationship are mountain of the Outliers abundance boreal showing portional vegetation. percentiles. the plots low-alpine 75th in Scatter the and show plot 4. in 25th boxes lowermost Figure uppermost the the the the represents in is is box lines 01 11 the bold number number of The Plot plot part ecotone. dots. lower the black and across by vary ergosterol, upper represented and status the factors mycorrhizal and edaphic on soil median, (b) based the the guilds, low-alpine groups fungal is dominant plant the 01 tree in (c) the number uppermost and (a) Plot how the illustrating dataset. is Boxplots ITS2 11 3. Figure number the plot distribution on level while based Class forest, classes birch (c) fungal mountain data. of vegetation. boreal 18S the Distribution the the on of in (d) based distribution plot ecotone data. 100 lowermost level the 18S phylum from across fungal (b) stretching groups the and taxonomic transect of Kingdom of (a) m abundances reads. 200 relative of line. the the number forest illustrating along birch represents Barplots mountain meters 11 2. the to 20 Figure above 1 every Numbers meters collected line. 100 Schema- were forest (c) to birch site. samples below mountain Lemonsjøen meters soil the the across at where from stretching line plots transects forest data the the studied map at the change with of vegetation view 3.4.14, abrupt tic (Har- the version Ustevatn of QGIS Photo Haglebu, (Jotunheimen), (b) north: in geonorge.no. Bessheim to drawn south Slidre), was (Vestre from (V˚ag˚a), Norway, map Domb˚as. Storlifjell south-central Lemonsjøen The (Hemsedal), all in and Skyrvedalen sites and Sel, Strandavatnet, sampled ET nine dangervidda), and The (a) HK 1. with Figure collaboration the in LRT, designed manuscript and work HK the legends lab and wrote Figure did AB LRT LN commented. LM, and data. and SM ET, the edited LRT, ET, analysed co-authors LRT. LM, LRT, other SB of fieldwork. thesis and conducted HK MSc SM and the LM, LM of ET, version LRT, and revised research a represent study This Contributions Author ,(18) ., ehmcl minima Leohumicola oteel humilis Mortierella otnru talus Cortinarius p,(6) sp., (14), , Amanita Meliniomyces eooaericae Pezoloma Venturiales Chaetothyriales (23) , p,(19) sp., . Phialocephala p ,(33) 3, sp. (11) , (37) , p,(15) sp., (28) , Helotiales Herpotrichiellaceae (2) , p,(7) sp., Hygrophorus uo abundans Mucor Herpotrichiellaceae Herpotrichiellaceae Lecanorales Cenococcum p,(24) sp., p,(20) sp., p,(38) sp., Helotiales Herpotrichiellaceae p,(3) sp., (29) , p,(12) sp., p,(8) sp., 14 eiimcsbicolor Meliniomyces p,(16) sp., Agaricales Helotiales p,(34) sp., < p,(25) sp., .5 r hw sarw ntedarm.Dark diagrams. the in arrows as shown are 0.05) otnru pseudocandelaris Cortinarius Chaetothyriales suooetlatristis Pseudotomentella Meliniomyces p,(30) sp., Luellia p,(4) sp., p,(21) sp., p,(26) sp., Helotiales otnru caperatus Cortinarius (39) , p,(9) sp., oioczm terricola Solicoccozyma p ,(35) 4, sp. uiomsrcei rosae Muriformistrickeria Chaetothyriales otnru armillatus Cortinarius p,(31) sp., (17) , (13) , Hyaloscypha Meliniomyces Meliniomyces Cortinarius (5) , p,(27) sp., (22) , Pilo- sp., , Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. Figure 1 (a) 0 3060km 01 Sub-alpine forest mountainbirch (c) (b) 15 02 03 04 05 Sampling plots Forest line 200m 06 07 08 09 Low-alpine vegetation 10 1 1 Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. (a) Figure 2 (c) 0.0 0.2 0.4 0.6 0.8 1.0 0.0 0.2 0.4 0.6 0.8 1.0 Fungi classes(18S) Kingdom (18S) 01 01 02 02 03 03 04 04 05 05 06 06 07 07 08 08 09 09 10 10 1 1 1 1 Archaeorhiz Mucorom Agar Ch Cr Sordar T Saccharom Pucciniom Glomerom Fungi unassigned Other Metaz Fungi Ciliophor Cercoz Apicomple Lobosa Conosa Other remellom yptom ytr icom idiom oa iom oa ycetes a ycetes ycetes ycetes ycetes ycetes ycetes xa ycetes ycetes om ycetes 16 (b) (d) 0.0 0.2 0.4 0.6 0.8 1.0 0.0 0.2 0.4 0.6 0.8 1.0 Fungi classes(ITS2) Phyla (18S) 01 01 02 02 03 03 04 04 05 05 06 06 07 07 08 08 09 09 10 10 1 1 1 1 Leotiom Agar Eurotiom Lecanorom Dothideom Archaeorhiz Xylonom Sordar Mor P T Mucorom Other Annelida Nematoda Ar Rotif Ascom Basidiom Mucorom Ch Cr Spirotr Filosa-Sarcomonadea Apicomple Other remellom eziz thropoda yptom ytr tierellom er icom om idiom a ichea iom ycota ycetes ycetes ycota ycetes ycetes ycota ycetes ycota ycetes ycetes ycetes xa ycota ycetes ycetes om ycetes ycetes Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. (a) (b) (c) 1000 2000 0.0 0.2 0.4 0.6 20 30 40 50 4 5 Figure 3 pH ErM plants(proportion) C:N r (#sequences) Root associatedascomycetes(RAA) 01 01 01 01 02 02 02 02 l l atio 03 03 03 03 l 04 04 04 04 05 05 05 l l 05 l 06 06 06 06 07 07 07 07 08 08 08 l 08 l 09 09 09 09 10 10 10 l 10 l 1 l 1 1 1 1 1 1 1 1000 2000 3000 4000 0.0 0.1 0.2 0.3 0.4 0.1 0.2 0.3 10 20 30 40 50 0 EcM plants(proportion) T %C Ectom 01 01 01 01 otal ergosterol(mg/gdryweight) l 02 02 02 02 l l l l ycorrhizal fungi 03 03 03 03 l 04 04 04 04 l l l l 05 05 05 05 06 06 06 06 l l l 17 07 07 07 07 l (#sequences) 08 08 08 08 09 09 l l 09 09 l 10 10 l 10 10 l 1 1 1 1 1 1 l 1 1 1000 2000 3000 150 200 250 300 350 0.4 0.8 1.2 1.6 0.0 0.2 0.4 0.6 %N T AM plants(proportion Saprotrophic fungi(#sequences) 01 01 01 otal P 01 l 02 02 02 02 03 µg/L 03 03 03 l l l 04 04 04 04 l l l 05 05 05 05 l l 06 06 06 06 l 07 07 07 07 l l l l l 08 08 08 08 l l l 09 09 09 09 l 10 10 l 10 10 l 1 1 1 1 1 l 1 1 1 Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary.

% carbon content 10 20 30 40 50 Figure 4 0.05 Ergosterol 0.10 0.15 (mg/g dryweight) 0.20 0.25 0.30 18

10 20 30 40 50 500 RAA (#sequences) 1000 1500 2000 2500 Posted on Authorea 14 Jan 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.161065834.45838723/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. (b) (c) (a)

GNMDS2 Figure 5 -1.0 -0.5 0.0 0.5 -0.4 -0.2 0.0 0.2 0.4 0.6 0.8 -0.5 0.0 0.5 -1.5 -1.0 1 10 09 08 07 06 05 04 03 02 01 1 -1.0 9 17 -1.0 33 24 34 ErM plants 32 31 37 8 20 30 -0.5 Ergosterol 26 2 3 2 -0.5 6 4 C 4 35 25 Ergosterol 1 3 1 -0.5 5 13 plants ECM 38 C 22 ErM plants 15 19 6 16 16 7 T 7 emp 0.0 29 0.0 N plants ECM Bedrock . 28 8 7 14 Precip GNMDS1 19 23 36 T Aspect 12 emperature . P 0.0 Slope 21 5 Precipitation 0.5 10 39 Bedrock 9 P Aspect 9 Slope 0.5 pH 18 1.0 0.5 40 AM plants Unassigned P Y Lichen Saprotrophs Ectom Root associatedascom 10 east ar AM plants asite pH ycorrhizal fungi 1.0 1.5 27 1.0 ycetes