1

Va ia ion in mo phological and chemical ai s o Medi e anean ee 1
oo s: linkage wi h lea ai s and soil condi ions 2
3
Teodo o Ma añón1*, Ca men M. Na a o-Fe nández1, Ma a Gil-Ma ínez1, Ma ía T. 4
Domínguez2, Paula Madejón1 and Ra ael Villa 3 5
6
1 IRNAS, CSIC, A enida Reina Me cedes 10, 41012 Se ille, Spain 7
2 Depa amen o de Mine alogía, C is alog a ía y Química Ag ícola, Uni e sidad de Se illa, 8
P o . Ga cía González s/n, 41012, Se ille, Spain 9
3 Á ea de Ecología, Facul ad de Ciencias, Uni e sidad de Có doba, Campus de Rabanales, 10
14071 Có doba, Spain 11
12
* Au ho o co espondence ([email p o ec ed]) 13
14
15
Au ho con ibu ion s a emen 16
TM concei ed he s udy, TM, CMNF and MTD conduc ed ieldwo k, CMNF and PM 17
measu ed mo phological and chemical ai s, TM, CMNF and MGM analysed he da a, TM 18
w o e he i s d a , TM, CMNF, MGM, MTD, PM and RV pa icipa ed in he in e p e a ion, 19
discussion and p epa a ion o he inal e sion. 20
21
Pos p in o : Plan and Soil (2020) h ps://doi.o g/10.1007/s11104-020-04485-5
2

Abs ac 22
Aims: Roo unc ions a e mul iple and essen ial o he g ow h and su i al o e es ial 23
plan s. The aim o his wo k was o analyse he main ends in he a ia ion o oo ai s, hei 24
coo dina ion wi h lea ai s and hei ela ionships wi h soil condi ions. 25
Me hods: We measu ed he a ia ion o 27 ine oo ai s ( i e mo phological, 20 chemical 26
and wo iso opic signa u es) in ees o se en species o a mixed plan a ion in a me al-27
con amina ed and emedia ed si e o Sou he n Spain. 28
Resul s: We ound e idences suppo ing he exis ence o a oo economics spec um (RES). 29
Howe e , o he dimensions we e iden i ied as being independen o he main RES: mainly he 30
a ia ion in he ca bon concen a ion, he accumula ion o ace elemen s associa ed wi h 31
ole ance o me al- ich soils, and he ac iona ion o δ15N as a ime-in eg a ed ai o 32
myco hizal-media ed nu i ion. In gene al, oo s and lea es we e unc ionally coo dina ed, 33
al hough mos o he ace elemen s showed s ong oo -lea disco dance. The soil condi ions 34
in e ac ed wi h he ine oo ai s in eedback p ocesses. The abili y o ee oo s o 35
accumula e ace elemen s and o educe hei ansloca ion o lea es is a desi able ai o he 36
phy o emedia ion o me al-con amina ed soils. 37
Conclusions: Roo s a e mul i unc ional. Unde s anding he a ia ions in he oo ai s o ees 38
will help us o p edic bo h he esponses o o es s o global changes, including soil 39
con amina ion, and he p o ision o soil-based ecosys em se ices. 40
41
Keywo ds 42
Roo ai s, Roo economics spec um, Roo -lea coo dina ion, T ace-elemen con amina ion, 43
Roo mul i unc ionali y 44
45
In oduc ion 46
3

Ad ances in ai -based plan ecology a e ocused on he analysis o unc ional ai s 47
ac oss indi iduals and species, o p edic eme gen p ope ies o communi ies and ecosys ems 48
(Ga nie e al. 2016; Lalibe é 2017). The e a e e olu iona y and biophysical cons ain s 49
limi ing he exis ing spec um o plan ai s (Reich 2014). In a global iew o he unc ional 50
di e si y o ascula plan s on Ea h (analysis o key ai s o mo e han 46,000 plan 51
species), mos o he ai a ia ion was concen a ed in a wo-dimensional spec um o plan 52
o m and unc ion: plan size and lea economics spec um (LES) (Díaz e al. 2016). The LES 53
e lec s he ade-o be ween esou ce acquisi ion and esou ce conse a ion. A one end o 54
he spec um, he e a e species wi h high pho osyn he ic and espi a ion a es, high ni ogen 55
(N) and phospho us (P) concen a ions, low lea mass pe a ea (LMA) and low lea longe i y. 56
A he o he end o he spec um, he e a e species wi h he opposi e ai s (W igh e al. 57
2004). 58
Al hough oo s a e essen ial o gans o e es ial plan s, no oo ai s we e conside ed 59
in he global s udy o Díaz e al. (2016). Roo unc ions a e mul iple and essen ial o plan 60
g ow h and su i al: hey include nu ien and wa e acquisi ion, ancho age, esou ce s o age 61
and suppo o symbio ic soil mic obes. A he ecosys em le el, hey con ibu e o soil 62
s uc u e and o he ca bon and nu ien cycles (E k an e al. 2018). The e o e, an 63
unde s anding o how oo ai s a y is undamen al o he comp ehension o plan unc ional 64
ecology. 65
Some s udies suppo he exis ence o a “ oo economics spec um” (RES), analogous 66
o he LES, wi h a ade-o be ween esou ce acquisi ion and conse a ion (Reich 2014; 67
Roume e al. 2016; de la Ri a e al. 2018a). Thus, plan s g owing in a ou able en i onmen s 68
would de elop ligh e oo s wi h a lowe d y ma e con en and highe speci ic oo leng h o 69
maximize esou ce acquisi ion. By con as , plan s g owing in ad e se o limi ing 70
en i onmen s would exhibi a esou ce conse a ion s a egy, de eloping dense oo s wi h a 71
4

highe d y ma e con en and lowe speci ic oo leng h. Howe e , he RES hypo hesis was 72
challenged by Weems a e al. (2016) who a gued ha oo ai s a e cons ained no only by 73
esou ce up ake, bu also by o he d i e s (like soil ex u e and chemis y), and ha he RES 74
hypo hesis does no inco po a e soil he e ogenei y and myco hizal symbiosis. Mo eo e , 75
se e al s udies (K ame -Wal e e al. 2016; Kong e al. 2019) ha e ound ha high SRL oo s 76
can be cons uc ed wi h any densi y, indica ing excep ions in he RES. 77
Resou ce acquisi ion is coupled and linked among plan o gans. Thus, as acquisi ion 78
and p ocessing o wa e and nu ien s by oo s would equi e as acquisi ion and p ocessing 79
o ca bon (C) by lea es (Reich 2014). A s ong coo dina ion be ween oo mo phology and 80
abo eg ound ai s was ound o a se o 80 woody species (de la Ri a e al. 2018a). Roo 81
ai s ha achie e nu ien conse a ion a ou issue longe i y and slowe g ow h a es, and 82
in consequence diminish nu ien equi emen s and amo iza ion o he cons uc ion cos s 83
(Poo e and Villa 1997; Villa e al. 2006; de la Ri a e al. 2016b, 2018a). Howe e , o he 84
s udies did no ind co ela ions be ween lea and oo ai s, sugges ing ha ade-o s in 85
di e en o gans ope a e independen ly and ha he lea - oo coo dina ion may depend on 86
speci ic limi ing ac o s in each habi a (Tjoelke e al. 2005; Kembel and Cahill 2011; 87
Fo unel e al. 2012). 88
Con as ing lea habi s in ees - ha is, e e g een e sus deciduous - a e usually 89
associa ed wi h di e en unc ional ai s. Fo example, deciduous species a e cha ac e ised 90
by acquisi i e ai s such as lowe LMA, highe a es o pho osyn hesis and espi a ion, and 91
highe nu ien concen a ions, in compa ison o e e g een species, which end o exhibi 92
mo e-conse a i e ai s (W igh e al. 2004; Villa e al. 2006; de la Ri a e al. 2018b). 93
Howe e , ew s udies ha e in es iga ed he di e ences in oo ai s be ween e e g een and 94
deciduous ees; o example, Ma inez e al. (2002) did no ind di e ences in oo C o N 95
concen a ions be ween deciduous and e e g een species o Que cus. 96
5

Mos ee oo s a e in ima ely associa ed wi h myco hizal ungi in a symbiosis ha is 97
c ucial o nu ien acquisi ion and ole ance o di e se s esses (d ough , hea y me als o 98
pa hogens), while he ungus ob ains ca bon compounds om he plan (Smi h and Read 99
2008). In ac , ha combina ion o oo and ungus (myco hiza) can be conside ed as he 100
unc ional abso p i e ai , in which ungal issues may ep esen up o 54% o he “ oo ” N 101
concen a ion (Ouime e e al. 2013). The deg ee o oo colonisa ion by myco hizal ungi 102
has p o ed o be a use ul plan ai o unde s and ecosys em p ocesses (Soudzilo skaia e al. 103
2015; Na a o-Fe nández e al. 2016; Lalibe é 2017). Fungal ai s, like he ype o hyphal 104
explo a ion, add mo e complexi y o he soil- ungus-plan ela ionships and esou ce 105
acquisi ion s a egies (Chagnon e al. 2013; Gil-Ma ínez e al. 2018; López-Ga cía e al. 106
2018). 107
Besides un a elling he RES, ano he esea ch challenge is o unde s and how 108
di e en d i e s o global change impac a sui e o oo ai s, and o p edic hei cascading 109
e ec s on soil-based ecosys em p ocesses (Ba dge e al. 2014). Roo ai s a e plas ic and 110
espond o physical soil limi a ions, he he e ogeneous dis ibu ion o soil wa e and nu ien s 111
and bio ic in e ac ions (Ba dge e al. 2014). Fo example, soil compac ion limi s he 112
o ma ion and pene a ion o hin oo s, inducing a lowe speci ic oo leng h (SRL) in ee 113
seedlings (Alameda and Villa 2012). Unde d y soil condi ions, plan s end o de elop 114
hinne oo s, wi h g ea e SRL and inc eased oo hai densi y, o imp o e wa e acquisi ion 115
(Comas e al. 2013; Olmo e al. 2014). Ni ogen deposi ion dec eases ine oo biomass, C:N 116
a io and ungal coloniza ion, while inc easing oo espi a ion (Li e al. 2015). 117
As a global change d i e , soil pollu ion may also p omo e he adjus men o oo 118
ai s in plan s. Fo ins ance, a high concen a ion o ace elemen s in soil o en educes oo 119
elonga ion and al e s oo a chi ec u e (Kahle 1993). Al hough, a a global scale, soil pollu ion 120
is one o he main h ea s o soils and he ecosys ems se ices p o ided by hem (Rod íguez-121

6
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Eugenio e al. 2018), i s e ec s on plan unc ional ai s ha e no been ully add essed. 122
Among he di e en soil pollu an s, hea y me als a e ele an s esso s, al e ing he plan -soil 123
in e ac ions (K umins e al. 2015). 124
In his s udy we analysed he a ia ion in mo phological and chemical oo ai s in 125
se en Medi e anean ee species, and explo ed he linkages wi h abo eg ound ai s and soil 126
condi ions in a he e ogeneously-pollu ed en i onmen (Guadiama G een Co ido , in SW 127
Spain). This a ea is a la ge-scale example o he phy o emedia ion o land con amina ed by a 128
mine-spill, wi h high concen a ions o me als. A mixed plan a ion o na i e ees and sh ubs 129
was se up a e cleaning and emedia ing he soil (Domínguez e al. 2008; Madejón e al. 130
2018a, b). This la ge-scale expe imen is an oppo uni y o explo e how he soil condi ions (in 131
his case, he concen a ion o hea y me als) a ec he oo ai s o di e en ee species 132
coexis ing in a simila en i onmen . We add essed he ollowing hypo heses: 133
1) Roo s o di e en ee species di e in hei unc ional ai s in acco dance wi h he 134
oo economics spec um (RES). Howe e , he e a e o he oo dimensions 135
(independen o RES) ha e lec oo mul i unc ionali y (Weems a e al. 2016). 136
2) Roo and lea ai s a e coo dina ed in acco dance wi h he plan economics spec um. 137
Fas plan g ow h depends on he coo dina ion o oo s and lea es, wi h oo s ensu ing 138
a wa e and nu ien supply su icien o main ain acquisi i e lea es wi h high 139
pho osyn he ic a es and high e apo a i e demand (Reich 2014). 140
3) Soil condi ions and me al con amina ion a ec oo ai s. Roo ai s a e plas ic and 141
espond o physical soil limi a ions, he he e ogeneous dis ibu ion o soil wa e and 142
nu ien s, bio ic in e ac ions (Ba dge e al. 2014) and soil pollu ion (Kahle 1993). 143
144
Ma e ial and me hods 145
S udy a ea 146
7

The s udy a ea is he Guadiama G een Co ido (Se ille, Spain). The clima e is 147
Medi e anean wi h mild, ainy win e s and ho , d y summe s. The a e age annual ain all is 148
450 mm and he mean annual empe a u e is 17 ºC, wi h a maximum o 33 ºC (in July) and a 149
minimum o 5 ºC (in Janua y). Fo mo e de ails, see he a ea desc ip ion in Domínguez e al. 150
(2008) and Madejón e al. (2018a). 151
The s udy a ea was a ec ed by a mine-spill (in Ap il 1998) ha pollu ed he soil wi h 152
ace elemen s. A e he spill, he soil was cleaned up, emedia ed and a o es ed wi h na i e 153
species o sh ubs and ees in mixed pa e ns o simula e a di e se o es (Madejón e al. 154
2018b). In a plo o abou 14 ha (37º 23.165´ N, 6º 13.668´ W) wi hin he emedia ed a ea we 155
andomly selec ed i e eplica es o se en ee species (35 ee samples in o al), wi h an 156
a e age dis ance o mo e han 100 me es be ween eplica es o he same species, esembling 157
a “common-ga den expe imen ”. The a ea was a o es ed in 2000, using seedlings (1-2 yea s 158
old) g own in a nea by nu se y. The ee species we e selec ed o his s udy acco ding o 159
hei con as ing lea habi s: deciduous species (Populus alba L., Cel is aus alis L. and 160
F axinus angus i olia Vahl) and e e g een species (Que cus ilex subsp. ballo a (Des .) 161
Samp., Olea eu opaea subsp. eu opaea a . syl es is (Mill.) Leh , Ce a onia siliqua L. and 162
Pinus pinea L.); he ea e we use only he genus name o simplici y. 163
The soil in he plo is o he Flu isol ype, being acidic (pH below 5) and nu ien -164
poo , wi h a loamy ex u e (abou 20% sand). In he spill-a ec ed and emedia ed soils he 165
esidual con amina ion by ace elemen s such as As, Cd, Cu, Pb and Zn was s ill high du ing 166
he s udy (16 yea s a e he spill). Howe e , he e was a low ans e a e o ace elemen s o 167
he abo eg ound pa s o he woody plan s (Domínguez e al. 2008; Madejón e al. 2018a, 168
2018b). 169
170
T ai measu emen s 171
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Roo s we e sampled (in la e au umn, Decembe 2014) a he indi idual ee le el, by 172
exca a ing he i s 20–30 cm o he soil adjacen o he ee unk base. We selec ed he ine 173
oo s (< 2 mm in diame e ) o he ai analysis. Al hough ine oo s a e composed by 174
abso p i e and anspo oo s (McCo mack e al. 2015), we assume ha , gi en he small 175
diame e o he selec ed oo s ( ange om 0.35 o 0.53 mm), mos o hem should be 176
abso p i e. 177
In he selec ed ine oo s we measu ed he ollowing mo phological ai s: speci ic oo 178
leng h (SRL, oo leng h pe uni o oo d y mass, m g-1), oo mass pe a ea (RMA, oo 179
mass pe uni o oo a ea, g m-2), oo mean diame e (RDI, mm), oo d y ma e con en 180
(RDMC, oo d y mass pe uni o wa e -sa u a ed esh mass, mg g-1) and oo issue mass 181
densi y (RTD, oo d y mass pe uni o oo olume, mg cm-3). We ollowed me hods in 182
Pé ez-Ha guindeguy e al. (2013) and de la Ri a e al. (2016a) o cha ac e ise hese a iables. 183
Howe e , in his s udy, we adop ed he ai RMA as an analogue o lea LMA, being he key 184
unc ional ai o oo s (see he a gumen s in a ou o using LMA in Poo e e al. 2009 and 185
de la Ri a e al. 2018b). The oo s we e scanned wi h an EPSON Pe ec ion V700 pho o 186
scanne a 1200 dpi. The leng h, diame e , a ea and olume o he oo s we e ob ained by 187
analysing he scanned oo samples wi h WinRHIZO 2009 so wa e (Regen Ins umen s Inc., 188
Quebec, Canada). The myco hizal ype associa ed wi h each ee species was assigned 189
acco ding o se e al sou ces (Ma emmani e al. 2003; Manau e al. 2015; Na a o-Fe nández 190
e al. 2016). We assigned he “ec omyco hizal ype” (ECM) o hose ee species (Pinus, 191
Populus and Que cus) ha o m symbio ic associa ions p edominan ly wi h ECM ungi, 192
al hough hey can also associa e wi h a buscula myco hizal (AM) ungi. In con as , ees o 193
he “a buscula myco hizal ype” (AM) a e exclusi ely associa ed wi h AM ungi (see Table 194
S1). 195
9

In he selec ed ees, mo phological ai s o ully-expanded lea es we e eco ded 196
ollowing he me hods o Pé ez-Ha guindeguy e al. (2013). Young, ully expanded lea es 197
s ill a ached o a po ion o s em o he p e ious yea we e collec ed om each indi idual 198
ee. These s ems wi h lea es we e s o ed in plas ic bags o p e en wa e loss and anspo ed 199
o he labo a o y, whe e hey we e main ained wi h he basal po ion o he s em subme ged in 200
wa e a 10 ºC o 24 h, in da kness, o allow comple e e-hyd a ion (de la Ri a e al. 2016a). 201
They we e sampled in ea ly au umn (Oc obe 2014), when we expec hem o ha e hei 202
maximum concen a ions o chemical elemen s (Madejón e al. 2004, 2006; Domínguez e al. 203
2008). We measu ed he lea mass pe a ea (LMA, lea d y mass pe uni o a ea, g m−2) and 204
lea d y ma e con en (LDMC, d y mass pe uni o wa e -sa u a ed esh mass; mg g-1). We 205
also measu ed he s em d y ma e con en (SDMC, d y mass pe uni o wa e -sa u a ed esh 206
mass; mg g-1) and s em wood densi y (SWD, d y mass di ided by he s em esh olume; mg 207
cm-3; based on he A chimedes p inciple, measu ing he olume o wa e displaced by 208
imme sion o he s em) in he sampled b anches and wigs. One o he F axinus ees su e ed 209
summe de olia ion and only had young lea es; he e o e, i was excluded om he lea ai s 210
da ase (n=34). 211
A subsample o he oo s and lea es collec ed om each ee was d ied and hen 212
g ound using a s ainless s eel mill, o chemical analyses. The N and C concen a ions and he 213
iso opic a ios o ni ogen (δ15N) and ca bon (δ13C) we e de e mined, in lea and oo samples 214
combus ed a 1020 ºC, using a con inuous low iso ope- a io mass spec ome y sys em. This 215
in ol ed a Flash HT Plus elemen al analyse coupled o a Del a-V Ad an age iso ope- a io 216
mass spec ome e ia a CONFLO IV in e ace (The mo Fishe Scien i ic, B emen, 217
Ge many); he analy ical measu emen e o s we e ± 0.2‰ o δ15N and ± 0.1‰ o δ13C. The 218
concen a ions o mac o- (P, K, Ca, Mg, S) and mic onu ien s (B, Co, Cu, Fe, Mn, Na, Ni, 219
Zn), as well as non-essen ial elemen s (As, Ba, Cd, Pb, S ), we e de e mined a e we 220
16

syl es is ha hicke oo s ha e low densi ies due o a hicke co ex, hus making hem less 371
cos ly o cons uc and mo e sui able o associa ion wi h myco hizal ungi, and enhancing 372
nu ien acquisi ion. Also, Kong e al. (2019) ound nonlinea oo ai ela ionships be ween 373
RDI, RTD and SRL, which can explain why SRL does no necessa ily con o m o he RTD-374
ela ed plan economics spec um in woody species. 375
The key mo phological ai s RMA and SRL, which a e indica o s o he oo up ake 376
po en ial, co ela ed wi h o he mo phological ai s and wi h some majo nu ien s (P, K, Ca 377
and Mg), which suppo s he up ake unc ion o hese ai s. Phospho us and K each he oo s 378
mainly by di usion om he bulk soil o he oo su ace (Lambe s e al. 2008), and he e o e 379
a nega i e co ela ion be ween hei up ake and RMA (bu a posi i e one wi h SRL) would be 380
expec ed. The co ela ion be ween RMA and oo nu ien s (o he han N) has been 381
o e looked p e iously, and his s udy is a ele an con ibu ion o suppo he RES. Despi e 382
he ac ha he e a e con adic o y esul s conce ning he SRL – oo N ela ionship (see 383
e iews in Reich 2014 and Weems a e al. 2016), based on ou da a we can concu wi h 384
Reich (2014) ha he RES exis s, al hough no as uni o mly and s ongly coo dina ed as he 385
LES. 386
This e idence suppo ing he RES as a main oo dimension does no p eclude he 387
exis ence o o he oo dimensions ep esen ing he mul i unc ionali y o oo s (Weems a e 388
al. 2016; Lalibe é 2017). 389
Fi s ly, in his s udy, oo C concen a ion was no co ela ed wi h he mo phological 390
oo ai s, bu i was nega i ely co ela ed wi h he concen a ion o oo N and 12 o he 391
chemical elemen s. Roo C concen a ion, oge he wi h oo b anching ai s (no measu ed 392
he e), de ined he second dimension in he PCA o 14 oo ai s o 96 woody species om 393
sub opical o es s in China (Kong e al. 2014). 394

17

Secondly, soil esou ces a e mul iple (wa e and nu ien s) and plan oo s di e in 395
hei up ake s a egies, such as myco hizas, N2- ixing symbioses, and P-abso bing clus e 396
oo s (Lambe s e al. 2008). The mic onu ien s Cu, Mn and Ni and he non-essen ial elemen 397
Pb exhibi ed a end ha was o hogonal o ha o he RES (Fig. 2). In pa icula , he Pb 398
concen a ion in oo s was co ela ed nega i ely wi h RMA and oo C, bu posi i ely wi h 13 399
o he elemen s. Among he oo ai s which con e imp o ed ole ance o ele a ed me al 400
concen a ions in soils is he abili y o bind ace elemen s o oo cell walls and accumula e 401
hem belowg ound; in his way, oo s may be ba ie s impeding he up ake o po en ially oxic 402
elemen s and hei ansloca ion o he lea es (Lambe s e al. 2008, Zhao e al. 2016). This 403
addi ional oo dimension ha con e s me al ole ance may be e y impo an o plan i ness 404
in me al- ich en i onmen s, like he s udy si e. 405
Thi dly, he N iso ope composi ion (δ15N) has been used o in e symbio ic up ake o 406
N by myco hizal ungi and i s ans e o plan s, due o he disc imina ion agains hea ie 15N 407
in hese p ocesses (Hobbie and Hobbie 2008; Hobbie and Högbe g 2012). In his s udy, oo 408
δ15N was a ela i ely-independen ai ; i was co ela ed nega i ely wi h he oo C 409
concen a ion and δ13C bu he e was no ela ionship wi h mo phological o chemical ai s 410
(wi h he excep ion o oo Co and Mn). Recen ly, Lalibe é (2017) has sugges ed he use o 411
he N iso ope composi ion in plan s as a ime-in eg a ed ai showing he myco hizal 412
in luence on N acquisi ion. Thus, we would expec lowe δ15N alues in he oo s o ees 413
associa ed wi h ECM ungi, which disc imina e agains 15N and p e e en ially ans e 14N o 414
hei hos plan s; while no o only sligh deple ion o 15N is expec ed o AM plan s (Hobbie 415
and Högbe g 2012; C aine e al. 2015). Howe e , we ound ha he δ15N alues in oo s o 416
ECM ees (mean=1.61, n=15) we e no di e en om hose in AM ype ees (mean=1.40, 417
n=20). The na u al abundance o 15N in plan s is no easy o in e p e because i is a single 418
18

esponse a iable wi h mul iple d i e s (i.e. clima e, myco hizal ungi, and mic obial 419
p ocessing; C aine e al. 2015). 420
421
Coo dina ion o oo and lea ai s 422
Fas plan g ow h depends on he coo dina ion o oo s and lea es, wi h he o me 423
p o iding enough wa e and nu ien s supply o main ain acquisi i e lea es wi h high 424
pho osyn he ic a es and high e apo a i e demand (Reich 2014). In gene al, we ound ha he 425
main oo a ia ion end (PCA axis 1) was signi ican ly co ela ed wi h he co esponding 426
lea a ia ion end (Fig. 4), suppo ing he exis ence o a plan economics spec um (Pé ez-427
Ramos e al. 2012, de la Ri a e al. 2016b, 2018a). 428
In pa icula , we ound signi ican co ela ions be ween mo phological oo ai s 429
(RMA and RDMC) and he analogous lea ai s (LMA and LMDC), suppo ing such oo -430
lea coo dina ion, as epo ed in o he s udies (Holdaway e al. 2011; de la Ri a e al. 2018a). 431
Howe e , he e a e excep ions: o ins ance, La ix decidua ees display acquisi i e lea ai s, 432
ypical o deciduous ees, bu conse a i e oo ai s, ypical o coni e s (Wi hing on e al. 433
2006; Weems a e al. 2016). 434
In his s udy, he C concen a ions in oo s and lea es we e posi i ely co ela ed. 435
Villa e al. (2006) also ound a posi i e co ela ion be ween oo and lea C in 16 woody 436
species. The C concen a ion is no mally high o species wi h s ong s uc u al de ences 437
(such as lignin o cellulose) (Poo e and Villa 1997) and, he e o e, wi h a conse a i e 438
s a egy (de la Ri a e al. 2016b). On he o he hand, he e was no a signi ican ela ionship 439
be ween RMA and he oo C concen a ion, al hough RMA was posi i ely co ela ed wi h he 440
C:N a io ( =0.55, p=0.001). The C:N a io e lec s he ela i e in es men s in s uc u e 441
(mainly ca bon) espec o cell me abolism (indica ed by ni ogen). Thus, plan o gans wi h a 442
19

highe C:N a io ep esen a conse a i e s a egy (Villa e al. 2006; de la Ri a e al. 2016c, 443
2018b). 444
Ou esul s do no i he p e iously- epo ed global end o oo and lea N 445
concen a ions, which a e highly co ela ed in woody species (n=89, =0.58, p<0.001; 446
Val e de-Ba an es e al. 2017). T ees wi h N- ich lea es and high pho osyn he ic a es a e 447
expec ed o ha e N- ich and exploi a i e ine oo s (Reich 2014). Howe e , his end can be 448
in luenced by he speci ic symbiosis ( ype o myco hiza) p esen . In his s udy, we ound a 449
signi ican co ela ion only when analysing he subse o ECM ee species ( =0.72, p=0.002). 450
Howe e , ee species in a symbiosis wi h AM ungi did no show his oo -lea N 451
ela ionship. The wo ypes o myco hizal ees ha e di e en nu ien economies: ECM ees 452
a e able o acqui e N om he soil o ganic ma e due o he g ea e enzyma ic capabili ies o 453
ECM ungi, while AM ees depend mos ly on ino ganic N (Phillips e al. 2013). In he N-454
limi ing condi ions o he s udy si e, we expec ECM ees o be mo e e icien a aking up 455
soil N, h ough he oo - ungi symbiosis, and ansloca ing i o hei lea es. Kong e al. 456
(2019) also ound di e en ela ionships o oo ai s depending o myco hizal ypes (ECM 457
e sus AM). Thus, o ECM species, hin oo s we e ela ed wi h highe oo N concen a ion, 458
bu he con a y o AM species. This could explain he posi i e ela ionship ound in ou 459
s udy be ween oo N and lea N only o ECM species. 460
The C iso ope composi ion in lea issues is widely used as a unc ional ai 461
ep esen ing he ime-in eg a ed measu emen o wa e -use e iciency. I is based on he 462
disc imina ion by pho osyn he ic enzymes agains he hea ie iso ope 13C du ing 463
pho osyn hesis, and depends on he a io be ween he in e nal and ai CO2 concen a ions, in 464
u n egula ed by s oma al opening (Seibd e al. 2008, Pé ez-Ha guindeguy e al. 2013). The 465
δ13C alues in oo s should e lec he iso opic signa u e o he ca bohyd a es syn hesised in 466
he lea es, al hough du ing he lea - oo ansloca ion some 13C en ichmen in oo s ( ela i e 467
20

o lea es) has been obse ed (Ce nusak e al. 2009). In his s udy, he δ13C alues in oo s and 468
lea es we e posi i ely co ela ed (Table 1), indica ing oo -lea coo dina ion. In gene al, long-469
li ed issues a e associa ed wi h a mo e-conse a i e use o esou ces and a highe e iciency 470
in wa e -use, usually e lec ed in hei highe δ13C alues (Reich 2014, de la Ri a e al. 471
2016b). Howe e , in his case, he δ13C alues in oo s o deciduous and e e g een ees we e 472
no di e en (Table S4). 473
The plan N iso ope composi ion e lec s mainly he soil sou ce o N, and also any 474
iso ope ac iona ion and N pool mixing (Robinson 2001). Al hough he oo alues o δ15N 475
did no show signi ican di e ences among species, when analysing he in a-plan 476
ac iona ion (i.e. δ15N oo – δ
15Nlea ) he e we e signi ican di e ences among species and 477
myco hizal ypes (Fig. S5). The deple ion o 15N in ECM ees may be ela ed o he 478
p e e en ial e en ion o 15N by he ECM ungal biomass (bu no by ha o AM ungi) and 479
he consequen ans e o 15N-deple ed N o he hos ees (C aine e al. 2015). 480
Nu i ional di e ences among ee species esul om he unc ional di e si y in 481
mechanisms o nu ien up ake om soil, nu ien equi emen s and long- e m nu ien use 482
e iciency (Lambe s e al. 2008). The coo dina ed a iabili y in P, Ca and Mg concen a ions 483
be ween oo s and lea es indica es ha hese nu ien s a e unde biological con ol, due o 484
hei impo ance o plan g ow h (Newman and Ha 2006; Geng e al. 2014; Zhao e al. 485
2016). 486
In con as , mos o he ace elemen s had a s ong disco dance be ween hei 487
concen a ions in oo s and lea es. The excess up ake o non-limi ing elemen s seems poo ly 488
egula ed by plan s, and he e o e hey exhibi high a iabili y (Ladanai e al. 2010). Plan s 489
end o accumula e ace elemen s in oo s, binding hem o cell walls as a de oxi ica ion 490
mechanism (Domínguez e al. 2009; Kaba a-Pendias 2011; Zhao e al. 2016). Howe e , some 491
ee species ha e a selec i e up ake and anspo o ce ain ace elemen s, accumula ing 492
21

hem in lea issues. No able examples a e he accumula ion o Cd and Zn in Populus lea es 493
(Madejón e al. 2004) and he accumula ion o Mn in Que cus lea es (Madejón e al. 2006), 494
bu no in hei oo s (Fig. S4). In soils con amina ed by ace elemen s, he adequa e selec ion 495
o plan species o phy os abilisa ion is essen ial. One o he main c i e ia is ha he selec ed 496
ee species con ol he mobili y o he ace elemen s, keeping hei oo o shoo 497
ansloca ion ac o s as low as possible, o a oid oxici y isks in he ophic web (Mendez and 498
Maie 2008; Bolan e al. 2011; Madejón e al. 2018b). 499
500
Roo ai s and soil condi ions 501
The e a e ecip ocal in e ac ions and eedbacks be ween oo s and soil. The soil 502
condi ions in luence oo ai s and plas ici y (Ba dge e al. 2014). In u n, oo s modi y he 503
hizosphe ic soil; o example h ough oo exuda es o inc ease nu ien up ake (Dako a and 504
Phillips 2002). He e, soil pH was signi ican ly ela ed o he oo mo phological ai s. On he 505
one hand, his indica es ha oo s wi h ai s indica i e o lowe explo a ion (highe RMA o 506
lowe SRL, as in Pinus and Que cus) could compensa e wi h highe p oduc ion o acid 507
exuda es o p omo e nu ien up ake, dec easing he soil pH (Dako a and Phillips 2002). On 508
he o he hand, his ela ionship be ween soil acidi y and a conse a i e oo s a egy could be 509
linked o he e ec s o he li e compounds o hese species (wi h a high C:N a io and high 510
LMA) on soil. The accumula ion o li e wi h a high C:N a io, such as ha o coni e ous 511
species, ends o ha e an acidi ying e ec on soil (Augus o e al. 1998; Sa iyildiz e al. 2005; 512
Alameda e al. 2012). As a consequence, soil pH usually dec eases a e he a o es a ion o 513
g asslands o o me ag icul u al lands wi h coni e ous species (Jug e al. 1999; Saue e al. 514
2007; Be h ong e al. 2012). The analysis o he amoun and quali y o oo exuda es would 515
be needed o elucida e he causes behind he obse ed ela ionship be ween soil pH and RMA, 516
besides he indi ec e ec s o li e ai s on soil chemis y. In any case, as sugges ed by 517

22

Lalibe é (2017), i would be wo h including oo exuda ion as a physiological ai o 518
ad ance in ai -based plan ecology. 519
In me al- ich soils low pH usually leads o a highe solubili y o hese elemen s and 520
he e o e o a high a ailabili y o oo s. In his ace-elemen pollu ed si e, we obse ed some 521
signi ican ela ionships be ween soil me al con en and some oo ai s; in pa icula , RDMC 522
was posi i ely co ela ed o he soil con en o Mn, Ni and Cd. One o he i s symp oms o 523
plan oxici y o soil me als is he inhibi ion o oo elonga ion (Kahle 1993; Wisniewski and 524
Dickinson 2003). O he esponses o me al oxici y a e: collapsing o oo hai s, inc emen s o 525
sube i ica ion and ligni ica ion, dec ease o essel diame e and s uc u al al e a ions o 526
hypode mis and endode mis (A duini e al. 1994; Ba celó and Poschen iede 2004). 527
Expe imen al exposu e o Que cus ilex oo s o Cd esul ed in a decline in ine oo 528
p oduc ion and in a educ ion in he leng h o ap oo s (Domínguez e al. 2009), linked o a 529
high capaci y o e ain Cd a he oo le el, likely by binding Cd o cell wall pec ins. Thus, he 530
links be ween pH, soil me al con en , and RDMC obse ed in his s udy could also indica e a 531
end owa ds a mo e conse a i e s a egy a he oo le el o p omo e he immobiliza ion o 532
hese me als in he hizosphe e, a oiding hei ansloca ion o he abo eg ound biomass. 533
The oo chemical ai s we e ela ed o he a ailabili y in he soil o some nu ien s 534
(K, Mn, Na and Zn), as expec ed. Howe e , he concen a ions o many o he elemen s in he 535
oo s we e ela i ely independen o he soil condi ions; his weak coupling be ween he soil 536
and plan concen a ions o chemical elemen s has been ound in o he s udies (Ladanai e al. 537
2010; Zhao e al. 2016). The up ake and accumula ion o nu ien s in oo s is a complex 538
p ocess which depends on nume ous ac o s - such as he ela i e alloca ion wi hin he plan , 539
he de elopmen al s age, he plan species and he en i onmen al condi ions (Lambe s e al. 540
2008). Mo e esea ch is needed o unde s and how hose ac o s a ec he ans e o ace 541
elemen s om he soil o he oo s. 542
23

A pa icula ly-in e es ing oo physiological ai is he po en ial o educe me al 543
a ailabili y in soil, by se e al mechanisms like p ecipi a ion o me als, hei complexa ion 544
wi h o ganic p oduc s, hei so p ion on o oo su aces o hei accumula ion inside oo 545
issues (Mendez and Maie 2008). The plan ing o ee species wi h highe phy os abilisa ion 546
po en ial would imp o e and emedia e me al-con amina ed soils (Madejón e al. 2018b). In 547
ac , one o he c i e ia used o selec he bes -sui ed ee species is o ha e a high 548
bioconcen a ion ac o ( oo :soil a io) o di e en me als, in pa icula o hose wi h 549
ha m ul e ec s (i.e. Cd and Pb) (Madejón e al. 2018b). 550
551
Conclusion 552
The e is inc easing in e es in ad ancing ou knowledge abou oo ai s because o hei 553
o en-o e looked bu essen ial con ibu ion o plan unc ional ecology. Ou esul s ein o ce 554
he exis ence o a oo economics spec um (RES) as he main de e minan o ine oo ai s 555
in Medi e anean ees, e en in soil con amina ed by hea y me als. Howe e , his s udy also 556
suppo s he idea o oo mul i unc ionali y and he impo ance o ine oo dimensions 557
independen o he RES; namely, oo ca bon concen a ion, ac iona ion o ni ogen iso opes 558
as a ime-in eg a ed ai o myco hizal-media ed nu i ion, and he abili y o bind ace 559
elemen s in oo cells (associa ed wi h ole ance o high le els o me als in soils). We ound 560
ha oo s and lea es we e unc ionally coo dina ed; howe e , mos o ace elemen s showed 561
s ong oo -lea disco dance. We also ound links be ween soil pH, soil me al con en , and 562
oo ai s (RDMC) p omo ing he immobiliza ion o me als in he hizosphe e. In summa y, 563
he hizosphe e is a complex en i onmen whe e soil, oo s and mic oo ganisms in e ac in 564
eedback p ocesses. An unde s anding o he mul i unc ionali y o oo ai s would help us o 565
p edic he o es esponses o global changes and he p o ision o soil-based ecosys em 566
se ices. 567
24

568
Acknowledgemen s 569
This wo k was inancially suppo ed by he Eu opean Union Se en h F amewo k 570
P og amme (FP7/2007–2013) (G an No. 603498- RECARE), he Spanish Minis y o 571
Science, Inno a ion and Uni e si ies (G an s No. CGL2014-52858-R-RESTECO, CGL2017-572
82254-R-INTARSU, and CGL2014-53236-R- ECO-MEDIT), and Eu opean FEDER unds. 573
MG-M was suppo ed by he Spanish Minis y o Economy and Compe i i eness (G an No. 574
BES-2015-073882), and MTD by he Uni e sidad de Se illa (Con a o de Acceso, V Plan 575
P opio de In es igación). We hank J. M. Mu illo and J.M. Aleg e o hei help in he ield 576
wo k, he IRNAS Analy ical Se ice o mul ielemen analyses o plan s and soil, and he 577
EBD-CSIC Labo a o y o S able Iso opes o de e mina ions o δ15N and δ13C. 578
579
Con lic o in e es 580
The au ho s decla e ha hey ha e no con lic o in e es . 581
582
Re e ences 583
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35

Table 1. Co ela ions be ween oo ai s and analogous lea ai s (n=34), and be ween oo
chemical ai s and soil a ailabili y o elemen s (n=35). Soil a ailabili y (CaCl2 ex ac ed) o
As and Pb we e below de ec able limi s. Pea son´s es coe icien and p alues a e
indica ed; signi ican alues (p<0.05) a e in bold. RMA: oo mass pe a ea; LMA: lea mass
pe a ea; LDMC: lea d y ma e con en ; RDMC: oo d y ma e con en .
Roo -lea Roo -soil
T ai p p
RMA/LMA 0.548 0.001 - -
LDMC/RDMC 0.371 0.031 - -
C 0.553 0.001 -0.110 0.528
N 0.018 0.920 -0.020 0.910
P 0.486 0.004 -0.111 0.526
K
0.289 0.098 0.379 0.025
Ca 0.826 <0.001 0.166 0.340
M
g
0.393 0.021 0.163 0.348
B 0.075 0.675 0.025 0.887
Co -0.250 0.154 0.380 0.024
Cu 0.087 0.623 -0.002 0.990
Fe 0.131 0.461 -0.073 0.678
Mn -0.023 0.897 0.357 0.035
Na 0.051 0.775 0.596 <0.001
Ni -0.070 0.695 -0.031 0.861
S 0.023 0.899 0.134 0.441
Zn -0.093 0.602 0.346 0.042
As -0.001 0.998 - -
Ba 0.516 0.002 -0.028 0.872
C
d
0.019 0.914 0.071 0.686
Pb 0.331 0.056 - -
S
0.774 <0.001 0.381 0.024
δ13C 0.391 0.022 -0.174 0.317
δ15N 0.165 0.351 0.437 0.009
36

Figu e legends
Figu e 1. Va ia ion among ee species in oo mass pe a ea (RMA, g m-2). Mean and SE
(n=5) ba s a e shown; le e s indica e signi ican di e ences be ween he ee species
(Tukey´s pos -hoc es ). Deciduous species a e ma ked in g ey and e e g een species in black.
Figu e 2. Resul s o he p incipal componen analysis o 27 oo ai s o se en ee species
(n=35). Sco es o ai a iables and ee samples a e ep esen ed in he plane de ined by i s
(PC1) and second (PC2) axes. Abb e ia ions o oo ai s a e: RMA: oo mass pe a ea;
RDMC: oo d y ma e con en , RDI: oo mean diame e ; RTD: oo issue densi y; SRL:
speci ic oo leng h; and o ee species names a e: C.a.: Cel is aus alis; C.s.: Ce a onia
siliqua; F.a.: F axinus angus i olia; O.e.: Olea eu opaea; P.a.: Populus alba; P.p.: Pinus
pinea; Q.i.: Que cus ilex. Symbol ills a e in g ey o deciduous and in black o e e g een
species.
Figu e 3. Co elog am ac oss mo phological and chemical oo ai s, o de ed acco ding o
hei co ela ion coe icien s. The s eng h and di ec ion o he co ela ions a e indica ed by
he ci cle size and he colou , shown in he igh side scale. RTD: oo issue densi y; RDMC:
oo d y ma e con en ; RMA: oo mass pe a ea; RDI: oo mean diame e ; SRL: speci ic
oo leng h.
Figu e 4. Co ela ion analysis be ween he main a ia ion ends (PCA axis 1 sco es) in lea
and oo , compa ing 24 analogous ai s ( =0.59, p=0.0003). Abb e ia ions o ee species
names a e: C.a.: Cel is aus alis; C.s.: Ce a onia siliqua; F.a.: F axinus angus i olia; O.e.:
Olea eu opaea; P.a.: Populus alba; P.p.: Pinus pinea; Q.i.: Que cus ilex. Symbol ills a e in
g ey o deciduous and in black o e e g een species.
37

Figu e 1

38

Figu e 2
39

Figu e 3
40

Figu e 4
41

7

Table S2. Resul s o he p incipal componen analyses (PCA) o he six key oo
ai s (see o dina ion in Figu e S2) and o all 27 ai s (see Figu e 2), indica ing
a iance explained by he h ee main axes and s anda dized ac o loading o each
oo ai . The highes sco es (in absolu e alue) o each axis a e ma ked in bold.
See main ex o abb e ia ions o ai names.
6 key ai s PCA All- ai s PCA
T ai s Axis 1
(57.3%)
Axis 2
(23.4%) Axis 3
(14.7%) Axis 1
(37.6%) Axis 2
(18.2%) Axis 3
(11.0%)
RMA 0.97 -0.06 0.05 0.61 0.61 0.13
SRL -0.87 -0.42 -0.04 -0.49 -0.58 -0.23
RDMC 0.87 -0.28 0.23 0.56 0.71 -0.01
RTD 0.60 -0.76 0.16 0.43 0.42 -0.31
RDI 0.68 0.71 -0.06 0.36 0.40 0.45
N -0.38 0.25 0.89 -0.65 0.15 -0.26
C 0.74 -0.53 0.34
Ca -0.62 0.07 -0.62
Mg -0.88 -0.23 0.09
K -0.35 -0.70 0.17
S -0.70 0.14 -0.05
P -0.45 -0.51 -0.11
As -0.86 0.31 0.04
B -0.31 -0.72 -0.12
Ba -0.69 0.24 -0.09
Cd -0.28 -0.31 0.69
Co -0.70 0.38 0.50
Cu -0.82 0.21 0.30
Fe -0.88 0.29 0.06
Mn -0.66 0.32 0.37
Na -0.27 -0.62 0.51
Ni -0.73 0.45 -0.08
Pb -0.88 0.29 -0.04
S -0.57 -0.01 -0.61
Zn -0.67 -0.10 0.40
δ13C -0.14 -0.35 -0.41
δ15N -0.17 0.54 0.21

8

Table S3. Mean alues and SE (n=5) o abo eg ound ai s o he s udied ee species (excep n=4 o lea ai s o F axinus). F-s a is ics om
one-way ANOVA es o Chi-squa e- alue om K uskal Wallis es (ma ked wi h Ksupe s ip ) a e shown, depending on da a no mali y and
homoscedas ici y. Signi ican le el is p < 0.05 (in bold). LDMC: lea d y ma e con en ; LMA: lea mass pe a ea; RDI: oo mean diame e ; SRL:
speci ic oo leng h; SDMC: s em d y ma e con en ; SWD: s em wood densi y; HEI: ee heigh ; CRP: c own p ojec ion a ea; LITT: li e
accumula ion on soil su ace.

9

Table S4. Compa ison be ween oo ai s o ees, acco ding o hei lea habi (deciduous o
e e g een); ai uni s a e like in Table S1. They ha e been anked by he signi icance le el
(ANOVA´s F and p) ma king he di e ence be ween lea habi s. Signi ican le el is p < 0.05
(in bold). SRL: speci ic oo leng h; RDI: oo mean diame e ; RMA: oo mass a ea; RDMC:
oo d y ma e con en ; RTD: oo issue densi y.
Roo ai Deciduous
(n=15)
E e g een
(n=20)
ANOVA
s a is ics
Mean SE Mean SE F p
B 20.21 1.29 11.77 0.58 42.4 <0.001
P 1054.3 53.2 680.5 32.2 40.0 <0.001
SRL 21.76 2.05 10.62 0.65 33.7 <0.001
RDI 0.37 0.02 0.50 0.01 27.9 <0.001
RMA 43.20 2.08 63.89 3.34 23.2 <0.001
RDMC 267.3 16.6 354.6 13.2 17.4 <0.001
Ca 18329.2 2619.7 7613.8 1391.3 14.9 <0.001
Mg 1835.4 99.7 1224.5 121.6 13.7 <0.001
S 34.15 4.10 15.29 3.17 13.7 <0.001
K 8583.7 865.9 5983.3 384.9 8.9 0.005
Zn 163.4 17.3 110.1 9.80 8.1 0.008
Pb 42.28 8.72 20.10 3.36 6.9 0.013
As 13.15 2.97 6.62 1.10 5.2 0.029
S 2419.9 237.4 1652.9 236.7 5.0 0.032
Fe 3739.8 696.0 2208.6 339.8 4.6 0.040
Ba 13.57 1.50 10.17 0.92 4.1 0.051
Ni 9.11 2.14 5.49 0.57 3.4 0.073
N 1.20 0.14 0.92 0.09 3.0 0.092
C 42.9 1.17 44.6 0.36 2.7 0.111
Cu 112.2 12.4 85.8 11.9 2.3 0.141
Mn 102.8 15.3 78.3 8.4 2.2 0.143
Cd 1.55 0.23 1.16 0.14 2.2 0.148
Na 786.5 172.5 520.5 93.0 2.1 0.157
RTD 474.3 25.3 511.5 24.4 1.1 0.305
δ13C -26.2 0.19 -26.6 0.25 1.0 0.314
Co 2.35 0.4 1.99 0.5 0.6 0.444
δ15N 1.40 0.2 1.55 0.2 0.2 0.664
10

Table S5. Mean alues and SE (n=5) o opsoil pa ame e s (0-10cm dep h) associa ed o he s udied ee species, and adjacen open si es,
o compa ison; pH, o ganic C, o al N, a ailable concen a ions o nu ien s and ace elemen s, and C and N iso ope a ios. F-s a is ics om
one-way ANOVA es o Chi-squa e- alue om K uskal Wallis es (ma ked wi h Ksupe sc ip ) a e indica ed, depending on da a no mali y and
homoscedas ici y (only soil samples unde ees we e compa ed). Signi ican le el is p < 0.05 (in bold).
11

Table S6. Co ela ions o oo mo phological ai s wi h soil chemical a iables (pH,
o ganic C, o al N, a ailable concen a ions o nu ien s and ace elemen s, and C
and N iso ope a ios). Signi ican alues (p<0.05) a e in bold. RMA: oo mass pe
a ea; SRL: speci ic oo leng h; RDI: oo mean diame e ; RDMC: oo d y ma e
con en ; RTD: oo issue densi y.
Soil
p ope y
Mo pho unc ional oo ai
RMA SRL RDI RDMC RTD
p p p p p
pH -0.51 0.002 0.35 0.040 -0.26 0.130 -0.57 <0.001 -0.42 0.013
C -0.33 0.052 0.22 0.210 -0.07 0.690 -0.35 0.040 -0.35 0.041
N -0.27 0.114 0.22 0.205 -0.07 0.704 -0.32 0.059 -0.30 0.076
P -0.16 0.361 0.18 0.303 -0.22 0.194 0.09 0.597 0.03 0.856
K -0.28 0.101 0.12 0.490 0.01 0.954 -0.41 0.014 -0.35 0.037
Ca -0.32 0.061 0.32 0.061 -0.24 0.161 -0.20 0.252 -0.20 0.253
Mg -0.40 0.016 0.27 0.123 -0.12 0.494 -0.29 0.095 -0.37 0.027
S 0.08 0.637 0.05 0.782 -0.14 0.420 0.32 0.058 0.25 0.142
B -0.21 0.235 0.11 0.537 -0.13 0.473 -0.12 0.484 -0.09 0.600
Ba 0.13 0.451 -0.22 0.197 0.25 0.147 0.12 0.485 -0.04 0.834
Cd 0.26 0.127 -0.11 0.519 0.07 0.691 0.36 0.036 0.30 0.081
Co 0.13 0.459 -0.04 0.799 0.003 0.985 0.32 0.061 0.17 0.325
Cu 0.04 0.814 0.06 0.727 -0.10 0.552 0.33 0.052 0.16 0.354
Fe -0.11 0.526 0.15 0.384 -0.22 0.211 0.19 0.261 0.08 0.632
Mn 0.28 0.100 -0.15 0.396 0.09 0.600 0.43 0.009 0.29 0.090
Na -0.17 0.337 -0.01 0.987 0.16 0.359 -0.41 0.015 -0.38 0.026
Ni 0.32 0.060 -0.15 0.389 0.10 0.587 0.45 0.006 0.32 0.057
S -0.38 0.023 0.23 0.178 -0.15 0.379 -0.36 0.036 -0.33 0.056
Zn 0.08 0.661 0.01 0.934 -0.07 0.683 0.31 0.067 0.19 0.271
δ13C 0.07 0.688 -0.04 0.825 -0.004 0.982 0.21 0.235 0.06 0.749
δ15N -0.39 0.020 0.20 0.262 -0.11 0.545 -0.19 0.271 -0.37 0.027
12

Figu e S1. Rank o ela i e a ia ion in oo ai s and hei analogue lea ai s,
measu ed as coe icien o a ia ion (CV in %), o se en species (n=35). R/LDMC:
oo o lea d y ma e con en ; R/LMA: oo o lea mass pe a ea; 13C: δ13C and
15N: δ15N.

13

Figu e S2. Resul s o he p incipal componen analysis o six key oo ai s in ees
o se en species (n=35). Abb e ia ions o oo ai s a e: RDI: oo mean diame e ;
RMA: oo mass pe a ea; RDMC: oo d y ma e con en ; RTD: oo issue densi y;
and SRL: speci ic oo leng h; species names a e: C.a.: Cel is aus alis; C.s.:
Ce a onia siliqua; F.a.: F axinus angus i olia; O.e.: Olea eu opaea; P.a.: Populus
alba; P.p.: Pinus pinea; Q.i.: Que cus ilex.
14

Figu e S3. Resul s o he p incipal componen analysis o 29 abo eg ound ai s in
ees o se en species (n=34). T ai s and ees a e o de ed in he plane de ined by
PCA i s and second axes. Abb e ia ion names o ee species a e: C.a.: Cel is
aus alis; C.s.: Ce a onia siliqua; F.a.: F axinus angus i olia; O.e.: Olea eu opaea;
P.a.: Populus alba; P.p.: Pinus pinea; Q.i.: Que cus ilex.
15

Figu e S4. Coo dina ion be ween oo and lea ai s in he h ee igu es o le
column, compa ed wi h disco dan ai s in he igh column (n=34). Co ela ions
be ween N in oo s and in lea es a e signi ican o Pinus ( ed diamond and solid line,
n=5) and Populus (blue iangles and dashed line, n=5) bu no o he o he species
and o all da a (black ci cles). Species-speci ic accumula ion o Cd in lea es o
Populus (blue iangles) and Mn in Que cus (g een squa es) a e shown. The alues
o co ela ion coe icien and signi icance a e shown in Table 1. LMA: lea mass pe
a ea; RMA: oo mass pe a ea.
16

Figu e S5. In a-plan ac iona ion o N iso ope (di e ence be ween δ15N oo and
δ15Nlea , in ‰), sepa a ing ec omyco hizal (whi e) and a buscula myco hizal (g ey)
ee species. Mean and SE (n=5, wi h he excep ion o F axinus n=4) a e shown;
di e en le e s mean signi ican di e ence be ween species by pos -hoc Tukey es .
The e a e signi ican di e ences be ween myco hizal ypes, =-4.4, p<0.0001.