ORIGINAL ARTICLE
Managing clima e change impac s on c ops: The in luence
o soil illage on a i icale c op unde wa e s ess condi ions
Paula Madejón
1
| Elena Fe nández-Boy
2
| Eng acia Madejón
1
|
Lau a Mo ales-Salme ón
2
| Ma ía Te esa Domínguez
2
1
Ins i u o de Recu sos Na u ales y
Ag obiología de Se illa (IRNAS), CSIC, Se illa,
Spain
2
Depa amen o de C is alog a ía, Mine alogía
y Química Ag ícola, Uni e sidad de Se illa,
Se illa, Spain
Co espondence
Paula Madejón, Ins i u o de Recu sos
Na u ales y Ag obiología de Se illa (IRNAS),
CSIC, Se illa, Spain.
Email: [email p o ec ed]c.es
Funding in o ma ion
Eu opean Regional De elopmen Fund
(FEDER), G an /Awa d Numbe : US-1260627;
Spanish Minis y o Science and Inno a ion,
G an /Awa d Numbe : PID2021-122628OB-
I00 WASTE4DROUGHT; Eu opean unds;
Minis y o Science and Inno a ion,
G an /Awa d Numbe : PRE2018-084467
Abs ac
Wa e limi a ions o ag icul u e will likely become c ucial in he nex decades in
some egions such as he Medi e anean basin wi h he cu en clima e change p o-
jec ions. In his con ex , ecen e idence sugges s ha he applica ion o conse a-
ion ag icul u e, which educes he equency and in ensi y o soil illage, could
con e a highe s abili y o ag icul u al sys ems agains clima e a iabili y. Howe e ,
no many expe imen s ha e add essed he in e ac ion be ween illage ype and he
esis ance o d ough in ain ed c ops. In his wo k, we e alua ed he esis ance o
d ough o i icale (T i icale hexaploide L.) c ops managed wi h di e en illage sys-
ems: adi ional illage (TT), educed illage (RT) and no illage (NT). A ain all exclu-
sion expe imen was ca ied ou in a ypical whea /legume Medi e anean o a ion in
SW Spain, in a long- e m expe imen es ablished in 2008 compa ing he h ee illage
sys ems. G ain yield and di e en a iables ela ed o plan ecophysiology, oo
de elopmen , biomass alloca ion and colonisa ion by a buscula myco hizal ungi
(AMF) we e e alua ed o e one c op cycle. Tillage ype had a signi ican in luence on
soil wa e s o age (SWS), such ha soils unde NT had, on a e age, a 16% g ea e
SWS han soils unde RT o TT. G ain yield was signi ican ly educed by ain all exclu-
sion, in pa icula in he TT, whe e d ough educed g ain yield by 31%. Gas exchange
da a also showed ha plan s in he TT sys em we e mo e sensi i e o d ough , such
ha maximum pho osyn hesis a es we e educed by 25% because o ain all exclu-
sion in his illage sys em. D ough had a nega i e impac on oo biomass ac oss he
h ee illage sys ems, especially in he RT, whe e a educ ion in he oo :shoo a io
was obse ed. The e ec o illage on myco hizal colonisa ion was mo e e iden
han he e ec o d ough ; in gene al, conse a ion illage sys ems (RT and NT)
ended o ha e highe alues o all AMF ai s compa ed o he TT. In summa y, he
NT sys em ended o exhibi mo e a ou able pe o mance in e ms o soil wa e
e en ion, g ain yield s abili y unde d ough condi ions and myco hizal symbiosis,
which sugges s enhanced esou ce use e iciency in his sys em.
KEYWORDS
d ough , ain ed c ops, ainou shel e s, soil wa e s o age, illage ypes, i icale
Recei ed: 27 Decembe 2023 Re ised: 9 Sep embe 2024 Accep ed: 10 Sep embe 2024
DOI: 10.1111/aab.12947
This is an open access a icle unde he e ms o he C ea i e Commons A ibu ion License, which pe mi s use, dis ibu ion and ep oduc ion in any medium,
p o ided he o iginal wo k is p ope ly ci ed.
© 2024 The Au ho (s). Annals o Applied Biology published by John Wiley & Sons L d on behal o Associa ion o Applied Biologis s.
Ann Appl Biol. 2025;186:143–156. wileyonlinelib a y.com/jou nal/aab 143
1|INTRODUCTION
Global clima e change has inc eased he isk o sho - e m ex eme
clima e s esses in ag icul u e. D ough is eme ging as one o he mos
signi ican abio ic s esses limi ing c op g ow h and de elopmen
(C uz de Ca alho, 2008; La i e e al., 2007).
Ac ually, global wa e sho age cu en ly limi s c op yields in mo e
han 70% o a able lands. These wa e limi a ions will likely become
mo e impo an in he nex decades in some egions, such as he Med-
i e anean basin, wi h p edic ions o educed ain all (IPCC, 2022). In
addi ion, inc emen s o ag icul u al ac i i ies in un e ile a eas o sa -
is y g owing demands o ood will likely inc ease he need o wa e
and nu ien inpu s in ag icul u al sys ems (Hema hilake &
Guna hilake, 2022). The e has been a signi ican ise in ai empe a-
u e (almos 1.2C in he las wo decades) and e apo anspi a ion in
he Medi e anean egion, and he e is a high con idence ha clima e
change has wo sened hea wa es and d ough s, which a e al eady
a ec ing ag icul u e in his egion ( e iewed in Ali e al., 2022).
Acco dingly, unde s anding and imp o ing plan su i al and
g ow h unde es ic ed wa e a ailabili y is o cen al ele ance in
con empo a y plan science, and pa icula ly o Medi e anean c ops
gi en he ad e se clima e change p ojec ions o his egion.
T i icale (T i icale hexaploide L.) is a c op species b ed om whea
(T i icum aes i um L.) and ye (Secale ce eale L.) h ough in e gene ic
hyb idisa ion and doubling o hyb id ch omosomes (Ayalew e al.,
2018). This c op is eme ging as an impo an sou ce o ood and ani-
mal eed owing o i s good nu i ional quali y, s ong s ess esis ance,
wide adap abili y and high yield. T i icale is a aluable s ess- ole an
ce eal and a po en ial gene ic esou ce o b eeding win e and sp ing
ce eals (Blum, 2014). In ac , i is known o i s highe esis ance unde
d ough s ess compa ed o whea (Giun a e al., 1993). Mo eo e ,
some au ho s epo ed ha unde di e en le els o d ough , i icale
yield showed a non-signi ican educ ion (8%), while du um whea
was signi ican ly dec eased in compa ison o he i iga ed con ol
(54%) (Ala ash e al., 2022). Majo objec i es o he imp o emen o
i icale c ops include, among o he s, inc easing g ain yield, sho ening
plan heigh , and imp o ing wa e use e iciency and i s ole ance/
esis ance o a ious bio ic and abio ic s esses such as d ough
(Me goum e al., 2019). The e o e, i is necessa y o s udy he e ec s
o wa e s ess on he physiological p ocesses o his c op plan o i s
bes managemen (Munjonji e al., 2017).
Soil managemen can play a cen al ole in wa e s o age capaci y
in c ops, pa icula ly in ain ed sys ems. In a scena io o clima e unce -
ain y and sca ci y o wa e esou ces, some s udies ha e sugges ed
ha conse a ion ag icul u e could inc ease he s abili y o ag icul-
u al sys ems agains clima e a iabili y (Puig-Si e a e al., 2022). In
his ega d, educed illage (RT) o no illage (NT) ha e been adop ed
in a id and semi-a id egions wi h posi i e e ec s on c op p oduc ion
by changing he soil en i onmen (Du e al., 2022). Inc eases o soil
wa e in il a ion and e en ion unde NT o RT a e o en ela ed o
imp o emen s in g ain yields and enhanced wa e use e iciency o
he c op, especially unde d ough condi ions (Du e al., 2022). Mul i-
ple s udies in he Medi e anean egion ha e concluded ha no- ill
and low illage no only inc ease soil mois u e bu also imp o e wa e
use e iciency (Can e o-Ma ínez e al., 2007; Fe nández-Ga cía
e al., 2013; Mo ell e al., 2011). Howe e , his e ec is no always
obse ed (Madejón e al., 2023).
Tillage can ha e a p o ound e ec on soil biological communi ies,
which may ha e some implica ions o he esis ance o c op plan s o
wa e s ess. Fo ins ance, se e al wo ks ha e shown ha NT o en
p omo es an inc ease in he abundance o ungi (Pane ie i
e al., 2020), including hose es ablishing myco hizal symbiosis wi h
plan s (B i o e al., 2021; Sebbane e al., 2023), and his could be ben-
e icial o plan wa e up ake unde d ough s ess condi ions (Abdel-
Fa ah & As a , 2012; Liu e al., 2015).
The objec i e o his s udy was o e alua e he po en ial impac
o d ough p edic ions o he Medi e anean egion on g ain yield in
a i icale c op, and o assess whe he his impac can be minimised by
he adop ion o conse a ion illage p ac ices. The hypo hesis o his
wo k is ha he ype o illage could ha e an in luence on he esis-
ance o i icale o d ough condi ions. The e o e, we could expec
ha conse a ion illage would inc ease he c op esis ance o educ-
ions o wa e inpu s, and ha his inc ease could be e i ied h ough
c op ecophysiological indices and changes in oo a chi ec u e. To es
hese hypo heses, a ain all exclusion expe imen was ca ied ou in a
ypical whea /legume Medi e anean o a ion in SW Spain, aking
ad an age o a long- e m expe imen es ablished in 2008. Di e en
a iables ela ed o plan ecophysiology, oo de elopmen and a bus-
cula myco hizal ungi (AMF) colonisa ion and p oduc i i y we e e al-
ua ed o e one c op cycle. The use o ainou shel e s o simula e
d ough condi ions is a p ac ical app oach o es he in luence o ill-
age on c op esis ance o d ough , as i helps o con ol expe imen al
a iables and allows o di ec compa ison be ween ea men s. Mo e-
o e , his is a long- e m expe imen pe mi ing he cap u e o he
cumula i e e ec s o di e en illage sys ems on soil p ope ies,
which can in luence he plan esponse o d ough . Finally, his expe i-
men could con ibu e aluable in o ma ion o ag icul u al p ac ices in
Medi e anean egions and beyond. I conse a ion illage is indeed
ound o enhance i icale d ough esis ance, his knowledge could
be used o guide a me choices o illage me hods o mi iga e he
e ec s o wa e sca ci y on c op yields.
2|MATERIALS AND METHODS
2.1 |Expe imen al a ea and desc ip ion o he
expe imen al design
The long- e m illage expe imen was es ablished in 2008 a he ag i-
cul u e expe imen al a m ‘La Hampa’o he ‘Ins i u o de Recu sos
Na u ales y Ag obiología de Se illa’(IRNAS-CSIC), loca ed a Co ia del
Río (Se ille, SW Spain) o compa e h ee illage sys ems: adi ional
illage (TT), educed illage (RT) and no illage (NT). Th ee eplica e
plo s o 300 m
2
we e delimi ed o each illage ype in a comple ely
andomised expe imen al design. See mo e de ails o he design o
he expe imen in p e ious wo ks (Madejón e al., 2023; Pane ie i
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e al., 2020). The illage ea men s we e es ablished in 2008 and con-
inue o be ca ied ou e e y yea in all he plo s. B ie ly, TT consis s
o mouldboa d ploughing wi h soil in e sion (25–30 cm deep) and
wo chisel ope a ions a 25 cm dep h (0.57 m sepa a ion be ween
chisels) ollowed by a disc ha owing o 12 cm dep h; RT ope a ion
consis s o one chisel ope a ion a 25 cm dep h ollowed by a disc
ha owing o 5 cm dep h. Finally, in NT sowing is done by di ec d il-
ling (no p e-sowing ope a ions a e ca ied ou ). Mo eo e , a leas
30% o he soil su ace unde NT and RT emains co e ed by c op
esidues om he p e ious c op, while 10–15 % o c op esidues om
TT a e le on he soil su ace a e ha es , hese emaining o e he
summe o be bu ied by ploughing in he au umn.The soil ex u e a
he si e is sandy clay loam, and i is classi ied as Typic Xe o lu en
(Soil Su ey S a , 2014). Soil chemical p ope ies measu ed be o e
he sowing o he i icale c op a e shown in Table S1. The clima e is
Medi e anean, wi h mild ainy win e s and ho and d y summe s.
Wea he condi ions du ing he s udy we e eco ded a a me eo ologi-
cal s a ion loca ed a he expe imen al a m (Figu e 1).
FIGURE 1 (a) P ecipi a ion
(blue ba s) and empe a u e ( ed
line) du ing he s udy pe iod. Soil
wa e s o age (SWS) in he 0–
40 cm laye unde di e en illage
and ain all condi ions, om
sowing da e o c op ha es
(mean alues ± SD), (b) No illage
(NT), (c) Reduced illage (RT) and
(d) T adi ional illage (TT). Rain all
exclusion ea men (EXC) is
shown wi h yellow ci cles and
con ol ea men (CONT) wi h
g een ci cles.
MADEJÓN ET AL.145
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In Oc obe 2020, a ain all exclusion expe imen was se up. Fo
his pu pose, ainou shel e s we e used. Each shel e co e s a soil
su ace o 6.25 m
2
(2. 5 m 2.5 m) and is made o ligh me allic s uc-
u e wi h six bands made o anspa en ac ylic ha exclude app oxi-
ma ely 30% o ain all inpu s, which is wi hin he ange o clima e
change p ojec ions o he Medi e anean egion (IPCC, 2022). A
each illage plo (nine plo s in o al, h ee pe illage sys em), wo ain-
ou shel e s we e es ablished ( ain all exclusion ea men ). Rainou
shel e s we e es ablished in No embe 2020 and we e o e he plo s
h oughou he expe imen . They we e only emo ed o illage and
plan ing, bu immedia ely a e hese asks hey we e placed in hei
co esponding plo s. Likewise, a each illage plo , wo a eas o equal
size (2.5 m 2.5 m) we e delimi ed on he g ound, subjec ed o en i-
onmen al p ecipi a ion condi ions (con ol ea men ). The numbe o
o al expe imen al plo s was 36 ( h ee illage sys ems wo ain all
le els six eplica es). Soil mois u e ac oss he 0–40 cm dep h was
pe iodically eco ded wi h a F equency Domain Re lec ome y (FDR)
p obe (Del a-T De ices) in ib eglass ubes ins alled a he cen e o
each plo in hal he eplica es ( h ee illage wo ain all le els
h ee eplica es).
A legume- i icale c op o a ion was es ablished a he expe i-
men . Tillage ope a ions desc ibed abo e o NT, RT and TT a e pe -
o med du ing he au umn be o e seed sowing. Vicia aba L., was he
c op es ablished in he p e ious season ( om No embe 2020 o May
2021; see de ails in Madejón e al., 2023), while i icale was cul i-
a ed o e he 2021–2022 season. Tillage was conduc ed in
Sep embe 2021. In he middle o No embe , in he NT ea men ,
sp ay wi h p e-eme gence glyphosa e (KARDA) a a a e o 6 L ha
1
was applied. Finally, in he middle o Decembe 2021, i icale, ‘Sale-
oso R-2’ a ie y, was sown wi h a densi y o 250 kg o seeds ha
1
.
A he beginning o Ap il 2022, due o he high amoun s o weeds, i
was necessa y o ea he plo s wi h a mix u e o wo he bicides
(KINVARA a a a e o 0.33 g ha
1
and MONOLITH a a a e o
2.25 L ha
1
).
2.2 |Plan ge mina ion and myco hizal oo
colonisa ion
Seedling eme gence was es ima ed 34 days a e sowing, when i was
obse ed ha he majo i y o seedlings had eme ged, by coun ing all
he ge mina ed seedlings in all he ainou shel e s and in he con ol
plo s.
A he end o Ma ch (28 h), h ee comple e plan s (including
oo s) we e sampled in a selec ion o 24 plo s (only in ou plo s o
each illage sys em wi h and wi hou ain all exclusion). Fo he
s udy o he colonisa ion by AMF, a sub-sample o seconda y oo s
(diame e less han 2 mm) was ob ained and ea ed ollowing he
s aining me hod de elopedbyVie heilige al.(
1998). The oo
ma e ial was ho diges ed wi h 10% (w/ ) KOH un il he oo s
we e discolou ed and hen s ained wi h 0.05% blue ink (Pelikan
4001) in inega . Fo all he measu es, 30 agmen s o he com-
posi e samples pe plo we e selec ed. They we e u he cu in o
1 cm pieces and examined unde a mic oscope (Olympus BX40).
AMF colonisa ion and he abundance o a buscules and esicles
we e calcula ed ollowing he me hod o T ou elo e al. (1986).
The a iables s udied we e: deg ee o colonisa ion by AMF, indi-
ca ed by he equency o myco hizae in he oo sys em (F%), he
in ensi y o myco hizal colonisa ion in he oo sys em (M%),
in ensi y o myco hizal colonisa ion in oo agmen s (m%), abun-
dance o esicles s uc u es used o s o e lipids and o he ele-
men s ( %) and abundance o a buscules, whe e ca bon and
nu ien exchange occu s (a%) (Smi h & Read, 2008).
2.3 |Ecophysiological measu emen s
In he middle o Ap il 2022 (19 h), measu emen s o gas exchange
we e conduc ed wi h a po able pho osyn hesis sys em (LI-6400, Li-
Co , Lincoln, NE, USA) o de e mine maximum a es o pho osyn hesis
(AN, max), s oma al conduc ance (g
s
, max), in e cellula concen a ion
o ca bon dioxide (Ci), and anspi a ion a e (E). Gas exchange was
measu ed be ween 10:00 and 13:00 h on heal hy, ully de eloped
young lea es in a selec ion o 18 plo s ( h ee plo s h ee illage sys-
ems wo ain all le els), measu ing h ee plan s pe plo . Measu e-
men condi ions we e se o 350 μmol ai s
1
, 430 μmol CO
2
ai
mol
1
, wi h a PAR o 1500 μmol pho ons m
2
s
1
.
A Scholande - ype p essu e chambe (PMS Ins umen Company,
Albany, OR, USA) was used o measu e lea wa e po en ial. Fo hese
measu emen s, one lea pe plan and wo plan s pe plo we e
selec ed. Fully de eloped lea es om he ou e pa o he plan can-
opy we e selec ed o hese measu emen s. The sampled lea es we e
s o ed in closed plas ic bags wi h humid il e pape and kep in a po -
able coole un il lea wa e po en ial was measu ed in he labo a o y
(Rod iguez-Domínguez e al., 2022).
2.4 |Plan ai s and chemical analyses
Th ee plan s (shoo s and oo s) om each plo (only in ou plo s o
each illage sys em wi h and wi hou ain exclusion, 24 plo s) we e
sampled a he end o Ma ch 2022, coinciding wi h he sampling o
myco hizal ai s. Samples we e eshly weighed, washed wi h dis-
illed wa e and d ied a 60C o a leas 48 h. Plan shoo s (s ems,
lea es, spikes) and oo s (p incipal and seconda y) we e d ied sepa-
a ely and weighed. Lea es mass ac ion (LMF), s em mass ac ion
(SMF), ui mass ac ion (Spike, FMF), oo mass ac ion (RMF) and
he shoo : oo a io we e eco ded.
To analyse mac o and mic onu ien con en in lea es, hese
we e diges ed by we oxida ion wi h concen a ed HNO
3
in a Digi-
p ep Ms. block diges e , and he ex ac s we e de e mined by
induc i ely coupled plasma spec opho ome y (ICP-OES: Va ian
ICP 720-ES, wi h axially iewed plasma). The accu acy o he ana-
ly ical me hod was de e mined using a plan e e ence ma e ial:
INCT-ONTL-5 (Tobacco lea es). Reco e y a es o e e ence plan
samples anged be ween 95% and 105%.
146 MADEJÓN ET AL.
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2.5 |Plan p oduc ion
A he end o May (31s ), when he spikes we e a ma u i y, he
abo eg ound pa o he biomass was collec ed wi hin wo quad a s
o 0.50 m
2
pe plo . These samples included bo h i icale plan s and
weeds. T i icale spikes we e u he sepa a ed om s ems. Plan
ma e ial (spikes and weeds) was weighed esh, and subsequen ly
d ied a 60C o 48 h. A e d ying he spikes, hei g ains we e sepa-
a ed om he cha using a Shelle (SAS GOD
E, model V 05-204).
G ain d y weigh pe plo was also eco ded.
2.6 |Da a analysis
Soil wa e s o age (SWS, mm), o he 0–40 cm laye s ( oo zone), was
calcula ed om soil wa e con en measu emen s (θ) h ough he soil
p o ile on 15 sampling da es be ween 17 Sep embe 2021 and
18 May 2022 ollowing apezoidal ules (Jia e al., 2013) acco ding o
he equa ion:
SWS¼50θ1þX
3
i¼1
50θiþθiþ1
ðÞ:
whe e θ
i
ep esen s olume ic mois u e (cm
3
cm
3
) in he i h soil
dep h (1: 10 cm; 2: 20 cm; 3: 30 cm; 4: 40 cm); he soil dep h in e al
was 10 cm.
Linea mixed models we e applied o es o he e ec o illage
and ain all exclusion (as well as hei in e ac ions) on SWS, seedling
eme gence and c op yield, myco hizal ai s and ecophysiological a -
iables, as well as on plan chemical ai s, using he nlme package in
R. The plo was included as a andom e m o accoun o epea ed
measu emen s o soil wa e con en . Also, he andom e m included
he spa ially nes ed design, gi en ha wi hin each illage plo , wo
con ol plo s and wo d ough plo s we e delimi ed. Valida ion o he
model assump ions was done by explo a ion o model esiduals.
When homogenei y o a iance was no me , a a iance coe icien
was in oduced in he model o accoun o he e oskedas ici y among
di e en ac o le els, using he a Iden unc ion o he nlme package
(Pinhei o & Ba es, 2000). No ans o ma ion o da a was equi ed o
sa is y he assump ions o he analysis.
Pea son's co ela ion analyses we e used o explo e he ela ion-
ships be ween c op p oduc ion and weed incidence, as well as among
myco hizal a iables and plan ecophysiological measu emen s.
3|RESULTS
3.1 |Soil wa e s o age: E ec o ain all exclusion
and illage
Tillage ype had a signi ican in luence on SWS (linea mixed model-
ling, F=4.10, p=.044), and soils unde NT had g ea e SWS han
soils unde RT o TT. These di e ences among he h ee illage
sys ems we e especially no iceable om Janua y 2022 o he end o
he expe imen in May (Figu e 1). F om Sep embe 2021 o May
2022, SWS alues we e, on a e age, 16% highe in soils unde NT
han in hose unde RT and TT.
The impac o ain all exclusion on SWS depended on he da e o
he sampling. The simula ed d ough educed SWS signi ican ly
(p< .05, F es ) in 5 ou o 15 sampling da es, especially in he sp ing
season (Figu e S1). The g ea es di e ences be ween con ol and
exclusion plo s we e obse ed on 4 No embe 2021 o NT
and 18 Ma ch 2022 o TT, when he exclusion ea men led o a
educ ion o SWS o 21 and 22 mm in he NT and TT ea men s on
a e age, espec i ely (Figu e S1). In he case o RT, he maximum di -
e ence was ound 1 mon h la e (a educ ion o 22 mm). These alues
ep esen ed educ ions o 20%, 26% and 21% o SWS ela i e o con-
ol plo s in he NT, RT and TT, espec i ely. A he end o he expe i-
men , he e was a signi ican illage ain all ea men in e ac ion,
so ain all exclusion led o a educ ion o SWS in he RT sys em, bu
no in he TT o he NT sys ems (Figu e S1).
3.2 |Ge mina ion and g ain p oduc ion: E ec s o
ain all exclusion and illage
Seedling eme gence was no in luenced by ei he illage o ain all
exclusion (Table 1).
In con as , c op p oduc ion, assessed by he measu emen o he
g ain yield (Figu e 2a), was signi ican ly educed by ain all exclusion
(F=5.2653, p=.0308), wi h yields obse ed in he exclusion ea -
men a ound 22% lowe compa ed o he con ol. Among he illage
sys ems, only TT exhibi ed signi ican di e ences in g ain yield
(F=5.59, p=.045) because o ain all exclusion (31% less in he
exclusion ea men compa ed o con ol), whe eas changes in g ain
yield because o ain all exclusion we e no signi ican in he conse -
a ion illage sys ems (F=0.526, p=.489, and F=0.769, p=.406
o NT and RT, espec i ely).
Despi e he di e en impac s o ain all exclusion, bo h NT and TT
showed simila g ain yield alues in he exclusion ea men . Conside ing
he scena io o educed ain all, bo h NT and TT main ained g ain yields
o 1614 and 1490 kg ha
1
, espec i ely (sligh ly highe on NT).
Wild g asses (weeds) we e also e alua ed because hey can a ec
he p ope de elopmen o he c op (Figu e 2b). The nega i e and sig-
ni ican co ela ion ( =0.57, p=.0003; n=36) be ween g ain
yield and biomass o wild g asses highligh s he compe i ion o
TABLE 1 Numbe o ge mina ed seeds acco ding o he wo
expe imen al ac o s, illage and ain all condi ions (mean alues ±
SE; N=6).
Tillage Con ol Exclusion
NT 115.2 ± 54.8 136.2 ± 57.1
RT 144.2 ± 72.8 150.0 ± 81.1
TT 115.2 ± 44.8 98.83 ± 20.8
Abb e ia ions: NT, no illage; RT, educed illage; TT, adi ional illage.
MADEJÓN ET AL.147
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esou ces (nu ien s, wa e and ligh ) be ween he c op and weeds.
The lowe p oduc ion o wild g asses because o he bicide applica ion,
which was mo e in ense in he NT sys em, likely con ibu ed o
imp o e c op yield by educing compe i ion o esou ces.
3.3 |E ec s o ain all exclusion and illage on
biomass alloca ion and oo myco hiza ion
Rain all exclusion and illage did no ha e any signi ican e ec s on he
shoo mass o indi idual plan s (F=0.279, p=.604; F=0.116, p=.891,
espec i ely). Rain all only had some impac on he pa e ns o biomass
alloca ion o he oo s in i icale plan s, especially in he RT sys em.
3.3.1 | Roo biomass ac ions
Rain all exclusion had a signi ican nega i e e ec on o al oo bio-
mass and p ima y oo biomass (F=5.550, p=.036 and F=6.896,
p=.017, espec i ely) (Figu e 3). Conside ing all he illage sys ems,
seconda y oo s we e no signi ican ly (p> .05, F es ) a ec ed by
ain all exclusion. Howe e , when dis inguishing be ween illage
ypes, he e was a clea educ ion in he g ow h o seconda y oo s
unde ain all exclusion in he RT sys em, wi h nea ly a 50% educ ion
in biomass. Rain all exclusion also a ec ed RMF, SMF, and he a io
o oo s o shoo s (F=26.01, p=.0003, F=19.31, p=.0009, and
F=19.31, p=.0009, espec i ely) (Figu e S3). Rain all exclusion led
o a educ ion in RMF and he oo :shoo a io, bu o an inc ease in
SMF. When conside ing each illage sys em sepa a ely, he inc ease
in he shoo mass ac ion was pa icula ly high in he RT ea men
(Figu e S3).
3.3.2 | Myco hizal colonisa ion
The ai s s udied o his pu pose we e: F%: F equency o myco hiza
in he oo sys em, M%: In ensi y o he myco hizal colonisa ion in
he oo sys em, m%: In ensi y o he myco hizal colonisa ion in he
FIGURE 2 (a) G ain yield and (b) biomass o wild g asses (weeds) in he h ee illage sys ems and in con ol and ain all exclusion plo s (mean
and SE). G een ba s indica e con ol plo s and yellow ba s exclusion plo s. The as e isk (*) indica es signi ican di e ences be ween he con ol
and exclusion ea men s o each illage sys em (p< .05, F es ).
FIGURE 3 Biomass o di e en oo ac ions o plan s (a) o al oo ; (b) p ima y oo ; (c) seconda y oo ) g owing in he h ee illage sys ems,
and in con ol and ain all exclusion plo s (mean and SD). The as e isk (*) indica es signi ican di e ences (p< .05, F es ) be ween he con ol and
he ain all exclusion ea men s o each illage sys em. DW: D y weigh .
148 MADEJÓN ET AL.
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oo agmen s, a%: A buscule abundance in myco hizal pa s o oo
agmen s, %: Vesicle abundance in myco hizal pa s o
oo agmen s.
The high a iabili y o AMF ai s was an impo an obse a-
ion in his expe imen . Rain all exclusion ea men had a signi i-
can e ec (F=13.97, p=.003) on he equency o myco hizal
in ec ion in he oo sys em (F%), being sligh ly highe in he plan s
unde he ainou shel e s han in con ol plo s. The in e ac ion
be ween ain all exclusion and illage was signi ican o his a i-
able (F=7.92, p=.008), so in TT d ough clea ly inc eased F%,
bu no inNTo RT.
The e ec o illage was mo e e iden on %, he esicle abun-
dance (F=5.79, p=0.049). Fo his a iable, plan s in he conse a-
ion illage sys ems (RT and NT) had highe alues compa ed o he TT
sys em (Figu e 4d), which was a common end o all AMF ai s,
including he abundance o a buscule (F=4.64, p=.072).
When only conside ing he con ol ea men , signi ican ly
(p< .05, F es ) highe alues o F% and % ( esicle abundance) we e
obse ed in bo h conse a ion illage sys ems (NT and RT) in
compa ison o TT. When conside ing he d ough plo s only, signi i-
can di e ences (p< .05, F es ) among illage sys ems we e es ic ed
o he abundance o a buscules (a%) be ween RT and TT.
3.4 |E ec s o ain all exclusion and illage on
plan ecophysiology and nu i ion
3.4.1 | Ecophysiological da a
The measu ed pho osyn he ic a iables included maximum a es o
pho osyn hesis (AN
max
), in e cellula concen a ion o ca bon dioxide
(Ci), s oma al conduc ance (g
s
) and anspi a ion a e (E). The main
esul s indica e a limi ed e ec o ain all exclusion o he illage ype.
When dis inguishing among illage sys ems, maximum pho osyn he-
sis a es (AN_
max
), in he TT sys em we e educed by 25% because o
ain all exclusion, al hough because o da a dispe sion, his e ec was no
signi ican (F=2.12, p=.188; Figu e 5a). Fo conse a ion illage sys-
ems (NT and RT), simila alues we e eco ded be ween exclusion and
con ol plan s. Mo eo e , AN
max
alues we e posi i ely and signi ican ly
co ela ed o s oma al conduc ance ( =0.556, p< .001, n=18).
Fo s oma al conduc ance (g
s
), he e we e no di e ences because
o ain all exclusion o illage. In e cellula concen a ion o ca bon
dioxide (Ci) was no a ec ed by he ain all exclusion, al hough he e
was a end owa ds lowe alues in he con ol ea men , especially
in he TT sys em, whe e a educ ion o 50% o Ci in con ol plan s
was obse ed in compa ison o plan s om exclusion ea men s. Ci
FIGURE 4 Pe cen ages o myco hizal colonisa ion in T i icale oo s in he di e en ea men s (mean alues ± SE). Da a in g een indica e
con ol plo s and yellow da a indica e exclusion plo s. Fo each illage, signi ican di e ences be ween con ol and exclusion a e ma ked wi h an
as e isk (p< .05, F es ).
MADEJÓN ET AL.149
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alues we e also posi i ely co ela ed wi h g
s
( =0.575, p< .001,
n=18). As wi h pho osyn hesis a es, ain all exclusion led o some
educ ion in E ac oss all illage sys ems, pa icula ly in RT and TT
(a ound 25% and 22% lowe , espec i ely), al hough his end was
no signi ican (F=1.04, p=.312). These alues we e also signi i-
can ly co ela ed wi h g
s
( =0.809, p< .001, n=18).
FIGURE 5 (a) Maximum a es o pho osyn hesis (AN
max
), (b) s oma al conduc ance (g
s max
), (c) in e cellula concen a ion o ca bon dioxide
(Ci), (d) and anspi a ion a e (E) and (e) lea wa e po en ial in he h ee illage sys ems in con ol (g een poin s) and exclusion ain all (yellow
poin s) plo s (mean and SE).
150 MADEJÓN ET AL.
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Lea wa e po en ial (Ψ
lea es
) alues we e no signi ican ly
a ec ed by ain all exclusion in any o he illage sys ems
(F=0.002, p=.968). Su p isingly, in he TT sys em, wa e po en-
ial alues we e sligh ly mo e nega i e in con ol lea es compa ed
o exclusion lea es, despi e a highe s anda d de ia ion in he
exclusion. O e all, Ψ
lea es
alues did no show a consis en
esponse o ain all exclusion ac oss he illage sys ems
(Figu e 5e).
3.4.2 | Mac o and mic onu ien s in lea es
In gene al, nu ien con en s in lea eswe emo ein luencedby
ain all exclusion han by illage (Table 2). Mac onu ien s in lea es
ended o p esen highe con en s in he con ol ea men han
unde ain all exclusion, wi h a signi ican dec ease in N con en s
because o d ough (F=9.14, p=.008). Also, he e was a end
o educ ion in lea P unde ain all exclusion, al hough his e ec
was ma ginally non-signi ican (F=3.48, p=.081). In con as ,
he e was an inc ease in lea K in he ain all exclusion ea men
(F=4.55, p=.049).
Fo he s udied mic onu ien s, he e we e no clea s a i-
s ical di e ences because o ain all exclusion, excep o Fe,
which p esen ed signi ican ly highe alues in con ol lea es
(F=14.27, p=.0018). Finally, we could no ind any di e ences
because o he illage sys em in he s udied mac o and
mic onu ien s.
The main chemical p ope ies o he soils aken jus be o e
he i icale sowing (Table S1)showedaninc easeinEC o all
h ee sys ems unde ain all exclusion, bu an inc ease in N o
he NT sys em only. Fo all illage sys ems, con en s o OM and
a ailable P we e highe in he exclusion ea men han in he
con ol (F=7.341, p=.019; and F=3.818, p=.074,
espec i ely).
4|DISCUSSION
4.1 |Soil wa e s o age
Tillage ype can a ec he dynamics o wa e in il a ion and e en ion
h ough he soil p o ile. In ou case, soils om NT showed highe wa e
s o age han soils om RT o TT. Thisisconsis en wi hp e iouswo k
conduc ed a he si e ha showed ha conse a ion illage me hods end
o imp o e wa e e en ion in he soil compa ed o TT p ac ices (López-
Ga ido e al., 2014). In addi ion, se e al ield s udies ha e epo ed a pos-
i i e impac o he NT and RT on soil mois u e, which ensu es he abso p-
ion and u ilisa ion o wa e by plan s (Wang e al., 2007; Zhang
e al., 2022). Al hough RT has also been p oposed as a po en ial s a egy
o inc ease SWS in hose wo ks, we could no obse e his e ec in his
s udy. These esul s could be ela ed o he ac ha NT p oduces su i-
cien c op esidues o p o ide mulching on he soil su ace, educing
losses ia e apo anspi a ion and con ibu ing o wa e conse a ion
(Kühling e al., 2017), which is mo e e iden a he i s cm o soil (in his
s udy SWS was es ima ed down o 40 cm dep h). Mo eo e , NT can
es o e mac opo e connec i i y and agg ega ion, inc easing p e e en ial
low om opsoil o dep h (S udley e al., 2008).
The ain all exclusion ea men had a s a is ically signi ican
impac on SWS on ce ain da es, especially du ing he sp ing season.
Al hough he magni ude o SWS educ ion (in ela ion o con ol plo s)
was simila among illage ypes, benea h he ainou shel e s (exclu-
sion ea men ), NT had he highes mean alues o SWS. This indi-
ca es ha NT p ac ices may ha e compensa ed o he educ ion o
ain all inpu s wi h a highe wa e e en ion wi hin he soil.
TABLE 2 Concen a ions o mac o and mic onu ien s in T i icale lea es acco ding o he wo ea men s, illage and ain all condi ions (mean
alues and SD in b acke s).
Tillage Rain all N Ca K Mg Na P S Cu Fe Mn Zn
% mg kg
1
NT Con ol 3.45 0.70 1.96 0.30 0.07 0.27 0.28 7.80 272.1* 57.10 27.55
(0.36) (0.11) (0.13) (0.06) (0.05) (0.02) (0.04) (0.80) (38.9) (5.58) (1.96)
Excl. 2.92 0.64 2.25 0.27 0.23 0.24 0.25 7.71 214.1 58.49 27.62
(0.48) (0.08) (0.21) (0.05) (0.33) (0.04) (0.03) (2.71) (26.7) (4.61) (3.31)
RT Con ol 3.15 0.69 2.13 0.29 0.10 0.25 0.25 6.96 247.2 61.87 24.25
(0.42) (0.09) (0.03) (0.05) (0.06) (0.03) (0.05) (0.94) (84.7) (13.37) (7.47)
Excl. 2.90 0.66 2.31 0.31 0.06 0.24 0.24 7.07 224.2 56.01 26.15
(0.54) (0.04) (0.35) (0.03) (0.04) (0.03) (0.06) (0.75) (53.4) (10.49) (6.67)
TT Con ol 3.51 0.68 2.17 0.29 0.11 0.28 0.29 7.43 231.0* 58.21 25.55
(0.11) (0.05) (0.12) (0.02) (0.14) (0.03) (0.01) (0.88) (12.4) (8.60) (3.39)
Excl. 3.32 0.62 2.19 0.28 0.03 0.26 0.27 8.02 201.2 61.40 30.45
(0.24) (0.10) (0.14) (0.04) (0.03) (0.02) (0.03) (0.77) (16.6) (18.16) (8.82)
No e: The as e isk (*) indica es signi ican di e ences be ween he con ol and he ain all exclusion ea men s o each illage sys em (p< .05, F es ).
Abb e ia ions: NT, no illage; RT, educed illage; TT, adi ional illage.
MADEJÓN ET AL.151
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