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First insights on Tuber borchii distribution in both natural forests and experimental truffle fields in Portugal
Conference Paper · October 2014
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Beatrice Belfiori
Italian National Research Council
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Italian National Research Council
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Available from: Marcos Morcillo
Retrieved on: 08 July 2016

First
insights
on
Tuber
borchii
distribu.on
in
both
natural
forests
and
experimental
truffle
fields
in
Portugal
Anabela
Marisa
Azul1,
João
Trovão1,
Marcos
Morcillo2,
Andrea
Rubini3,
Alessandro
Trusso3,
Beatrice
Belfiori3,
Francesco
Paolocci3,
Helena
Freitas1,
and
Claudia
Riccioni3
1Centre
for
FuncAonal
Ecology,
Department
of
Life
Sciences.
University
of
Coimbra,
3001-­‐401
Coimbra,
Portugal.
E-­‐mail:
amjrazul@ci.uc.pt
2Micologia
Forestal
&
Aplicada,
Rbla.
Arnau
6
local
D,
Vilanova
i
la
Geltrú,
08800
Barcelona,
Spain
3
CNR
(NaAonal
Research
Council)
-­‐
InsAtute
of
Biosciences
and
BioResources
(IBBR),
Via
della
Madonna
Alta,
130
-­‐
06128
Perugia,
Italy
AIM
To
monitor
the
distribuAon
and
dynamics
of
T.
borchii
in
naAve
forests
and
experimental
truffle
trials
established
in
Portugal
(Alentejo
region).
BACKGROUND
Among
the
most
valuable
ectomycorrhizal
fungi
belonging
to
Tuber
spp.,
Tuber
borchii,
is
becoming
an
increasingly
popular
fungus
in
the
marketplace.
Thanks
to
a
good
adaptaAon
to
mulAple
environments,
T.
borchii
is
also
culAvated,
in
both
naAve
and
non
endemic
areas.
As
all
symbioAc
species,
this
fungus
can
improve
mineral
nutriAon
and
stress-­‐tolerance
of
the
host
plant.
The
scienAfic
relevance
of
T.
borchii
as
a
model
species
for
studying
plant-­‐fungus
symbioses
is
also
recognized.
MaAng
type
(MAT)
genes
governing
the
sexual
reproducAon
(i.e.
frucAficaAon)
have
been
recently
idenAfied
unveiling
that
this
is
a
heterothallic
fungus
(MarAn
et
al
2012
European
Patent
EP2426215).
PRELIMINARY
RESULTS
REFERENCES
Azul
et
al.
2010
Mycorrhiza
20,
73-­‐88
MarAn
F
et
al.
2012
European
Patent
EP2426215
Murat
et
al.
2013
New
Phytol
199:176-­‐187
Rubini
et
al.
2011
New
Phytol
189:710-­‐722
Rubini
et
al.
2014
Mycorrhiza
24,
S19-­‐S27
ACKNOWLEDGEMENTS
Bilateral
Agreement
between
FCT-­‐MCTES
(Portuguese
FoundaAon
for
Science
and
Technology)
and
CNR
(Italian
NaAonal
Research
Council),
and
VIDT
19118
Co-­‐funded
by
COMPETE-­‐
FEDER-­‐EU,
for
financial
support.
All
faciliAes
at
the
farm
units,
Freixo,
Breijinho,
and
Quinta
de
Sousa.
FUTURE
RESEARCH
During
Summer
2014
a
consistent
number
of
new
T.
borchii
ECMs
has
been
found
and
collected
in
the
same
natural
areas.
This
material
is
now
under
analysis
to
assess
whether
the
distribuAon
pajern
of
T.
borchii
ECMs
with
different
maAng
type
is
biased
on
their
hosts
as
it
has
been
recently
demonstrated
to
occur
on
host
plants
colonized
by
T.
melanosporum
(Rubini
et
al.
2011,
2014;
Murat
et
al.
2013).
CONCLUSIONS
Our
data
show
that,
at
least
in
experimental
truffle
grounds,
T.
borchii
ECMs
of
different
maAng
type
can
coexist
under
the
same
host-­‐plant.
This
pajern,
if
supported
by
further
data,
disAnguishes
this
species
from
the
valuable
black
truffle
T.
melanosporum,
where
a
biased
distribuAon
of
the
two
maAng
types
on
single
host
plants
was
evidenced,
suggesAng
intraspecific
compeAAon
(Rubini
et
al.
2011;
Murat
et
al.
2013).
It
could
be
very
interesAng
to
invesAgate
whether
or
not
such
a
dissimilarity
in
the
strain
dynamics
of
the
two
species
might
be
at
the
basis
of
their
very
different
ecological
requirements
and
pajern
of
geographic
distribuAon.
Tuber
borchii
SAMPLING
SITES
1
Natural
forests
(NF),
corresponding
to
naAve
Mediterranean
oak
woodlands
dominated
by
Quercus
suber
L.
(cork
oak,
Qs).
2
Experimental
field
trial
1
(EFT1),
with
plantlets
of
Pinus
pinea
L.
(stone
pine,
Pp)
inoculated
with
T.
borchii,
introduced
in
areas
dominated
by
Qs
and
Pp
(in
spring
2010).
3
Experimental
field
trial
2
(EFT2)
inoculaAon
in
situ
of
Qs
and
Pp
young
trees
with
spores
of
T.
borchii
in
oak
woodlands
(spring
2011).
METHODS
Samplings
were
performed
in
summer
2013.
Four
distant
plots,
two
in
the
NF,
and
two
in
EFT1
and
EFT2,
and
5
trees
per
plot,
were
selected.
Around
each
tree,
roots
and
soil
samples
were
collected
from
different
points.
Ectomycorrhizae
(ECM)
and
soil
samples
were
molecularly
analyzed
by
PCR-­‐amplificaAon
of
the
ITS
region
and
maAng
type
locus.
T.
borchii
ECMs
were
present
in
both
NF
and
EFTs
(Fig.
6),
with
higher
frequency,
up
to
50%
mycorrhiza.on,
in
EFT1
(Table
1),
and
≤5%
in
EFT2.
In
NF,
T.
borchii
ECMs
were
more
irregularly
found.
Moreover,
preliminary
data
revealed
that
under
the
same
host
plant,
T.
borchii
ECMs
of
opposite
ma.ng
type
can
coexist
in
experimental
grounds.
EFT
2
Pp
aTbor
aECM
iECM
nECM
Abundance
≤5%
75-­‐85%
12-­‐17%
<5%
EFT
2
Qs
aTbor
aECM
iECM
nECM
Abundance
≤5%
70-­‐80%
15-­‐20%
<5%
EFT
1
Pp
aTbor
aECM
iECM
nECM
Abundance
≤50%
45-­‐50%
40-­‐45%
≤5%
Interes.ngly,
although
T.
borchii
ECMs
were
infrequently
found
in
natural
forest,
almost
all
of
the
soil
samples
revealed
the
belowground
presence
of
mycelium
of
T.
borchii
and
a
dominance
of
one
ma.ng
type
over
the
other.
Figure
2.
above
experimental
field
trial
1,
plantlets
of
Pinus
pinea
inoculated
with
T.
borchii
(spring
2010);
below
experimental
field
trial
2,
inoculaAon
in
situ
of
young
trees
of
Quercus
suber
and
P.
pinea
with
spores
of
T.
borchii
(spring
2011).
Figure
3.
Natural
forest
–
Herdade
do
Freixo,
Montemor-­‐o-­‐Novo,
Portugal,
represenAng
oak
woodlands
dominated
by
Quercus
suber.
Figure
4.
EFT1
–
Herdade
do
Brejinho,
Grândola,
Portugal.
Figure
5.
EFT2
–
Quinta
de
Sousa,
Montemor-­‐o-­‐Novo,
Portugal.
Tabela
1.
Abundance
of
T.
borchii
ECM
in
EFT1
and
EFT2.
aTbor
=
acAve
T.
borchii
ECM;
aECM
=
total
acAve
ECM,
iECM
=
total
inacAve
ECM,
nECM
=
total
non
mycorrhizal
roots
(see
Azul
et
al.
2010)
Natural
forests
cork
oak
woodlands
Experimental
truffle
trials
Figure
1.
DistribuAon
of
naAve
Mediterranean
oak
woodlands
dominated
by
Quercus
suber
in
Portugal
with
the
sampling
sites
located
in
the
Alentejo
region.
1
2
3
4
5
6
7
8
9
10
11
12
13
500bp
1Kb
Figure
6.
MulAplex
PCR
with
specific
primers
of
both
maAng
types
on
ECMs
collected
under
the
same
host
plant
(site
EFT1).
Lane
(L)
2:
posiAve
control,
MAT1-­‐2-­‐1.
L13:
posiAve
control,
MAT1-­‐1-­‐1.
L3-­‐11:
each
lane
represent
a
single
ECM.
L12:
negaAve
control.
L1:
Gene
Ruler
DNA
ladder
mix
(Fermentas).