REVIEW New Phytol.

(2000), 147, 179187

Research review
The impact of elevated CO
#
and global
climate change on arbuscular mycorrhizas:
a mycocentric approach
A. H. FITTER*, A. HEINEMEYER \i P. L. STADDON
Department of Biology, University of York, PO Box 373, York YO10 5YW, UK
Received 4 November 1999; accepted 6 March 2000
si\nv
Arbuscular mycorrhizal (AM) symbioses are a potentially important link in the chain of response of ecosystems
to elevated atmospheric [CO
#
]. By promoting plant phosphorus uptake and acting as a sink for plant carbon, they
can alleviate photosynthetic down-regulation. Because hyphal turnover is likely to be fast, especially in warmer
soils, they can also act as a rapid pathway for the return of carbon to the atmosphere. However, most experiments
on AM responses to [CO
#
] have failed to take into account the dierence in growth of mycorrhizal and non-
mycorrhizal plants; those that have done so suggest that AM colonization of roots is little altered by [CO
#
],
although this issue remains to be resolved. Very little is known about the eects of other factors of global
environmental change on mycorrhizas. These issues need urgent attention. It is also necessary to understand the
potential for the various AM fungal taxa to respond dierentially to environmental changes, including carbon
supply and soil temperature and moisture, especially because of the dierential abilities of plant and fungal species
to migrate in response to changing environments. Indeed, there is a need for a new approach to the study of
mycorrhizal associations, which has been too plant-centred. It is essential to regard the fungus as an organism
itself, and to understand its biology both as an entity and as part of a symbiosis.
Key words: elevated [CO
#
], temperature, mycorrhizal function, diversity.
1noii1 o
Most (probably 90%; Smith & Read, 1997) plants
form mycorrhizal associations. About two-thirds of
these plants are symbiotic with arbuscular mycor-
rhizal (AM) fungi (Fitter & Moyersoen, 1996).
Except in boreal and some temperate forests and in
heathlands, AM symbiosis is the normal state of the
root systems of most plant species (Read, 1991).
This article concentrates on the AM symbiosis,
although many of the arguments apply equally to
other mycorrhizal associations, notably the ecto-
mycorrhizal and ericoid mycorrhizas. However,
these other associations are taxonomically and func-
tionally distinct from the AM symbiosis (Fitter &
Moyersoen, 1996), and it is dangerous to assume that
experimental conclusions obtained from one can be
applied to another.
The fungi involved in AM symbioses are members
of a single order, the Glomales, all of which are
obligate symbionts. Because the AM association
functions on the basis of carbon (C) xed in
*Author for correspondence (tel j44 1904 432814; fax j44 1904
434385; e-mail ahf1!york.ac.uk).
photosynthesis moving from plant to fungus, there is
an obvious need to understand what impact an
increasing atmospheric [CO
#
] will have on mycor-
rhizal symbioses. Any such eect is, of course,
indirect as far as the fungus is concerned. The
fungus exists in an environment rich in CO
#
: both
inside the root and in the soil, [CO
#
] is greater than
current atmospheric [CO
#
]. However, if, for
example, elevated atmospheric [CO
#
] increased the
supply of xed C to roots, this might promote the
growth of the fungus. In consequence, services
performed by the fungus might be enhanced, the
best characterized of which is phosphate uptake,
although resistance to drought and pathogens and
increased uptake of other mineral nutrients are also
known to occur (Newsham et al., 1995).
A positive feedback loop can be envisaged, in
which plants respond to elevated [CO
#
] by increased
C xation followed by the transfer of more carbon
to their root systems; consequently, mycorrhizal
fungi might grow more and capture more phosphate
(or perform other functions better). The plant would
thus be relieved of a potential deciency that might
otherwise restrict its ability to respond to elevated
180 REVIEW A. H. Fitter et al.
Atmospheric
CO
2
Respiration Photosynthesis
Leaf
carbon
P
i
and
P
o
Root
carbon
P
o
P
i
Plant
P
i
t
1/2
>30d
t
1/2
>10d
Fungus
carbon
t
1/2
?
>5d
P
i
P
o
Rapid turnover soil C
Slow turnover soil C
Fungus
Soil
Fig. 1. Feedback mechanisms in the response of mycor-
rhizal symbiosis to elevated atmospheric [CO
#
]. Fluxes of
C into and through the biota are shown in red, respiration
and decomposition that returns CO
#
to the atmosphere in
black, and movements of P in blue. Green arrows represent
controls on P movements and transformations by plant
and fungal carbohydrate status, and on photosynthesis by
leaf P status. Values of t
"
#
represent typical or estimated
half-lives of leaves, roots and hyphae that determine rates
of transfer of structural C to the soil organic matter pool.
P
i
and P
o
refer to inorganic and organic P; no distinction
is shown between the various components of plant and
fungal C pools.
[CO
#
] (Fig. 1), as suggested by the alleviation of
photosynthetic down-regulation (Staddon et al.,
1999b). The availability of C for metabolic processes
(Fig. 1) determines key processes in the phosphorus
(P) metabolism of the plant (uptake, transport and
utilization). In return, the P status of the leaf
determines photosynthetic rate. This positive feed-
back is eventually constrained by other deciencies
(e.g. nitrogen (N) or water), and is therefore only
likely to be important as a control on the processes
shown in Fig. 1 if P is a limiting factor. Nevertheless,
because AM fungi represent an interface in the
soilplant system displayed in Fig. 1, their potential
ability to regulate plant response to global change is
one key reason that their responses need to be
understood.
The other major feature of interest is that
mycorrhizal fungi are a link in the chain of transfers
by which C moves from plant to soil (Staddon et al.,
1999d). They can therefore potentially inuence C
cycling rates. Some of the structural C that is
transferred to a symbiotic AM fungus is used to
construct the extraradical mycelium. This network
of ne hyphae almost certainly has a faster turnover
rate than that of either roots or, especially, shoot
material (Fig. 1). This fungal C could therefore be
part of a rapid pathway in the C cycle that returns C
to the atmosphere. By contrast, some fungal com-
pounds might be resistant to microbial attack and
enter a slow pool of recalcitrant soil C, thus
decelerating the cycle (Treseder & Allen, 2000). A
consequence could therefore be the accumulation of
either more or less C in soils. The important
consequences of an increase in atmospheric [CO
#
]
for mycorrhizal functioning could therefore be
changes in the way in which they promote plant
growth and\or changes in rates of C cycling. Both of
these are indirect eects, determined by the
responses of plants to [CO
#
], and illustrated by the
top set of connections in Fig. 2. Increased C xation
can increase C availability to the fungus, thus
promoting fungal ability to provide P to the plant
and improve plant growth. However, other aspects
of climate change might have important direct eects
on AM fungi. If soils warm, the growth of the fungi
might be aected; for example, some fungi might
become active at times of year when they are
currently dormant, or they might respond to dis-
turbance at a dierent rate. Similarly, changes in soil
moisture content (both drier and wetter conditions
are predicted depending on location, vegetation and
circumstance) would almost certainly have large but
as yet unpredictable eects on AM fungi, as
illustrated by the lower sets of connections in Fig. 2.
However, our understanding of the basic biology of
these organisms is so limited that it is dicult to
make useful predictions about these direct impacts.
In a warmer climate, for example, C xation by
plants might increase, with the same impacts as
elevated [CO
#
] directly; alternatively, C xation or
fungal growth might be adversely aected, or there
might be qualitative eects on the structure of the
fungal community or on its temporal pattern (phe-
nology). The wide range of possibilities leads to the
complexity of the potential outcomes of global
change shown in Fig. 2. One important linkage also
shown here is that increased fungal growth, while
promoting P uptake, also increases the C demand by
the fungus, which might act as a regulator on these
interactions.
This article seeks to build on recent advances in
our understanding of the responses of mycorrhizal
associations to environmental change, and to suggest
that a conceptual shift is needed, in which the fungus
itself becomes the focus of experimental investi-
REVIEW Mycorrhizas and global change 181
C
Fixation
Plant
growth
Interface
Fungal
growth
P uptake
C to
fungus
C demand
by fungus
?
?
?
Changes to
fungal taxa
phenology
etc.
C to
fungus
P uptake
?
Drought
Temp.
[CO
2
]
?
Fig. 2. Direct and indirect eects of elevated atmospheric [CO
#
] on mycorrhizal fungi. Upward arrows
represent increases, and downward arrows decreases, in the specied processes. Direct eects of soil
temperature and moisture on fungi aect fungal growth and community structure in unknown ways and feed
back on the plant community. Indirect eects act through plant C xation. Both benecial and deleterious
eects on plant and fungal growth are potentially self-reinforcing, through positive feedback loops. Regulation
of these loops might be achieved via alterations in C demand by the fungus.
gation, rather than the plant alone or even the
symbiosis. We have made no attempt to provide a
comprehensive review of the literature; rather, our
goal is to point to changes in experimental approach
that we believe are needed, highlighting a small
number of recent studies that illuminate this point.
inni1 s1\1i oi ioviiioi
Impact of elevated atmospheric [CO
#
] on AM
colonization
Numerous workers have grown plants at ambient
(usually 350 l l
") and elevated (variously 500, 600,
610, 700 and 710 l l
") atmospheric [CO
#
] (hereafter
aCO
#
and eCO
#
respectively) and have measured the
degree of colonization of the roots by AM fungi
(reviewed by Staddon & Fitter, 1998). Some of these
studies have been of single plants in pots, and some
have been of whole communities exposed to eCO
#
in
open-topped chambers or free-air CO
#
enrichment
(FACE) rings. The results have been inconclusive.
Although decreases in colonization are rarely seen,
increases and null responses are about equally
frequent. In addition, most workers have examined
the morphological structures of the fungi (hyphae,
arbuscules and, where appropriate, vesicles) sep-
arately, and again have reported almost all possible
combinations of increase, decrease and no response.
Staddon & Fitter (1998) argued that this was due to
a serious aw in such experiments: because eCO
#
is
known to aect (typically increase) plant growth, if
plants are grown for the same length of time in aCO
#
and eCO
#
, any comparison between them is of plants
of dierent sizes and probably dierent growth
stages. Because the nature and degree of mycorrhizal
colonization of roots are not a plant characteristic,
but are instead highly dependent on plant condition,
consequent changes in colonization would be
expected. Those observed might therefore merely be
artefacts of comparing dissimilar plants (Staddon,
1998).
In a series of time-course experiments, in which a
sequence of harvests allowed the comparison of
plants of similar sizes and developmental stages by
covariance analysis, Staddon and co-workers could
nd no evidence that mycorrhizal colonization by an
isolate of Glomus mosseae was aected by growing
plants in eCO
#
(Fig. 3). This result was true whether
internal (Staddon et al., 1998) or external (Staddon
et al., 1999a) colonization was examined and was
robust across a range of 10 dierent plant species of
contrasting life histories and growth rates (Staddon
et al., 1999c).
There are very few other published studies in
which a proper allometric analysis of mycorrhizal
responses has been made, and it is therefore dicult
to be sure of the generality of these ndings. Rouhier
& Read (1998) used three harvests in a study of
Plantago lanceolata; they found no eect of eCO
#
at
the rst harvest but increases in colonization at the
later two. However, Staddon et al. (1998) showed
that these responses could have been due to growth
eects, which was the conclusion that ONeill et al.
182 REVIEW A. H. Fitter et al.
60
40
20
0
5 6 7 8 9 10
Log
e
plant biomass (mg)
%

R
o
o
t

l
e
n
g
t
h

c
o
l
o
n
i
z
e
d
(a)
(b) 7
6
5
4
3
5 6 7 8 9 10
Log
e
plant biomass (mg)
E
x
t
e
r
n
a
l

m
y
c
o
r
r
h
i
z
a
l

h
y
p
h
a
l

l
e
n
g
t
h
(
m
m

g

1

s
o
i
l
)
Fig. 3. The percentage of root length of Plantago lanceolata
colonized by Glomus mosseae (a) and the external hyphal
length (b) are unaected by growth at elevated (650 l l
" ;
diamonds and broken line) as opposed to ambient (400 l
l
" ; squares and solid line) [CO
#
], if the eect of [CO
#
] on
plant growth is taken into account. Lines are tted
regressions and do not dier between treatments. Data
from Staddon et al. (1999a).
(1991) came to in their three-harvest study of
Liriodendron tulipifera. However, Rillig & Allen
(1999) reported unpublished data that do not
conrm the ndings of Staddon and co-workers,
and Sanders et al. (1998) found a ve-fold increase in
extraradical mycelium in eCO
#
although there was
only a twofold increase in both root biomass and
intraradical mycelium. Until further work has been
completed, it therefore remains impossible to state
what the likely eects of eCO
#
will be on mycorrhizal
symbioses. We would urge mycorrhiza researchers
to ensure that they allow for the direct eects of
eCO
#
on plant growth in all future experiments.
Mechanisms
Because it is certain that the plant must mediate any
eect of eCO
#
on an AM fungus, we need rst to
understand the control of AM colonization. The key
event here is the transfer of C from plant to fungus,
because it is the fungus that is the obligate partner,
and it is that C supply which it cannot obtain
elsewhere. Unfortunately, our knowledge of this
central relationship is poor. It is often assumed that
the arbuscule is the site of both P transfer from
fungus to plant and C transfer from plant to fungus.
Although that assumption is probably correct for P,
it is likely not to be so for C. Histochemical evidence
and the fact that hyphae can colonize roots eectively
even when producing no arbuscules both suggest
that the intercellular hyphae might be at least in part
the location where C is lost from plant cells and is
absorbed by fungal membranes (Smith & Smith,
1996).
If C and P transfer are spatially dislocated, there is
no basis for the assumption that they are meta-
bolically linked. The fungi might merely be eective
scavengers for C that leaks or is exported across plant
cell membranes, able to acquire it in competition
with the reabsorption mechanisms of the plant cells.
The control might therefore be the rate of that loss
from plant cells. It is well established that if plants
are grown under conditions of reduced irradiance or
high P supply, mycorrhizal colonization declines.
This eect is highly variable in terms of the
quantitative relationship between the environmental
factor and the response, but it is nevertheless
qualitatively consistent. The eect of low light is
consistent with a non-specic control mechanism, in
which carbohydrate concentrations control the rate
of eux from plant cells. The eect of high P could
be explained on the basis that it would increase
metabolic activity, and hence demand for carbo-
hydrate, incidentally reducing eux. However, such
an explanation should apply equally to N and to P,
suggesting a more specic mechanism.
Syvertsen & Graham (1999) conducted a detailed
analysis of the response of two Citrus spp. (Citrus
aurantium and C. sinensis) to eCO
#
and a high P
supply and, importantly, measured plant carbo-
hydrate pools. High P, eCO
#
and mycorrhizal
inoculation (with Glomus intraradices) all increased
plant growth, and high P reduced mycorrhizal
colonization; the eect of eCO
#
on colonization was
weak. All three factors had large impacts on leaf and
root starch concentrations. Leaf starch concen-
trations were low in all P and mycorrhiza treatments
at aCO
#
, but greatly increased at eCO
#
except in non-
mycorrhizal plants at low P. At high P and aCO
#
,
plants had lower root-starch concentrations (Fig. 4).
Mycorrhizal colonization decreased root starch only
at low P and eCO
#
, conditions under which the C
assimilation rate (A) was most stimulated. These
results show that the large C cost imposed by
mycorrhizal colonization on the plant (estimated at
1020% of C xed in photosynthesis in well
colonized plants) decreases root carbohydrate stores
and stimulates photosynthesis. However, if the eect
is due to mycorrhizal fungi absorbing C that leaks
passively across cell membranes in the root, this does
not necessarily result in an increase in C supply to
REVIEW Mycorrhizas and global change 183
30
20
10
0
(a)
Low P
aCO
2
eCO
2
aCO
2
eCO
2
High P
20
10
0
(b)
Low P
aCO
2
eCO
2
aCO
2
eCO
2
High P
F
i
b
r
o
u
s

r
o
o
t

s
t
a
r
c
h
(
m
g

g

1
)
A
C
O
2
(
l
m
o
l

m

2

s

1
)
Fig. 4. (a) Colonization of roots of Citrus aurantium by the
mycorrhizal fungus Glomus intraradices (closed bars)
decreases root starch concentration only at low P supply
rates and principally under elevated (710 l l
") [CO
#
]
compared with ambient [CO
#
] (aCO
#
: 350 l l
") and non-
mycorrhizal plants (open bars). (b) The impact of
mycorrhizal colonization on C assimilation rate (A
CO
#
)
exactly mirrors that on root starch, with AM colonization
increasing A
CO
#
at low P and eCO
#
, suggesting the
alleviation of down-regulation of photosynthesis owing to
the more active root sink. Data from Syvertsen & Graham
(1999).
the fungus. It merely shows that root carbohydrate
stores decline as fungi compete eectively with plant
cells for apoplastic sugars, resulting in an export of
leaf carbohydrate and relief of the limitation that leaf
starch imposes on photosynthesis (Fig. 1). An
eventual consequence might be an increased C
supply to the fungus, but this is determined by the
dynamics of sugars in the root apoplast and is not a
necessary consequence of eCO
#
.
However, this result might not be universal : a
study of ectomycorrhizas found no correlation
between root starch and sugar concentrations and
the percentage of ne root tips colonized by
mycorrhizal fungi (Lewis et al., 1994). There is an
urgent need for quantitative studies of C uxes in
mycorrhizal plants under varying [CO
#
].
Fungal diversity
Much work on mycorrhizas has involved the use of
single isolates of AM fungi, often well known
laboratory organisms such as Glomus mosseae. Rillig
& Allen (1999) highlighted this as a feature of the
series of experiments by Staddon et al. (1998,
1999a,b,c) that suggested that the symbiosis was
largely unresponsive to eCO
#
. The potential for
fungal species to behave dierently from each other
has indeed received too little attention, partly
because of the inadequacy of our understanding of
fungal taxonomy. For example, many early papers
on AM function give names for fungal species used
that are impossible to verify. Most laboratories now
use a number of well dened isolates, not all of which
have been identied; increasingly, culture collections
such as INVAM (Bentivenga & Morton, 1994) are
able to supply well characterized cultures and this
might lead to a situation in which it is possible to
make direct comparisons between the work of the
various active research groups. As yet, however, the
number of taxa involved is unknown, although c. 150
have been reliably described, and the ecology and
physiology of most of these are equally obscure. If
they dierentiate in respect of their response to
environmental factors, as is likely from ecological
rst principles, then changing environmental factors
such as temperature, precipitation or N deposition
might alter fungal communities (Fig. 2).
Some fungi seem very much better at supplying P
to host plants than others (Pearson & Jakobsen,
1993). If improved P acquisition is viewed as the
main benet that plants gain from the symbiosis,
what is it that the other fungi do that keeps them in
the community? There are four main possibilities.
1. They are ineective mutualists that plants cannot
recognize as such, and so they are able to colonize
plants as extensively as more eective fungi.
2. They are eective mutualists with plant species
other than those that have been studied, giving a
system of specic interactions between plant and
fungal species.
3. They are eective mutualists in conditions other
than those that have been studied: most workers
use standard laboratory or growth-chamber con-
ditions and treated or articial growth media,
whereas soils are often colder (or occasionally
hotter), wetter (or drier), physically dicult (e.g.
compacted) and more biologically active (both
animals and microorganisms).
4. They perform functions other than P uptake:
fungal taxa that transport P poorly to roots might
be better at protecting them from pathogens or
drought, for example. It is reasonable to imagine
that the fungal attribute that best promotes P
acquisition might be an extensive external my-
celium, whereas that which best provided drought
resistance would be an intense development of
fungal mycelium in the rhizosphere, so binding
roots eectively to soil, and that which best gave
protection against pathogens would be an ex-
tensive internal mycelium. These traits might
therefore be mutually exclusive.
Given this level of uncertainty about the role of
fungal diversity, and with the knowledge that it is
not uncommon to nd 10 or more species of AM
fungi in the roots of plants in a single community
(Helgason et al., 1998), it is not surprising that we
cannot predict the consequences of an indirectly
acting environmental variable such as eCO
#
on the
184 REVIEW A. H. Fitter et al.
120
100
80
60
40
20
0
Glomus intraradices
Glomus entunicatum
Acaulospora denticulata
Scutellospora calospora
C
h
a
n
g
e

a
t

e
C
O
2

(
%
)
I
n
t
e
r
n
a
l
h
y
p
h
a
e
A
r
b
u
s
c
u
l
e
s
E
x
t
e
r
n
a
l
h
y
p
h
a
e
S
p
o
r
e
s
Fig. 5. Growing mycorrhizal plants of Artemisia tridentata
at elevated [CO
#
] (700 l l
") results in distinctive changes
in fungal growth parameters depending on the species of
AM fungus involved. Data are percentage change in
mycorrhizal fungal growth (internal and external hyphae,
arbuscules and spores) at eCO
#
compared with aCO
#
(350
l l
"). Data from Klironomos et al. (1998).
symbiosis. There is evidence that eCO
#
aects fungal
taxa dierentially. First, it has been shown that eCO
#
can aect mycorrhizal and non-mycorrhizal fungi
dierently (Klironomos et al., 1997). Klironomos et
al. (1998) also showed that mycorrhizal fungi
themselves can respond dierently to eCO
#
. They
grew the shrub Artemisia tridentata with four AM
fungi, two species of Glomus (G. intraradices and G.
etunicatum), Acaulospora denticulata and Scutello-
spora calospora in pots of an articial growth medium
in growth chambers. This was a single harvest-pot
experiment, but eCO
#
had little eect on plant
growth, so the usual strictures about comparability
are less important. The two Glomus spp. promoted
shoot growth more than did the other two fungi
(there were no non-mycorrhizal controls), but they
promoted root growth only under eCO
#
. The
impacts of eCO
#
on A. denticulata and S. calospora
are therefore especially interesting, because plants
colonized by them were apparently identical in aCO
#
and eCO
#
. [CO
#
] had no eect on either the internal
mycelium or spore production by A. denticulata and
S. calospora, but did increase external mycelium
production (Fig. 5). By contrast, eCO
#
had no eect
on the external mycelium of the two Glomus spp.,
but it did increase the frequency of arbuscules and
internal hyphae, and also spore production, albeit on
larger root systems. These results demonstrate that
growing plants in high atmospheric [CO
#
] might
have dierential impacts on the fungal taxa with
which they associate, but we are a long way from an
understanding of the functional implications.
Furthermore, the reported eects of eCO
#
on AM
fungi are strongly inuenced by soil nutrient avail-
ability, especially N (Klironomos et al., 1996, 1997).
This led Klironomos et al. (1997) to conclude that
eCO
#
could signicantly alter the community struc-
ture of the plantsoil system towards either a more
mutualistic-closed, mycorrhizal dominated food web
under high [CO
#
] and low N availability, [or] a more
opportunistic-open, saprobe\pathogen dominated
one under high N availability. N deposition is
another increasingly important component of global
change, which will have large impacts on AM fungi.
Future studies therefore need to be alert to the
possibility that outcomes can be determined by the
selection of isolates. Generalizations about mycor-
rhizal behaviour need to be based on studies with
multiple taxa.
1ii noii oi iio\i v oioov
Our ability to predict the impact of rising atmos-
pheric [CO
#
] on mycorrhizal symbioses is strongly
limited by our poor grasp of the biology of the fungal
partner. The great bulk of research on the symbiosis
has used a limited number of often poorly dened
isolates. Most studies have fallen into one of two
categories: pot experiments with single plants and an
inoculum of the fungus, or eld experiments with
natural communities comprising several to many
plant species and an unknown number of largely
unidentied fungi. Both of these approaches have
characteristics that limit their value.
In pot experiments, the soil initially contains no
mycelium, except for those fragments that are added
as inoculum. As the plant grows, so must the fungus,
to create the extraradical mycelium with which it
forages in soil. This requires the transfer of large
amounts of C (and possibly other elements) from
plant to fungus, so creating the carbon drain
phenomenon that has been shown repeatedly. Such
experiments therefore represent systems far from
any equilibrium: both plant and fungus are estab-
lishing themselves. The nearest analogous eld
system is arable agriculture. In most natural and
many agricultural situations, the plants already exist
and are replacing rather than creating structure, and
this, critically, is also true of the fungus. Estimates of
the quantity of extraradical mycorrhizal mycelium
vary widely (Smith & Read, 1997, Table 2.3, p. 66),
but 100 m m
" of root is a reasonable average. Even
allowing for a dierence in radius of 10100-fold
between hyphae and roots, this is a substantial
quantity and most pot experiments must be in a
phase of mycelial development when harvested,
unless they are maintained for a long time. Because
the various fungal taxa have distinct patterns of
extraradical growth, it is unsurprising if they respond
dierently to a change in C supply in the roots, but
these responses might be quite dierent from those
that would be shown by an existing mature my-
celium, for which the C was required largely for
maintenance rather than for growth.
REVIEW Mycorrhizas and global change 185
2
1
0
1
14 15 16
(a)
(b)
1
0
1
2
3
23 24 25
R
o
o
t

s
h
o
o
t

1
3
C
Root
13
C
Fig. 6. Carbon transport into mycorrhizas of plants of the
C
%
grass Cynodon dactylon linked by a mycorrhzal network
to plants of the C
$
herb Plantago lanceolata (a) in ambient
CO
#
(380 l
") and (b) in elevated CO
#
(610 l l
"). (a) In
aCO
#
, C. dactylon has "$C of c. k15=. More negative
values are due to c transport via mycorrhizal hyphae from
P. lanceolata, which has "$C of c. k30=. Root "$C
represent both plant and fungal tissue. The dierence
between root and shoot "$C signals is linearly related to
the root "$C, showing that all transported C stays in the
roots, even after clipping of the shoots at harvest 1
(squares) and the subsequent regrowth of the shoots
before harvest 2 (triangles). Most C. dactylon plants at
aCO
#
have a fraction of the C in their roots derived from
P. lanceolata. Regression analysis showed that the degree
of movement was a function of mycorrhizal hyphal coloni-
zation of roots. (b) Growing the plants at eCO
#
(in which
the C. dactylon signal was c. k24= and the P. lanceolata
signal c. k40=, due to the dierent "$C of the added
CO
#
) had no eect on C transport. From Fitter et al.
(1998), with permission.
These characteristics of AM fungi mean that
conventional pot experiments are inappropriate for
studies on C budgets, symbiotic eectiveness and
other high-level phenomena, although they might
still be ideal arenas for gaining basic data on fungal
physiology, for example the response to soil tem-
perature. An analogy would be that caged birds
might be usable for studies of basic physiology but
not of breeding behaviour.
Field experiments suer from a quite dierent
problem. Here the fungal mycelia are in place but
their identity is almost always unknown, except in
the most general of senses. Many workers have
identied spores extracted from eld soils, but Clapp
et al. (1995) showed that there might be no
correlation between spore collections and the fungi
identied within the roots by DNA-based methods.
At some level this problem might be unimportant. If
the concern is merely to quantify rates of C ux from
roots to soil, or turnover rate of hyphal C in soil
(Rillig & Allen, 1999), the identity of the fungal
species can temporarily be ignored. However, pre-
diction is made dicult if the fungal taxa do not all
respond in the same manner to changing environ-
mental conditions.
Field experiments nevertheless have the great
merit of studying the mature fungus, which com-
prises a mycelium in soil that colonizes a large (and
typically unknown) number of root systems. Fitter et
al. (1998) have argued that the behaviour of AM
fungi must be studied from this perspective, on the
basis of an examination of the movement of C
between root systems linked by a common mycor-
rhizal network. Using the stable C isotope "$C and
mixtures of C
$
(Plantago lanceolata) and C
%
(Cynodon
dactylon) plants, which discriminate dierently
against this isotope in photosynthesis, they found
that as much as 40% (although typically 10%) of
the C in the root system of a plant could be derived
from a neighbouring plant. This C could only have
moved through mycorrhizal hyphae linking the root
systems. However, the C always stayed in the roots
and was never transported to the shoots, even when
the shoots were cut and made to regrow, using stored
reserves from the roots (Fig. 6). This was strong
evidence that the C remained at all times in fungal
structures. Moreover, the great variation in move-
ment of C between plants was best explained by
variations in fungal structures in the roots, with C
moving into roots with many vesicles and out of
roots predominantly colonized by hyphae. Because
vesicles are thought to be C storage structures and
hyphae could be the sites of C acquisition by the
fungus, this result leads to a view of the fungus as an
organism with its own C dynamics and growth
pattern, transporting C from some parts of its
mycelium and into other parts where it is building
reserves. To test whether plants could control these
C dynamics through variation in C supply in the
roots, Fitter et al. (1998) replicated this experiment
under aCO
#
and eCO
#
. The increased C supply in
the roots of plants under eCO
#
did not result in any
greater movement out of those root systems, again
supporting the view that fungal behaviour is more
important than variations in plant nutrition in
determining fungal response.
Our viewpoint on mycorrhizas has shifted mark-
edly from one in which the plant alone was studied,
with often detailed measurements of plant per-
formance, and a perfunctory assessment of fungal
colonization, to one in which the pair of organisms is
perceived as an integrated symbiosis. However, as
ONeill et al. (1991) have pointed out and as has
recently been reiterated by Rillig & Allen (1999), this
is itself but a step on the path to a proper viewpoint.
They have argued for a hierarchical framework
incorporating plant host, plant population, plant
community, functional group and ecosystem, so as to
be able to answer questions about nutrient uptake,
the control of plant community structure or C ux in
186 REVIEW A. H. Fitter et al.
ecosystems. However, in such an approach it is still
essential to study the fungus as an organism. It is
essential that we not only recognize the existence of
mycorrhizal fungi, but also study them directly as
complete mycelia, including both the extensive
extraradical phase and the numerous intraradical
elements. This is dicult because a single mycelium
can extend for a considerable distance, but the use of
DNA-based techniques now permits the reliable
identication of fungal genotypes in the eld, both in
soil and in roots.
We need to understand how these organisms
respond to changes in their biotic and abiotic
environments. For example, one of the few clear
predictions about global change is that there will be
widespread increases in temperature. To our knowl-
edge, the only published work on the eect of
climate change in a broad sense on arbuscular
mycorrhizas is that of Monz et al. (1994). Although
they provided no data on plant responses to the
altered environmental conditions ([CO
#
], tempera-
ture and precipitation), they did nd that an increase
in temperature of 4mC above ambient decreased the
percentage root length colonized (RLC) by AM
fungi in Pascopyrum smithii after both 2 and 4 yr.
They also found that increased precipitation altered
RLC in both P. smithii and Bouteloua gracilis but not
in a consistent or predictable manner, and that eCO
#
increased RLC in B. gracilis but not in P. smithii.
There are very few other studies on the eects of
soil temperature on mycorrhizas. Barley was not
colonized by Glomus etunicatum at 10mC but was at
15mC (Baon et al., 1994). Was this because of eects
on the plant or on the fungus? The former seems
more likely, because the mycorrhizal colonization of
roots of bluebells (Hyacinthoides non-scripta) reaches
a peak in a woodland in Yorkshire, UK, in
midwinter, when soil temperatures are 5mC
(Merryweather & Fitter, 1995). Nevertheless,
colonization of roots of Plantago lanceolata by
Glomus mosseae (but not extraradical hyphal growth
per unit root length) was greater at 20mC than at
12mC, even allowing for the eects of temperature on
plant growth (A. Heinemeyer, unpublished; Fig. 7).
Some fungi might be more sensitive to temperature
than others, oering an axis for niche dierentiation
in the fungal community. Such a situation would
also ensure that changes in soil temperature have
profound impacts on the composition of that com-
munity, with consequent eects on the plant popu-
lations.
There is also a clear lack of research on the eects
of other atmospheric pollutants such as NH
$
and O
$
on mycorrhizas in general (Cairney & Meharg,
1999). Pe! rez-Soba et al. (1995) showed that both
NH
$
and O
$
had negative eects on ectomycorrhizal
colonization and that these were not alleviated by
eCO
#
. In practice, however, there will be interactions
between all these components of the changing global
50
40
30
20
10
0 20 40 60 80
(a)
Days after planting
L
R
C

(
%
)
25
20
15
10
5
0 0.2 0.4 0.6 0.8 1.0
T
o
t
a
l

L
R
C

(
m
)
Total plant biomass (g)
(b)
Fig. 7. (a) The percentage of the root growth of Plantago
lanceolata colonized by the fungus Glomus mosseae (L
R
C)
increases more rapidly at 20mC (closed diamonds, solid
line) than at 12mC (open diamonds, broken line). (b) The
total length of colonized root in the same experiment is
higher at 20mC (closed diamonds, solid line) than at 12mC
(open diamonds, broken line), irrespective of the eect and
temperature on plant biomass. Lines are best-t
regressions. Data from A. Heinemeyer (unpublished).
environment, and other factors not yet mentioned,
such as habitat fragmentation (the eects of which
will be dependent on the distribution and dispersal
abilities of both plants and fungi). It will be
impossible for researchers to adopt the largely
empirical approach that has so far characterized
much ecological work on mycorrhizas. Rather, we
must develop predictive models based on a proper
understanding of the biology of the organisms
involved, and test the explicit predictions of these
models.
\ioviiioii1s
We thank Owen Atkin and Angela Hodge for their
comments on the manuscript. Much of the work reported
here was funded by the Natural Environment Research
Council.
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