New Phytologist - May 1986 - FINLAY - THE STRUCTURE AND FUNCTION OF THE VEGETATIVE MYCELIUM OF ECTOMYCORRHIZAL PLANTS

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New Phytol. i\9S6) 102, H3-\56 ^43
THE STRUCTURE AND FUNCTION OF THE

VEGETATIVE MYCELIUM OF
ECTOMYCORRHIZAL PLANTS
L TRANSLOCATION OF i*C-LABELLED
CARBON BETWEEN PLANTS INTERCONNECTED BY
A COMMON MYCELIUM
BY R. D . F I N L A Y AND D . J. READ
Department of Botany, University of Sheffield, Sheffield, SIO 2TN, UK
{Accepted 23 December 1985)
SUMMARY
It is increasingly evident that in natural plant communities the vegetative mycelia of ectomy-
corrhizal fungi can form networks of hyphal interconnections which link the root systems of
their host plants in both intra- and inter-specific combinations. Root observation chambers have
been used to examine the development of these mycelial networks and to assess their functional
significance as pathways for the transfer of assimilate between individuals in a range of host-fungus
associations. Carbon transfer between plants of Pinus spp. is significantly increased by the
presence of mycelial connections and preliminary evidence suggests that such transfer may be
enhanced where concentration gradients are induced by shading. The significance of these
experimental results is discussed in relation to nutrient cycling processes in natural ecosystems.
Key words: Assimilate transfer, ectomycorrhizal fungi, Pinus spp., nutrient cycling.
INTRODUCTION
Studies of the structure and function of individual mycorrhizal plants or roots have
provided much information concerning their basic physiology but much less is
known about the way in which these mutualistic associations function in natural
plant communities. In such situations the roots of a number of difTerent plant
species may grow in close proximity to each other, forming mycorrhizal associations
with a range of fungal species. Both vesicular-arbuscular mycorrhizal (VAM) and
ectomycorrhizal fungi show low levels of host specificity and one consequence of
this is that hyphae growing from root to root and plant to plant form a mycelial
network capable of connecting both intra-specific and inter-specific combinations
of their host plants. The functional significance of such a network, in terms of its
capacity to act as a direct pathway for inter-plant nutrient transfer, has been
considered by a number of workers (Bjorkman, 1960; Woods & Brock, 1964; Hirrel
& Gerdemann, 1979; Chiariello, Hickman & Mooney, 1982); however, only
indirect evidence of such a pathway is available from these studies. Reid & Woods
(1969) were able to demonstrate transfer of ^*C between adjacent seedlings of Pinus
taeda L. infected with Thelephora terrestris (Ehrh.) Fr., but only when the root
systems were artifically connected by wrapping a mycelial strand from the recipient
around a mycorrhizal root of the donor.
The delicate nature and inaccessibility of these mycelial systems make non-
destructive investigation of their biology in the field almost impossible and
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14698137, 1986, 1, Downloaded from https://nph.onlinelibrary.wiley.com/doi/10.1111/j.1469-8137.1986.tb00603.x by EVIDENCE AID - BELGIUM, Wiley Online Library on [18/03/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
144 R- ^- FiNLAY AND D . J. READ
necessitates the use of laboratory observation chambers which permit in situ

examination of the development and function of mycelial networks in appropriate
combinations of host plants, fungi and soils and under conditions resembling, as
closely as possible, those found in the field. Such observation chambers have been
developed in our laboratory and used to demonstrate inter-plant transfer of current
assimilate in both VAM (Francis & Read, 1984) and ectomycorrhizal (Brownlee
et al., 1983) plants. In both types of system the mycelia spreading from mycorrhizal
roots rapidly initiate infection in uninfected plants and a network of hyphal
interconnections between plants is thus established. Francis & Read (1984) have
used autoradiography to provide direct qualitative evidence that ^*C transfer
between VAM plants occurs primarily by this hyphal pathway, and quantitative
data from their experiments indicate that the magnitude of such transfer is strongly
influenced by the shading of 'receiver' plants, suggesting that the transfer of
assimilate occurs along concentration gradients influenced by source-sink relation-
ships. Qualitative evidence of ^^C transfer between Pinus sylvestris L. plants via
mycelial strands of the ectomycorrhizal fungus Suillus bovinus (Fr.) O. Kuntze has
been presented by Brownlee et al. (1983) and preliminary quantitative data have
been presented by Read, Francis & Finlay (1985). In this paper we present
quantitative evidence of such a transfer capability in a range of ectomycorrhizal
associations together with data relating to ^^C translocation rates, hyphal growth
rates, and the influence of source-sink relationships on inter-plant transfer.
M A T E R I A L S AND M E T H O D S
Synthesis of mycorrhizas and establishment of observation chambers
Ectomycorrhizas were synthesized aseptically in 250 ml Erlenmeyer flasks
containing an autoclaved peat-vermiculite (1:4 v/v) mixture moistened with
modifled Melin-Norkrans solution (MMN) and inoculated with pure cultures of
niycorrhizal fungi, isolated from fruit bodies. After the fungus had colonized the
flasks, aseptically germinated seedlings oi Pinus sylvestris and Pinus contorta Dougl.
ex Loud were added. The flasks were maintained in a growth chamber and exposed
to a 16 h day, with day temperature of 15 °C, night temperature of 10 °C and
irradiance of 38 W m~^. Mycorrhiza formation usually occurred within eight
weeks, after which time the infected seedlings were transferred to observation
chambers.
Chambers were constructed from square plates of transparent perspex or
'Darvic' (ICI) sheet with a thickness of 15 mm. Three chamber sizes were used,
12x12 cm (small), 20 x 20 cm (medium) and 40 x 40 cm (large). Perspex spacers
were glued to the outer margins of the basal plate of each chamber so that the upper
plate rested on these rather than the peat surface. They were of sufficient thickness
to enable a layer of non-sterile, milled peat or forest soil between 3 and 6 mm deep
to be placed on the base of each chamber. The peat was moistened with a dilute
(1:10 v/v) solution of MMN without a carbon source and flattened to a level just
below that of the spacers so that there was a small gap between the peat surface
and the chamber lid. The lid and base of each chamber were held together using
3 cm long clips cut from plastic document-binding spines. The chambers were
wrapped in aluminium foil and kept in an upright position inside plastic plant
propagators which were then transferred to a growth room maintained under the
conditions described above, and supplied with dilute carbon-free M M N solution
as required to maintain moisture levels.
Carbon transfer between ectomyeorrhizal plants 145
Carbon transfer experiments in mycorrhizal and non-mycorrhizal chambers

Mycorrhizal and non-mycorrhizal plants, of similar size, were transferred to
medium-sized observation chambers. In mycorrhizal chambers extensive mycelial
colonization of the upper surface of the peat took place within four to six weeks,
after which time one or two uninfected four week old pine seedlings of the same
or a related species were introduced into all chambers 4 to 8 cm on either side of
the central plant. Where the roots of uninfected seedlings made contact with the
spreading mycelium, infection of their lateral roots was initiated within 3 d. After
12 to 14 weeks, further differentiation and spread of the mycelium had taken place
and the plants in chambers with mycorrhizas were interconnected by a network
of mycelial strands.
Associations of the type described above were used to measure the capacity of
mycelial strands to act as pathways for carbon transport between plants (Experi-
ments la and Ib). The shoot system of the older, central plant in each chamber was
enclosed in a perspex box and exposed to ^^COg produced by the addition of 10%
lactic acid to NaH^^COg. Different amounts of ^^COa and different feeding periods
were used in different experiments, the details of which are summarized in Table
1. Chambers were maintained in the illuminated growth chamber throughout the
feeding period after which the 'donor' shoot was removed. In selected chambers
the distribution of label was examined autoradiographically by opening the
chamber, covering the peat surface with thin acrylic film (Melinex, ICI) and
freezing the chamber prior to incubation at sub-zero temperature in contact with
X-ray film. Quantitative determination of isotope distribution was obtained by
grinding plant roots, including mycorrhizas, and shoots in liquid nitrogen,
Table 1. Summary of experimental treatments in six different experiments

Feeding Amount
Fungal Donor Receiver Replicate period of "CO2
Experiment species plant plant chambers (d) QiCi)
Intraspecific transfer
1 (a) Non-mycorrhizal Pinus contorta Pinus contorta 4 3 50
Suillus granulatus Pinus contorta Pinus contorta 8 3 50
Suillus bovinus Pinus contorta Pinus contorta 8 3 50
Inter-specific transfer
1 (b) Non-mycorrhizal Pinus contorta Pinus sylvestris 4 4 10
Suillus bovinus Pinus contorta Pinus sylvestris 5 4 10
Non-mycorrhizal Pinus sylvestris Pinus contorta 4 4 10
Suillus bovinus Pinus sylvestris Pinus contorta 7 4 10
Shading effects
Suillus granulatus Pinus contorta Pinus contorta 4 5 50
2 (a)
shaded/unshaded
Suillus granulatus Pinus contorta Pinus sylvestris 4 5 50

2(b)
shaded/unshaded
Pisolithus Pinus sylvestris Pinus contorta 6 7 50

2(c)
tinctorius shaded/unshaded
Suillus bovinus Pinus sylvestris Pinus radiata 4 7 120

2(d)
shaded/unshaded
All experiments were carried out in chambers containing peat except in 2 (c) where colliery spoil was used.
146 R. D . F I N L A Y AND D . J. READ
digesting aliquots of the extract in NCS tissue solubilizer and adding liquid
scintillant prior to counting on a Packard Liquid Scintillation counter.
Shading experiments
In experiments 2a, b, c and d the effect of shading on carbon transfer was
investigated by loosely covering the plant on one side of each chamber with
aluminium foil for 9, 19, 28 and 56 d respectively before feeding. Shading was
continued throughout the feeding period and the plants in each chamber were then
analysed in the manner described above.
Measurement of fungal growth rates

The growth of Suillus bovinus and Pisolithus tinctorius (Mich, ex Pers) Coker
& Couch in chambers similar to those mentioned previously was followed by
tracing the position of the leading edge of each mycelial fan onto the lid of the
observation chamber at weekly intervals for six weeks. These growth patterns were
photocopied onto white paper and then digitized using a microcomputer interfaced
to a graphics tablet. Sequential measurements of the area of each chamber
colonized by fungal mycelium were made using the computer and these data were
used to calculate daily rates of mycelial spread. Manual measurements were also
made at weekly intervals to calculate rates of strand extension.
Measurement of ^*C translocation rates in mycelial strands

Translocation rates of ^*C in mycelial strands were made using large observation
chambers extensively colonized by well-developed mycelial fans. The lid of each
chamber was carefully removed and the peat surface wrapped in 'Melinex' to
prevent desiccation and disruption of the mycelium. One plant in each chamber
was fed with ^^COg as described above and the movement of label was followed
using a windowless ^ detector connected to a Packard radiochromatogram scanner
with ratemeter and chart recorder. The detector was mounted in a thick sheet of
perspex and covered by a metal shield with a central aperture (1-5x10 mm). It
could thus, with appropriate positioning, detect activity in small, localized regions
of the mycelium without being influenced by the background levels in roots. The
detector was placed over a mycelial strand and the passage of label was followed
by moving the detector through a series of positions away from the root, recording
the time taken for the activity to rise above the background level at each of the
positions.
Statistical analysis
In experiments la and lb data relating to statistically non-independent plants
within a chamber were combined to obtain mean values for each chamber. The
number of replicate chambers in each experiment is indicated in Table 1. Data
were In transformed before being subjected to one way analysis of variance. The
significance levels of these F tests are indicated above the histograms in Figures
5 to 7. In experiments 2a, b, c and d, because of the inherently high variation
between chambers, individual chambers were treated as experimental blocks and
the shading treatment was assigned randomly to half of each chamber, the other
half remaining fully illuminated. Data from statistically non-independent plants
were, as before, combined to obtain mean values and after In transformation these
data were subjected to a one way analysis of variance for a fully randomized block
design.
Carbon transfer between ectomyeorrhizal plants I47
RESULTS
Mycelial growth and strand extension

Rates of mycelial growth and strand extension for two species, Suillus bovinus
and Pisolithus tinctorius, are displayed in Table 2. Colonization of individual
chambers by different fungal strains was variable but the mean growth rates were
similar for both species. Growth of mycelial fans across peat surfaces was rapid
(91 to 15-9 cm^ d"^) but colonization of the bulk volume of peat occurred more
slowly making extrapolation of these figures to three dimensions impossible.
Strand extension occurred at 24 to 4-3 mm per day, a rate similar to that measured
for Rhizopogon luteolus growing in associations with Pinus radiata (Skmner &
Bowen, 1974).
Table 2. Rates of mycelial spread and strand extension in Suillus bovinus and
Pisolithus tinctorius grown in association with Pinus contorta plants
Rate of mycelial spread Mycelial strand extension
Fungus (cm^ d"!) (mm d"")
Suillus bovinus (Strain A) 11 7 ± 27 4-3 ± 1 1

Suillus bovinus (Strain B) 9-1 ± 39 31 ± 0-7
Pisolithus tinctorius 15-9 ±5-9 24 + 0-6
Growth chambers were maintained at a day temperature of 20 °C for 16 h per day and a night temperature
of 15 °C for 8 h p e r day.
95 % confidence intervals are shown.
The high density of hyphae at the advancing mycelial front (Fig. 1) resulted in
an intensive exploitation of soil micro-habitats. Aggregation of hyphae behind the
front to form mycelial strands led to a reduced intensity of hyphal colonization
in the already exploited soil (Fig. 2) and the maintenance of arterial structures
which, with progressively greater age, had fewer and fewer hyphal contacts with
the soil [Fig. 3(a), (b)], except in certain localized areas which remained heavily
colonized by fine hyphae [Fig. 4(a)]. In these sites proliferation of short hyphae
led to the formation of dense patches of mycelium which made close contact with
the substrate [Fig. 4(b)]. Autoradiographic analysis of these patches revealed that
they were strong sinks for the carbon being translocated from the host root [Fig.
4(c), (d)]. Intensive investment of carbon in these areas suggests that they may have
been areas of nutrient enrichment. The concept of selective exploitation of soil
heterogeneity has been described in relation to distribution of roots and VA
mycelium by St John, Coleman & Reid (1983) and is discussed further in the
context of ectomyeorrhizal infection in the following paper (Finlay & Read, 1986).
Measurement of ^^C translocation rates

Long distance translocation of ^^C in the fungal mycelium of infected plants
was measured in large observation chambers with actively growing mycelial fans.
Activity was detected in the root system of the fed plant within 3 h of the exposure
of the shoot system to ^^COg. Translocation of the label within mycelial strands
was detected over distances of 10 cm within 30 min suggesting translocation
velocities in excess of 20 cni h^^.
148 R. D. F I N L A Y AND D . J. READ
Fig. 1. Mycelial fans of Suillus bovinus developing in a root chamber containing interconnected
host plants of Pinus contorta and Pinus radiata. Exploitation of fresh peat is occurring four weeks
after a fully colonized, medium sized chamber has been transferred into a large chamber. The high
density of mycelium behind the hyphal front is evident.
Transfer of ^^C in mycorrhizal and non-mycorrhizal chambers

Autoradiographs of systems with mycelial interconnections between plants
[Fig. 4(a), (b)] show that the myceUa provided direct pathways for the translocation
of ^*C compounds over distances exceeding 20 cm, both to the actively growing
hyphal fronts of mycelial fans and to the mycorrhizal roots of plants incorporated
into the mycelial network. Newly formed mycorrhizal roots of 'receiver' plants
acted as major sinks for the carbon. Thus, current assimilates of fed ' donor' plants
were widely distributed to other plants with access to the same mycelium.
Quantitative determination of the distribution of radioactivity within intra-
specific associations oi Pinus contorta plants (Fig. 5) showed that significantly larger
amounts of label occurred in both shoots and roots of plants grown in mycorrhizal
chambers than in those of non-mycorrhizal plants where mycelial connections were
Carbon transfer between ectomycorrhizal plants 149
Fig 2 A chamber similar to that shown in Figure 1, but photographed eight weeks after transfer
of a medium sized chamber into a larger one and with Pinus sylvestris and Ptnus contorta as host
plants. The shape of the mycelial fan is still evident but the hyphal density is reduced and strand
differentiation is more advanced.
Fie 3 (a) Mycelial strand showing diffuse hyphal growth along the margin which maintains contact
with the substrate, (b) Reduction of external hyphal development in older parts of a mycelial
strand.
absent. Levels of radioactivity in the peat surrounding the roots of 'donor' plants
in both mycorrhizal and non-mycorrhizal chambers did not differ significantly
from background levels, further indicating that mycelial connections provided the
major pathway for carbon transfer to 'receiver' plants over the time scale of the
experiments reported here.
R. D. FINLAY AND D . J. READ
V I
Fig. 4. (a) Chamber containing mycorrhizal Pinus contorta plants interconnected by fungal
mycelium. A 'donor' plant is being fed with "COg. Note residual patches of intense hyphal
colonization, (b) Autoradiograph of chamber shown in (a) revealing extensive transfer of labelled
carbon through the mycelium. Particularly high levels of radioactivity are seen in the mycorrhizal
roots of receiver plants and in the mycelial patches within the peat, (c) Higher magnification of
mycelial patches developing in the peat, (d) Autoradiograph of area shown in (c).
In chambers where inter-specific combinations of host plants were connected

by a common mycelium the distribution of labelled compounds followed a similar
pattern. Significantly larger amounts of label accumulated in the roots of plants
grown in mycorrhizal chambers than in those of plants in non-mycorrhizal
chambers (Fig. 6) where levels of activity in 'receiver' plants were not significantly
Carbon transfer between ectomycorrhizal plants
P<0-00\
3000 Non-mycorrhizal
01
E Suillus bovinus
E
d.
2000 Suillus granulatus
P<0-05
>
o
'•X3
1000
P<0-05
Shoots
n Roots
Fig. 5. The distribution of radioactivity in roots and shoots of mycorrhizal and non-mycorrhizal
Pinus contorta plants grown in association with 'donor' plants of the same species. Donor plants
were fed with 50 fiCi of NaH^^COs for 72 h. Significance levels refer to differences between
mycorrhizal and non-mycorrhizal treatments and are based on analysis of variance of In transformed
d.p.m. data.
Pinus contorta-^ Pinus sylvestris Pinus sylvestris—* Pinus contorta

1000-
Non-mycorrhizal
•01
\A^i\ Suillus bovinus
E
d
500 P<0-00\
8
o
Fig 6 The distribution of radioactivity in mycorrhizal and non-mycorrhizal roots of Pinus contorta
and Pinus sylvestris grown in association with 'donor' plants of the opposite species. In each
combination the 'donor' plant is the first named species. Donor plants were fed with 10/*Ci of
NaH^COj for 96 h. Significance levels refer to differences between nJiycorrhizal and non-
mycorrhizal treatments and are based on analysis of variance of In transformed d.p.m. data.
different from background levels. Typically 0 2 to 1-0% of the labelled assimilate

in the fed 'donor' plants was transferred to each neighbouring mycorrhizal
seedling within 3 to 5 d compared with 0-01 to 0-02% to each non-mycorrhizal
seedling.
The effect of shading on ^^C transfer

In experiment 2a, where interconnected Pinus contorta plants were mfected
with Suillus granulatus, significantly higher amounts of label per unit dry weight
152 R. D . FiNLAY AND D . J. R E A D
/°<O-OOI Unshaded
NS
_° 20 JMi Shaded
I
E
ro
o
E NS
d.
d
10
o_
g
o
ir rt
Suillus granulatus Pisolithus tinctorius Suillus bovinus
Fig. 7. The distribution of radioactivity in roots of shaded and unshaded Pinus plants grown in
association with donor plants infected with different ectomycorrhizal fungi. Full treatment details
are given in Table 1. Significance levels refer to differences between shaded and unshaded
treatments based on blocked analysis of variance of In transformed d.p.m. data. Vertical bars
indicate standard errors.
were transferred to roots of shaded seedlings than to those of unshaded seedlings

(P < 0-01) (Fig. 7). In experiments 2b, c and d, although the roots of shaded plants
had consistently higher mean levels of label per mg dry weight than those of
unshaded plants, the individual levels of activity were highly variable, particularly
in the shaded plants, and the differences between shaded and unshaded treatments
were not statistically significant. The results of experiment 2a, however, support
the hypothesis that movement of label between plants occurs along concentration
gradients and that source-sink relationships may be influenced by diflFering levels
of irradiance of the host shoot.
In the present study only small amounts of label were translocated to the shoots
of' receiver' plants. An attempt was made to determine partitioning of label within
the root system by separately measuring the levels of activity in the sheathed lateral
roots and in the older main root axes. Distribution of activity was highly variable
but between 30 and 60 % of the total root label was found in the older axes. Since
even these structures may have a Hartig net such partitioning does not necessarily
mean that transfer from fungus to host has occurred but demonstrates that activity
first arriving in the infected laterals can be redistributed to older tissue whether
it is of host or fungal origin. Detailed microautoradiographic analysis of carbon
transfer from fungus to host tissue is at present being carried out.
DISCUSSION
The root systems of many of the tree species which are normally ectomycorrhizal
are differentiated into long roots, which have the potential for indefinite extension,
and short roots which have a relatively restricted life-span and limited powers of
extension growth. The latter structures, the apices of which are frequently
ensheathed in mycorrhizal mycelium, have long been recognized as the ' feeding
roots' of the tree (Harley, 1969). It is established from studies of both beech
(Clowes, 1951) and pine (Robertson, 1954) that in the mature root system infection
Carbon transfer between ectomycorrhizal plants i53
of the short roots arises from inoculum already established on the long root, but
these studies do not show how infection takes place in newly emerged seedling root
systems. Robertson showed that spores could act as a source of inoculum and many
pot experiments have shown that infection can arise through contact with mycelial
fragments in the soil. Such experiments, however, do not adequately simulate the
field situation. In the forest the majority of seeds of a given tree fall within close
proximity of the parent and thus germinate in an environment already containing
mycorrhizal roots supporting compatible fungi. Mason et al. (1983) have recognized
a distinction between fungi which colonize seedling roots at an 'early stage', and
which are frequently not strand formers, and those which occur later in the
'succession', which normally form extensive strand systems. However, in labora-
tory studies (Brownlee et al., 1983) and in natural forest soils (Fleming, 1983)
it has been shown that strand forming fungi growing from established sites of
infection can be the most vigorous colonists of developing seedling roots. The
previously infected roots act as a food base, providing a very high inoculum
potential sensu Garrett (1960). Mycelia of fungi such as S. bovinus spread rapidly
from infected roots and the high density of hyphae at each mycelial front provides
an efficient means of exploiting the soil for available nutrients, water and
uninfected roots. Aggregation of hyphae in older parts of the mycelium leads to
reduced hyphal colonization of soil which has already been exploited and the
formation of mycelial strands which interconnect plants and have the potential to
act as conduits for the long distance translocation of nutrients. It therefore seems
likely that if seedlings germinate in the forest they will be infected by strand-
forming rather than ' early stage' fungi and as a result will become integrated into
the large mycelial network which these fungi provide. Comparative studies of the
role of strand-forming and non-strand-forming fungi in the infection process and
in the facilitation of nutrient transfer are at present being undertaken.
The pattern of hyphal advance discussed above is not unique to ectomycorrhizal
fungi but is very similar to that seen in such wood-rotting fungi as Serpula
lacrimans (Jennings, 1982). Growth of the vegetative mycelium is, in both cases,
supported by carbon translocated from the food base and it is interesting that a
similar spectrum of carbon compounds, in which trehalose predominates (Soder-
strom, Finlay & Read, 1986), is found in the mycelium. The present study
confirms the observation of Brownlee et al. (1983) that, in addition to being a major
source of inoculum, the mycelial system provides conduits through which carbon
flows from an established food base to newly infected roots. Measured rates of
movement of ^^C-labelled compounds through strands to hyphal tips exceed
20 cm h"i and are comparable with the rates of transport through mycelial strands
of Serpula lacrimans (Wulf. ex Fr.) measured by Brownlee & Jennings (1982).
Assimilate moves from the host root to the advancing mycelial front of the
heterotroph and, as in the case of Serpula, it seems likely that transport occurs as
a result of a pressure driven flow, the sink for carbon being provided by growth
and wall building at the hyphal front. Clearly, without net carbon flow, the
heterotroph, which has very limited saprotrophic capability, would be unable to
spread through the soil.
When hyphae in the mycelial front make contact with an uninfected root,
mycorrhizal infection takes place. Such an event will inevitably influence the
polarity of carbon movement in that sector of the strand system since the new
infection will normally provide access to an additional source of assimilate. Under
these circumstances, when equilibrium has been achieved, net flow of carbon to
154 R- D . FiNLAY AND D . J. READ
the new infected root will not necessarily occur unless there is a gradient of carbon
concentration arising, for example, from a lower assimilate supply from the new
host. Experiments of the type reported here establish that assimilate transfer
between mycorrhizal hosts is significantly greater than that between those in
chambers lacking mycelial connections but do not unequivocally demonstrate the
occurrence of net flow of assimilate to the 'receiver' root. The presence of
functional mycelial connections, however, provides the potential for flow along
gradients so that areas of low substrate availability can be supplied with assimilate.
Factors such as irradiance are likely to determine assimilate supply to roots in
nature and the experimental data suggest that manipulation of sink size by shading
'receiver' plants can lead to enhanced carbon transfer. The situation is thus
similar to that observed by Francis & Read (1984) in VA mycorrhizas, though in
the case of ectomycorrhizas the response to shading is more variable and occurs
over a longer time period. There are a number of possible explanations for the
relatively high variability observed in ectomycorrhizal plants. The pattern of
infection of 'receivers' is itself highly variable within the individual chambers.
This is inevitable as infection is a random process, the level of which depends on
the rates of development of 'donor' and 'receiver' plants and the direction of
spread of mycelial fans. In addition the relative growth rates of the coniferous
ectomycorrhizal hosts are lower than those of the herbaceous species used by
Francis and Read so that respiratory depletion of reserves and establishment of
concentration gradients will take place over longer periods of time.
The precise resolution of the mycelial strands and even of individual hyphae
obtained in the autoradiographs indicates that the transfer of carbon is direct and
that little leakage of labelled assimilate occurs into the growing medium. The
pathway of transfer is thus similar to that found by Francis & Read (1984) who
reported that in VA mycorrhiza assimilate transfer occurred through clearly
defined inter-plant hyphal pathways. Transfer of carbon between mycorrhizal pine
and non-mycorrhizal Tradescantia plants has recently been reported by Taber &
Taber (1985). In the absence of a mutually compatible heterotrophic symbiont it
must be assumed that such transfer occurred by a process of leakage and
re-absorption. It is well known that actively growing roots release some carbon
in the form of exudates and sloughed tissues and such material is inevitably
available for absorption by the microbial populations of the rhizosphere and even
by autotroph tissues. The difference between such leakage and the direct transfer
process is, as originally pointed out by Reid & Woods (1969), that the materials
being transported through mycorrhizal mycelia are retained within the symbiotic
system and a far tighter cycling of carbon is thus obtained.
The demonstration that carbon can be transferred from host to host by way of
the mycorrhizal mycelium and that polarity of movement can be influenced by
shading raises a number of important issues. Bjorkman (1942, 1970) showed that
shading reduced levels of mycorrhizal infection in pine seedlings and suggested
that this reduction was attributable to a lowered internal carbon concentration in
the shaded plants. Marx, Hatch & Mendicino (1977) later confirmed that variation
in the sucrose content of the short roots could account for 85 % of the variation
in intensity of mycorrhiza formation. Harley & Smith (1983) point out that these
results are in accordance with the fact that the fungus depends upon its host for
carbohydrate compounds. These results and conclusions are, however, based upon
experiments in which individual seedlings were grown with inoculum that was not
attached to a food base. The inoculum potential was therefore low. In the more
Carbon transfer between ectomycorrhizal plants 155
natural circumstance of seedlings growing in association with infected adults the
potential for transfer of carbon from mature plants would be expected to reduce
fungal dependence upon the internal carbon concentration of the new host and
enable infection to occur.
The ecological significance of the formation of mycelial connections between
plants has been discussed by Read, Francis & Finlay (1985) who point out that
the newly revealed carbon pathway might be critically important for the survival
of seedlings growing in the shade of adult overstorey plants. Naturally regenerating
seedlings can spend a number of years in a suppressed condition below an adult
population before opening ofthe canopy permits sustained positive net assimilation
rates. Survival during this period, when the plant is living below its compensation
point, might well be dependent upon sustained or seasonal supply of carbon from
the illuminated overstorey. In addition, since, on infection, the seedling becomes
integrated into a network of mycelial strands it is potentially able to receive mineral
nutrients absorbed from soil at a considerable distance from its own root system.
The following paper reports an analysis of the uptake and distribution of
phosphorus in such a mycelial network.
ACKNOWLEDGEMENTS
We wish to thank the Natural Environment Research Council for financial

support, and Mr G. Woods for photographic assistance.
REFERENCES
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New Phytologist - May 1986 - FINLAY - THE STRUCTURE AND FUNCTION OF THE VEGETATIVE MYCELIUM OF ECTOMYCORRHIZAL PLANTS

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New Phytologist - May 1986 - FINLAY - THE STRUCTURE AND FUNCTION OF THE VEGETATIVE MYCELIUM OF ECTOMYCORRHIZAL PLANTS

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14698137, 1986, 1, Downloaded from https://nph.onlinelibrary.wiley.com/doi/10.1111/j.1469-8137.1986.tb00603.x by EVIDENCE AID - BELGIUM, Wiley Online Library on [18/03/2023].

THE STRUCTURE AND FUNCTION OF THE

{Accepted 23 December 1985)

necessitates the use of laboratory observation chambers which permit in situ

Carbon transfer experiments in mycorrhizal and non-mycorrhizal chambers

Table 1. Summary of experimental treatments in six different experiments

Suillus granulatus Pinus contorta Pinus sylvestris 4 5 50

Pisolithus Pinus sylvestris Pinus contorta 6 7 50

Suillus bovinus Pinus sylvestris Pinus radiata 4 7 120

Measurement of fungal growth rates

Measurement of ^*C translocation rates in mycelial strands

Mycelial growth and strand extension

Suillus bovinus (Strain A) 11 7 ± 27 4-3 ± 1 1

Measurement of ^^C translocation rates

Transfer of ^^C in mycorrhizal and non-mycorrhizal chambers

In chambers where inter-specific combinations of host plants were connected

Pinus contorta-^ Pinus sylvestris Pinus sylvestris—* Pinus contorta

different from background levels. Typically 0 2 to 1-0% of the labelled assimilate

The effect of shading on ^^C transfer

were transferred to roots of shaded seedlings than to those of unshaded seedlings

We wish to thank the Natural Environment Research Council for financial

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