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Action Potential-Like Activity Found in Fungal Mycelia Is Sensitive to

Stimulation

Article in The Science of Nature · August 1995

DOI: 10.1007/BF01167867

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of analytical procedures will support the
understanding of sorption phenomena on
natural and anthropogenic humic sub-
stances in soil, sediments, and water. This
knowledge is important for a precise
description of transport processes in the
environment and may also be of relevance
for developing remediation strategies.
Received June 1 and August 17, 1994
Naturwissenschaften 82, 30-31 (1995) @ Springer-Verlag 1995
Action Potential-Like Activity Found
in Fungal Mycelia Is Sensitive to Stimulation
S. Olsson and B.S. H a n s s o n
Department of Ecology, Lund University, S-223 62 Lund
Basidiomycetes, especially wood decom-
posers, are known to exhibit coordinated
responses of the whole mycelium during
the search for suitable substrates [1]. Sig-
nal transmission between different parts
of the mycelium seems at times very rapid
and long-range [2]. Some basidiomycete
mycelia are among the largest and maybe
oldest organisms and can cover many
hectares [3]. Thus, it appears that some
means of fast communication between
mycelial parts might exist. Since the
fungal mycelium is a network of filamen-
tous, interconnected cells with a cyto-
plasmic continuum enveloped in a com-
mon membrane, electrical signaling simi-
lar to that which occurs in nerve cells
should, in theory, be possible for the fast
communication within the mycelium. In
the present study, the wood decomposers
Pleurotus ostreatus (Jacquin: FrO Kum-
mer and Arrnillaria bulbosa (Barla) Kile
& Watling were investigated for the possi-
ble occurrence of action potentials in the
mycelium and the possibility that elec-
trical signaling might take place in fungal
mycelia.
The isolates were Ams01 for A . bulbosa
and Po2 for P ostreatus, obtained from
Dr. S. Gray, University of Luton, UK. The
Ares01 was a tissue isolate from a basi-
diocarp found on Merlewood Grounds,
Merlewood Research Station, UK. The
Po2 was originally obtained from Dr. P.
Markham, Kings College, London.
30
1. P6rschmann, J., Stottmeister, U.: Chroma-
tographia 36, 207 (1993)
2. Frimmel, E H., Christman, R,E (eds.):
Humic Substances and Their Role in the
Environment. Chichester: Wiley 1988
3. Perdue, E.M., Gjessing, E.T. (eds.): Organic
Acids in Aquatic Ecosystems. Chichester:
Wiley 1990
4. Kopinke, E-D., P6rschmann, J., Stott-
meister, U.: Environ. Sci. Technol. (submit-
ted)
A. bulbosa and P. ostreatus were grown
on malt extract agar (Oxoid) in 9 cm
diameter Petri dishes at 20 °C. Two drops
of ethanol were added at the edge of the
A. bulbosa plates to stimulate the forma-
tion of rhizomorphs. One- to 2-month-
old cultures, covering the whole or most
of the plates, were selected for measure-
ments. A glass microelectrode filled with
2 M KC1 was inserted among the mycelial
strands. The tip of the electrode was
moved downward by a Huxley style mi-
cromanipulator, and the tip of the elec-
trode was vibrated electrically to facilitate
penetration of the hyphae. Upon entering
a hypha, potentials could be recorded and
stored on a Vetter 8-channel video re-
corder, adapted for storage of neurophysi-
ological data. The signal was amplified by
an AxoClamp amplifier (Axon Instru-
ments) and visualized on a Tektronix
digital storage oscilloscope. For analysis,
the potentials were printed on an elec-
trostatic printer (Gould). The reference
electrode was inserted into the agar
medium.
The probing of undifferentiated hyphae
was not successful in either of the two
species. The electrode slid off the hyphae,
probably due to the relatively tough cell
walls. The rhizomorph tissue of A . bul-
bosa was easier to probe since there was
no possibility for individual cells to slide
off the tip of the electrode. For P.
ostreatus strands with looser tissue, stab-
Naturwissenschaften 82 (1995)
5. Wershaw, R.L.: ibid. 27, 814 (1993)
6. Hawthorne, S.B., Miller, D. J., Pawliszyn,J.,
Arthur, C.L.: J. Chromatogr. 603, 185
(1992); Potter, D.W., Pawliszyn,J.: Environ.
Sci. Technol. 28, 298 (1994)
7. Kopinke,E-D., Remmler, M.: in preparation
8. Daylight Chemical Information Systems,
Inc., Irvine, CA, USA: Pomona 92 Database
1992; Howard, T.H.: Chemfate Database.
Syracuse Research Corp. 1993
bing of a hyphal strand with the electrode
by using the micromanipulator coarse
control was a successful method for pene-
trating the cell wall.
Spontaneous action potential-like activity
was found in both fungi (Fig. 1). The
spontaneous activity was relatively even,
but could vary between 0.5 and 5 Hz. The
recorded potentials had an amplitude of
5 - 50 mV and displayed varying duration,
Fig. 1. Spontaneous action potential-like activ-
ity in P. ostreatus (A) and A. bulbosa (B). Ver-
tical scale bar 20 mV, horizontal scale bar I s
25-
[]
20-
"~ 15-
i0- 0
[]
5-
0
current ~nA~
Fig. 2. Effect of current injections through the
microelectrode. Currents from -0.4 to 0.1 nA
were tested. The frequency of potentials was
substantially increased by injection of negative
currents
© Springer-Verlag 1995
from approximately 20 ms up to 500 ms.
The resting potential was between - 7 0
and - 1 0 0 inV. The frequency of "action
potentials" was increased by injecting
negative current, and decreased by inject-
ing positive current, through the electrode
(Fig. 2). When a drop of 2.5 M sulfuric
acid, 2% malt extract, or water was added
to the mycelia 1 - 2 cm from the electrode,
the frequency of the "action potentials"
increased after approximately 30 s, sug-
gesting that the message regarding the
stimulus was propagated at a speed of
approximately 0.5 m m s -~. In a few
preparations, the frequency after stimula-
tion increased to over 20 Hz. A similar in-
crease in frequency was observed when a
fresh piece (5 x 5 x 5 ram) of wood (beech
wood), the normal substrate of the
fungus, was placed on A. bulbosa
mycelium 1 - 2 cm from the electrode, but
not when it was replaced by a piece of
Perspex plastic of equal size and weight.
The change in activity was reversible, so
that when the fresh-wood stimulus was
removed, the activity decreased, and was
subsequently regained when the stimulus
was reintroduced (Fig. 3). The other
stimuli could not be tested in this way,
since they could not be removed, once in-
troduced. When the piece of wood was
suspended 1 - 2 mm above the mycelium,
no effect was seen, thus ruling out the
possibility that the observed effect was
due to volatile action at the site of the
microelectrode.
The occurrence of frequent potentials in
fungal mycelia is a new observation and
might be a special phenomenon for basi-
diomycete fungi. The rate of the spon-
taneous firing was very similar to that
recorded from sensory neurons and cen-
tral nervous neurons involved in sensory
systems in classic animal preparations.
The phenomenon of spontaneous firing
potentials is usually interpreted as a way
for the sensory system to be prepared for
incoming stimuli. Spontaneous "action
potential'like activity in fungi has been
recorded before for the fungus Neuro-
spora erassa, but the activity was very low
(0.005 Hz), and it could not be induced to
change its spontaneous activity by current
injections [4]. The fungus tested was not
one in which the mycelium persists for a
long time in the natural environment, so
the need for a communication system
could be questioned in this case.
Current injection modified the cell activi-
ty (Fig. 2). The negative current also in-
creased the amplitude of the potentials.
These results indicate that the ions and
ion channels involved in the buildup of
fungal action potentials are different
from those involved in classical action po-
tential elicitation in animal nerve cells.
There, a negative current injection in-
hibits the activity of neurons, while a
positive current injection depolarizes the
neuron to the threshold value for action
potential initiation, and action potentials
are eventually fired.
The sensitivity to external stimuli is ex-
citing, since it means that the fungus can
use electrical signaling to send messages
between different mycelial parts, mes-
sages conveying information regarding
food sources, injury, local conditions
within the fungus, or the presence of
other individuals around it. The findings
reported here do not rule out pressure as
a vehicle for signal transmission [2], since
more than one mechanism can be operat-
ing simultaneously. Different mechanisms
of communication might also be present
in different fungi.
If the potentials recorded in this investiga-
tion are used for communication in A.
bulbosa mycelia in nature, where the
mycelia can cover several hectares and
weigh tens of tons [3], these mycelia
would be the largest known electrically
communicating biological systems.
We thank Dr. S. Anton for her help dur-
ing the experiments. This work was sup-
ported by grants from the Swedish Natu-
ral Science Research Council to SO and
BSH.
Received June 6 and August 15, 1994
1. Rayner, A.D.M., Griffith, G.S., Wildman,
H.G., in: Shape and Form in Plants and
Fungi, p. 293 (D. S. Ingram, ed.). London:
Academic Press 1994
2. Rayner,A. D. M., Griffith, G.S., Ainsworth,
A.M., in: The Growing Fungus (N.A.R.
Gow, G.M. Gadd, eds.). London: Chapman

I00
!!]!!.W!I!!!]!!_IJ.W!!!!!J!_I!!]!;!!!!!]!II and Hall (in press)
3. Smith, M.L., Bruhn, J.N., Anderson, J.B.:
Nature 356, 428 (1992)
4. Slayman, C.L., Long, W. S., Gradmann, D.:
Biochim. Biophys. Acta 426, 732 (1976)

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Naturwissenschaften 82 (1995) © Springer-Verlag 1995 31

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