You are on page 1of 9

Mycological Society of America

The Influence of Humidity upon Transpiration and Growth in Psilocybe cubensis


Author(s): Edmond R. Badham
Source: Mycologia, Vol. 77, No. 6 (Nov. - Dec., 1985), pp. 932-939
Published by: Mycological Society of America
Stable URL: http://www.jstor.org/stable/3793305 .
Accessed: 21/11/2013 22:23
Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at .
http://www.jstor.org/page/info/about/policies/terms.jsp

.
JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of
content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms
of scholarship. For more information about JSTOR, please contact support@jstor.org.

Mycological Society of America is collaborating with JSTOR to digitize, preserve and extend access to
Mycologia.

http://www.jstor.org

This content downloaded from 130.194.20.173 on Thu, 21 Nov 2013 22:23:32 PM


All use subject to JSTOR Terms and Conditions

Mycologia,77(6), 1985, pp. 932-939.


? 1985, by The New York Botanical Garden, Bronx,NY 10458
THE

INFLUENCE
AND

OF
GROWTH

HUMIDITY
IN

UPON

PSILOCYBE

Edmond

TRANSPIRATION
CUBENSIS

R. Badham1

DepartmentofBiological Sciences, Lehman College,


The City University
ofNew York,Bronx,New York 10468
ABSTRACT
The influenceof humidityupon individual basidiocarps of of Psilocybecubensis was
studied using an environmentallycontrolledwind tunnel and a computerprogramwhich
helped to model growthand development.Regressionmodels were developed which were
able to explain 77% of the variation in the transpirationrate and 68% of the variation in
correlatedwith
growthrate. Transpirationand growthof this mushroomwere significantly
thehumidityoftheair. The fastestgrowthand thelowesttranspiration
occurredat thehighest
humidities.No inhibitionof growthwas detectedat 0 pascals VPD (100% RH). Misting
acceleratedgrowthand transpirationwhilelighthad no effect.Althoughhumiditywas a very
importantfactorinfluencingtranspirationand growth,the size and shape of the mushroom
were also importantin water relations.The finalwater contentof basidiocarps with thin
lowerthatof thickstipesor those withlargersurfacearea-to-volumeratioswas significantly
stipedmushroomsor those withsmall surfacearea-to-volumeratioseven whengrownunder
equal humidity.Growthratesunderconditionswhichpromotedthe highestlevels of hydration of the basidiocarp were rapid (up to estimated4% increasein dryweightper h).
Key Words: transpiration,humidity,growth,mushroom.
Few precise studies of the influence of humidity on mushroom growth have
of fleshy mushrooms in nature is frequently
been conducted. The appearance
associated with the presence of moisture. Indeed, the fact that most fleshy mush?
rooms come up after rains and grow in, or very near, moist substrates suggests
that they may be adapted to humid habitats.
Most research on the influence of humidity on mushroom growth has been
conducted with commercially important species. For the growth of Agaricus bi?
sporus (J. Lange) Pilat, Edwards (1978) recommends 90% RH for wind flows of
50-100 mm/sec and 80% RH for air flows of 25 mm/sec in the temperature range
15-16 C. For Pleurotus sp., Kurtzman and Zadrazil (1982) report that humidities
lower than 60% RH would be expected to cause rapid drying of the substrate
while localized condensation would be expected to occur with humidities of 95%.
Kurtzman and Chang-Ho (1982) recommend between 78 and 92% RH for Volvariella volvacea (Bull. ex Fr.) Sing., depending upon the ventilation. Badham
(1983) found that high relative humidities (> 90% RH) and low wind speeds (< 100
mm/sec) favored the growth of Psilocybe cubensis (Earle) Sing. in the laboratory
and natural environment. Many authors emphasize the importance of minimizing
a build-up of carbon dioxide despite the maintenance of high humidity.
Transpiration in several species of mushrooms was studied by Zoberi (1981).
He found that evaporative water loss decreased with an increase in atmospheric
humidity. Within the range of his experiments, the water content ofthe substrate
did not have a great influence on the transpiration rate. A precise laboratory study
that considered the influence of humidity on growth and transpiration of basid?
iocarps was that of Plunkett (1956). He showed that the woody mushroom Po?
lyporus brumalis Pers. ex Fr. grew poorly at high humidities while the growth of
the fleshy mushroom Collybia velutipes [Flammulina
velutipes (Fr.) Karst.] was
1Presentaddress: Carolina Fungi, Inc, 2736 Lakeview Dr.,
Raleigh,NorthCarolina 27609.
932

This content downloaded from 130.194.20.173 on Thu, 21 Nov 2013 22:23:32 PM


All use subject to JSTOR Terms and Conditions

Badham:

Humidity

933

enhanced. He estimated that growth was arrested in C velutipes at transpiration


rates exceeding 1.4 mg/cm2/h while P. brumalis was able to survive rates exceeding
9 mg/cm2/h. Schiitte (1956) stated that transpiration decreases with the age ofthe
and that there is "very considerable"
basidiocarp
transpiration in Clitocybe sp.
under saturated conditions. Gruen (1983) has demonstrated that fruitbodies with?
in a flush compete for resources and that their final lengths and weights depend
on their rank in the flush.
Perhaps one ofthe reasons for the lack of research regarding the influence of
humidity on mushrooms is the difficulty of measurement, especially of high humidities in confined areas. Many researchers use "per cent relative humidity"
although this is one of the least desirable measures of humidity since it does not
take into account the influence of temperature (Oosting, 1956). The preferred
measure of humidity is vapor pressure deficit (VPD) with SI units of pascals (1
pascal = 10-2 mbars). An alternative is to measure water loss from a moist surface,
as Flegg (1974) has suggested. This method compensates
to some degree for
differential air flow which also has an influence upon the evaporation of water.

MATERIALS AND METHODS


The strain of P. cubensis used was (ATCC
36462). Cultures were grown on
sterilized brown rice (6 g/15 ml deionized water) in 60 x 15 mm Petri dishes
until mycelia covered the substrate. The mycelial mats (and substrate) were then
removed and placed in 130 mm pie tins with vermiculite, 30 ml of deionized
water was added, and the containers were covered with glass. All mushrooms
were allowed to develop to at least 10 mm in height under deep Petri plates (80 x
100 mm) and 12 h photoperiod of 11.2 W/m2 (2685 lux) "cool white" fluorescent
The deep Petri plate was removed (to allow for
light prior to experimentation.
aeration) when the cultures were weighed daily, and any loss in weight was made
up with distilled water. Dry weights were determined to the nearest mg after
drying in an oven at 60 C for 24 h. In each test culture only the largest basidiocarp
of the first flush was used for experiments. The others were removed at the
beginning of the test period and the water content of the second largest (second
rank) was determined at that time to be used as an indication of the initial per
cent water ofthe experimental mushroom (first rank). The water content of these
two mushrooms from the same mycelium would be very similar because their
size, age, and stage of development was nearly the same and they had been exposed
to the same environmental conditions.
The cultures were placed in a wind tunnel (Badham,
1982) under constant
wind speed and variable humidity. Certain mushrooms were misted with distilled
water (2 ml) on all sides prior to the experiment. The pie tins were sealed with
aluminum foil and tape (except where the mushroom protruded) so that weight
loss could be attributed to water loss from the mushroom. During the experiments,
fluorescent light (cool white, 1.4 W/m2 or 216 lux) illuminated certain basidiocarps
while others developed in darkness. Wind velocity was constant at 120 ? 30 mm/
see and was measured with a TSI hot wire anemometer (TSI, P.O. Box 43394,
St. Paul, MN 55164). For descriptions ofthe variables see Table I. Experiments
were conducted at 27.2 ? 1.1 C for 10.7 ? 2.6 h. Temperature and humidity
were measured with a dual thermocouple,
digital thermometer (#871, Omega,
Box 4047, Stamford, CT 06907) using 0.0762 mm NiCr-NiAl
thermocouples.
For humidity measurements, one thermocouple was embedded in a cotton string
wicked to a bottle of distilled water and ventilated at wind speeds greater than
100 mm/sec (Slavik, 1974). As in Badham (1984) an Apple?
BASIC program

This content downloaded from 130.194.20.173 on Thu, 21 Nov 2013 22:23:32 PM


All use subject to JSTOR Terms and Conditions

Mycologia

934

Table I
Experimental variables*
Variable

Description

Transpirationratebd
Waterreductionrate8

paper cylindermade froma single


sheet of filterpaper rolled to 12
mm diam, 150 mm height,and
placed in 48 mm tall vial of water
Weightloss frombasidiocarp divid?
ed by the square root of the sur?
face area per h
(Per cent waterof the second rank
minus finalper cent waterof the
experimentalbasidiocarp) per h

Units

ml/mm/h

Mean

S.D.

1.1

0.68
(xlO-3)

%/h

0.47

0.39

a n = 89, non-mistedmushroomsonly.
bVariable is normallydistributed.
c
Startingand finalvalues were averaged.
d See Badham (1984).
c Used as a measure of shape withoutbeing influencedby size, see Vogel (1981).
f
Normallydistributedforevaporativedemands <0.2 ml/h.
*n = 75.
(available from the author) calculated many derived variables as well as RH and
VPD [based on the wet and dry bulb temperatures tables of Unwin (1980)].
of 6 mushroom dimensions were made at the beginning and
Measurements
end of 165 experiments by following the procedure of Badham (1984). During
development the shape ofthe basidiocarp of P. cubensis corresponds to common
geometric solids. Therefore, the surface area, volume, and stage of maturity could

This content downloaded from 130.194.20.173 on Thu, 21 Nov 2013 22:23:32 PM


All use subject to JSTOR Terms and Conditions

Badham:

Humidity

935

be estimated for fruitbodies of different sizes. Since the square root ofthe surface
area is linearly correlated with dry weight (r = .85, P < .001; Badham, 1984),
change in surface area implies a similar change in dry weight. If "per cent change"
is used then small basidiocarps
may be compared directly with larger ones. In
this study "per cent change in the square root of the surface area per hour" was
used as a non-destructive measure of growth while "weight loss per square root
ofthe surface area per hour" was used as a measure of transpiration. For statistical
analyses I used The Statistical Analysis System (SAS, Box 8000, Cary, NC 27511),
Sokal and Rohlf (1981), or Zar (1984).
RESULTS AND DISCUSSION
of Psilocybe cubensis,
Although light may be necessary for the development
under these conditions no influence of light was detected on any variable. Illu?
cultures showed no significant differences for four
minated and non-illuminated
important variables listed in Table II. Therefore, illuminated and non-illuminated
cultures were treated identically for further analyses.
Misting the basidiocarps did result in significant differences in many variables
(Table II). Growth rate, transpiration, and final water content were increased by
misting. Even though the set of misted mushrooms was grown under slightly lower
mean humidity (due to chance) its growth rate was significantly greater than that
of its non-misted counterpart. This indicates that cell hydration may have been
a limiting factor for growth. Since misting seemed to have a significant influence
on several variables, only non-misted mushrooms were considered for further
analyses.
To indicate the important variables which influenced transpiration and growth,
a correlation matrix was determined for these two variables versus all other vari?
ables (Table III).
transpiration rate was highly correlated with hu?
Effects on transpiration. ?The
was
a
There
midity.
highly significant linear relationship between evaporative
demand and transpiration at evaporative demands lower than 0.2 ml/h (Fig. 1).
at evaporative demand greater than 0.2 ml/h or VPD
Most of the basidiocarps
greater than 475 pascals failed to grow and had final water contents less than 80%.
Two other measures of humidity (RH and VPD) were also significantly correlated
with the transpiration rate (Table III). The transpiration rate ranged from 0.03 x
10"3 to 3.2 x 10"3 ml/mm/h. There was no significant correlation between the

a t-testcomparison of illuminatedand non-illuminatedbasidiocarps and comparison of misted


and non-mistedbasidiocarps.
b ns = P > 0.05, * = 0.01 < P < 0.05, ** = 0.001 < P < 0.01, *** = P < 0.001.

This content downloaded from 130.194.20.173 on Thu, 21 Nov 2013 22:23:32 PM


All use subject to JSTOR Terms and Conditions

Mycologia

936

Table III
Spearman correlation matrix8

an =
bns
=n =

89
= P > 0.05, * = 0.01 < P < 0.05, ** - 0.001 < P < 0.01, *** = P < 0.001.
75.

transpiration rate of the fruitbodies and the water content of the mycelial mat.
The evaporative demand ofthe air was significantly correlated with the final water
content ofthe basidiocarp (r = ?.77, P < .001).
Estimates ofthe highest transpiration rate under which this mushroom would
be able to develop may be made by extrapolating the 10 h experiments to the
amount of time necessary for development. If one assumes that mushrooms start
at 90% hydration (the mean ofthe second rank), and that growth is greatly reduced
by 85% hydration (the mean ofthe growth rate of mushrooms of less than 85%
final water content was 17% that ofthe mushroom with greater than 85% water),
and that it takes 48 h to grow from stage 1 to stage 4 (see Fig. 5 in Badham,
1984), then one can calculate a maximal water reduction rate of 0.1%/h and a
transpiration rate of 0.5 x 10~3 ml/mm/h. This transpiration rate would occur
at an evaporative demand of 0.015 ml/h and a VPD of 58 pascals. This maximal
transpiration rate corresponds exactly to what Plunkett (1956) estimated for Collybia velutipes.
In addition to water vapor loss, basidiocarps
lose weight due to the loss of
metabolic gases such as C02. This may explain some ofthe variability in Fig. 1,

Fig. 1 (upper). Influenceoftheevaporativedemand oftheair upon transpiration.


For definitions
of "evaporative demand" and "transpiration"see Table I. R2 = .77, P < 0.001, n = 82. Best fitline:
transpirationrate (ml/mm/h)= 0.01 x evaporativedemand (ml/h)]+ 0.0003.
Fig. 2 (lower). Importantfactorsforpredictingbasidiocarp growthrate in the mushroomP.
cubensis and theirrelationshipto actual growthrate. For the definitionof "growth" see Table I.
Growth may be estimated with a multiple linear regressionusing the basidiocarp water content,
transpirationrate,the heightto width ratio, and humidityas independentvariables. R2 = .68, P <
0.001, n = 77. Best fitline: growthrate (%/h) = {[0.157 x final water content(%)] - [1179 x
transpirationrate (ml/mm/h)]- (0.061 x height/width
ratio) + [0.004 x VPD (pascals)]} - 11.11.

This content downloaded from 130.194.20.173 on Thu, 21 Nov 2013 22:23:32 PM


All use subject to JSTOR Terms and Conditions

Badham:

0.03
Evaporative

937

Humidity

0.09

0.15

Demand

(ml/h)

co

o
u
O

01

-u

kl
Qm

12

-2-10
Actual

Growth

3
1%/h)

This content downloaded from 130.194.20.173 on Thu, 21 Nov 2013 22:23:32 PM


All use subject to JSTOR Terms and Conditions

938

Mycologia

especially at the lowest evaporative demands. If one assumes a C02 loss of 0.001
g per 1.0 g of mushroom per h (Vedder, 1978), the average C02 loss would equal
approximately 3% ofthe average water loss for my experimental conditions.
many variables had significant correlations, no
Effects on growth rate. ?Although
normal variable explained more than 38% of the variation in growth rate. All
variables relating to the humidity of the air stream and the dimensions or water
content of the basidiocarp
were significantly correlated with the growth rate.
Further statistical analyses were conducted to determine which combinations
of
variables explained most of the variation in growth rate. A computer program
chose four variables among all normal variables as those giving
(SAS, MAXR)
the largest R2 value. They were (i) the per cent water of the basidiocarp,
(ii)
transpiration rate, (iii) height/width ratio, and (iv) the humidity. Sixty-eight per
cent ofthe variation in growth rate was explained in a multiple linear regression
using these variables (Fig. 2). When the values for these variables are put into
the best fitline equation, the expected growth rate will be obtained. If the equation
explained all of the variation in growth rate, then all of the points would fall on
a line with a slope of 1 and an intercept of 0. Negative growth occurred when
mushrooms were exposed to drying conditions which caused the basidiocarp to
shrivel and decrease in surface area. Each of these variables is significantly cor?
related individually with the growth rate even when the other three variables are
statistically held at their means.
Further analyses were conducted to determine the influence of mushroom
dimensions upon the final water content. Non-misted mushrooms were divided
into three groups according to their stipe diam. The final water content of the
mushrooms with the thinnest and thickest stipes were compared with a t-test
(Table IV). Although there was no significant difference in the humidity under
which the two groups were grown, there was a significant difference in the final
water content and the growth rate. Mushrooms with the thinnest stipes had sig?
nificantly lower final water contents and growth rates. A similar procedure was
conducted with the surface/volume ratio. The mushrooms with the largest surface/
volume ratios had lower final water contents, transpiration rates, and growth rates
at similar humidities. The reason for this may have to do with the pathway for
water loss. Thin-stiped mushrooms
or those with large surface/volume ratios
Table IV
Influence of mushroom dimensions

a t-testcomparison of smallest mushroom stipe diam (<4.1 mm) with largeststipe diameters
(>4.76 mm).
bns = P > 0.05, * = 0.01 < P < 0.05, ** = 0.001 < P < 0.01, *** = P < 0.001.
c t-testcomparisonof smallestadjusted surface/volume
ratios(<460) withlargestsurface/volume
ratios (>610).

This content downloaded from 130.194.20.173 on Thu, 21 Nov 2013 22:23:32 PM


All use subject to JSTOR Terms and Conditions

Badham:

Humidity

939

would be expected to have a shorter pathway and a greater concentration gradient


for water loss than thick-stiped basidiocarps
or those with small surface/volume
ratios. This may explain why dimensional differences result in lower final hydration levels.
An interesting correlation was found between the dry weight of the mycelial
mat and the growth rate (Table III). This could indicate that the translocation of
dry matter from the substrate was a limiting factor in the growth rate. Dry mat
Cultures
weight is inversely correlated with the dimensions of the basidiocarp.
with large basidiocarps have proportionally smaller mat weights. There is also an
inverse relationship between mushroom size and growth rate. Further study is
needed to determine if the dry mat/growth rate correlation is an artifact of this
intercorrelation, or whether it plays a significant role in determining the growth
rate.
There was a significant correlation between stage of development
and the
growth rate. However, whether this correlation was primarily the result of differ?
ences in maturity or changes in the simple physical shape has not been addressed
studied fell within a narrow range
adequately because most of the basidiocarps
of stages (95% between stage 1 and 3). This precluded appropriate statistical
analysis.
ACKNOWLEDGMENTS
The author is grateful to Dwight T. Kincaid for help with statistical analysis,
to Y. Schechter for the use of his lab, and to Sigma Xi for an equipment grant.
LITERATURE CITED
Badham, E. R. 1982. Tropisms in the mushroomPsilocybecubensis.Mycologia 74: 275-279.
-.
1983. Culturalstudieson the mushroomPsilocybecubensis.Ph.D. Dissertation,University
Microfilms.Ann Arbor,Michigan.
1984. Modeling growth,development,transpirationand translocationin the mushroom
Psilocybecubensis.Bull TorreyBot. Club 111: 159-164.
Edwards,R. L. 1978. Cultivationin Westerncountries:growingin houses. Pp. 299-336. In: Biology
and cultivationof edible mushrooms.Eds., S. T. Chang and W. A. Hayes, Academic Press, New
York.
Flegg, P. B. 1974. The measurementof evaporative loss in relationto water managementduring
croppingof Agaricusbisporus.Mushroom Sci. 9(1): 285-292.
Gruen,H. E. 1983. Effectsof competitionamong Flammulina velutipesfruitbodieson theirgrowth.
Mycologia 75: 604-613.
Kurtzman,R. H., Jr.,and Y. Chang-Ho. 1982. Physiologicalconsiderationsforcultivationof Vol?
variella mushrooms. Pp. 138-166. In: Tropical mushrooms:biological nature and cultivation
methods.Eds., S. T. Chang and T. H. Quimio. Chinese Univ. Press, Hong Kong.
and F. Zadrazil. 1982. Physiologicaland taxonomicconsiderationsforcultivationof Pleu?
-,
rotusmushrooms.Pp. 299-348. In: Tropical mushrooms:biologicalnatureand cultivationmeth?
ods. Eds., S. T. Chang and T. H. Quimio. Chinese Univ. Press, Hong Kong.
Oosting,H. J. 1956. The studyofplant communities.W. H. Freemanand Co., San Francisco.440 p.
Plunkett,B. E. 1956. The influenceof factorson the aeration complex and lightupon fruitbody
formin pure culturesof an agaric and a polypore.Ann. Bot. (London) 20: 563-586.
Schiitte,K. H. 1956. Translocationin the fungi.New Phytol.55: 164-182.
New York. 449 p.
Slavik, B. 1974. Methods ofstudyingplant waterrelations.Springer-Verlag,
Sokal, R. R., and F. J. Rohlf. 1981. Biometry.W. H. Freeman and Co., San Francisco. 859 p.
Unwin,D. M. 1980. Microclimatemeasurementfor ecologists.Academic Press, New York. 97 p.
Vedder,P. J. C. 1978. Modern mushroomgrowing.StanleyThornes,Cheltenham,England. 420 p.
Vogel, S. 1981. Life in movingfluids. Willard Grant Press, Boston, Massachusetts.352 p.
Zar, J. H. 1984. Biostatisticalanalysis. Prentice-Hall,Englewood Cliffs,New Jersey.718 p.
Zoheri. M. H. 1981. Transnirationin mushrooms.MushroomSci. 11C2V217-232
Accepted forpublicationMarch 27, 1985

This content downloaded from 130.194.20.173 on Thu, 21 Nov 2013 22:23:32 PM


All use subject to JSTOR Terms and Conditions

You might also like