Research

Spatial and temporal abundance of mycelial mats in

Blackwell Publishing, Ltd.

the soil of a tropical rain forest in Mexico and their

effects on the concentration of mineral nutrients in
soils and fine roots
Roger Guevara and Iliana Romero
Instituto de Ecología, AC Departamento de Biología de Suelos, Apartado Postal 63, Código Postal 91000, Xalapa, Veracruz México

Summary

Author for correspondence:
R. Guevara
Tel: +52 2288421800 ext 4405
Fax: +52 2288187809
Email: roger@ecologia.edu.mx
Received: 20 November 2003
Accepted: 16 March 2004
doi: 10.1111/j.1469-8137.2004.01099.x
• Here, we investigated the spatial and temporal abundance of mycelial mats in
a tropical rain forest to determine their effects on the concentration of mineral
nutrients in soils and fine roots.
• Mats were marked and followed over three seasons. Fine-root mass and the
concentration of mineral nutrients in both soils and roots were determined for
mat-associated soils and for a control group.
• Mats were more abundant in the dry season than in the wet season. The con-
centration of mineral nutrients in soils and fine roots increased from the rainy season
to the end of the dry season. Mats appeared to affect the concentration of phosphorus,
potassium and calcium, and the carbon : nitrogen and nitrogen : phosphorus ratios
in soils and roots during all seasons.
• Mats appeared to compete with plants for certain minerals. This could be part of
‘bottom up’ effects that may influence underground herbivory, as well as the above-
ground concentrations of mineral nutrients in plants. Mats are relevant to
understanding soil biodiversity and the potential feedback paths between the soil and
above-ground subsystems.

Key words: Chajul, saprotrophic fungi, life forms, mineral nutrient uptake, southern
Mexico.

© New Phytologist (2004) 163: 361–370

have been, to our knowledge, centred on temperate forests;

Introduction
Mycelia of saprotrophic and mycorrhizal fungi occur in eco-
systems as various life-form types: diffuse networks, thick and
dense mycelial mats, cords and rhizomorph systems (Cooke
& Rayner, 1984). Such hyphal forms in saprotrophic fungi
are used to decompose substrates and can be considered as
strategies for gaining access to food. Mycelial life forms not
only differ morphologically, but also have marked behavioural
and physiological differences that determine, to a great extent,
the diversity of their interactions with other organisms (e.g. with
fungivores and plants roots) and their effects on the physical
environment (e.g. soil chemistry).
Mycelial mats occur on soils associated with certain tem-
perate and tropical forest. However, studies of these life forms
and particularly on Hysterangium setchellii, an ectomycor-
rhizal fungus that profoundly affects soil biology.
Saprotrophic mycelial mats are characteristic of tropical
rain forest soils in the Montes Azules Biosphere Reserve, Chi-
apas, Mexico. These mats are frequently observed, during all
seasons, at the interface between the loose, leaf litter layer and
the actual soil. In this study, we investigated the spatial and
temporal abundance of mycelial mats, as well as their associ-
ation with fine-root mass and the concentration of mineral
nutrients in soils and roots.
Some of the effects of mycelium mats on soil biology that
have been documented are available for Douglas Fir (Pseudotsuga
menziesii ) stands in Oregon where more than 10% of the
soil surface is directly influenced by mycelial mats of H.

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362 Research
setchellii (Cromack et al., 1979). For example, there is a study
showing that fungivores (such as springtails, oribatid
mites, nematodes and amoebae) are more abundant in
mycelial mat soils than in control soils (Cromack et al., 1988).
In addition, it has been shown that Douglas Fir seedlings are
strongly associated with mats of H. setchellii and Gauteria
monticola (Griffiths et al., 1991a), and that H. setchellii
mats favour nitrogen (N) (Griffiths et al., 1991b), phospho-
rus (P), aluminium (Al), zinc (Zn), iron (Fe), copper (Cu),
boron (B), manganese (Mn), potassium (K), calcium (Ca)
and magnesium (Mg) (Entry et al., 1992) mineralization dur-
ing the decomposition of Douglas Fir needles (Entry et al.,
1991a) and the stems of White fir, Abies concolor (Entry et al.,
1991b). Despite all this evidence, and to the best of our
knowledge, no information on the distribution and effects of
mycelial mats on soil biology has been generated for other
forest ecosystems.
In order to investigate the spatial and temporal abundance
of mycelial mats, as well as their association with fine-root
mass and the concentration of mineral nutrients in soils
and roots we have made the following considerations.
(1) It is widely known that water availability and temperature
(Cooke & Whipps, 1993) determine, to a great extent, the
distribution and metabolic activity of mycelia. Also, it is
known that microenvironmental conditions in forest soils
vary considerably, even at the spatial scale of a few square
meters. This variation can be caused by subtle differences
in the proximity of watercourses, soil depth and slope, and
canopy shading (Guevara et al., 2002). Therefore, we hypo-
thesize that seasonal changes in weather conditions might
have differential effects on the abundance of mycelial mats
when considering different portions of a landscape unit.
There is evidence for some tropical forests that, during the
rainy season, lowland soils tend to be water saturated, if
not partly flooded, whereas soils belonging to higher ground
maintain conditions more favourable for aerobic metabolism.
By contrast, throughout the dry season, water availability
in soils associated with higher elevations is less than that
in lowland soil because of the proximity of water sources
(Meinzer et al., 1999). Thus, we expect to observe a high
abundance of mats in lowland areas and a low abundance of
mats on higher ground during the dry season. The opposite
pattern is anticipated during the rainy season, when lowland
soils are water saturated. (2) Mycelial mats actively decompose
leaf litter and presumably increase rates of local mineraliza-
tion. However, there is evidence that trees respond positively
to changes in the concentration of mineral nutrients in the
soil (i.e. P, N and K) by increasing their fine-root mass when
soils are enriched (Cuevas & Medina, 1988; Persson, 1990;
Raich et al., 1994; McGrath et al., 2001). Therefore, when
compared with a soil control group, we hypothesize that
mat-associated soils will show: (1) greater fine-root mass, and
(2) larger concentrations of mineral nutrients in soil and fine
roots.
Materials and Methods
Study site
This study was conducted at the Chajul field station, in the
southern part of the Montes Azules Biosphere Reserve in
the state of Chiapas, Mexico. The protected area covers c.
13 000 km2 (Arriaga et al., 2000). Around the field station,
the dominant vegetation is mature tropical rain forest on
lowland areas and small hills (80–200 m above sea level). The
average annual temperature (Fig. 1) is c. 25°C and the annual
rainfall is c. 2214 mm.
This study was conducted within an area of 5 km2 on two
landscape units, each of which included riparian, lowland and
high ground. The dominant tree species in the riparian and
lowland areas were: Andira inermis (Wright) DC., Bravaisia
integerrima (Spreng.) Standl., Calophyllum brasiliense Camb,
Coccoloba barbadensis Jacq., Cordia sp., Dendropanax arboreus
(L.) Decne. & Planch., Diospyros digyna Jacq., Guarea spp.,
Pachira aquatica Aubl., Pithecellobium arboreum (L.) Urban,
Platymiscium yucatanum Standley, Quararibea funebris (La
Llave) Vischer, Sapindus saponaria L., Scheelea liebmannii
Becc., Spondias mombin L., Tabebuia rosea (Bertol) DC. and
Vatairea lundellii (Standl.) Killip. In the high ground areas,
the dominant tree species were: Alchornea latifolia Sw., Ber-
noullia flammea Oliv., Brosimum alicastrum Sw., Cedrela
odorata L., Cupania sp., Cymbopetalum penduliflorum (Dunal)
Baill, Dialium guianense (Aublet) Sandw., Guatteria anomala
R. E. Fr., Lonchocarpus sp., Manilkara zapota (L.) van Royen,
Pseudolmedia oxyphyllaria Donn. Sm., Quararibea funebris
(La Llave) Vischer, Schizolobium parahybum (Vell.) Blake,
Sterculia apetala ( Jacq.) Karst., Swietenia macrophylla King,
Terminalia amazonia (Gmel.) Exell and Vatairea lundelli
(Standl.) Killip.
Fig. 1 Monthly average rainfall (20 yr of records, solid line) and
mean temperature (13 yr of records, dashed line) from Agua Azul
weather station (16°45′ N and 90°46′ W) c. 75 km north of the
Chajul Field Station, Mexico. The tinted area indicates the dry season.
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Mycelial mat abundance and size
In each of the three main seasons, dry (March), early rainy
( June) and late rainy season (October), we recorded the
number of mycelial mats in 30 randomly selected plots
(different plots for each season), each plot measuring 4 m2
(2 m × 2 m). Plots were distributed so as to encompass
existing landscape heterogeneity. Ten plots were located in
riparian vegetation next to the Lacan-Tu river, 10 plots were
assigned to the lowland ground at least 250 m away from the
river, and 10 plots established on high ground with mild
slopes (10–12°).
In addition, we marked each mat with a flexible plastic
straw (0.5 cm diameter and 50 cm long) that was partly
buried (20 cm) at the centre of each mat, and then recorded
the straw’s position within each plot. During the early rainy
season and the late rainy season, we recorded the presence
or absence of previously marked mats.
To investigate seasonal changes in the size of mycelial mats,
we randomly selected one mat from each plot containing two
or more mats, and also evaluated all mats from plots with only
a single mat. We recorded largest and shortest diameters for
each mat in order to calculate surface areas.
Soil and fine-root sampling
Each season we randomly selected 12 mats. We collected a soil
core (10 cm depth from a 15 × 15 cm square, surface area
225 cm2) from the centre of each of these mats. As a control,
we collected a second soil core 30 cm away from each mat.
From the top 5 cm of each soil core, we manually removed all
roots and debris, and then saved the soil samples for chemical
analysis. The rest of the soil cores were washed in a sieve to
recover roots. These roots were then added to those already
manually removed, and then all root samples were stored in
paper bags and air-dried. In the laboratory, fine roots < 2 mm
diameter, and those between 2 mm and 5 mm diameter, were
separated and oven-dried at 55°C for 72 h, and then weighed.
Chemical analyses of soils
For soil analysis, we followed the methods described in Etchevers
(1984). Soil pH was measured in a standard solution, 1 : 2 of
soil in distilled water. Total carbon was estimated based
on the methods of Walkley and Black (Nelson & Sommers
1982). One gram of soil sample was transferred to 250 ml
Erlenmeyer and added 10 ml of 1 N K2Cr2O7and 10 ml of
concentrated H2SO4. After 30 min, 50 ml of deionized water,
3 ml of concentrated H3PO4 and 0.5 ml of 1% defenilamina
indicator (Aldrich, 11,276–3) were added. Then, titrated
with 1  FeSO4 solution up to a green colour endpoint.
Total N was estimated using the micro-Kjeldahl method.
Soil samples, 200 mg, were digested with 4 ml of a digestion
mixture (0.42 g of selenium, 14 g Li2SO4 350 ml of 30% H2O2
© New Phytologist (2004) 163: 361–370 www.newphytologist.org
Research 363
and 420 ml of concentrated H2SO4) at 360°C for 2 h; enough
water was then added to make 100 ml. An aliquot from of the
clear solution of the digest was mixed with 25 ml of alkali
mixture (500 g NaOH and 25 g of sodium thiosulphate in
water, made up to 1000 ml) and distilled. A total of 25 ml of
the distillate was collected in 50 ml beaker containing 5 ml
of boric acid indicator solution (20 g boric acid in water
and 15 ml of pH 4.5 indicator solution diluted to 1000 ml)
and titrated with 0.01  H2SO4. (Bremmer, 1965; Bremer &
Mulvaney, 1982).
For phosphorous, 1 g of soil was placed in porcelain cru-
cibles and heated, in a muffle, up to 550°C. The temperature
was maintained at 550°C for 1 h, before allowing the samples
to cool. After cooling the organic P in the samples was extracted
with 0.5  H2SO4 and then phosphorus was determined
using the ammonium molybdate assay (Olsen & Sommers,
1982). Available P was carried out using NH4F and assayed
using the ammonium molybdate method (Bray & Kurtz,
1945; Olsen & Sommers, 1982a).
Calcium, magnesium, and potassium were extracted from
the soil by mixing 10 ml of 1  ammonium acetate (pH 7.0)
with a 10 g soil sample. The filtered extract was analysed with
an inductively coupled plasma atomic emission spectrometer
(Lanyon & Heald, 1982) for calcium and magnesium and by
flame photometry for potassium (Knudsen et al., 1982).
Chemical analyses of fine roots
Total nitrogen was estimated using the method of micro-
Kjeldahl. A 150 mg sample of fine roots was digested with
4 ml of a digestion mixture (0.42 g of selenium, 14 g Li2SO4
350 ml of 30% H2O2 and 420 ml of concentrated H2SO4) at
360°C for 2 h; enough water was added to make 100 ml. The
digest was then distilled and titrated as described earlier
(Bremer & Mulvaney, 1982).
For P, K, Ca, and Mg, we predigested 500 mg of fine roots,
finely ground in 4 ml of 65% HNO3 and 2 ml of 70% HClO4,
overnight. The samples were then heated to 150°C for c.
45 min and then to 210°C for c. 60 min. Then the digest
was made up to 10 ml with deionized water. Phosphorus
was determined using the ammonium molybdate assay
(Saunders & Williams, 1955 as modified by Walker & Adams,
1958). Calcium and Mg were quantified by inductively
coupled plasma atomic emission spectrometry (Allen, 1971)
and K was quantified by flame spectrometry (Knudsen et al.,
1982).
Statistical analyses
All statistical analyses were done with  5.0.1 (SAS
Institute Inc., 1999). To test whether mat abundance and mat
size varied spatially and seasonally, we used a two-way 
with the factors being season (three levels: dry, early rainy and
late rainy season) and location (three levels: riparian, lowland
364 Research
and high ground). In the case of mat abundance, the number
of mycelial mats per plot (i.e. the response variable) was
transformed by adding 0.5 to the observed value and then
taking the square root, in order to normalize residuals
(Zar, 1996). For nutrient concentration in roots and soil, we
used a  test. Because the sample design was paired, we
performed the analysis on the per cent differences between
pairs [(M − C )/C × 100] (C = control, M = mat) in order to
account for nonassociated spatial variability in the presence
of mycelial mats. Although the response variable was per cent
difference, there were no lower or upper limits because potential
values could range from plus to minus infinity. All data
were considered independent, as percentages were calculated
separately for each pair of observations. For this analysis, the
factor associated with season included three levels (dry, early
rainy and late rainy season). We used Fisher’s protected, least-
significant differences test to compare seasons. In addition,
we tested differences between observed means and zero (i.e.
whether mycelial mats affected chemical content). To do this,
we used the Dunnett, post hoc comparison test (Dunnett, 1955;
Zar, 1996), establishing the control as a set of 21 observations
with mean and variances equal to zero. The number of
zero observations in the control group was approx. n(k − 1)
(k = the number of means to be compared, including the
control; n = the number of observations made during each
season) as recommended by Dunnett (1955). Similar pro-
cedures (i.e. per cent differences and post hoc mean comparisons,
but based on a one-way  test) were used to address
whether mycelial mats affected fine-root mass.
Results
Abundance and size of mats
The microscopic analysis of mats revealed the presence of
clamped-connections (fibula), indicating that these mats were
basidiomycetes. We observed, on a few occasions, the fruiting
bodies of different species of Marasmius and Collybia growing
from mat-colonized substrates. However, a more detail molecular
study will be needed to establish taxonomic identities.
We observed a total of 411 mats: 181 in the dry season, 175
in the early rainy season and 55 in the late rainy season.
Spatially, the largest number of mats observed was associated
with high ground (175 mats), whereas the lowest number
(104 mats) was observed in riparian soil; 132 mats were
observed in lowland soils. On average, mycelial mats covered
a small percentage of the soil surface area (Table 1) (usually
less than 1%, but varying from zero to 15%). In general, mats
were short lived. Only c. 2% (four mats) of the mats recorded
in the dry season persisted until the late rainy season. About
14% (49 mats) of the mats persisted over two consecutive
seasons. About 77% (139 mats) and 96% (168 mats) of the
mats observed in the dry and early rainy season, respectively,
were recorded only once.
Table 1 Average percentage of soil surface covered by mycelium
mats and range of coverage (in brackets)
Riparian Low land High ground
Dry 0.6 (0 –3.3) 1.0 (0 –3.1) 2.1 (0 –7.9)
Early rainy 0.7 (0 –6.4) 1.2 (0 –11.5) 1.9 (0 –15.3)
Late rainy 0.3 (0 –2.5) 0.4 (0 –5.7) 0.1 (0 – 0.8)
n = 10.
The abundance of mycelium mats varied significantly among
seasons (F = 3.5; df = 5, 81; P = 0.0003, Fig. 2a). The abund-
ance of mats was higher in the dry season (mean ± standard
error, 2.5 ± 0.5 mats per plot) than in the early rainy season
(1.8 ± 0.5) and late rainy season (1.5 ± 0.5). The same pattern
was separately observed in the high ground (Fig. 2b) and the
lowland (Fig. 2c). Also, mycelium mats were differentially
distributed in landscape units and these varied among
seasons. In the dry season (Fig. 2d) the number of mats per
plot observed in the riparian area (1.4 ± 0.71) was signific-
antly lower than the numbers observed in the high ground
(3.0 ± 0.71) and in the lowland (3.6 ± 0.71). A similar pattern
was observed for the early rainy season (Fig. 2e). In addition,
the mycelial mats recorded on high ground in the dry season
(352.4 ± 43.1 cm2) were significantly larger (F = 2.67, df =
4,132, P = 0.023) than those found in the riparian vegetation
(135.5 ± 61.0 cm2) in the dry season and also were larger than
those located on high ground during the late rainy season
(53.1 ± 83.2 cm2).
Fine-root mass
On average, over twice the mass of fine roots, less than 2 mm
in size, was encountered in the late rainy season (mean ± SD,
3.79 ± 1.37 g ) when compared with the dry (1.55 ± 0.70)
and early rainy seasons (1.82 ± 0.86) (F = 36.5, df = 2, 75,
P = 0.0008). The observed value for fine roots < 5 mm observed
in the late rainy season (1812 g m−2) was close to values reported
for some Amazon Caatinga forests in Venezuela – 1721–2879
g m−2 (Klinge & Herrera, 1978) – and similar to values asso-
ciated with other tropical forests (Edwards & Grubb, 1982;
Cavalier, 1992).
The  test on per cent differences for fine-root mass
between pairs showed no significant effects (F = 2.5; df = 3,56;
P = 0.044) for season, and none of the observed mean differ-
ences was significantly different from zero (in other words,
mycelial mats showed no statistical effect on fine-root mass).
Seasonal patterns of mineral nutrients
Soil pH varied from 4.9 to 5.8 among soil samples, and there
were no significant differences between seasons. Mineral nutrients
in the soil (Wilks’ lambda = 0.57, F = 34.99, df = 16, 176,
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 Research 365

Fig. 2 Seasonal and spatial variation in the

abundance of mycelium mats per plot:
(a) overall seasonal variation; (b) seasonal
variation in the high ground; (c) seasonal
variation in the low land; (d) spatial variation
in the dry season; and (e) spatial variation in
the early rainy season. *, P < 0.05.

Table 2 Summary of the ANOVA tests for mineral nutrients of soil and fine roots

Soil Seasonal Effects of mycelium mats analysis

analysis
SS-season SS-error F Power SS-factor SS-error F Power

C 995.9 400000 0.12ns > 0.99 0.107 2.682 0.9ns 0.23

P-total 38.95 19.76 93.63*** > 0.99 3.616 4.511 17.6*** > 0.99
P-ext 0.002 0.012 7.92*** > 0.96 5 105.47 1.0ns 0.26
N 96.07 1145 3.98* 0.76 0.682 6.737 2.2ns 0.53
K 36.72 11.58 150.62*** > 0.99 1.324 0.879 33.1*** > 0.99
Ca 582.7 1336 20.72*** > 0.99 9.559 4.703 44.7*** > 0.99
Mg 15.3 42.41 17.14*** 0.98 0.301 2.358 2.8* 0.65
C:N 402.9 1729 11.07*** C:N 0.141 4.438 0.7ns 0.18
N:P 2806 2204 60.46*** > 0.99 9.314 11.847 17.3*** > 0.99
Fine roots
C 51562 20758 93.1*** > 0.99 0.008 0.051 2.93* 0.68
P 99.73 43.67 85.6*** > 0.99 2.935 0.639 85.74*** > 0.99
N 45.99 383.4 4.5* 0.76 0.141 1.04 2.53ns 0.59
K 42.03 136 11.6*** > 0.99 3.604 0.789 85.27*** > 0.99
Ca 1997 1670 44.8*** > 0.99 2.197 0.645 63.58*** > 0.99
Mg 37.1 104.9 13.3*** > 0.99 0.669 7.825 1.6ns 0.39
Na 0.35 0.66 19.9*** > 0.99 4.678 23.537 3.71*** 0.78
Fe 315 133.6 88.4*** > 0.99 13.01 106.47 2.28ns 0.54
C:N 318.1 1292 9.2*** > 0.98 0.259 1.139 4.24** 0.84
N:P 89438 13088 256.3*** > 0.99 22.11 16.954 24.35*** > 0.99

C, carbon; Ca, calcium; Fe, iron; K, potassium; Mg, magnesium; N, nitrogen; P, phosphorus; P-tot, total phosphorus;P-ext, readily available
phosphorus; SS, sum of squares.
ns
, *, **, ***, indicate P > 0.05, P < 0.05, P < 0.01 and P < 0.001, respectively.

P = 0.00065; Roy’s greatest difference = 5.89, F = 65.54,
df = 8, 89, P = 0.0032) and fine roots (Wilks’ lambda = 0.1,
F = 52.11, df = 20, 132, P = 0.00056; Roy’s greatest root =
29.4, F = 196.8, df = 10, 67, P = 0.00093) showed significant
differences in concentrations among seasons (Table 2).
In soil samples total phosphorus (P-tot), readily available
phosphorus (P-ext), K, Ca and Mg had high concentrations
in the early rainy season followed by a significant decrease in
concentration by the late rainy season (Table 3 and Fig. 3a).
The same pattern was observed for the C : N ratio. The con-
centration of P-ext, K and Ca in the dry season did not differ
significantly from the concentrations observed in the early
rainy season, whereas the concentration of P-tot in the dry
season was lower than the concentration observed in the early

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366 Research

Table 3 Summary of the mineral nutrient concentration (mg g−1) in the overall seasonal analysis for soil and fine roots and per cent differences
((M − C )/C × 100, where M is mat soil and C is control soil) in mineral nutrient concentration between mat-associated samples and control
samples of soil and fine roots

Soil Overall seasonal analysis Effects of mycelium mats analysisa

Early rainy Late rainy Early rainy Late rainy

Dry Mean 95% CI Mean 95% CI Mean 95% CI Dry Mean 95% CI Mean 95% CI Mean 95% CI

C 306.98 17.75 309.0 17.66 301.77 48.72 7.48 7.48 2.31 7.77 8.92 13.19
P-total 1.92 0.36 2.19 0.36 0.81 0.14 − 44.81 3.75 − 40.49 3.59 8.48 18.07
P-ext 0.023 0.009 0.026 0.007 0.014 0.004 10.65 149.25 189.08 152.28 60.48 75.65
N 14.18 1.17 11.51 1.07 13.50 2.60 0.21 4.53 12.30 8.41 21.84 21.85
K 2.37 0.28 2.26 0.28 1.09 0.11 −27.34 4.48 −26.64 2.7 3.38 7.69
Ca 15.05 1.68 16.12 2.15 10.76 2.42 −22.93 3.61 −31.37 3.51 59.06 18.47
Mg 2.34 0.17 3.45 0.21 2.76 0.51 −1.62 5.76 7.68 4.14 13.81 12.79
C:N 21.95 1.83 27.38 2.50 23.25 2.76 7.56 7.34 −7.78 10.24 − 0.37 17.17
N:P 8.07 1.53 5.69 1.10 17.45 3.68 83.71 16.64 91.17 21.97 25.50 26.82
Fine roots
C 512.10 8.01 479.99 15.66 542.17 5.74 2.15 1.89 3.07 3.55 1.13 1.54
P 2.18 0.62 2.82 0.61 0.23 0.04 −56.64 8.96 − 42.98 5.01 − 39.16 6.23
N 18.41 1.57 16.71 1.35 16.77 1.21 3.73 13.81 −10.22 8.69 − 4.71 10.40
K 3.07 0.97 2.05 0.60 3.83 0.81 − 58.20 9.77 −49.72 12.59 − 42.39 4.30
Ca 15.34 3.95 19.76 3.40 7.78 1.13 − 49.83 11.26 −31.63 6.27 − 28.61 4.94
Mg 4.05 0.96 3.94 0.56 2.58 0.60 −28.55 27.32 −11.55 12.42 − 10.55 42.03
Na 0.26 0.07 0.11 0.04 0.11 0.06 −30.50 23.22 35.30 84.42 − 40.60 30.21
Fe 3.37 0.98 5.83 1.12 0.97 0.28 −29.85 31.32 −13.92 16.21 97.61 183.74
C:N 28.25 2.26 29.27 2.97 32.83 2.29 1.88 11.82 17.20 11.81 8.96 11.45
N:P 11.15 4.64 6.53 1.27 78.34 11.15 169.27 74.57 58.57 13.28 58.02 13.77

C, carbon; Ca, calcium; Fe, iron; K, potassium; Mg, magnesium; N, nitrogen; P, phosphorus; P-tot, total phosphorus; P-ext, readily available
phosphorus. aA negative mean indicates that mycelial mats lowered the concentration of minerals in compared with the control samples.

rainy season but significantly higher than the concentration
recorded for the late rainy season. The N concentration showed
a different seasonal pattern: N concentration in the early rainy
season was low and significantly different from the observed
concentrations in the dry and late rainy season. Similarly, the
N : P ratio was highest in the late rainy season (17.45) and
decreased significantly by the dry and early rainy seasons.
In fine roots, the observed seasonal patterns of the concen-
tration of mineral nutrients were similar to those found in soil
samples (Table 3 and Fig. 3b). Concentrations of P, Ca and
Mg in fine roots were highest in the early rainy season fol-
lowed by a decrease in concentration by the late rainy season.
By contrast, the concentration of N in fine roots also showed
a pattern similar to that observed in soil samples, with the
highest concentration in the dry season and the lowest con-
centration in the early rainy season. The N : P ratio in fine
roots was highest in the late rainy season and low in the dry
and early rainy season.
Mycelium mats effects
Mycelial mats had a significant effect on the observed concen-
tration of mineral nutrients in soil (Wilks’ lambda = 0.05,
F = 10.8, df = 27, 170, P < 0.001; Roy’s greatest root = 6.6,
F = 44.1, df = 9, 60, P < 0.001) and fine roots (Wilks’ lambda
= 0.1, F = 52.11, df = 20, 132, P < 0.001; Roy’s greatest root
= 29.4, F = 196.8, df = 10, 67, P < 0.001) (Table 2).
Overall, mycelium mats had a negative effect on the
concentration of mineral nutrients in soil (Table 3 and Fig. 4a)
and fine roots (Fig. 4b). Soils affected by mycelium mats had
a significantly lower concentration (a reduction ranging from
25% to 45%) of P-tot, K and Ca in the dry and early rainy
season than concentrations observed in control soils. Myce-
lium mats lowered the concentrations of P, K and Ca in fine
roots by nearly 50% compared with concentrations in control
fine roots and increased the N : P ratio by > 80% in soil and
from c. 50% to c. 160% in fine roots compared with control
samples.
Discussion
Our data showed that soil mineral properties varied greatly over
the year. Low concentrations of mineral nutrients occurred
during the rainy season, while fine-root mass was twice that
observed in dry and early rainy seasons. This is in accord with
the observations of Lodge et al. (1994) and Roy and Singh
(1994) that, during the rainy season, uptake by plants and
microorganisms, together with lixiviation, lowers the concen-
tration of mineral nutrients in the soil. It appears that the
mineral nutrient contribution of microorganisms builds up

www.newphytologist.org © New Phytologist (2004) 163: 361–370


Fig. 3 Seasonal effects in the concentration of mineral nutrient (C,
carbon; Ca, calcium; K, potassium; Mg, magnesium; N, nitrogen;
P, phosphorus; P-tot, total phosphorus; P-ext, readily available
phosphorus) in (a) soil and (b) fine roots showed as a per cent
reduction from the highest observed concentration. All differences
shown were statistically significant.
in the soil during the dry season, as plants are not very actively
engaged in mineral nutrient uptake. However, from the
beginning of the rainy season nutrient uptake by plants begins
in earnest and, accordingly, a decrease in the concentrations
of mineral nutrients in the soil can be perceived (Lodge et al.,
1994). A similar seasonal trend is also observed in the con-
centration of mineral nutrients (P, Fe, Ca and Mg) in fine roots,
a pattern congruent with soil mineral nutrient availabilities
recorded at our study site, as well as with the published
evidence (Aerts & Chapin, 2000; Harrington et al., 2001).
Our data also show that this forest, like other tropical forests,
is a strongly P-limited ecosystem (Klinge, 1976; Fölster &
Huber, 1984; Sim & Nykvist, 1991; Bloomfield et al., 1993;
Hondersmann, 1995; Murach et al., 1995; Priest et al., 1999;
Campos & Dirzo, 2003) as was shown by the observed N : P
ratio values in fine roots in the late rainy season (c. 78), which
was much higher than 16, a value suggested as indicative of a
P-limited environment (Koerselman & Meuleman, 1996).
The evidence gathered in this study showed that mycelial
mats had a strong effect on soil chemical properties and on the
© New Phytologist (2004) 163: 361–370 www.newphytologist.org
Research 367
Fig. 4 Effects of mycelium mats on the concentration of mineral
nutrients (C, carbon; Ca, calcium; K, potassium; Mg, magnesium; N,
nitrogen; P, phosphorus) in (a) soil and (b) fine roots shown as the per
cent difference between soil samples associated with mycelium mats
and control soils. All shown differences were statistically significant.
concentration of mineral nutrients found in fine roots. The
average (0.9%) and range of variation (0–15.0%) for soil
surface area covered by mycelium mats at the study site were
low compared with the average of 15.0% and range of 4.3–
27.4% reported for a Douglas Fir stand (Cromack et al., 1979).
Such a contrast suggests that mycelial mats in this tropical rain
forest in southern Mexico have little effect on soil biology.
However, three factors have to be taken into account in
order to better understand the meaning of these contrasting
figures. (1) The estimates of Cromack et al. (1979) were based
only on three plots, so that the reported figures would certainly
be inaccurate. (2) Our observations are from a highly diverse,
tropical forest. By contrast, the Cromack et al. (1979) data are
from a secondary growth (45- to 60-yr-old), Douglas Fir
stand, representing a managed woodland where the density
368 Research
of host trees, and very likely that of their associated fungi,
had been unnaturally increased. (3) Cromack et al. (1979)
reported that mycelial mats of H. setchellii were long-lived,
whereas the mycelial mats at our study site were short-lived
(less than 4 months) with high turnover rates. Therefore, it is very
likely that, by using our approximation of a ‘snapshot’ every
3 months, we have underestimated the soil surface that is directly
affected by mycelial mats in the tropical rain forest at Chajul.
In relation to the distribution of mycelial mats, our data did
not support the initial prediction that large-scale patterns of
water availability might determine the distribution of mats.
Contrary to our prediction, the abundance of mycelial mats
was low in the rainy season. Also, in the dry season the
abundance of mats was high on high ground, far removed
from water sources, whereas it was low in the riparian vegeta-
tion. These findings disagree with published evidence that
indicates higher fungal activity near water sources (Worthen
& McGuire, 1990) and during the wet season. Contrary
to large-scale seasonal patterns, it seems likely that microscale
weather conditions could determine the abundance of myce-
lial mats. We have unpublished evidence that indicates that
mycelial mats are closely associated with large, buttressed trees
that are more abundant on high ground than in riparian
vegetation. Such large trees might favour a humid soil micro-
climate because of shading effects (canopy and trunk shadow),
low wind exposure and a thick leaf litter layer.
Another factor affecting the distribution of mycelial mats
might be the trophic activity of fungivores. Such activity has
already been reported to influence the distribution of mycelia
(Newell, 1984a; Newell, 1984b; Guevara et al., 2002). This,
together with the evidence that fungivores are more abundant
in soil associated with mycelial mats of H. setchellii than in
adjacent soil (Cromack et al., 1988), suggests that fungivory
could play an important role in the spatial and temporal
distribution of mycelial mats. This is a line of research that
deserves further investigation.
Contrary to our initial hypothesis, mycelial mats seem to
compete, by interference, with fine roots. Although we did
not observe differences in fine-root mass between mat soils
and the control soil, we did detect a strong effect of mats on
the concentration of mineral nutrients (P, K and Ca) and on
the N : P ratio in both soils and fine roots. The case of P is of
particular interest since in an overall P-limited ecosystem,
mycelium mats form patches in which the availability of P for
plants is markedly lowered when compared with control
samples. In addition mycelium mats showed high turnover
rates. Therefore, mycelium mats may contribute significantly
to the soil heterogeneity that plants are facing in relation to
the mineral nutrient uptake.
The same effect on the concentration of mineral nutrients
in soils and roots was observed over the three seasons, but the
strongest effect occurred during the dry season. We have no
data to weigh the relative importance of seasonal water avail-
ability, or to assess the contribution of biological processes,
such as fungivory, in shaping seasonal competition gradients
for mineral nutrients between mycelial mats and fine roots. As
discussed above, fungivory could be an important factor lim-
iting the abundance and size of mycelial mats. Therefore, we
might hypothesize that as fungivory increases, the competitive
ability of mycelial mats should decrease, thus favouring an
increase in concentrations of mineral nutrients in fine roots.
This hypothesis is congruent with published evidence show-
ing that, in a tropical cloud forest, fungivory is higher in the
rainy season than during the dry season (Guevara & Dirzo,
1999). It is also consistent with our own observations that
fungivorous soil arthropods are more abundant during the
rainy season than in the dry season.
Overall, this study has shown that saprotrophic, mycelial
mats in a tropical rain forest at the Chajul field station have a
strong effect on soil and root chemical contents. The spatial
and temporal dynamics of mycelial mats partly contribute to
soil heterogeneity at various scales and have the potential to
affect other biotic elements, such as the richness and diversity
of soil arthropods (Cromack et al., 1988; Guevara et al.,
2002). Furthermore, since mycelial mats alter the chemical
content of fine roots, this may represent part of a ‘bottom up’
effect that modifies underground and above-ground plant
tissue quality and herbivory (van Tol et al., 2001; Preisser,
2003). Evidence suggests that changes in the P content of
plant tissue from 16% to 23% may significantly affect the
levels of herbivory in a tropical dry forest in Mexico (Campos
& Dirzo, 2003). Therefore, it is plausible that the observed
differences in the P concentration (32% to 56%) between
mycelium-mat associated fine roots and control roots may
have significant effects on underground herbivory. These are
aspects deserve further experimental investigation, since they
are relevant to understanding the diversity of organisms and
process in the soil and may be part of feedback paths between
soil and above-ground subsystems.
Acknowledgements
We thank the staff in Chajul Field Station for facilitating our
fieldwork. A very special thanks to Soraida Irisson who per-
formed all chemical analysis. CONACYT (I32791-N) and the
Department of Soil Biology, INECOL sponsored this project.
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