Soil Biology & Biochemistry 50 (2012) 33e39

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Soil Biology & Biochemistry

journal homepage: www.elsevier.com/locate/soilbio

Soil biota effects on soil structure: Interactions between arbuscular mycorrhizal

fungal mycelium and collembola
Md. Rezaul Karim Siddiky a, Josef Kohler a, Marco Cosme b, Matthias C. Rillig a, *
a
Institut für Biologie, Plant Ecology, Freie Universität Berlin, Altensteinstr. 6, D-14195 Berlin, Germany
b
Institut für Biologie, Functional Biodiversity, Freie Universität Berlin, D-14195 Berlin, Germany

a r t i c l e i n f o a b s t r a c t

Article history: Soil aggregation is an important ecosystem process mediated by soil organisms. Collembola and
Received 3 December 2011 arbuscular mycorrhizal (AM) fungi are major soil biota representing different functional groups, and are
Received in revised form known as two key promoters of soil aggregation. Although several studies have experimentally
29 February 2012
demonstrated that AM fungi and, more recently, collembola affect soil structure, there is no study
Accepted 1 March 2012
Available online 16 March 2012
investigating how both soil organisms affect soil aggregation excluding the influence of plant roots,
another important driver of soil aggregation. Considering the importance of AM fungi and collembola in
terrestrial ecosystems, here we asked if both organisms have any influence on soil aggregation when
Keywords:
Arbuscular mycorrhizal fungi
roots are not present.
Hyphae In order to examine this question we conducted a completely factorial greenhouse study manipulating
Root the presence of both collembola and AM fungi and excluded the roots of Plantago lanceolata using
Soil structure a 38 mm nylon screen compartment. We quantified soil aggregation as water stable soil aggregates in four
Soil aggregation size classes in the hyphal compartment and monitored a number of other explanatory variables,
Microarthropods including AM (and non-AM) fungal soil hyphal length.
Collembola The soil in the hyphal compartment showed greater soil aggregation with larger mean weight
diameter when collembola were present, and a similar result was found in the presence of AM fungi,
compared to control treatments. Moreover, combined presence of both AM fungi and collembola resulted
in a non-additive increase of soil aggregation.
Our study clearly indicated that collembola can enhance soil aggregation, that they can partially
complement effects of AM fungi, and that these effects are independent of roots.
Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction particular for macroaggregates (>250 mm) (Tisdall and Oades,

Soil structure, as the three dimensional matrix of pore and
solid spaces, is an ecosystem property essential for facilitating
water and gas exchange, carbon storage, nutrient cycling, resistance
to erosion and other functions (e.g., Six et al., 2000; Coleman et al.,
2004). Soil aggregation, the process leading to soil structure, is thus
a principal ecosystem process, which can be directly or indirectly
controlled by various soil biota in a given environmental setting
(e.g. soil organic matter, texture, climate) (Bronick and Lal, 2005;
Rillig and Mummey, 2006).
Aggregation of soil is the result of various binding agents,
where plant roots and fungal hyphae play an important role, in
* Corresponding author. Tel.: þ49 (0) 30 838 53165; fax: þ49 (0) 30 838 53886.
E-mail address: matthias.rillig@fu-berlin.de (M.C. Rillig).
1982), as opposed to microaggregates, which are stabilized by
more permanent binding agents. Indeed, several studies have
highlighted the important contribution of arbuscular mycorrhizal
(AM) fungi to soil aggregation (Miller and Jastrow, 1990; Jastrow
and Miller, 1998; Jastrow et al., 1998; Rillig and Mummey, 2006).
AM fungi are considered a key functional component in the soil
(Rillig, 2004; Smith and Read, 2008), serving as an important link
within the plantesoil continuum (Wilson et al., 2009), and are
recognized as key promoters of soil aggregation (Piotrowski et al.,
2004; Rillig and Mummey, 2006; Chaudhary et al., 2009). The
evidence for AM fungal contribution to soil aggregation is strong, as
provided by a wide range of field observational studies (Rillig et al.,
2002a, b), field experiments (Wilson et al., 2009), mechanistic
greenhouse experiments (Thomas et al., 1993; Bearden and
Petersen, 2000; Piotrowski et al., 2004; Hallett et al., 2009; Bedini
et al., 2009), and more recently, by tests with exclusion of all
other biota (Rillig et al., 2010).

0038-0717/$ e see front matter Ó 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.soilbio.2012.03.001
34 Md.R.K. Siddiky et al. / Soil Biology & Biochemistry 50 (2012) 33e39
By comparison, a lot less direct experimental data exists for soil
animals in regard to soil aggregation, with the exception of earth-
worms, which are well-documented in their effects on soil struc-
ture (Bossuyt et al., 2005, 2006; Davidson and Grieve, 2006; Kavdir
_
and Ilay, 2011). Soil microarthropods, in particular collembola, are
ubiquitous soil animals (Petersen and Luxton, 1982) and can
influence a range of ecosystem processes (e.g., Finlay, 1985; Wardle
and Bardgett, 2004). Lussenhop (1992) hypothesized that collem-
bola could contribute to soil aggregation through their fecal pellets,
which are typically 30e90 mm in diameter (Rusek, 1975). Recently,
studies in our lab have provided the first direct experimental
evidence that collembola are capable of enhancing soil macro-
aggregation (Caruso et al., 2011; Siddiky et al., 2012). In these
studies, the combined presence of collembola and AM fungi led to
a higher level of macroaggregation than the individual effects of
collembola or AM fungal presence, which also individually signifi-
cantly enhanced soil aggregation (Siddiky et al., 2012). These
previous greenhouse studies were conducted in the presence of
plant root systems, i.e. plant roots were not experimentally isolated
from the added AM fungi or collembola. Thus it is not clear to what
extent the observed positive responses in soil aggregation to AM
fungi, collembola or their combination were mediated by indirect
effects via the roots.
Roots are widely appreciated agents of soil aggregation
(Tisdall and Oades, 1982; Miller and Jastrow, 1990; Jastrow et al.,
1998; Rasse et al., 2000; Six et al., 2004). Entanglement of soil
particles by roots may directly promote macroaggregates (Tisdall
and Oades, 1982; Miller and Jastrow, 1990; Jastrow et al., 1998).
Moreover, roots contribute to soil aggregate stabilization by
releasing organic materials (e.g., rhizodeposition), serving as
aggregate binding agents (Morel et al., 1991). Moreover, roots can
also indirectly influence soil aggregation through alteration of
microbial communities (Morel et al., 1991) or modification of the
soil water status (Reid and Goss, 1982). A number of studies (e.g.,
Monroe and Kladivko, 1987; Materechera et al., 1994) also reported
that the penetration of roots into macropores can result in
a decrease of macroaggregates (up to 50%). Irrespective of the
mechanism(s) involved, effects of roots, and their possible inter-
actions with the biota under investigation, need to be excluded in
any attempt to isolate effects of AM fungi or collembola.
Nevertheless, few previous studies have tried to experimentally
separate the influence of AM fungal hyphae and their host
plant roots on soil aggregation (e.g., Thomas et al., 1993; Bearden
and Petersen, 2000; Hallett et al., 2009). To disentangle the
influence of AM fungal hyphae and collembola on soil aggregation
from the effects of plant roots, in the present study we excluded
the roots by using a screen that permits only the passage of AM
fungal hyphae. We hypothesized that in the absence of plant root
effects, collembola would reduce AM fungal abundance, and
therefore would indirectly lead to a decrease in soil aggregation.
However, in addition to these indirect effects, collembola may have
direct positive effects on soil aggregation. In order to test these
hypotheses, we conducted a factorial greenhouse experiment
manipulating the presence of both AM fungi and collembola in
a volume of soil from which roots were excluded.
2. Materials and methods
2.1. Experimental design and greenhouse experiment
We conducted a 2  2 factorial greenhouse experiment where
ten replicates were set up for each combination of the four treat-
ments, Collembola (present, absent) and AM fungi (present,
absent), with a total of 40 experimental units (pots); the full,
balanced factorial design allowed us to test for two effects and
their interactions.
We used a sandy soil collected from an experimental field of
Freie Universität Berlin. The site is a meadow, and the soil an
Albic Luvisol. The soil properties were: sand ¼ 74%, silt ¼ 18%
and clay ¼ 8%; 6.9 mg/100 g P (calciumeacetateelactate);
5.0 mg/100 g K (calciumeacetateelactate); 0.12% N (total); 1.87%
C (total) (analyses conducted by LUFA Rostock Agricultural Analysis
and Research Institute, Germany; and using an Euro EA C/N
analyzer, HEKAtech GmbH, Wegberg, Germany). The soil was
chosen due to its high mycorrhizal inoculum potential and general
responsiveness to biota in terms of soil aggregation (Rillig et al.,
2010). Soil was sieved (10 mm) prior to use to remove stones and
roots. In order to reduce soil fertility, the soil was thoroughly mixed
with sand (70% soil with 30% sand). Following that, the soil was
steamed at 90  C (4 h) to eliminate AM fungi and collembola.
Pots (4 L) were divided in two compartments as shown in Fig. 1.
The compartments were set up using a solid round mesh tube
(height 20 cm; diameter 7 cm) made of 0.5 mm aluminum net
(supplied by: BAHAG AG, Bauhaus Handelsges., D-68167 Man-
nheim, Germany) covered with a 38 mm plastic screen (obtained
from: SEFAR AG, CH-9410 Heiden, Switzerland). The screened tube
was open at the top to allow the plant to grow within. This mesh
size allows only the passage of AM fungal hyphae between
compartments, but not of roots. The outer compartment is desig-
nated as hyphal compartment. We closed the bottom of mesh tubes
with plastic tape (2 mm thickness) and sealed with silicon
(supplied by BAHAG AG, Bauhaus Handelsges., D-68167 Mannheim,
Germany) to prevent plant roots from penetrating the hyphal
compartment during the experiment. The inner compartment is
Plantago lanceolata
Collembola
38 µm mesh tube
Root compartment
190 mm
Hyphal compartment
120 mm
Fig. 1. Schematic representation of root compartment set vertically in the center in
each experimental unit (pot). Each compartment was open at the top to receive a plant,
the side wall was covered by a 38 mm nylon mesh, and the bottom was closed with
plastic tape sealed with silicone. Inside the compartment, a plant grows inoculated or
not with AM fungi, while outside there was addition or not of collembola. The 38 mm
mesh prevents the roots (which have larger diameters) from growing outside the
comportment, allowing only the AM fungal mycelium to grow into the hyphal
compartment. The rim of the hyphal compartment (height from soil surface: 10 cm)
prevented collembola from entering the root compartment.
 Md.R.K. Siddiky et al. / Soil Biology & Biochemistry 50 (2012) 33e39
designated as root compartment. Both compartments were filled
with soil to the same level (3.0 kg soil per pot).
To establish the AM fungal treatment, the soil was thoroughly
mixed with an AM fungal spore inoculum. Soil for inoculum was
also collected from the same experimental field, and this inoculum
was produced according to the method of Klironomos (2002). The
inoculum produced from 300 g of soil was added to the root
compartment of each pot assigned to the treatments with presence
of AM fungi. We also collected a microbial wash produced from
non-sterile soil solution (same soil as used in the experiment)
filtered through a 20 mm sieve; in order to equilibrate the microbial
communities between the treatments, the filtrate was similarly
added to the root compartment in each pot assigned to the treat-
ments without AM fungi. We selected as plant species Plantago
lanceolata, a plant frequently used in mycorrhizal studies, and
which occurs at the grassland site from which the soil was taken.
We initially added two seedlings per pot, but after 1 week we
thinned to one plant (per pot). The plants were then grown in the
greenhouse for a period of 22 weeks. After confirming AM fungal
mycelial growth during the 10th week, we added collembola to the
hyphal compartment; the collembola treatment consisted of 80
Proisotoma minuta (Collembola: Isotomidae) individuals per pot.
This euedaphic species has a cosmopolitan distribution in Europe
and globally, frequently occurs in large numbers, and is often
associated with decaying organic matter (Fjellberg, 2007). These
collembola were reared in our lab before starting the greenhouse
experiment (laboratory culture since 2005; originally isolated from
northern Germany). The average air temperature in the greenhouse
was 22  C. During the vegetation period we watered the experi-
ment using tap water with pH of 7.7, and all pots received the same
amount of water (300 ml/pot) about every three days. The position
of pots was re-randomized once a week.
2.2. Plant and fungal measurements
After harvesting, plant shoots and roots were dried at 40  C for
72 h and then weighed to determine dry weight. We confirmed the
presence of AM fungal structures by measuring root colonization
following ink staining (Vierheilig et al., 1998) at 200 magnification
(at least 120 intersects per sample) as described by Rillig et al.
(1999). We also measured AM and non-AM fungal soil hyphal
length from a 4.0 g soil subsample from the outside compartment
of each pot, using an aqueous extraction/filtration method
(Jakobsen et al., 1992) followed by microscopic quantification of
hyphae at 200 (Rillig et al., 1999).
2.3. Microarthropods extraction
For determination of collembola abundance, a subsample of
150 g of soil from each pot was taken during harvest, and micro-
arthropods were extracted using a modified Macfadyen apparatus
(Macfadyen, 1961). The extraction was performed for 2 weeks;
during this extraction period the temperature was gradually
increased from 25  C to a maximum of 40  C. Afterwards collem-
bola were counted and the total abundance per pot calculated.
2.4. Water-stable aggregates measurement
Soil collected from the hyphal compartment, and soil prior to
the beginning of the experiment, was air-dried at 25  C for 10 days,
and then passed through a 4 mm screen before further analysis. Soil
stability was quantified as abundance of water stable aggregates
(WSA) using a series of stacked sieves (modified from Kemper and
Rosenau, 1986). We immersed a stack of sieves (2 mm, 1 mm,
0.5 mm, 212 mm; with the smaller sieve size at the bottom) in
35
a bucket filled with water. 50 g of air-dried soil were re-wetted by
capillary action (10 min), placed on the top of the sieve, and then
the sieves were moved up and down in the water (3 cm) for 3 min,
always with the surface of the top sieve (2 mm) completely
immersed in water. Material remaining on each sieve was crushed
and then passed through that sieve again to separate coarse matter
(sand and particulate organic matter) from soil. The soil and coarse
matter fractions from each sieve, and the material passing through
the smallest sieve, were collected and dried at 80  C for 48 h. The
mean weight diameter (MWD), coarse matter (CM) and the fraction
of water stable aggregate (WSA) in each size classes were calculated
as described in Barto et al. (2010).
2.5. Statistical analyses
The water stable aggregates (WSA; 212e500 mm, 0.5e1.0 mm,
1.0e2.0 mm and 2.0e4.0 mm) were analyzed by two-way multi-
variate analysis of variance (MANOVA, using the Pillai-Bartlett trace
statistic), followed by two-way univariate analysis of variance
(ANOVA), using collembola and AM fungi as categorical factors.
Data on initial aggregate size distribution prior to the experiment
were not included in these models. Data on percentage of total
WSA, mean weight diameter (MWD), AM and non-AM fungal
hyphal length in soil, root and shoot dry biomass were analyzed by
two-way univariate ANOVA only. Data on collembolan abundance
and AM fungal root colonization were analyzed by one-way
univariate ANOVA with collembola and AM fungi as categorical
factor, respectively (i.e., we did not observe any contamination in
non-collembola and non-AM fungal pots, and ANOVA is not valid
with zero variances). We conducted the full 2-factorial analyses
with hyphal lengths, since these were not expected to reach zero
abundance (decomposition of hyphae takes several months, and is
much reduced in sterilized soils; Rillig et al., 2010).
We tested residuals for normality (Shapiro test) and homoge-
neity of variance (Bartlett test); and when data deviated from
ANOVA assumptions we used appropriate transformations (Quinn
and Keough, 2002), specifically arcsine and square-root trans-
formations. Tukey’s honestly significant difference (HSD) test was
used to conduct post hoc analyses. All statistical analyses were
performed with the R software version 2.8.1 (R Development Core
Team, 2008).
3. Results
3.1. Demonstration of treatment effectiveness
During harvesting no roots were found outside the root
compartment. The treatment applications were successful for both
soil biota factors, AM fungi and collembola. The AM fungal inocu-
lated pots had roots colonized to around 60% with AM fungal
structures (Fig. 2i), whereas non-inoculated treatments contained
no recognizable AM fungal structures in the roots.
Extraction of collembola from the hyphal compartment soil in
the pots where P. minuta was added yielded between 350 and 450
individuals at the end of the experiment (Fig. 2ii), indicating pop-
ulation increase from the initial 80 individuals. No collembola were
extracted from the pots where no P. minuta individuals were added.
Furthermore, we did not find any animals during extraction of soil
from the root compartment either.
3.2. Soil aggregation
The MANOVA results showed that the interaction between both
factors significantly (P < 0.001; Pillai ¼ 0.460; F ¼ 7.02; df ¼ 4, 33)
affected WSA in the hyphal compartment soil. Moreover, there
36 Md.R.K. Siddiky et al. / Soil Biology & Biochemistry 50 (2012) 33e39

Fig. 2. Effects of collembola on AM fungal root colonization (i), and effects of AM fungi on collembola abundance (total number per pot) in the soil (ii). Interaction between AM fungi
and collembola on AM fungal hyphal soil length (iii), and non-AM fungal soil hyphal length (iv). Interaction between AM fungi and collembola on root biomass (v), and shoot
biomass (vi). Mean  SE, N ¼ 10. For F statistics see Table 1. Bars with the same letter are not different according to Tukey’s honestly significant difference (HSD) test, P > 0.05.

were significant main effects of AM fungi (P < 0.001; Pillai ¼ 0.961;
F ¼ 204.77; df ¼ 4, 33) and collembola (P < 0.001; Pillai ¼ 0.937;
F ¼ 121.81; df ¼ 4, 33) on WSA as well. This interaction, however,
was not detected for all classes in our experimental set up. The
subsequent univariate ANOVAs showed that only the two larger
classes (2e4 mm and 1e2 mm) were significantly affected by the
combined presence of both soil organisms (Table 1). These size
classes (WSA 2e4 mm and 1e2 mm) were increased significantly
by the combined addition of AM fungi and collembola, in
comparison to their individual presence or the control (Fig. 3). The
two smaller classes (212e500 mm and 0.5e1 mm) were still
significantly affected by AM fungi and collembola as main factors
(Table 1, Fig. 3). Furthermore, the combined presence of both
organisms resulted in a significant increase of percentage of total
WSA (see Supplementary online material Fig. S1; Table 1) in the
hyphal compartment, and a similar result was found for the MWD
(see Supplementary online material, Fig. S2). For both parameters,
the addition of just AM fungi or collembola alone resulted in
a significant increase (Figs. S1 and S2). When both organisms
were added, this increase was higher in comparison to the single
addition of AM fungi or collembola (Figs. S1 and S2). Collembola
effects were similar to those of AM fungi in term of percentage of
 Md.R.K. Siddiky et al. / Soil Biology & Biochemistry 50 (2012) 33e39 37

Table 1 just collembola compared to the control treatment (Fig. 2iv). The
F statistics of two-way ANOVA with arbuscular mycorrhizal fungi (AMF) and two-way ANOVA shows that both AM and non-AM fungal param-
Collembola as categorical factors. The respective univariate ANOVAs per class of
WSA are shown (see text for MANOVA results). Data on percentage of total WSA,
eters were significantly affected by the interaction between AM
mean weight diameter (MWD), AM and non-AM fungal soil hyphal length, root and fungi and collembola (Table 1). The presence of collembola alone or
shoot dry biomass were analyzed by two-way ANOVA. Data on AM fungal root combined with AM fungi drastically reduced the non-AM fungal
colonization and collembola abundance in soil were analyzed only by one-way hyphal length, close to zero (Fig. 2iv). Throughout our experiment,
ANOVA with collembola and AM fungi as categorical factor, respectively. Signifi-
we observed that AM fungal hyphal length was several-fold higher
cant F values are in bold and followed by * and *** corresponding to P < 0.05 and
0.001, respectively. Non-significant F values are followed by ns. than non-AM fungal hyphal length (Fig. 2iii and iv).
We did not detect a significant interaction between AM fungi
AMF Coll AMF  Coll Residuals
and collembola on root or shoot dry weight (Table 1). However, as
df 1 1 1 36 main factor, AM fungi significantly increased these two plant
212e500 mm F 169.58*** 95.91*** 1.40 ns
0.5e1 mm F 178.25*** 108.86*** 0.61 ns
parameters compared to the control treatment (Table 1; Fig. 2v and
1e2 mm F 79.05*** 44.47*** 17.04*** vi). Root dry weight was increased by about 28% in the presence of
2e4 mm F 306.26*** 183.09*** 10.54*** AM fungi, while no significant effect was observed in the presence
Percentage of total WSA F 720.92*** 297.93*** 56.14*** of just P. minuta (Fig. 2v). In the presence of just AM fungi
MWD F 507.45*** 285.80*** 41.16***
shoot biomass increased significantly by 34%, while again collem-
AM hyphal length F 6.34* 216.77*** 8.92*
Non-AM hyphal length F 36.91*** 522.49*** 145.12*** bolan presence did not have an effect (Fig. 2vi).
Root dry biomass F 36.15*** 5.80* 1.52 ns
Shoot dry biomass F 43.76*** 1.17 ns 0.01 ns 4. Discussion
df 1 1 e 18
AM root colonization F e 15.72*** e We showed for the first time that the interaction between
Collembola abundance F 41.64*** e e two major, widespread soil biota groups, AM fungi and collembola,
can enhance soil aggregation. A previous study (Siddiky et al., 2012)
had already demonstrated similar patterns, but included effects
of plant roots. Here we clearly show that these effects are inde-
total WSA (Fig. S1). Comparison of end-point soil aggregation pendent of the mediation by plant roots.
measurements with values from the onset of the study (Fig. 3) In absence of plant root effects P. minuta did not strongly reduce
showed that, overall, soil aggregation increased during the study. AM fungal hyphal abundance (Table 1; Fig. 2iii). Likely as a conse-
quence of this, soil aggregation was not reduced compared to
3.3. Other parameters control treatments (Fig. 3), complementing result of previous
studies with the same soil but with root effects included (Caruso
We observed that AM fungal colonization of the roots was et al., 2011; Siddiky et al., 2012). These studies were conducted
significantly increased in the pots where collembola were added to using different collembola species (i.e. Folsomia candida, Sinella
the hyphal compartment (Table 1; Fig. 2i). On the other hand, coeca) than the one used in the present study, highlighting that this
collembola abundance was significantly increased in the pots may be a consistent effect for collembola. Also, experimental
inoculated with AM fungi (Table 1; Fig. 2ii). addition of P. minuta in this study increased soil macroaggregation
AM fungal soil hyphal length was significantly reduced by compared to the non-inoculated controls, without influence of
approximately 6% by the presence of collembola compared to AM plant roots. In fact, the effect size of adding collembola was
fungal treatment (Fig. 2iii). By comparison, non-AM fungal soil comparable to that of the much better documented response to
hyphal length greatly decreased by about 98% in the presence of AM fungi.
Even the combined addition of P. minuta and AM fungi resulted
in a positive but non-additively increased soil aggregation
compared to their individual presence. In addition to soil aggre-
gation, AM fungal colonization rate and collembolan abundance
consistently increased when both of this soil biota co-occurred,
which supported our previous findings (Siddiky et al., 2012).
Collembola greatly reduced non-AM fungal abundance in our
study, likely due to the intense grazing of P. minuta on this hyphal
type (Fitter and Garbaye, 1994). Even though AM fungi are
consumed by collembola, including the species we used here
(Thimm and Larink, 1995), they do not typically prefer AM fungal
hyphae when given a choice (Klironomos and Kendrick, 1996;
Klironomos and Ursic, 1998), and these soil animals mostly feed
on non-AM fungi (Moore et al., 1987; Crossley et al., 1992; Gange,
2000), supporting our findings. Comparison of control and
pre-experiment soil aggregate size distribution (Fig. 3), which was
very similar, indicates that non-AM fungal hyphae were likely not
important contributors to soil aggregation; they would have also
been present in the control.
In our study, the single addition of AM fungi or collembola
induced a 30 and 32% increase of total WSA compared with the
Fig. 3. Effects of interaction between collembola (Coll) and AM fungi (AMF) on water control, respectively. Their combined presence, however, induced
stable aggregates (WSA) in 4 size classes. Pre-Exp refers to the initial soil aggregate size
distribution prior to the experiment. Mean  SE, N ¼ 10. For F statistics see Table 1. Bars
an increase of 40%. Thus, these effects were far from being additive.
within the same response variable (WSA class) with the same letter are not different However, we should consider the shifts in abundances induced
according to Tukey’s honestly significant difference (HSD) test, P > 0.05. between organisms. The feeding by collembola reduced the
38 Md.R.K. Siddiky et al. / Soil Biology & Biochemistry 50 (2012) 33e39
AM fungal hyphal length in the soil by 6%, which likely reduced
their contribution to aggregation (unless there were large changes
in species composition with functional consequences). But the
presence of AM fungi increased the collembola population by
20%. Although feeding by collembola could reduce AM fungal
contribution to aggregation, as initially hypothesized, the greater
gain in collembola population may be the mechanism by which
both organisms together can increase aggregation. There may
also be mechanisms beyond such changes in population size, but
related to mechanistic complementarity: collembola may process
organic matter, for example by adding fecal pellets, and fungal
hyphae can enmesh developing aggregates and potentially
produce biochemical binding agents (Rillig and Mummey, 2006).
Macroaggregation, and thus soil porosity has been suggested to
present AM fungal mycelium in the soil with clear growth advan-
tages (Rillig and Steinberg, 2002; Drew et al., 2003). In the case of
collembola, there could be as well an ecological advantage of
increased aggregation. Collembola avoid narrow pores to protect
their wax coat against damage (Choudhuri, 1961; Heisler and
Kaiser, 1995), therefore larger aggregates (and therefore larger
pore spaces) may indirectly enhance collembolan growth and
performances as well.
5. Conclusions
The trophic interaction between collembola and AM fungi did
not lead to a reduction of soil aggregation. We observed that both
soil organisms can contribute independently to the soil aggregation
process, likely through partially complementary mechanisms,
extending our previous findings obtained in the presence of plant
roots (Caruso et al., 2011; Siddiky et al., 2012). Our study contrib-
uted decisively to understanding the organismal component of the
soil aggregation process by disentangling the effect of AM fungal
hyphae, collembola, and their interaction from the mediation of
the root system. Further investigations, for example on soil aggre-
gation dynamics, are now needed to advance our knowledge about
the precise mechanisms by which these soil organisms improve
aggregation.
Acknowledgments
We thank Dr. Tancredi Caruso for helpful discussions and
Mr. Mahmud Maighul for greenhouse assistance. MRKS was funded
by BRAC University, Bangladesh, and this study was supported by
DFG and Freie Universität Berlin, Germany.
Appendix. Supplementary material
Supplementary data related to this article can be found online at
doi:10.1016/j.soilbio.2012.03.001.
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