European Journal of Soil Biology 45 (2009) 455–458

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European Journal of Soil Biology

journal homepage: http://www.elsevier.com/locate/ejsobi

Earthworms enhance plant regrowth in a grassland plant diversity gradient

Nico Eisenhauer a, *, Alexandru Milcu b, Alexander C.W. Sabais c, Stefan Scheu a
a
Georg August University of Göttingen, J.F. Blumenbach Institute of Zoology and Anthropology, Berliner Str. 28, 37073 Göttingen, Germany
b
NERC Centre for Population Biology, Division of Biology, Imperial College London, Silwood Park Campus, Ascot, Berkshire, UK
c
Darmstadt University of Technology, Institute of Zoology, Schnittspahnstr. 3, 64287 Darmstadt, Germany

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

Article history: Knowledge of the role of decomposers in the plant diversity–productivity relationship is scarce. In the
Received 27 April 2009 framework of the Jena Experiment, we observed regrowth of grassland plant communities varying in
Received in revised form plant species and functional group richness three weeks after mowing. We investigated earthworm
9 June 2009
subplots and subplots with reduced earthworm density in order to explore if earthworms enhance plant
Accepted 16 June 2009
Available online 3 July 2009
regrowth and if earthworm effects depend on plant diversity. Earthworms significantly enhanced each of
Handling editor: Stefan Schrader the plant regrowth parameters (plant coverage and maximum and average height of the vegetation)
suggesting that particularly fast growing species, such as grasses, benefit from earthworm activity.
Keywords: However, the average height of the vegetation was not affected in 16-species mixtures suggesting
Above- and belowground interactions compensation of the impact of earthworms on plant regrowth in complex plant communities.
Decomposers Ó 2009 Elsevier Masson SAS. All rights reserved.
Diversity–productivity relationship
Grassland
Mowing
The Jena Experiment

1. Introduction with mostly beneficial impacts on fast growing plants to the

It has been repeatedly shown that plant productivity increases
with diversity [1,3,14]. Although decomposers are known to
profoundly impact nutrient cycling and, thereby, resource avail-
ability for plants [2,21,24], knowledge on the role of decomposers
for the diversity–productivity relationship is scarce [11]. Earth-
worms are important decomposers in many terrestrial ecosystems
[5,6,17] and recent laboratory studies suggest that they not only
affect plant growth but also competitive interactions between plant
species [7,25]. Furthermore, in the field it has been shown that
earthworms indeed enhance plant productivity. However, effects
did not depend on the diversity of the plant community although
plant community biomass was only slightly increased in the most
diverse plant communities (16-species mixtures) [11]. Moreover,
earthworms increased the invasibility of grassland plant commu-
nities [9]. Remarkably, earthworm impacts were most pronounced
in intermediate diverse plant species mixtures showing that effects
of earthworms on the plant community might differ between plant
diversity levels. Further, previous greenhouse [7,8,16,25] and field
studies [9,11] indicated that earthworms change the competition
between grassland plant species and/or plant functional groups
* Corresponding author. Tel.: þ49 0 6151 164299; fax: þ49 0 6151 166111.
E-mail address: eisenhauer@bio.tu-darmstadt.de (N. Eisenhauer).
expense of slower growing ones. Since Central European mesobiont
grasslands are typically mown twice a year, the question arises if
earthworms also affect plant regrowth after mowing and if this
may vary with plant diversity. For studying the impact of earth-
worms on plant regrowth in a plant diversity gradient, we observed
plant recovery in earthworm subplots and subplots with reduced
earthworm density three weeks after mowing in the framework of
the Jena Experiment [20]. We hypothesized that (1) earthworms
enhance plant regrowth and (2) earthworm impacts depend on the
diversity of the plant community.
2. Materials and methods
2.1. Study site
The present study was performed on the field site of the Jena
Experiment, a large grassland experiment investigating the role of
plant diversity for element cycling and trophic interactions. Mean
annual air temperature is 9.3  C and annual precipitation is 587 mm
(measured 3 km south of the field site) [15]. The soil is an Eutric
Fluvisol [12] and the site had been used before as an arable field
for the last 40 years. In May 2002, a pool of 60 native plant species
was used to establish a gradient of plant species (1–60) and func-
tional group richness (1–4) in plots of 20  20 m (see [20] for
detailed experimental design). Using above- and belowground

1164-5563/$ – see front matter Ó 2009 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejsobi.2009.06.001
456 N. Eisenhauer et al. / European Journal of Soil Biology 45 (2009) 455–458

morphological traits (growth form, canopy height, rooting depth
and capacity for clonal growth), phenological traits (occupancy of
seasonal niches, life cycle and seasonality of foliage) and N2 fixation
ability, plant species were aggregated into four plant functional
groups: grasses (16 species), small herbs (12 species), tall herbs (20
species), and legumes (12 species). Experimental plots (n ¼ 82)
were mown twice a year (in June and September), as is typical for
hay meadows, and weeded twice a year to maintain the target
species composition. Plots were assembled into four blocks
following a gradient in soil characteristics, each block containing an
equal number of plots of all plant species and plant functional
group richness levels.
density manipulations was measured via the soil surface activity of
L. terrestris which is known to bury plant seeds irrespective of seed
size and shape [8,18]. In May 2006 (after six earthworm density
manipulation campaigns), we performed a seed dummy experi-
ment to determine earthworm soil surface activity [9]. While
earthworm soil surface activity in earthworm subplots did not
differ from that in control subplots, it was decreased considerably
in earthworm reduction subplots (38%) [9]. Four years after
establishment earthworm densities were saturated as indicated by
similar earthworm soil surface activities in earthworm and control
subplots [9]. Thus, we continued earthworm extraction campaigns
but did not add any L. terrestris individuals after spring 2006.

2.2. Earthworm density manipulations 2.3. Plant regrowth parameters

Starting in September 2003, earthworm densities were manip-
ulated within the 1 (15 replicates), 4 (16 replicates) and 16 plant
species plots (14 replicates; 45 plots altogether). On each plot, two
randomly selected subplots of 1  1 m were used to establish the
treatments ‘‘earthworm’’ and ‘‘earthworm reduction’’ (90 subplots
altogether). Subplots were located around the core area of each plot
[20] and enclosed with PVC shields aboveground (20 cm) and
belowground (15 cm) to reduce the escape or colonization of
earthworms. Earthworm subplots received 25 adult individuals of
Lumbricus terrestris L. per year (average individual fresh weight
with gut content 4.10  0.61 g). We chose this anecic earthworm
species due to its role as key decomposer in temperate grassland [5]
and since the pre-treatment of experimental plots [20] primarily
decreased anecic earthworm densities [19]. Further, two earth-
worm extraction campaigns were performed per year (spring and
autumn) in the earthworm reduction subplots by electro-shocking
in order to decrease earthworm densities. A combination of four
octet devices (DEKA 4000, Deka Gerätebau, Marsberg, Germany)
was used [23]. Since this method is known to depend on weather
conditions [10], we always performed earthworm extraction
campaigns during mild and humid weather periods. In each
subplot, earthworm extraction was performed for 35 min,
increasing the voltage from 250 V (10 min) to 300 V (5 min), 400 V
(5 min), 500 V (5 min), and 600 V (10 min) [10]. Extracted earth-
worms were removed, identified, counted and weighed in the
laboratory. The earthworm community consisted of five species
belonging to two functional groups, anecics (L. terrestris L.) and
endogeics (Aporrectodea caliginosa Savigny, Aporrectodea rosea,
Allolobophora chlorotica Savigny and Octolasion tyrtaeum Savigny).
The most abundant earthworm species was Ap. caliginosa, whereas
L. terrestris had the highest biomass [19]. The success of earthworm
Plant regrowth in the earthworm subplots (1 m2 each) was
investigated three weeks after mowing in September 2008 by
determining plant community coverage and maximum and average
height of the vegetation. Plant community coverage was deter-
mined by visual estimation in intervals of 5% ([%]; modified after
[4]). The maximum height of the vegetation was determined by
measuring the perpendicular distance from the soil at the base to
the highest point reached with all parts in the natural position by
the highest plant individual using a ruler ([cm]; [13]). Further, the
average height of the plant community was determined by visual
estimation of the mean height of all plant individuals in their
natural position ([cm]; [13]). All three plant regrowth parameters
were determined by two people independently and the respective
means were calculated per subplot.
2.4. Statistical analysis
Data were log-transformed (log10[xþ1]) to improve normal
distribution and homogeneity of variance. Means presented in text
and figures represent non-transformed data. Split plot ANOVA
(GLM, type I sum of squares) was used to analyze the effects of
block, plant species richness (SR), plant functional group richness
(FR), and presence of grasses (GR), small herbs (SH), tall herbs (TH),
legumes (LE), earthworms (EW), EWSR and EWFR on plant
community coverage and the maximum and average height of the
vegetation in a hierarchical order (SAS 9.1, SAS Institute Inc., Cary,
USA). F-values given in the text refer to those where the respective
factor (and interaction) was fitted first [22]. Block was always fitted
first (in order to exclude the variance of soil abiotic parameters
from the analysis), followed by SR and FR. Then, the effects of
presence/absence of certain plant functional groups were

Table 1
ANOVA table of F- and P-values on the effects of Block, plant species richness (SR), plant functional group richness (FR), presence/absence of grasses (GR), small herbs (SH), tall
herbs (TH) and legumes (LE), and earthworms (EW) on the coverage of the plant community [%], and on the maximum and average vegetation height [cm].

D.f. Vegetation coverage Max. vegetation height Av. vegetation height

F-value P-value F-value P-value F-value P-value

Block 3 0.68 0.5726 2.36 0.0897 4.28 0.0120
SR 2 7.09 0.0028 [ 0.22 0.8013 0.11 0.8993
FR 3 2.85 0.0529 0.73 0.5412 0.19 0.9010
GR 1 9.73 0.0038 [ 7.77 0.0089 [ 29.61 <0.0001 [
SH 1 2.91 0.0975 5.49 0.0255 Y 18.30 0.0002 Y
TH 1 0.66 0.4219 0.60 0.4431 1.56 0.2211
LE 1 0.74 0.3951 0.00 0.9835 0.23 0.6367
Plot 32 8.37 <0.0001 5.71 <0.0001 5.45 <0.0001
EW 1 5.64 0.0227 [ 10.25 0.0028 [ 12.08 0.0013 [
EW  SR 2 0.19 0.8270 0.57 0.5728 3.68 0.0345
EW  FR 3 0.09 0.9647 0.79 0.5078 1.09 0.3631
Error 38

Significant effects (P < 0.05) are given in bold; error terms are given in italics. D.f. ¼ degrees of freedom, [ ¼ increase (with increasing plant diversity or in presence of the
respective factor), Y ¼ decrease.
 N. Eisenhauer et al. / European Journal of Soil Biology 45 (2009) 455–458
calculated followed by Plot, EW, EWSR and EWFR. Plant
community treatments (SR, FR, GR, SH, TH and LE) were tested
against the variance between plots (n ¼ 45; to avoid pseudor-
eplication) and earthworm treatments (EW, EWSR and EWFR)
against the variance between subplots (n ¼ 90).
3. Results
Plant community coverage increased significantly with plant
species richness from monocultures (65%) to 4-species (71%) and
16-species mixtures (84%; Table 1; Fig. 1A). Further, there was
a tendency towards higher plant community coverage in mixtures
containing two, three, and four plant functional groups than in
mixtures with one plant functional group (Table 1). Plant
community coverage was increased significantly in presence of
grasses (þ27%) and in earthworm subplots (þ6%; Table 1). The
Fig. 1. Variations in (A) coverage of the plant community as affected by plant species
richness (1, 4, 16) and earthworms (earthworm reduction subplots [-ew] and earth-
worm subplots [þew]). Variations in (B) maximum and (C) average height of the plant
community as affected by plant species richness and earthworms. Means with stan-
dard error (boxes) and standard deviation (error bars); circles indicate outliers and
asterisks indicate extremes.
457
effect of earthworms did not differ between plant diversity treat-
ments (Table 1, Fig. 1A). The maximum and average height of the
vegetation was not affected by plant species (Table 1; Fig. 1B and C)
and plant functional group richness (Table 1). However, the
maximum and average height of the vegetation was increased in
presence of grasses (þ23% and þ41%) and decreased in presence of
small herbs (9% and 20%). Moreover, both parameters increased
significantly in earthworm subplots (þ13% and þ15%). Earthworm
effects on the maximum height of the vegetation did not depend on
plant diversity (Table 1; Fig. 1B). However, the average height of the
vegetation was only increased in earthworm subplots in the 1- and
4-plant species mixtures whereas it was not affected in 16-species
mixtures (Table 1; Fig. 1C).
4. Discussion
Each of the three plant regrowth parameters observed signifi-
cantly increased in subplots with higher earthworm density con-
firming hypothesis (1). Probably, this was due to fragmentation and
incorporation of litter into the soil [19], stimulation of microbial
activity and/or nutrient availability for plants [5,11,17]. In most
studies (79%) investigating the response of plants to presence of
earthworms, plant shoot biomass was significantly increased in
presence of earthworms (review of 67 studies by Scheu [21]). The
main mechanisms underlying earthworm impacts on plant perfor-
mance presumably are the change of soil structure, the minerali-
zation of nutrients, hormone-like effects, dispersal of growth
stimulating microorganisms and dispersal of micro- organisms
antagonistic to root pathogens [21]. Moreover, recent studies indi-
cate that earthworms also drive plant competition and plant
community assembly [7,25]. Results of the present study underline
the important function of earthworms in grasslands as they also
impact plant recovery after mowing. Particularly fast growing plant
species, such as grasses, might be promoted by earthworm activity,
probably resulting in a competitive advantage against slower
regrowing species. Indeed, presence of grasses increased the
maximum and average height of the plant community in the present
study. This is in line with recent studies showing that earthworms
foster the competitive strength of grasses at the expense of herbs
and legumes [7,14,25]. Moreover, enhanced recovery of the plant
community might also have been due to an increased number and
biomass of plant seedlings in earthworm subplots [9]. However,
earthworm density manipulation in the present study only reduced
earthworm densities temporarily (rather than eliminating earth-
worms; [9]); the observed effects therefore are minimum effects.
Although these results are in line with recent greenhouse experi-
ments, they contrast field observations by Eisenhauer et al. [11]
suggesting a loose mutualistic relationship between earthworms
and legumes. However, the present study indicates that grasses
might benefit from frequent mowing and the presence of earth-
worms; thus, the effect of earthworms on plant competition under
semi-natural conditions deserves further attention.
Impacts of earthworms on the plant communities were either
shown to vary with [9] or to be independent of the diversity of plant
communities [11]. Taking these results into account and since
earthworms rely on the availability and quality of plant residues
[18,19], we hypothesized that the impacts of earthworms on plant
recovery are modified by plant diversity (2). Our results confirmed
this hypothesis in part as the average height of the vegetation was
the sole variable affected by the interaction between earthworms
and plant species richness. Earthworms enhanced plant regrowth
in monocultures and 4-species mixtures but not in 16-species
mixtures. This supports the findings of Eisenhauer et al. [9] that
earthworm effects are less pronounced in diverse plant commu-
nities. Presumably, mechanisms such as niche partitioning or
458 N. Eisenhauer et al. / European Journal of Soil Biology 45 (2009) 455–458
facilitation have stronger immediate impact on plant regrowth at
higher diversity levels.
Acknowledgements
This research was supported by the German Science Foundation
and was conducted in the framework of the Jena Experiment. We
thank C.M. Pusch, T. Keil, V. Hörsch and A. Roos for the help during
earthworm extractions. Comments of two anonymous referees
improved the manuscript.
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