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

¨ttingen, J.F. Blumenbach Institute of Zoology and Anthropology, Berliner Str. 28, 37073 Go ¨ttingen, Germany Georg August University of Go 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
b

a r t i c l e i n f o
Article history: Received 27 April 2009 Received in revised form 9 June 2009 Accepted 16 June 2009 Available online 3 July 2009 Handling editor: Stefan Schrader Keywords: Above- and belowground interactions Decomposers Diversity–productivity relationship Grassland Mowing The Jena Experiment

a b s t r a c t
Knowledge of the role of decomposers in the plant diversity–productivity relationship is scarce. In the framework of the Jena Experiment, we observed regrowth of grassland plant communities varying in plant species and functional group richness three weeks after mowing. We investigated earthworm subplots and subplots with reduced earthworm density in order to explore if earthworms enhance plant regrowth and if earthworm effects depend on plant diversity. Earthworms significantly enhanced each of 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. However, the average height of the vegetation was not affected in 16-species mixtures suggesting compensation of the impact of earthworms on plant regrowth in complex plant communities. Ó 2009 Elsevier Masson SAS. All rights reserved.

1. Introduction 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 availability for plants [2,21,24], knowledge on the role of decomposers for the diversity–productivity relationship is scarce [11]. Earthworms 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 communities [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

with mostly beneficial impacts on fast growing plants to the 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 earthworms 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 functional group richness (1–4) in plots of 20 Â 20 m (see [20] for detailed experimental design). Using above- and belowground

* Corresponding author. Tel.: þ49 0 6151 164299; fax: þ49 0 6151 166111. E-mail address: eisenhauer@bio.tu-darmstadt.de (N. Eisenhauer). 1164-5563/$ – see front matter Ó 2009 Elsevier Masson SAS. All rights reserved. doi:10.1016/j.ejsobi.2009.06.001

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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. 2.2. Earthworm density manipulations Starting in September 2003, earthworm densities were manipulated 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 Â 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 Æ 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 earthworm 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 ¨ tebau, Marsberg, Germany) octet devices (DEKA 4000, Deka Gera 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 earthworms 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

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 experiment 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 (À38%) [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. 2.3. Plant regrowth parameters 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 determined 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), EWÂSR and EWÂFR 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 F-value Block SR FR GR SH TH LE Plot EW EW Â SR EW Â FR Error 3 2 3 1 1 1 1 32 1 2 3 38 0.68 7.09 2.85 9.73 2.91 0.66 0.74 8.37 5.64 0.19 0.09 P-value 0.5726 0.0028 0.0529 0.0038 0.0975 0.4219 0.3951 <0.0001 0.0227 0.8270 0.9647 [ [ Max. vegetation height F-value 2.36 0.22 0.73 7.77 5.49 0.60 0.00 5.71 10.25 0.57 0.79 P-value 0.0897 0.8013 0.5412 0.0089 0.0255 0.4431 0.9835 <0.0001 0.0028 0.5728 0.5078 Av. vegetation height F-value 4.28 0.11 0.19 29.61 18.30 1.56 0.23 5.45 12.08 3.68 1.09 P-value 0.0120 0.8993 0.9010 <0.0001 0.0002 0.2211 0.6367 <0.0001 0.0013 0.0345 0.3631

[ Y

[ Y

[

[

[

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.

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calculated followed by Plot, EW, EWÂSR and EWÂFR. Plant community treatments (SR, FR, GR, SH, TH and LE) were tested against the variance between plots (n ¼ 45; to avoid pseudoreplication) and earthworm treatments (EW, EWÂSR and EWÂFR) 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

effect of earthworms did not differ between plant diversity treatments (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 (À9% and À20%). 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 significantly increased in subplots with higher earthworm density confirming 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 performance presumably are the change of soil structure, the mineralization of nutrients, hormone-like effects, dispersal of growth stimulating microorganisms and dispersal of micro- organisms antagonistic to root pathogens [21]. Moreover, recent studies indicate 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 earthworms; [9]); the observed effects therefore are minimum effects. Although these results are in line with recent greenhouse experiments, 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 earthworms; 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 communities. Presumably, mechanisms such as niche partitioning or

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 earthworm subplots [þew]). Variations in (B) maximum and (C) average height of the plant community as affected by plant species richness and earthworms. Means with standard error (boxes) and standard deviation (error bars); circles indicate outliers and asterisks indicate extremes.

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