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Plant and Soil 208: 1–8, 1999. © 1999 Kluwer Academic Publishers. Printed in the Netherlands.


Earthworm responses to plant species’ loss and elevated CO2 in calcareous grassland
J.G. Zaller1,2,∗ and J.A. Arnone III1,3
Institut, Universität Basel, Schönbeinstrasse 6, CH-4056 Basel, Switzerland. Present addresses: Center, Utah State University, 5205 Old Main Hill, Logan, UT 84322-5205, USA; 3 Desert Research Institute, Biological Sciences Center, 7010 Dandini Blvd., Reno, NV 89512, USA
2 Ecology 1 Botanisches

Received 21 January 1998. Accepted in revised form 29 October 1998

Key words: carbon dioxide enrichment, earthworm electro-sampling, earthworm populations, Lumbricidae, plant species diversity, plant-animal interactions, soil ecology

Abstract The objectives of this study were: (1) to quantify the effects of plant species’ loss from designed calcareous grassland communities at a field site in northwestern Switzerland on the size and composition of earthworm communities, and (2) to evaluate how exposure of plant communities to elevated atmospheric CO2 might alter the effects of plant species’ loss on earthworm communities. We non-destructively censused earthworm communities in each of 24 1.2 m2 experimental plots in autumn 1996 when soils were wet and earthworms were active. Each plot contained an experimental plant community with 31, 12 or 5 native plant species (eight plots each). Half of the plots in each species treatment were exposed to ambient CO2 concentrations (350 µL CO2 L−1 ) and half to elevated CO2 (600 µL CO2 L−1 ) using screen-aided CO2 control. The study was conducted in the fourth year after community establishment and the third year of CO2 treatment as part of a long-term study on the interactive effects of plant species’ loss and elevated CO2 on grassland communities. The size (density and biomass) of earthworm communities declined linearly when the number of plant species in the community was reduced from 31 to 5 species (e.g. 32 ± 1 g m−2 to 23 ± 2 g m−2 ) due mainly to a decline in the endogeic worm species Allolobophora rosea which was the most abundant of nine earthworm species observed (nearly half of all worms in each plot). However, no changes in the relative contribution of individual species or the three main earthworm ecological groups (anecics, endogeics, epigeics) to the entire earthworm community were observed with declining number of plant species. The responses of earthworm communities to plant species’ loss appear to reflect changes in community fine root biomass in the topsoil (e.g. declining worm biomass with declining fine root biomass) observed in parallel studies conducted at this site. Further the results of this study demonstrate that a loss of plant species’ from these calcareous grassland communities may also alter the age structure of earthworm communities, but not significantly influence their diversity or composition. Our data also indicate that rising atmospheric CO2 may not greatly impact the size and composition of worm communities or alter the effects of plant species’ loss on earthworm communities. Therefore, the disappearance of plant species from these native grasslands, as a result of ever increasing human activities, may be expected to lead to reductions in the size of earthworm communities and the ecosystem services they provide.

Introduction Human activities leading to loss of plant species from natural terrestrial ecosystems, and activities contribut∗

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ing to the rise of atmospheric CO2 levels, are expected to alter the functioning of these systems globally (e.g. Chapin et al. 1997, Hooper and Vitousek, 1997; Körner, 1995; Peters and Lovejoy, 1992; Schulze and Mooney, 1993; Wedin and Tilman, 1990). Soil heterotrophs are key members of these ecosystems who,

2 as decomposers and nutrient cyclers, not only depend on plants as primary producers but who also feed back to affect plants (e.g. Coleman and Crossley, 1996). Thus, any changes in the community of primary producers, resulting from either species’ loss or induced by elevated CO2 , must affect soil heterotrophs, and thus the functioning — and perhaps even the stability (e.g. McNaughton, 1977; Tilman et al., 1996) and resilience (e.g. Tilman, 1996) — of the ecosystem. However, experimental data needed to test these assertions are scant, and even less is known about how plant species’ loss and rising atmospheric CO2 will interact. In calcareous grasslands of central Europe, earthworms are among the most important soil heterotrophs (70 – 80% of soil zoomass). At our field site, they are responsible for an annual turnover of about 16 t soil (dry mass) ha−1 and 70 kg N ha−1 under current ambient CO2 concentrations, and up to 35% more in grassland plots maintained under elevated atmospheric CO2 (Zaller and Arnone 1997). Increased activity under elevated CO2 appears mainly to be due to enhanced carbon inputs to the soil (cf. Zaller and Arnone, 1998a) but also to CO2 -induced increases in soil moisture (Zaller and Arnone, 1997). It is uncertain, however, whether increased activity was due to larger earthworm communities (as reported by Yeates et al. (1997) in managed New Zealand pasture land), or simply enhanced feeding and burrowing activity. Loss of plant species from these calcareous grasslands may impact earthworm communities in two important ways: first, via changes in net primary productivity (NPP; sensu Tilman, 1996); and second via the elimination of certain plant species to which earthworms are particularly attracted (because of the palatability of their litter or because of the plant species’ effect on stand microclimate; see Zaller and Arnone 1998b). In the designed plant communities established at our site in which the number of native plant species present was reduced in three steps from the present level of 31 to 12 to 5 (Leadley and Körner, 1996), plant species’ loss did not affect net ecosystem carbon uptake (NEC; Stocker et al., 1998) or net aboveground biomass production (P. Leadley, personal communication). This suggests that plant species’ loss may not affect NPP significantly. Fine root biomass in the topsoil, on the other hand, declined significantly with reductions in plant species (P. Leadley, personal communication) suggesting that carbon supply to soils may decline with elimination of plant species from communities. Consequently, the size or activity of earthworm communities may also be reduced by plant species’ loss. Further, the functional composition of earthworm communities, viewed at either the species or ecological-group level (anecic, endogeic and epigeic species), may even be altered. In field plots where species’ number was manipulated, elevated CO2 caused several important effects relevant to our study of earthworm communities. First, soil moisture was higher in all plots maintained at high CO2 (P. Niklaus, unpublished data; Zaller and Arnone, 1997) theoretically improving the environment for earthworms. Second, NEC (Stocker et al., 1998) was markedly greater in communities maintained under elevated CO2 , than in plots maintained at ambient CO2 , but plant biomass only slightly higher (P. Leadley, pers. comm.) indicating an increase in carbon inputs to the ecosystem. The objectives of our study were to quantify the effects of a loss of plant species and elevated atmospheric CO2 on (1) the overall size (numbers/density and biomass) of earthworm communities, (2) individual earthworm species’ responses (density and biomass), (3) the representation of species and the three main earthworm ecological groups, (4) the earthworm species’ diversity, and (5) the age structure of earthworm communities.

Materials and methods Study site and experimental design This study was conducted as part of an ongoing, longterm study designed to evaluate the effects of plant species composition and elevated CO2 on the ecology of calcareous grasslands (e.g. Körner, 1995; Leadley and Körner, 1996). The study site is located near the village of Nenzlingen south of Basel on a 20◦ southwest-facing slope in NW-Switzerland (520 m, 47◦ 27’ N, 7◦ 34’ E). Average annual precipitation is about 900 mm, average annual air temperature between 8.5–9.0 ◦ C (Ogermann et al., 1994). Soils are classified as a Rendzina (pH is about 6.5, bulk density of the top soil 1.1 g/cm3 ), with a well developed, rockfree, loamy topsoil and a rapid transition at 10–15 cm depth to the underlying scree material (Ogermann et al., 1994). Up to 1993 this calcareous grassland had been used as an extensively managed pasture, mainly for cattle. The overall experimental setup and design is described in detail in Leadley and Körner (1996). Briefly, the topsoil from 24 hexagonal plots (1.2 m2 ) was

Table 1. Number of plant species per functional group used to construct grassland communities. Half of the communities were exposed to ambient atmospheric CO2 concentrations (350 µL CO2 L−1 ) and half to elevated CO2 (600 µ CO2 L−1 ); n = 4. Abstracted from Leadley and Körner (1996) Functional groups Number of plant species High Medium Low 8 17 6 31 2 7 3 12 1 2 2 5

Graminoids Non-legume forbs Legumes Total number of spp.

removed in the spring of 1993, homogenized and returned to the plots. Most plant species used to establish communities were propagated from seed collected from the site. Some species were vegetatively propagated using local material. Communities were created by planting individuals from either 31, 12 or 5 species on a hexagonal grid with a distance of 3.8 cm between individuals (590 individuals m−2 ). The representation of each of the three functional groups — graminoids (55% of all individuals planted), non-legume forbs (30%) and legumes (15%) — was held constant across all plots, with only the number of species in each functional group varying (Table 1). Since March 1994, four plots of each community type have been maintained round the clock (except from December to early March during winter months) to elevated CO2 concentrations (600 µL CO2 L−1 ) and half to ambient CO2 concentrations (350 µL CO2 L−1 ) using screen-aided CO2 control (SACC: 50-cm-tall transparent open-top, open-bottom chambers; Leadley et al., 1997). Plots were arranged in a randomized complete block design with four blocks. Earthworm sampling Earthworms in each plot were censused in October 1996 when soils were wet and earthworms were active (i.e. based on surface cast production) using the socalled ‘Octet’ electro-sampling method (Thielemann, 1986). We chose to use this method for two main reasons. First, we needed to insure that neither earthworms nor plants in the community would be damaged by repeated sampling required to census earthworm communities. Second, this non-destructive method has been shown to provide comparable estimates of earthworm community size and composition to other more

conventional sampling methods (hand-sorting and formaline infiltration methods), as long as earthworms are sampled at times when they are active and when soil moisture is sufficient (Vetter, 1996). Electro-sampling involved pushing eight steel rods (65 cm long × 6 mm dia.) into the ground in the centre of each plot along a circumference of a 60 cm diameter circle at equal distances. Electrical voltage (300–500 V DC) was applied over the entire length of the rods in 4-second pulses supplied pairwise to opposing rods in the ‘Octet’. Each successive electrical voltage was applied to the next pair of opposing rods. The portable transformer (DEKA 4000, Deka Gerätebau, Germany) providing electrical pulses was powered by a 12 V car battery. All earthworms driven to the soil surface within 40 min were collected and stored in cool water. Earthworms were sorted by species (Bouché, 1972) and age class (adult: well developed clitellum; subadult: developed puberculum, but no clitellum; juvenile: puberculum and clitellum absent), assigned to one of three ecological groups (epigeics, anecics, endogeics; Bouché, 1977), and were counted and weighed. Epigeic earthworm species live in or near the surface litter feeding primarily on coarse particulate organic matter. Endogeic species live within the soil profile and predominantly feed on soil and associated organic matter, whereas anecic species live in vertical burrow systems and are believed to feed primarily on surface litter which they pull into their burrows (e.g. Bouché, 1977; Edwards and Bohlen, 1996). Statistical analysis The General Linear Model-ANOVA procedure in the SYSTAT statistical package (SYSTAT, v. 5.2.1, Systat Inc., Evanston, IL, USA; Wilkinson et al., 1992) was used to test the effects of Plant Species Number (df = 2), CO2 (df = 1), Block (df = 3) and Plant Species Number × CO2 (df = 2) on: community earthworm density and biomass, the density and biomass of individual earthworm species common to all plots (six species), the relative contribution of individual species and ecological groups to community density and biomass, earthworm community diversity (Shannon index, see below), and the contribution of earthworm age classes to earthworm community density and biomass. Relative contribution data were arcsine-transformed prior to statistical analyses (Sokal and Rohlf, 1981). The Shannon diversity index (cf. Zar, 1996) was calculated for each plot using e as logarithmic base to assess the effects of treatments on

4 earthworm community diversity. In the interpretation of our results we considered P < 0.05 as significant and 0.05 < P ≤ 0.10 as marginally significant. Treatment means given in the text and figures are presented with their standard errors (±SE). effect of elevated CO2 appeared to be a reduction in A. rosea biomass. Neither the loss of plant species from communities nor elevated CO2 affected the diversity of earthworm communities(Table 2).

Results Total density and biomass of earthworm communities declined linearly with declining plant species number (density: from 106 ± 11 to 73 ± 20 worms m−2 ; biomass: from 31.8 ± 1.3 to 22.5 ± 2.3 g fresh mass m−2 ), however the decline in density was only marginally significant (P = 0.108; Figure 1). Elevated CO2 had no apparent effects on either earthworm community density or biomass (Figure 1). We found nine species of earthworms across all treatments, six of which occurred in all plots. Anecic species (Lumbricus terrestris L., Nicodrilus longus Ude, Nicodrilus nocturnus Ude) accounted for 35% of total earthworm community biomass. Endogeic species (Octolasion cyaneum Sav., Allolobophora rosea Sav., A. chlorotica Sav., Nicodrilus caliginosus Sav.) represented the largest ecological group with 62% of total earthworm biomass, while epigeic species (Lumbricus castaneus Sav., Dendrobaena mammalis Ger.) represented the smallest fraction with 3% of total biomass. Nicodrilus nocturnus, and Lumbricus terrestris were present in only two of the 24 plots, and Dendrobaena mammalis in only one. Allolobophora rosea was the most abundant earthworm species in all plots (49% of all worms averaged across treatments), followed by Octolasion cyaneum (19%), Allolobophora chlorotica (11%), Nicodrilus longus (10%), Lumbricus castaneus (6%) and Nicodrilus caliginosus (5%) (Figure 2). Overall, the density and biomass of individual earthworm species and earthworm ecological groups was affected very little by either plant species’ loss or elevated CO2 (Figure 2). Consequently, the contribution of individual earthworm species, and earthworm ecological groups, remained largely unaffected by these treatments (P = 0.40, data not shown). Plant species’ loss appeared to only impact the endogeic species Allolobophora rosea and Nicodrilus caliginosus as indicated by a decline in their populations (density and biomass; Figure 2). The decline in these two species, but especially in A. rosea, contributed substantially to the overall decline in earthworm community biomass described above. The only significant

Table 2. Shannon diversity index of the earthworm community as a function of the number of plant species in a community and atmospheric CO2 concentrations (ambient CO2 ; 350 µL CO2 L−1 ; elevated CO2 ; 600 µL CO2 L−1 ; mean ± SE, n = 4) CO2 Number of plant species in communities 31 12 5 1.27 ± 0.08 1.25 ± 0.05 1.28 ± 0.08 1.28 ± 0.10 1.50 ± 0.15 1.31 ± 0.12

Ambient Elevated

The age structure of all earthworm communities was hour-glass-shaped, where 80% of all worms viewed across treatments were juveniles, 2% subadults, and 18% adults (Figure 3). Reductions in the number of plant species affected only the contribution of subadult earthworms to the entire earthworm community, resulting in a linear decrease from 5% in communities with 31 plant species to 1% in communities with 5 plant species. Elevated CO2 did not alter this pattern, nor did it affect the contribution of adult worms to the community. However, the effects of declining number of plant species on the contribution of juvenile earthworms to the worm community were different in communities maintained at ambient and elevated CO2 (Figure 3).

Discussion The reduction in earthworm community biomass and density by about 30% with reduction in the number of plant species from 31 to 5 in these calcareous grassland communities four years after plant communities were established paralleled reductions in root biomass observed in the topsoil of these communities with increasing species’ loss (P. Leadley, pers. comm.). This suggests that plant detritus made available to earthworms via the fine roots (e.g. rhizodeposition or support of rhizosphere microorganisms consumed by earthworms) regulates earthworm community size (density and biomass). The apparent linear decrease in worm community size with reductions in the number of plant species (Figure 1) suggests


Figure 1. Density (a) and biomass (b) of earthworm communities as a function of the number of plant species present in a community (31, 12, and 5 species) and the atmospheric CO2 concentration (ambient CO2 : 350 µL CO2 L−1 , and elevated CO2 : 600 µL CO2 L−1 ). Mean ± SE, n = 4. P values derive from ANOVAs with Plant Species Number (Pl), CO2 concentration (CO2 ), and Block as factors.

that each plant species plays an equal role (perhaps via its contribution to plant community fine root biomass) in sustaining earthworm communities as long as the contribution of plant functional types to the plant community is held constant. Significantly increased ecosystem carbon uptake by all communities maintained under elevated atmospheric CO2 at our site (Stocker et al., 1998; but also reported in other studies: Hungate et al., 1997; Tate and Ross, 1997), as well as increased soil moisture under elevated CO2 (P. Niklaus, unpublished data; Zaller and Arnone, 1997), had no apparent effect on the overall size of earthworm communities, even after three years of CO2 treatment. This is surprising because we have observed dramatic increases in earthworm activity (surface casting) in calcareous grassland ecosystems maintained under elevated CO2 (Arnone et al., 1998; Zaller and Arnone, 1997), including in the plots containing 31 plant species used in the present study (G. Hofer, unpublished data). Taken together these data suggest that elevated CO2 only stimulates the activity of earthworms and does not necessarily alter the size of their populations (see also Yeates et al., 1997). Additionally, the lack of any significant differences in topsoil fine root biomass between communities maintained at ambient and elevated CO2 (P. Leadley, pers. comm.), further supports the notion that some property associated with fine roots is important for maintaining earthworm communities.

Although plant species’ loss and elevated CO2 had relatively little effect on the composition of earthworm communities at either the ecological-group or species level, their suppressive effects on endogeic species, particularly A. rosea, may have wide ecological ramifications. This is because earthworm communities in the plots we studied are made up mostly (62%) of species from this ecological group known to be important in the establishment and maintenance of soil structure (e.g. Bolton and Phillipson, 1976; Scheu, 1987; Springett, 1983). The absence of any treatment effects on earthworm community diversity suggests that reductions in the diversity of communities of primary producers may not affect the diversity of associated heterotrophs and that rising CO2 may not alter this. The overall age structure of earthworm communities censused in autumn 1996, with almost four times more juveniles than subadults and adults together, was typical of that exhibited by soil-inhabiting invertebrates (e.g. Edwards and Bohlen, 1996; Gerard, 1967; Lee, 1985). In general, the fact that both plant species’ loss and elevated CO2 affected the age structure of earthworm communities suggests that these two important global changes may also affect earthworm population and community dynamics. Thus, the results of this study demonstrate that a loss of plant species’ from these calcareous grassland communities may: (1) result in reductions in the size of earthworm communities, due to reductions in certain endogeic species; and (2) alter the age structure of


Figure 2. Density and biomass of earthworm species in grassland communities with different plant species numbers (31, 12, and 5 species) and atmospheric CO2 concentration (ambient CO2 : 350 µL CO2 L−1 , open symbols; elevated CO2 : 600 µL CO2 L−1 , filled symbols). Mean ± SE, n = 4. Earthworm species arranged by ecological group. Note the use of different scales on Y-axes. P values derive from ANOVAs with Plant Species Numbers (Pl), CO2 concentration (CO2 ), and Block as factors. Treatment effects at the P < 0.10 are shown in bold-faced type.

7 earthworm communities; however (3) not significantly influence the diversity or composition of earthworm communities. Our data also indicate that rising atmospheric CO2 may not greatly impact the size and composition of worm communities or alter the effects of plant species’ loss on earthworm communities. Therefore, the disappearance of plant species from these native grasslands, as a result of ever increasing human activities, may be expected to lead to reductions in the size of earthworm communities and the ecosystem services they provide.

Acknowledgements We are grateful to Paul Leadley and Pascal Niklaus for maintaining the field CO2 enrichment facility, and to Christian Körner and P. N. for reviewing an earlier version of the manuscript. Beate Kittl and Monika Keller helped with earthworm sampling. This work was funded by grants from the Swiss National Science Foundation to J. A. A. III (NF 3100-042401.94/1) and Ch. K., J. A. A. III and Bernhard Schmid (NFSPPU 5001-035214). J. A. A. III greatly appreciates the generous support of the Treubel-Fonds of Basel, Switzerland.

Arnone J A III, Zaller J G and Bohlen P J 1998 How earthworms alter the response of calcareous grassland communities to elevated CO2 . Global Change Biol., in preparation. Bolton P J and Phillipson J 1976 Burrowing, feeding, egestion and energy budgets of Allolobophora rosea (Savigny) (Lumbricidae). Oecologia 23, 225–245. Bouché M B 1972 Lombriciens de France. Ecologie et Systématique. INRA Publ. 72–2. INRA, Paris. Bouché M B 1977 Ecologie et paraecologie: Peut-on apprecier le role de la faune dans les cycles biogeochimiques. In Soil Organisms as Components of Ecosystems. Eds. U Lohm and T Persson. Ecol. Bull. (Stockholm) 25, 157–163. Chapin F S III, Walker B H , Hobbs R J, Hooper D U, Lawton J H, Sala O E and D Tilman 1997 Biotic control over the functioning of ecosystems. Science 277, 500–502. Coleman D C and Crossley D A Jr 1996 Fundamentals of Soil Ecology. Academic Press, San Diego. Edwards C A and Bohlen P J 1996 Biology and Ecology of Earthworms. 3rd ed. Chapman & Hall, London. Gerard B M 1967 Factors affecting earthworms in pastures. J. Anim. Ecol. 36, 235–252. Hooper D U and Vitousek P M 1997 The effects of plant composition and diversity on ecosystem processes. Science 277, 1302–1305. Hungate B A, Holland E A , Jackson R B, Chapin F S III, Mooney H A and Field C B 1997 The fate of carbon in grasslands under carbon dioxide enrichment. Nature 388, 576–579.

Figure 3. Age structure of earthworm communities (e.g. contribution of juveniles, subadults, and adults to the total density of earthworm communities) in grassland communities as a function of plant species number (31, 12, and 5 species) and atmospheric CO2 concentration (ambient CO2 : 350 µL CO2 L−1 , elevated CO2 : 600 µL CO2 L−1 ). Mean ± SE, n = 4.

Körner C 1995 Biodiversity and CO2 : Global change is under way. Gaia 4, 234–243. Leadley P W and Körner C 1996 Effects of elevated CO2 on plant species dominance in a highly diverse calcareous grassland. In Carbon dioxide, populations, and communities. Eds. C Körner and F A Bazzaz. pp. 159–175. Academic Press, San Diego. Leadley P W, Niklaus P, Stocker R and Körner C 1997 Screen-aided CO2 control (SACC): a middle ground between FACE and opentop chambers. Acta Œcol. 18, 207–219. Lee K E 1985 Earthworms. Their ecology and relationships with soils and land use. Academic Press, Sydney, New York. McNaughton S J 1977 Diversity and stability of ecological communities: a comment on the role of empiricism in ecology. Am. Natur. 111, 515–524. Ogermann P, Spycher B, Schaub D and Sollberger R 1994 Die Landschaftsstruktur im Raum Nenzlingen — geoökologisch gesehen. Regio Basiliensis 35, 91–100. Peters R and Lovejoy T 1992 Global Warming and Biodiversity. Yale University Press, New Haven, USA. Scheu S 1987 The role of substrate feeding earthworms (Lumbricidae) for bioturbation in a beechwood soil. Oecologia 72, 192–196. Schulze E-D and Mooney H A 1993 Biodiversity and ecosystem function. Ecological Studies 99. Springer Verlag, Berlin, Germany. Sokal R R and Rohlf F J 1981 Biometry. The principles and practice of statistics in biological research. 2nd ed. Freeman & Company, New York. Springett J A 1983 Effect of five species of earthworms on some soil properties. J. Appl. Ecol. 20, 865–872. Stocker R, Schmid B, Körner C and Leadley P W 1998 A field study of the effects of elevated CO2 and plant species diversity on ecosystem level gas exchange in a planted calcareous grassland. Global Change Biol. (In Press). Tate K R, Ross D J 1997 Elevated CO2 and moisture effects on soil carbon storage and cycling in temperate grasslands. Global Change Biol. 3, 225–235. Thielemann U 1986 Elektrischer Regenwurmfang mit der OktettMethode. Pedobiol. 29, 296–302. Tilman D 1996 Biodiversity: population versus ecosystem stability. Ecology 77, 350–363. Tilman D, Wedin D and Knops J 1996 Productivity and sustainability influenced by biodiversity in grassland ecosystems. Nature 379, 718–720. Vetter F 1996 Methoden zur Regenwurm-Extraktion. Vergleich der Formalin-, Senf- und Elektromethode. BUWAL (Bern), UmweltMaterialien 62, 46 pp. Wedin D A and Tilman D 1990 Species effects on nitrogen cycling: a test with perennial grasses. Oecologia 84, 433–441. Wilkinson L, Hill M A and Vang E 1992 Systat: Statistics. Systat Inc., Evanston, IL, USA. Yeates G W, Tate K R and Newton P C D 1997 Response of the fauna of a grassland soil to doubling of atmospheric carbon dioxide concentration. Biol. Fertil. Soils 25, 307–315. Zaller J G and Arnone J A III 1997 Activity of surface-casting earthworms in a calcareous grassland under elevated atmospheric CO2 . Oecologia 111, 249–254. Zaller J G and Arnone J A III 1998a Earthworm and soil moisture effects on the productivity and structure of native calcareous grassland communities. Soil Biol. Biochem. (In press). Zaller J G and Arnone J A III 1998b Interactions between earthworm casts and plant species in a calcareous grassland under elevated CO2 . Ecology (In press). Zar J H 1996 Biostatistical Analysis. 3rd ed. Prentice-Hall, Englewood Cliffs, USA. Section editor: E Garnier