Ecosystem Level Impacts of Invasive Acacia saligna in the South African Fynbos

S. G. Yelenik,1,2,3 W. D. Stock,4 and D. M. Richardson5

Abstract Recent efforts to clear invasive plants from the fynbos of South Africa forces managers to think about how N2-fixing invasives have altered ecosystem processes and the implications of these changes for community development. This study investigated the changes in nitrogen (N) cycling regimes in fynbos with the invasion of Acacia saligna, the effects of clear-cutting acacia stands on soil microclimate and N cycling, and how altered N resources affected the growth of a weedy grass species. Litterfall, litter quality, soil nutrient pools, and ion exchange resin (IER)-available soil N were measured in uninvaded fynbos, intact acacia, and cleared acacia stands. In addition, a bioassay experiment was used to ascertain whether the changes in soil nutrient availability associated with acacia would enhance the success of a weedy grass species. Acacia plots had greater amounts of litterfall, which had higher concentrations of N. This led to larger quantities of organic matter, total N, and IER-available N in the soil. Clearing acacia stands caused changes in soil moisture and temperature, but did not result in differences in IER-available N. The alteration of N availability by acacias was shown to increase growth rates of the weedy grass Ehrharta calycina, suggesting that secondary invasions by nitrophilous weedy species may occur after clearing N2-fixing alien species in the fynbos. It is suggested that managers use controlled burns, the addition of mulch, and the addition of fynbos seed after clearing to lower the levels of available N in the soil and initiate the return of native vegetation. Key words: Acacia saligna, available N, biological invasions, invasive alien plants, litter quality, nitrogen cycle, nitrogen fixation, nutrient dynamics, restoration, secondary invasions.

Introduction Over the past decade ecologists have become increasingly interested in the feedbacks between changes in ecosystem processes and changes in community structure that accompany the invasions of alien plant species (Vitousek 1990; Hobbie 1992; Vinton & Burke 1995). Changes in species composition due to alien plant invasion may change ecosystem functions such as nutrient cycling, water availability, and fire regime, which can then lead to further changes in community structure by inducing secondary invasions of weedy species (DAntonio & Vitousek 1992; Maron & Connors 1996; Mack et al. 2001). Vitousek et al. (1987) suggest that an invasive species is most likely to change the properties of ecosystems when it offers a novel biological process. Within low-nutrient ecosystems invasions of nitrogen (N)-fixing plant species have thus been the obvious focus of studies testing for changes in ecosystem function. N2-fixing invasive alien plant spe1 Institute for Plant Conservation, University of Cape Town, Rondebosch 7701, South Africa. 2 Present address: Department of Evolution, Ecology, and Marine Biology, University of California, Santa Barbara, CA 93106, U.S.A. 3 Address correspondence to S. G. Yelenik, email yelenik@lifesci.ucsb.edu 4 Centre for Ecosystem Management, Edith Cowen University, Joondalup 0627, Western Australia. 5 Institute for Plant Conservation, University of Cape Town, Rondebosch 7701, South Africa.

2004 Society for Ecological Restoration International

cies in Hawaii, California, and South Africa have been shown to cause shifts from low to high N-cycling regimes (Vitousek et al. 1987; Vitousek & Walker 1989; Stock et al. 1995; Maron & Connors 1996; Pickart et al. 1998; Maron & Jefferies 1999). Changes in soil N can then feedback to alter community dynamics. For example it was shown that the increase in soil N due to the invasive plant Lupinus arboreus in California led to invasions of European annual grasses after large-scale lupine mortality (Maron & Connors 1996). These feedback cycles are of particular interest to restoration ecologists. After the large-scale clearing of invasive plants, changes in N cycling and secondary invasions may hinder efforts to re-establish native species (Clarke 1997; Pickart et al. 1998; Zink & Allen 1998). The species-rich fynbos vegetation of the Cape Floristic Region of South Africa, one of the most nutrient-poor regions (Read & Mitchell 1983) of all of its Mediterranean counterparts, might be highly susceptible to the impacts of N2-fixing invaders. N cycling in the natural vegetation of the fynbos is a slow process (Stock & Allsopp 1992). The vegetation consists largely of slow-growing sclerophyllous shrubs that have slowly decomposing leaf litter due to high levels of lignins and soluble phenolic compounds and low levels of tissue N (Mitchell et al. 1986; Witkowski 1991a). Within this ecosystem, soil characteristics, including N, determine community structure, species presence or absence (Richards et al. 1997a, 1997b), and, possibly,

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speciation patterns (Cowling et al. 1992). In addition, some native fynbos species have shown decreases in growth rates with increased levels of N or P (Lamb & Klaussner 1988; Witkowski 1989). Because of these characteristics, shifts to high N cycling in soils may affect community structure and function. Although native N2 fixers exist in the fynbos, they are short lived in the post-fire community, usually distributed patchily, and do not deposit enough of their N-rich litter to make an impact on soil N status (Cocks & Stock 2001). Invasive alien N2-fixing Acacia species can form tall, dense stands that regenerate rapidly after each fire (Milton & Hall 1981). The resulting acacia-dominated community is persistent (Richardson & Cowling 1992) and has constant, large inputs of N-rich litter. Acacia cyclops and A. saligna (rooikrans and blue-leafed wattle, respectively), which produce more litter with a higher leaf N concentration than native fynbos shrubs, elevate total soil N and organic matter (Witkowski 1991a). Stock et al. (1995) found that changes in soil N pools lead to higher rates of net N mineralization in A. cyclops, but not A. saligna stands. Because the sites for A. saligna and A. cyclops were on different soil types, the lack of increased N mineralization in A. saligna stands in that study might reflect differences in species or differences in pre-invasion soils (Stock et al. 1995). Research surrounding the restoration of fynbos vegetation is highly relevant due to new environmental legislation regarding invasive plants. The Working for Water Programme, launched in 1995, aims to remove waterdemanding invasive alien trees from waterways and catchment areas, thereby simultaneously avoiding the need for future dams and creating over 35,000 jobs for previously unemployed people (van Wilgen et al. 1998). It is estimated that 1.7 million ha, containing about 15 invasive woody species, will be cleared by the year 2015 (van Wilgen et al. 1998). Although Acacia spp. are only a part of this pool, large areas of land invaded by N2-fixing invasive aliens will be cleared. The high levels of N in the soils, however, may cause problems for restoration, because native species may not be able to grow under these enriched conditions. Clearing large areas of acacia stands may increase N mineralization due to soil microclimates that change after the tree canopy is removed. Increases in soil temperatures as small as 510 C can stimulate microbial communities, resulting in greater quantities of available N (Nadelhoffer et al. 1991; Verburg et al. 1999). Increased N mineralization after acacia clearing may exacerbate the detrimental effects of high N cycling for native vegetation recolonization. Already increases in alien grass abundance have been noted after the clearing of A. saligna (Holmes & Cowling 1997). Within Riverlands Nature Reserve, land cleared of A. saligna 12 years ago has high cover of the grass Ehrharta calycina Smith (personal observation). Although native, this species is not fynbos dominant at any time in the post-fire succession of acid sandplain lowland fynbos

(Hoffman et al. 1987) and is generally found as a colonizer of highly disturbed areas. To explore ecosystem shifts and, at the same time, better inform managers about how to restore indigenous vegetation back to degraded land, this study investigated changes in N cycling with invasions of A. saligna (Labill.) Wendl. (acacia). The research was aimed at answering the following questions: (1) Does acacia increase the rates of N cycling through high N inputs in acid sand lowland fynbos? (2) Will the clearing of acacia stands further increase net N mineralization rates in soils due to altered soil microclimates? (3) Will the growth of weedy grass species be enhanced by higher levels of N in soils?

Materials and methods

Study Site

All fieldwork was performed on Riverlands Nature Reserve (33 320 S; 18 350 E), 63 km north of Cape Town, South Africa. The native vegetation is broadly classified as acid sandplain lowland fynbos (Moll et al. 1984), some of which is located on seasonal wetlands. The climate is Mediterranean, with hot dry summers and cool wet winters. Riverlands soils are similar to those of Pella, the Fynbos Biome Projects intensive study site, consisting of well-drained aeolian acidic sands approximately 12 m deep in the Constantia and Clovelly forms (Lambrechts & Fry 1988) according to the South African binomial classification (MacVicar et al. 1997). Riverlands, which is 1,300 ha in size, hosts about 400 native plant species, 41 of which are critically rare and endangered (Kilian 1995). Riverlands consists of a farm that was purchased from Transnet (formerly South African Railways) in 1985 by Cape Nature Conservation because of its high concentration of rare plants. Owing to its history, it consists of a patchwork landscape including invasive alien vegetation, fallow fields, and uninvaded native fynbos communities. In 1995, 300 ha was covered in dense alien vegetation, mostly Acacia saligna. By examining aerial photographs from 1960, 1968, 1977, and 1988, it was estimated that acacias established before 1968, dispersing into undisturbed fynbos from trees planted on the neighboring property. In 1998, the Working for Water Programme started clearing A. saligna and hopes to have it eradicated by 2004.
Design

To investigate the effects of acacia and the effects of clearing acacia stands we compared N cycling in acacia thicket, cleared acacia thicket and pristine fynbos, all on the same soil type. Each treatment had three different plots, and each plot had four sub-plots. Sub-plots were 3 3 3 m and, except for the fynbos treatment, randomly located. In each fynbos plot, sub-plots were centered around individuals of the dominant reseeding proteoid

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species (or the species with the closest life history to A. saligna). The target species were: plot 1, Protea scolymocephela L. Reichard aged approximately 45 years within a Restionaceae-dominated community; plot 2, Leucospermum parile (Salisb. ex Knight) aged approximately 45 years within a Proteaceae-dominated community; and plot 3, L. parile aged approximately 56 years in a Proteaceaedominated community. For each of the three acacia thicket plots, an adjacent 18 3 4.5m space was cleared of 100% of the A. saligna trees using chain saws and axes by staff of the Working for Water Programme in December 1998. Leaf litter was not removed. Within each cleared plot we established 3 3 3m sub-plots with a 0.75 m buffer all the way around.

Biomass, Litterfall, and N Return to Soil

To measure litterfall four mesh-bottom litter traps (20 3 16 cm) were placed in each of the fynbos and acacia thicket sub-plots in February 1999. Litterfall was not measured in cleared acacia plots because they were clear-cut, and there was no significant regeneration to litterproducing trees within the time of the study. Litter was allowed to collect in the traps for 3 months at a time, after which the litter was collected, dried for at least 48 hr at 80 C, and weighed. For acacia plots, litter was separated into leaves, pods, and flowers. For fynbos plots litter was separated into leaves and flowers. All litter was coarse ground through a 1 mm mesh. Once the separate fractions were weighed, average proportions of different litter components for all treatments were calculated and mixed such that 10 g of coarse ground material was achieved. Subsamples of the pre-mixed fractions were ground to a fine powder using a mixer mill (Retsch Inc., Haan, Germany) and analyzed for total N and total P. Total N was determined using a Kjeldahl digest with 0.1 g plant material and subsequent colorimetric determination of ammonium using the same procedures as described in Stock & Lewis (1986). Total P was determined using 0.1 g plant material as per the methods of Murphy & Riley (1962).

slot for the bags. In this way one anion and one cation resin bag were placed next to each other beneath 5 cm of relatively undisturbed soil. These were taken out and sealed into clean plastic bags after approximately 30 days, at which time newly regenerated resin bags were placed into the same slot within the plot. Resin bags were replaced monthly to quantify N availability for one full year. Once removed from the field, resin bags were kept in a freezer and eluted within 48 hr. The bags were cleaned by brushing off dirt, pulling off small roots, and rinsing briefly with distilled water. Each bag was then shaken separately with 50 mL of 5% HCl for 30 min. The eluates were stored in glass vials at 4 C until ammonium (NH41) and nitrate (NO3) analyses were performed. NH41 (from cation resin bags) was determined using an indophenol blue method and NO3 (from anion resin bags) was determined with a coppercadmium reduction followed by a nitrite analysis by the GriessIlosvay method (Stock 1983). Absorption efficiencies were calculated for both anion and cation resin bags by shaking the bags for 24 hr in 50 mL of standard solutions made with known concentrations of KNO3 or (NH4)2SO4. Four bags were shaken individually in each of seven values of total N ranging between 0 and 40 mg/mL. The range was based on maximum values found for ammonium and nitrate in resin bag eluates. The resin bags were squeezed free of excess standard solution, rinsed in distilled water, and shaken in 5% (v/ v) HCl for 30 min. Eluates were analyzed for ammonium and nitrate as above, and field results were then transformed according to the resultant calibration curves.
Soil Descriptions

Inorganic N Availability

N availability was assessed for each of the three treatments using ion exchange resin (IER) bags (Gibson 1986; Richards et al. 1997b). Resin bags were constructed by adding 5 g of wet Amberlite resin (anion, IRA-400 or cation, IR-120 plus, Rohm and Haas Inc., New Germany, South Africa) to nylon stockings cut to 5 cm2 and sealed with a glue gun. Bags were regenerated by rinsing alternately with distilled water and shaking with 5% HCl for 30 min three times, and finally washing them three times with distilled water. In the field holes were dug at the center of the plot or beneath the target plant, and a spatula was used to lift the soil at the side of the hole to create a

Soil samples for total N, total P, pH, organic matter, Na1, Mg11, Ca11, and K1 were taken with a 7 cm deep, 5 cm diameter soil core directly adjacent to, but without disturbing, the resin bags. One core was taken per plot, air dried and sieved to 2 mm before analysis. Total N and total P were analyzed as above with 1 and 2 g of soil material, respectively. To measure pH, 20 g of soil material from each plot was shaken individually in 50 mL 0.01M CaCl2.2H2O for 30 min. The resulting slurry was measured with a WTW pH meter (Hoskin Scientific, Vancouver, BC, Canada). Percent organic matter was quantified as percent mass lost after combustion of soils at 420 C in a muffle furnace. Soil samples were sent to Matrolab (Brakenfell, South Africa) for analysis of Na1, Mg11, Ca11, and K1 using flame photometry and atomic absorption. Soil moisture was recorded in all three treatments on each date that resin bags were switched by collecting approximately 40 g of soil into one screw-top glass vial per plot. Soils were weighed, oven dried at 105 C for 48 hr and reweighed to obtain percent soil moisture. Soil temperature was also measured on each date that resin bags were switched except May and November (due to equipment failure) using a Fluke (Everett, WA, U.S.A.)

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thermocouple placed in the soil at 5 cm in the acacia thicket and acacia cleared plots only.
Glasshouse Bioassay

were compared using one-way ANOVA with post hoc Tukey tests.

Soil temperature (C)

A soil bioassay was conducted using the grass Ehrharta calycina to compare growth rates and N uptake of a weedy species on fynbos versus acacia soils. Ehrharta calycina was chosen due to its prominence in acacia stands that had been cleared 1 year earlier at Riverlands (personal observation). Although E. calycina is a native grass it generally establishes on disturbed soils and is not an abundant plant in mature or early successional acid sandplain lowland fynbos (Hoffman et al. 1987). It is also considered an invasive alien in Australia, New Zealand, and the U.S.A. (Milberg & Lamont 1995). Within each of the 12 sub-plots in the fynbos and cleared acacia treatments, three sets of bulked soils were taken in mid-October, giving 36 replicates per soil type. Each bulked sample was obtained by coring eight times with a 7 cm deep, 5 cm diameter core. In the fynbos, cores were taken in open spaces under the assumption that grasses would recruit away from large bush canopies and dense root mats. Soils were air dried, sieved to 2 mm, and placed into small plastic pots 10 cm high and 12 cm in diameter. Soil samples from each pot were analyzed for total N (as above) before the start of the bioassay experiment. Ehrharta calycina seed (mission variety) was obtained from Agricol (Brackenfell, South Africa) and 1020 individuals added to each pot. Starting in mid-December 1999 the pots were placed in a random formation within the glasshouse at the University of Cape Town and given 50 mL of tap water once every day. One week after seed germination, pots were weeded such that only the largest individual remained. Thereafter all germinating shoots were weeded. After 45 days plants were harvested, separated into root and shoot material, and washed free of remaining dirt. The material was dried at 80 C for 48 hr, weighed, and ground with a mixer mill (Retsch Inc.). Due to the stunted growth of E. calycina on fynbos soils, the three replicates per sub-plot were bulked (yielding n 5 24 per treatment) before grinding to obtain sufficient material for total N analysis. Plant material was analyzed for total N as described above.
Statistical Analyses

Results
Abiotic Factors

Maximum and minimum average monthly temperatures during the study period (25.58 5.25 and 11.67 3.76 C, [means standard deviations], respectively) were similar to average values over the last 40 years (24.08 5.10 and 12.27 3.35 C, respectively). Rainfall during the study period was 91.1 mm lower and peaked later than data averaged over the last 40 years. Soil moisture content increased in all treatments in May, the first month in which the sample date occurred after the winter rains began (Fig. 1a). The soils remained moist until November, when the seasonal rain ended. Averaged over the course of the year, intact acacia stands had significantly higher average soil moisture than fynbos, and cleared acacia stands were intermediate between the two (Fig. 1a). Averaged over the year, soil temperatures at 5 cm depth were warmer in cleared than uncleared acacia stands (Fig. 1b).

12 % Soil moisture 10 8 6 4 2 0

a
a Fynbos b Acacia Cleared acacia ab

35 30 25 20 15 10 5 0

a Acacia Cleared acacia b

Before analysis, data from the four sub-plots were averaged to give three-plot observations in each treatment (fynbos, acacia, and cleared acacia). All variables were analyzed using one-way analysis of variance (ANOVA). All data were ln transformed to correct for normality and homoscedasticity except percentage data (soil moisture and organic matter), which were arcsine (square-root) transformed. For all ANOVA analyses, post hoc Tukey tests were used to determine the significant differences between treatments. Data from the bioassay experiment

Figure 1. (a) Monthly soil moisture content within fynbos, acacia, and cleared acacia plots. Letters represent significant differences for one-way ANOVA with data averaged over the year (F[2,6] 5 6.16, p 5 0.05) with post hoc Tukey tests. (b) Monthly soil temperature data for plots with intact acacia and cleared acacia stands. Letters represent significant differences for one-way ANOVA using data averaged over the year (F[1,4] 5 67.9, p 5 0.05).

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ar ch Ap ril M ay Ju ne Ju ly Au Se gu pt st em O ber ct N obe ov e r D mb ec er em Ja be nu r Fe ary br ua ry

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Inorganic N (mg N m2 month1)

Biomass, Litterfall, and N Return to Soil

900 Fynbos a Acacia b Cleared acacia b

Litterfall was four times greater in acacia than fynbos plots (Fig. 2) and contained twice as much N as the fynbos litter. N return to the soil (quantified as litterfall 3 litter N concentration) was 10 times greater in acacia than fynbos plots. Phosphorus was significantly lower in acacia litter than fynbos litter (F[1,4] 5 7.7, p 5 0.05).

600

300

0
M ar ch Ap ril M ay Ju ne Ju Au ly Se gus t pt em O ber ct ob N ov er e D mb ec er em b Ja er nu Fe ar br y ua ry
3,000 2,500

Net N Mineralization

Calibration of resin bags showed that NH41 was absorbed according to the equation: absorbed N 5 0.74 3 actual N 1 0.41. In this equation, actual N represents the amount of N (g N/mL) with which the resin bag was treated and absorbed N represents the amount of N (g N/mL) measured in the HCl in which the resin bags were shaken. Similarly NO3 was absorbed according to the equation: absorbed N 5 0.44 3 actual N 1 0.38. Total annual IER-available N was greater in both acacia treatments than in the fynbos, with no difference between cleared and uncleared acacia (Figs. 3 and 4a). The same patterns of significance held for total annual IER-available NH41, but not NO3 (Fig. 4a). The two acacia treatments pulsed in both forms of inorganic N with the first rains of the season (late April) and then dropped back to lower levels (Fig. 3).

Figure 3. Monthly total inorganic available N (NH41 1 NO3) for fynbos, intact acacia, and cleared acacia plots. Letters represent significant differences for one-way ANOVA with post hoc Tukey tests for data summed over the year (F[2,6] 5 16.0, p 5 0.05).

Glasshouse Bioassay

The soils collected for use in the glasshouse bioassay experiment showed that total N was higher in soils from acacia stands than from fynbos soils (F[1,22] 5 44.0, p 5 0.05). After 45 days of growth, total biomass of E. calycina was greater for plants grown on acacia than

mg N m2 y1

Soil Descriptions

Fynbos Acacia Cleared acacia b b

Uncleared and cleared acacia soils were significantly higher than fynbos soils in total N and percentage organic matter (Fig. 4b). Total P (F[2,6] 5 2.4, p 5 0.17) and pH (F[2,6] 5 0.8, p 5 0.47) showed no statistical differences between treatments, although total P was somewhat higher in acacia than fynbos plots (14.0 0.9 and 10.1 0.9, mean 1 SE, respectively).

2,000 1,500 1,000 a 500 0

Ammonium 4

Nitrate c

Total N

b
b

200 150 100 Fynbos Acacia

2 a 1 a 0 % Organic matter Soil total N mg N/g soil b b

50 0

*
Litterfall g m2 y1

*
Plant N mg N/g plant N return mg N m2 y1

Figure 2. Litterfall, plant tissue nitrogen (N), and N return to soil (litterfall 3 plant tissue N) for fynbos and acacia plots. * represents significant differences for one-way ANOVA (litterfall: F[1,4] 5 18.3, p 5 0.05; plant N: F[1,4] 5 10.3, p 5 0.05; and N return: F[1,4] 5 21.1, p 5 0.05).

Figure 4. (a) Available inorganic N in the forms of ammonium (NH41), nitrate (NO3), and total N (NH41 1 NO3) for fynbos, intact acacia, and cleared acacia stands. Letters represent significant differences for one-way ANOVA (ammonium: F[2,6] 5 12.4, p 5 0.05, nitrate: F[2,6] 5 4.16, p 5 0.07; and total N: F[2,6] 5 16.0, p 5 0.05) with post hoc Tukey tests. (b) Soil organic matter and soil total N data for fynbos, intact acacia, and cleared acacia plots. Letters represent significant differences for one-way ANOVA (organic matter: F[2,6] 5 9.8, p 5 0.05; soil total N: F[2,6] 5 44.3, p 5 0.05) with post hoc Tukey tests.

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0.7 0.6 0.5 Grams 0.4 0.3 0.2 0.1 0

Fynbos Acacia

Total biomass
Figure 5. Total biomass of Ehrharta calycina seedlings grown in soils taken from fynbos and acacia plots. Letters represent significant differences for one-way ANOVA (F[1,22] 5 66.6, p 5 0.05).

fynbos soil (Fig. 5). The total N content of each plant shoot (aboveground biomass, mg N/ plant) was greater on plants from acacia (means 1 SE 5 9.5 1.2) than fynbos (2.0 0.4) soils (F[1,22] 5 37.2, p 5 0.05) although shoot N concentration (mg N/g plant) was not significantly different between acacia (29.2 2.1) and fynbos (30.5 1.3) soil types (F[1,22] 5 0.28, p 5 0.59). Discussion The N2-fixing alien species investigated in this study, Acacia saligna, appears to be able to alter N-cycling regimes through long-term (31 years) invasions within the otherwise low-nutrient soils characteristically associated with the fynbos vegetation of the Cape Floristic Region. These findings concur with the observations of Witkowski (1991a) and Stock et al. (1995) who showed the pronounced enrichment effect of other invasive Acacia species, although they were not able to show feedbacks into inorganic N mineralization for A. saligna. The higher availability of N in acacia soils led to higher growth rates of a weedy grass species, leading us to believe that enrichment of soils due to invasive alien acacias may lead to long-term problems for restoration of native vegetation. The altered N regimes found in this study may have longlasting impacts given the residence times of organic N in soils. For example, Maron & Jefferies (1999) predicted that elevated levels of N in soils underneath Lupinus arboreus would take about 25 years to reduce by half. In our study, as with the earlier ones, the impact of the alien N2-fixing plants stems from higher litterfall rates which, coupled with higher tissue N concentrations, results in more N being returned from the aboveground biomass to the soil. Increased levels of organic matter and soil total N under acacia stands led to consistently higher levels of IER-available inorganic N (NH41 and NO3) than in native fynbos stands where IER N availability remained

low and relatively constant throughout the year. The pulse effects in acacia stands are probably due to a mixture of the seasonal moisture stimulating mineralization of the high soil organic N pool in acacia stands by microbes and the dependence of resin bags on water for transferring ions from the soil to the resin (Lajtha 1988). Acacia litter contained less P than fynbos litter, although due to higher litter inputs under acacia there was a trend toward higher concentrations of P in the soils of acacia stands. This, as well as the higher concentrations of the cations Ca11, Mg11, and K1 (S. Yelenik, unpublished data), suggests that the acacias are also concentrating nutrients other than N in the top layers of the soil (Witkowski 1991a). Elevated N under invasive N2-fixing plants may be quite general. Other systems that have been investigated include Myrica faya in Hawaiian montane tropical forests (Vitousek et al. 1987; Vitousek & Walker 1989), L. arboreus in coastal Californian sand dunes (Maron & Connors 1996; Pickart et al. 1998; Maron & Jefferies 1999), and Acacia cyclops in South African strandveld (Stock et al. 1995). All four species investigated increased the inputs of N into the soil through higher litterfall rates in combination with N-rich litter. These inputs led to higher levels of organic and available N in all of the systems. It was originally hypothesized that the clearing of acacia stands would increase the levels of available inorganic N by increasing temperatures within the soil and stimulating soil microbes to mineralize N. Despite overall statistical differences in temperature, mineralization rates did not differ between cleared and intact acacia treatments. Although temperature effects on N mineralization have been shown with relatively small changes (Nadelhoffer et al. 1991), one possible explanation for a lack of temperature effect in cleared acacia stands may be that optimum temperatures for NH41 and NO3 mineralization were never reached (Alexander 1977). It also may be that N mineralization is water limited across all sites. It was also hypothesized that acacia soils would promote the growth of weedy species, and data from our study support this. The higher N status of acacia soils results in higher growth rates of Ehrharta calycina and a release from nutrient limitation, as shown by the higher biomass. Ehrharta calycina exhibited a logistic growth curve (r2 5 0.40, p  0.01, n 5 69, data not shown), which flattened to give a somewhat constant biomass yield at about 0.7 mg N/g soil, well above the maximum amount of total N (0.25 mg N/g soil) found in any of the Riverlands fynbos sites. This suggests that growth of E. calycina on fynbos soil is N limited and helps to explain its lack of abundance in the habitat (Hoffman et al. 1987) even in fynbos patches close to E. calycina seed sources at reserve boundaries (S. Yelenik, unpublished data). Ehrharta calycina therefore may have a competitive advantage over slow-growing (Stock & Lewis 1984; Witkowski et al. 1990), shade intolerant (Cowling & Gxaba 1991) fynbos seedlings in enriched soils and might therefore change overall community dynamics after acacia invasions have been cleared.

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Just as important as secondary invasions by grasses may be the re-establishment of acacia seedlings from the large seedbanks created through the long-term invasion (Milton & Hall 1981), especially given the competitive advantage A. saligna seedlings have over native fynbos species (Witkowski 1991b). The secondary invasion of exotic grasses after acacia removal might be aided by the winter germination of these species. Ehrharta calycina (veldtgrass), Briza maxima (quaking grass), Avena fatua (wild oat), Lolium perenne (rye grass), and Cynodon dactylon (bermuda grass) germinate with the concomitant winter pulse in N availability and soil water that occur in acid sandplain lowland fynbos soils after invasions by A. saligna. This trend was also noted by Maron and Jefferies (1999) when investigating secondary invasions by alien grasses after massive die-offs of the invasive N2-fixing plant L. arboreus in California. It should be noted, however, that the results from this experiment were not achieved under strict Mediterranean-climate, winter watering conditions, and the results should be viewed tentatively. Managers will now be left with the task of returning soils to pre-invasion levels of N so that native species can recolonize. The most common practice, burning an area after alien clearing (Holmes & Richardson 1999), can volatilize large quantities of organic N (Stock & Lewis 1986). However, burning may still leave high N levels due to larger amounts of standing litter and soil organic matter in acacia stands (Wan et al. 2001). A different strategy might be to add a mulch with a high C : N to help immobilize the excess N in the soil. Although the total N content of the soil would not change, less of the N would be in plant-available inorganic forms, potentially keeping weedy grasses and forb species from invading. Soil amendments with high C : N ratios have been used previously in restoration projects to lower the levels of plantavailable inorganic N, decrease the abundance of alien species, and increase the abundance of native species ro k et al. (Wilson & Gerry 1995; Zink & Allen 1998; To 2000). In conclusion, A. saligna was able to increase the rates of N cycling within the otherwise low-nutrient soil of the fynbos. Mechanistically, this was done through greater amounts of litter with higher N concentrations, resulting in more N being returned to the soil from the aboveground biomass and an increase in inorganic N availability. Although this effect was not exacerbated by the removal of acacia stands, this alteration of ecosystem function may aid in secondary invasions of weedy grasses after acacia stands are cleared for restoration purposes. Acknowledgments We thank P. Holmes, K. Wienand, R. Cowling, W. Bond and three anonymous reviewers for help with the project and manuscript. Willem de Vlam, Deon Rousseau, and the Working for Water crew are thanked for access to and

help within the Riverlands Nature Reserve. This project would not have been possible without help in the field and laboratory from S. Archibald, N. Bergh, W. Foden, and A. Roberts. Funding for this work was provided by the University of Cape Town Research Committee and National Research Foundation (W. D. Stock and D. M. Richardson) and the Institute for Plant Conservation (D. M. Richardson).

LITERATURE CITED
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