Sabine Hilt

Searching for allelopathic effects of submerged macrophytes on phytoplankton—state of the art and open questions

Allelopathy, here defined as biochemical interactions between aquatic primary producers, has always been intriguing as a process explaining the dominance of certain plant or algal species over others. Negative chemical interference has been invoked as one of the steering mechanisms behind mutual dominance of either submerged macrophytes or phytoplankton in shallow eutrophic lakes. Yet, despite much effort, convincing evidence for allelopathic interactions in situ is still missing. Also, laboratory approaches often lack reality. Inspired by a series of talks at the Shallow Lakes 2005 meeting in Dalfsen, the Netherlands, we argue that there is circumstantial but strong evidence that allelopathic interference between submerged macrophytes and phytoplankton may indeed exist in aquatic ecosystems despite the problems associated with research in this field. We first discuss experimental approaches combining laboratory and field studies, based on examples presented at this meeting. We then discuss the impact of nutrient status of both producing and target organism and biotic factors such as herbivory or pathogens that might affect allelopathy. Further topics are the potential seasonality of effects and the species-specificity of certain allelochemicals. We conclude with some thoughts why a final proof for allelopathy in situ might remain difficult or even inaccessible in some cases, and why we nevertheless should not abandon this idea.

In Situ Allelopathic Potential of Myriophyllum Verticillatum (Haloragaceae) against selected Phytoplankton Species

The potential allelopathic impact of Myriophyllum verticillatum L. under in situ conditions was determined in a series of field and laboratory experiments. Coexistence experiments were performed in a lake dominated by M. verticillatum (Van Goor) Meffert where we exposed three unialgal phytoplankton cultures in dialysis tubes to macrophyte exudates regularly during the vegetated period. Plant content and exudation of polyphenolic compounds were determined, and the inhibitory activity of polyphenol‐containing extracts was tested in bioassays with cyanobacteria. To account for possible resource interference, we monitored growth and photosynthesis of phosphorus‐limited and unlimited cyanobacterium Limnothrix redekei in dialysis tubes exposed to M. verticillatum in aquaria.

Seasonal and interannual dynamics of polyphenols in Myriophyllum verticillatum and their allelopathic activity on Anabaena variabilis

In 4 successive years, we investigated the seasonal and interannual variability of the total polyphenolic pool and of the individual polyphenolic compounds in Myriophyllum verticillatum, as well as their allelopathic activity in a small eutrophic lake. We tested whether nutrient availability explained interannual and seasonal changes in the production of polyphenols. There were no strong interannual variations in plant tissue carbon, nitrogen and phosphorus concentrations, while total phenolic compounds (TPC) significantly differed between years, especially in apical meristems (range: 38 122 mg g 1 dry weight (DW)). Seasonal patterns, with maxima between May and July, changed between years. Partially confirming the carbon nutrient balance hypothesis sensu Bryant et al. [Bryant, J.P., Chapin III, F.S., Klein, D.R., 1983. Carbon/nutrient balance of boreal plants in relation to vertebrate herbivory. Oikos 40, 357 368], we found correlations between TPC and the C/N (carbon/nitrogen) ratio in some but not all years, especially in apical meristems. Plant tissue phosphorus content accounted also for the variability in TPC in some years. Crude extracts of apical meristems always inhibited the growth of Anabaena variabilis, used as a target cyanobacterium. Plant TPC concentration and allelopathic activity were significantly correlated in all years except in 2005. Bioassay directed fractionation of M. verticillatum extracts coupled with LC MS analyses of the respective fractions revealed several isomers of HHDP di and tri galloylglucose apparently responsible for the allelopathic effects. The individual active compounds revealed a more distinct seasonal pattern compared to the pool of phenolic compounds in M. verticillatum, with a clear maximum in May, the ecologically most relevant period for inhibitory effects of submerged macrophytes on phytoplankton.

Can submerged macrophytes influence turbidity and trophic state in deep lakes? Suggestions from a case study.

Feedback between submerged macrophytes and water transparency stabilizing the clear, macrophyte-dominated regime has been described so far for shallow lakes. Based on data of total phosphorus (TP) concentrations, underwater light supply, phytoplankton and submerged macrophyte abundance from narrow, stratified Lake Scharmützelsee (mean depth: 9 m, retention time: 16 yr) of the period 1994-2006 we hypothesize that submerged macrophytes may influence transparency and trophic state in deep lakes. The lake was characterized by summer epilimnion TP concentrations of 38 to 57 mug L(-1), turbid water due to mass development of cyanobacteria, and low abundance of few submerged macrophyte species until 2003. Thereafter, a sudden increase in water transparency was followed by a rapid submerged macrophyte colonization of the littoral down to about 5 m depth corresponding to the depth of a light supply of 3 E m(-2) d(-1). Initially, this recolonization was probably a consequence of decreased turbidity. We argue that the increase of submerged macrophyte coverage from < 10% in 1994 to 2003 to about 24% in 2005-2006 has contributed to the stabilization of the clear-water regime during the subsequent years. This is supported by the fact that earlier shifts to clear-water regimes in 1994 and 2000 without a significant spread of submerged macrophytes were not stable. We discuss potential mechanisms that may have resulted in a positive effect of plants on transparency such as P uptake and immobilization by the dominant rootless macrophyte species Nitellopsis obtusa and Ceratophyllum demersum and other macrophyte-related mechanisms such as increased zooplankton grazing and allelopathy.

Recovery of Potamogeton pectinatus L. stands in a shallow eutrophic lake under extreme grazing pressure

In shallow lakes, submerged macrophytes contribute to the stabilization of the clear water state. If lost, a number of mechanisms prevent re-colonization. Lake Muggelsee (730 ha) lost its submerged vegetation due to increasing eutrophication and switched to phytoplankton dominance in 1970. After the reduction of nutrient loading in 1990, Potamogeton pectinatus L. started re-colonizing the lake. During the following years, it spread at a mean rate of 2.5 ha per year to all available areas <80 cm depth. Between 1993 and 1999, decreasing maximum biomass indicated hampered growth. Exclosure experiments revealed that herbivory reduced the aboveground biomass by more than 90%. Both waterfowl and fish were found to contribute to the grazing pressure despite a low abundance of the known herbivorous fish species and waterfowl in spring and summer. Protection of stands against grazing resulted in higher biomass of shoots, whereas shoot and tuber density did not change. Both shading by phytoplankton and periphyton, as well as grazing pressure, prevented the submerged vegetation of Lake Muggelsee from developing back to a dense zone that contributed to the reduction of turbidity.

Whole-lake experiments reveal the fate of terrestrial particulate organic carbon in benthic food webs of shallow lakes.

Lake ecosystems are strongly linked to their terrestrial surroundings by material and energy fluxes across ecosystem boundaries. However, the contribution of terrestrial particulate organic carbon (tPOC) from annual leaf fall to lake food webs has not yet been adequately traced and quantified. In this study, we conducted whole-lake experiments to trace artificially added tPOC through the food webs of two shallow lakes of similar eutrophic status, but featuring alternative stable regimes (macrophyte rich vs. phytoplankton dominated). Lakes were divided with a curtain, and maize (Zea mays) leaves were added, as an isotopically distinct tPOC source, into one half of each lake. To estimate the balance between autochthonous carbon fixation and allochthonous carbon input, primary production and tPOC and tDOC (terrestrial dissolved organic carbon) influx were calculated for the treatment sides. We measured the stable isotope ratios of carbon (delta13C) of about 800 samples from all trophic consumer levels and compared them between lake sides, lakes, and three seasons. Leaf litter bag experiments showed that added maize leaves were processed at rates similar to those observed for leaves from shoreline plants, supporting the suitability of maize leaves as a tracer. The lake-wide carbon influx estimates confirmed that autochthonous carbon fixation by primary production was the dominant carbon source for consumers in the lakes. Nevertheless, carbon isotope values of benthic macroinvertebrates were significantly higher with maize additions compared to the reference side of each lake. Carbon isotope values of omnivorous and piscivorous fish were significantly affected by maize additions only in the macrophyte-dominated lake and delta13C of zooplankton and planktivorous fish remained unaffected in both lakes. In summary, our results experimentally demonstrate that tPOC in form of autumnal litterfall is rapidly processed during the subsequent months in the food web of shallow lakes and is channeled to secondary and tertiary consumers predominantly via the benthic pathways. A more intense processing of tPOC seems to be connected to a higher structural complexity in littoral zones, and hence may differ between shallow lakes of alternative stable states.

A regime shift from macrophyte to phytoplankton dominance enhances carbon burial in a shallow, eutrophic lake

Ecological regime shifts and carbon cycling in aquatic systems have both been subject to increasing attention in recent years, yet the direct connection between these topics has remained poorly understood. A four-fold increase in sedimentation rates was observed within the past 50 years in a shallow eutrophic lake with no surface in- or outflows. This change coincided with an ecological regime shift involving the complete loss of submerged macrophytes, leading to a more turbid, phytoplankton-dominated state. To determine whether the increase in carbon (C) burial resulted from a comprehensive transformation of C cycling pathways in parallel to this regime shift, we compared the annual C balances (mass balance and ecosystem budget) of this turbid lake to a similar nearby lake with submerged macrophytes, a higher transparency, and similar nutrient concentrations. C balances indicated that roughly 80% of the C input was permanently buried in the turbid lake sediments, compared to 40% in the clearer macrophyte-dominated lake. This was due to a higher measured C burial efficiency in the turbid lake, which could be explained by lower benthic C mineralization rates. These lower mineralization rates were associated with a decrease in benthic oxygen availability coinciding with the loss of submerged macrophytes. In contrast to previous assumptions that a regime shift to phytoplankton dominance decreases lake heterotrophy by boosting whole-lake primary production, our results suggest that an equivalent net metabolic shift may also result from lower C mineralization rates in a shallow, turbid lake. The widespread occurrence of such shifts may thus fundamentally alter the role of shallow lakes in the global C cycle, away from channeling terrestrial C to the atmosphere and towards burying an increasing amount of C.

Extending one-dimensional models for deep lakes to simulate the impact of submerged macrophytes on water quality

Submerged macrophytes can stabilise clear water conditions in shallow lakes. However, many existing models for deep lakes neglect their impact. Here, we tested the hypothesis that submerged macrophytes can affect the water clarity in deep lakes. A one-dimensional, vertically resolved macrophyte model was developed based on PCLake and coupled to SALMO-1D and GOTM hydrophysics and validated against field data. Validation showed good coherence in dynamic growth patterns and colonisation depths. In our simulations the presence of submerged macrophytes resulted in up to 50% less phytoplankton biomass in the shallowest simulated lake (11?m) and still 15% less phytoplankton was predicted in 100?m deep oligotrophic lakes. Nutrient loading, lake depth, and lake shape had a strong influence on macrophyte effects. Nutrient competition was found to be the strongest biological interaction. Despite a number of limitations, the derived dynamic lake model suggests significant effects of submerged macrophytes on deep lake water quality. Existing models were innovatively combined to study macrophyte effects in deep lakes.Submerged macrophytes can significantly affect the water clarity in deep lakes.This effect depends on lake geometry, depth, and nutrient loading.

COMPARISON OF METHODS TO DETECT ALLELOPATHIC EFFECTS OF SUBMERGED MACROPHYTES ON GREEN ALGAE1

Detecting allelopathic inhibition of phytoplankton by submerged macrophytes in an ecologically meaningful way is not easy. Multiple‐approach investigations from a laboratory scale to the ecosystem level have been recommended to overcome the shortcomings of individual methods. Whether results of different methods are qualitatively or quantitatively comparable has not yet been tested. Here, we compare the sensitivity of the green algae Desmodesmus subspicatus (Chodat) E. Hegewald et Ant. Schmidt and Stigeoclonium helveticum Vischer to the allelopathic effect of the submerged macrophyte Myriophyllum verticillatum L. The following three approaches were used: (i) coincubation of algae in dialysis membrane tubes in a lake inside and outside a M. verticillatum stand, (ii) coincubation of algae in dialysis membrane tubes in aquaria with and without M. verticillatum, and (iii) single additions of tannic acid (TA), an allelopathically active polyphenol present in this macrophyte, to the algae cultures. For each method, fluorescence‐based (chl a, PSII activity) and particle‐based (cell count, biovolume) parameters were compared after 48 h of incubation. Results revealed quantitative and qualitative differences between methods. Algae incubated in dialysis membrane tubes in aquaria showed a strong decrease in all parameters under the influence of macrophytes. In situ measurements were influenced by adverse growth conditions for the test algae and only detected significant reductions for biovolume. Single additions of TA induced a strong reduction of fluorescence‐based parameters similar to aquarium results, but an increase in the cell count. Even the qualitative transfer of laboratory results to field conditions thus requires caution and a proper selection of parameters.

Impact of water-level fluctuations on cyanobacterial blooms: options for management

Climate change can promote harmful cyanobacteria blooms in eutrophic waters through increased droughts or flooding. In this paper, we explore how water-level fluctuations affect the occurrence of cyanobacterial blooms, and based on the observations from case studies, we discuss the options and pitfalls to use water-level fluctuations for lake and reservoir management. A drawdown in summer causes an increase in retention time and increased water column nutrient concentrations and temperature of shallow water layers, which may lead to severe cyanobacterial blooms. This effect can potentially be counteracted by the positive response of submerged macrophytes, which compete for nutrients with cyanobacteria, with a higher chance of cyanobacterial blooms under eutrophic conditions. The balance between dominance by submerged macrophytes or cyanobacteria is temperature sensitive with stronger positive effects of drawdown as inhibition of cyanobacterial blooms expected in colder climates. Complete drying out reduces the amount of cyanobacteria in the water column after refilling, with lower water nutrient concentrations, lower fish biomass, lower abundance of cyanobacteria, higher transparency, and higher cover of submerged plants compared to lakes and reservoirs that did not dry out. Water-level rise as response to flooding has contrasting effects on the abundance of cyanobacteria depending on water quality. We conclude that water-level fluctuation management has potential to mitigate cyanobacterial blooms. However, the success will depend strongly on ecosystem properties, including morphometry, sediment type, water retention time, quality of inlet water, presence of submerged vegetation or propagules, abundance of fish, and climate.