Absolute configuration and biosynthesis of pahayokolide A from Lyngbya sp. Strain 15-2 of the Florida Everglades

Pahayokolides A-D are cytotoxic cyclic polypeptides produced by the freshwater cyanobacterium Lyngbya sp. strain 15-2 that possess an unusual β-amino acid, 3-amino-2,5,7,8-tetrahydroxy-10-methylundecanoic acid (Athmu). The absolute configuration of pahayokolides A-D was determined using advanced Marfey’s method. It was also confirmed that a pendant N-acetyl- N-methyl leucine moiety in pahayokolide A was absent in pahayokolides B and pahayokolides C-D were conformers of pahayokolide A. Feeding experiments indicated that the biosynthesis of the Athmu sidechain arises from leucine or α-ketoisovalerate, however could not be further extended by three rounds of condensation with malonate units. Putative four peptide and one unique polyketide synthetases in Lyngbya sp. strain 15-2 were identified by using a PCR method and degenerate primers derived from conserved core sequences of known NRPSs and PKSs. Identification of one unique KS domain conflicted with the logic rule that the long side chain of Athmu was assembled by three rounds of ketide extensions if PKSs were involved. A gene cluster (pah) encoding a peptide synthetase putatively producing pahayokolide was cloned, partially sequenced and characterized. Seven modules of the non-ribosomal peptide synthetase (NRPS) were identified. Ten additional opening reading frames (ORFs) were found, responsible for peptide resistance, transport and degradation. Although the predicted substrate specificities of NRPS agreed with the structure of pahayokolide A partially, the disagreement could be explained. However, no PKS gene was found in the pah gene cluster.

Cascading ecological effects of low-level phosphorus enrichment in the Florida Everglades

Few studies have examined long-term ecological effects of sustained low-level nutrient enhancement on wetland biota. To determine sustained effects of phosphorus (P) addition on Everglades marshes we added P at low levels (5, 15, and 30 µg L-1 above ambient) for 5 yr to triplicate 100-m flow-through channels in pristine marsh. A cascade of ecological responses occurred in similar sequence among treatments. Although the rate of change increased with dosing level, treatments converged to similar enriched endpoints, characterized most notably by a doubling of plant biomass and elimination of native, calcareous periphyton mats. The full sequence of biological changes occurred without an increase in water total P concentration, which remained near ambient levels until Year 5. This study indicates that Everglades marshes have a near-zero assimilative capacity for P without a state change, that ecosystem responses to enrichment accumulate over time, and that downstream P transport mainly occurs through biota rather than the water column.

Landscape patterns of periphyton in the Florida Everglades

Periphyton is an abundant and ubiquitous feature of the Florida Everglades, often forming thick mats that blanket shallow sediments and submersed plants. They are considered to be primary ecosystem engineers in the Everglades by forming and stabilizing soils, controlling concentrations of nutrients and gases, and supplying food and structure for other organisms. Distribution patterns are related to underlying physicochemical gradients as well as those hydrologic changes imposed by water management. Because communities respond rapidly to environmental change, their use has been advocated to provide indication of system degradation or restoration. The authors review studies on the distribution of periphyton in the Everglades, highlighting major findings relevant to water management, and also areas where additional exploration is necessary.

Carbon isotopic composition of cypress trees from South Florida and changing hydrologic condition

δ13C values were determined from cypresstree rings from two different study areas in SouthFlorida. One site is located in the Southeastern Everglades Marsh, where pond cypress (Taxodium ascendens) was sampled from tree islands (annual tree rings from 1970 to 2000). Bald cypress (Taxodium distichum) trees were sampled at the other site, located along the Loxahatchee River in a coastal wetland (decadal tree rings from 1830 to 1990). The isotopic time series from both sites display different, location-specific information. The pond cypressisotopic time series has a positive correlation with the total amount of annual precipitation, while the bald cypress data from the Loxahatchee River study area had two different records dependent on the level of saltwater stress. In general, for terrestrial trees growing in a temperate environment, water stress causes an increase in water-use efficiency (WUE) resulting in a relative 13C enrichment. Yet, trees growing in wetland settings in some cases do not respond in the same manner. We propose a conceptual model based on changes in carbon assimilation and isotopic fractionation as controlled by differences in stomatal resistance (water stress) and mesophyll resistance (biochemical and nutrient related) to explain the isotopic records from both sites. With further work and a longer time series, our approach may be tested, and used to reconstruct change in hydroperiods further back in time, and potentially provide a baseline for wetland restoration.

Rapid responses of vegetation to hydrological changes in Taylor Slough, Everglades National Park, Florida, USA

We analyzed the dynamics of freshwater marsh vegetation of Taylor Slough in eastern Everglades National Park for the 1979 to 2003 period, focusing on cover of individual plant species and on cover and composition of marsh communities in areas potentially influenced by a canal pump station (‘‘S332’’) and its successor station (‘‘S332D’’). Vegetation change analysis incorporated the hydrologic record at these sites for three intervals: pre-S332 (1961–1980), S332 (1980–1999), post-S332 (1999–2002). During S332 and post-S332 intervals, water level in Taylor Slough was affected by operations of S332 and S332D. To relate vegetation change to plot-level hydrological conditions in Taylor Slough, we developed a weighted averaging regression and calibration model (WA) using data from the marl prairies of Everglades National Park and Big Cypress National Preserve. We examined vegetation pattern along five transects. Transects 1–3 were established in 1979 south of the water delivery structures, and were influenced by their operations. Transects 4 and 5 were established in 1997, the latter west of these structures and possibly under their influence. Transect 4 was established in the northern drainage basin of Taylor Slough, beyond the likely zones of influence of S332 and S332D. The composition of all three southern transects changed similarly after 1979. Where muhly grass (Muhlenbergia capillaris var. filipes) was once dominant, sawgrass (Cladium jamaicense), replaced it, while where sawgrass initially predominated, hydric species such as spikerush (Eleocharis cellulosa Torr.) overtook it. Most of the changes in species dominance in Transects 1–3 occurred after 1992, were mostly in place by 1995–1996, and continued through 1999, indicating how rapidly vegetation in seasonal Everglades marshes can respond to hydrological modifications. During the post-S332 period, these long-term trends began reversing. In the two northern transects, total cover and dominance of both muhly grass and sawgrass increased from 1997 to 2003. Thus, during the 1990’s, vegetation composition south of S332 became more like that of long hydroperiod marshes, but afterward it partially returned to its 1979 condition, i.e., a community characteristic of less prolonged flooding. In contrast, the vegetation change along the two northern transects since 1997 showed little relationship to hydrologic status.

Hurricane disturbance and recovery of energy balance, CO2 fluxes and canopy structure in a mangrove forest of the Florida Everglades

Eddy covariance (EC) estimates of carbon dioxide (CO2) fluxes and energy balance are examined to investigate the functional responses of a mature mangrove forest to a disturbance generated by Hurricane Wilma on October 24, 2005 in the Florida Everglades. At the EC site, high winds from the hurricane caused nearly 100% defoliation in the upper canopy and widespread tree mortality. Soil temperatures down to -50 cm increased, and air temperature lapse rates within the forest canopy switched from statically stable to statically unstable conditions following the disturbance. Unstable conditions allowed more efficient transport of water vapor and CO2 from the surface up to the upper canopy layer. Significant increases in latent heat fluxes (LE) and nighttime net ecosystem exchange (NEE) were also observed and sensible heat fluxes (H) as a proportion of net radiation decreased significantly in response to the disturbance. Many of these impacts persisted through much of the study period through 2009. However, local albedo and MODIS (Moderate Resolution Imaging Spectro-radiometer) data (the Enhanced Vegetation Index) indicated a substantial proportion of active leaf area recovered before the EC measurements began 1 year after the storm. Observed changes in the vertical distribution and the degree of clumping in newly emerged leaves may have affected the energy balance. Direct comparisons of daytime NEE values from before the storm and after our measurements resumed did not show substantial or consistent differences that could be attributed to the disturbance. Regression analyses on seasonal time scales were required to differentiate the storm's impact on monthly average daytime NEE from the changes caused by interannual variability in other environmental drivers. The effects of the storm were apparent on annual time scales, and CO2 uptake remained approximately 250 g C m-2 yr-1 lower in 2009 compared to the average annual values measured in 2004-2005. Dry season CO2 uptake was relatively more affected by the disturbance than wet season values. Complex leaf regeneration dynamics on damaged trees during ecosystem recovery are hypothesized to lead to the variable dry versus wet season impacts on daytime NEE. In contrast, nighttime CO2 release (i.e., nighttime respiration) was consistently and significantly greater, possibly as a result of the enhanced decomposition of litter and coarse woody debris generated by the storm, and this effect was most apparent in the wet seasons compared to the dry seasons. The largest pre- and post-storm differences in NEE coincided roughly with the delayed peak in cumulative mortality of stems in 2007-2008. Across the hurricane-impacted region, cumulative tree mortality rates were also closely correlated with declines in peat surface elevation. Mangrove forest-atmosphere interactions are interpreted with respect to the damage and recovery of stand dynamics and soil accretion processes following the hurricane.

Physiological responses of red mangroves to the climate in the Florida Everglades

This manuscript reports the findings of physiological studies of red mangrove (Rhizophora mangle L.) conducted from June to August 2001 and from May to June 2003 in the Florida Everglades. In situ physiological measurements were made using environmentally controlled gas exchange systems. The field investigations were carried out to define how regional climate constrains mangrove physiology and ecosystem carbon assimilation. In addition, maximum carboxylation and photosynthetic active radiation (PAR) limited carbon assimilation capacities were investigated during the summer season to evaluate whether ecophysiological models developed for mesophyte plant species can be applied to mangroves. Under summertime conditions in the Florida Everglades, maximum foliar carbon dioxide (CO2) assimilation rates reached 18 μmol CO2 m−2 s−1. Peak molar stomatal conductance to water vapor (H2O) diffusion reached 300 mmol H2O m−2 s−1. Maximum carboxylation and PAR‐limited carbon assimilation rates at the foliage temperature of 30°C attained 76.1 ± 23.4 μmol CO2 m−2 s−1 and 128.1 ± 32.9 μmol (e−) m−2 s−1, respectively. Environmental stressors such as the presence of hypersaline conditions and high solar irradiance loading (>500 W m−2 or >1000 μmoles of photons m−2 s−1 of PAR) imposed sharp reductions in carbon assimilation rates and suppressed stomatal conductance. On the basis of both field observations and model analyses, it is also concluded that existing ecophysiological models need to be modified to consider the influences of hypersaline and high radiational loadings on the physiological responses of red mangroves.

Controls on mangrove forest-atmosphere carbon dioxide exchanges in western Everglades National Park

We report on net ecosystem production (NEP) and key environmental controls on net ecosystem exchange (NEE) of carbon dioxide (CO2) between a mangrove forest and the atmosphere in the coastal Florida Everglades. An eddy covariance system deployed above the canopy was used to determine NEE during January 2004 through August 2005. Maximum daytime NEE ranged from −20 to −25 mmol (CO2) m−2 s−1 between March and May. Respiration (Rd) was highly variable (2.81 ± 2.41 mmol (CO2) m−2 s−1), reaching peak values during the summer wet season. During the winter dry season, forest CO2 assimilation increased with the proportion of diffuse solar irradiance in response to greater radiative transfer in the forest canopy. Surface water salinity and tidal activity were also important controls on NEE. Daily light use efficiency was reduced at high (>34 parts per thousand (ppt)) compared to low (ppt) salinity by 46%. Tidal inundation lowered daytime Rd by ∼0.9 mmol (CO2) m−2 s−1 and nighttime Rd by ∼0.5 mmol (CO2) m−2 s−1. The forest was a sink for atmospheric CO2, with an annual NEP of 1170 ± 127 g C m−2 during 2004. This unusually high NEP was attributed to year‐round productivity and low ecosystem respiration which reached a maximum of only 3 g C m−2 d−1. Tidal export of dissolved inorganic carbon derived from belowground respiration likely lowered the estimates of mangrove forest respiration. These results suggest that carbon balance in mangrove coastal systems will change in response to variable salinity and inundation patterns, possibly resulting from secular sea level rise and climate change. Citation: Barr, J. G., V. Engel, J. D. Fuentes,

Periphyton responses to eutrophication in the Florida Everglades: Cross-system patterns of structural and compositional change

We examined periphyton along transects in five Everglades marshes and related compositional and functional aspects to phosphorus(P ) gradients caused by enriched inflows. Results were compared to those of a P-addition experiment in a pristine Everglades marsh. While the water total P (TP) concentration was not related to P load in the marshes or experiment the concentration of TP in periphyton was strongly correlated with the distance from the P source. Increased P concentration in periphyton was associated with a loss of biomass,p articularly of the calcifying mat-forming matrix, regardless of the growth form of the periphyton (epiphytic, floating,or epilithic). Diatom species composition was also strongly related to P availability, but the TP optima of many species varied among marshes. Enriched periphyton communities were found 14 km downstream of P inputs to one marsh that has been receiving enhanced P loads for decades, where other studies using different biotic indicators show negligible change in the same marsh. Although recovery trajectories are unknown, periphyton indicators should serve as excellent metrics for the progression or amelioration of P-related effects in the Everglades.

Spatial scale and abundance patterns of large fish communities in freshwater marshes of the Florida Everglades

Anthropogenic habitat alterations and water-management practices have imposed an artificial spatial scale onto the once contiguous freshwater marshes of the Florida Everglades. To gain insight into how these changes may affect biotic communities, we examined whether variation in the abundance and community structure of large fishes (SL . 8 cm) in Everglades marshes varied more at regional or intraregional scales, and whether this variation was related to hydroperiod, water depth, floating mat volume, and vegetation density. From October 1997 to October 2002, we used an airboat electrofisher to sample large fishes at sites within three regions of the Everglades. Each of these regions is subject to unique watermanagement schedules. Dry-down events (water depth , 10 cm) occurred at several sites during spring in 1999, 2000, 2001, and 2002. The 2001 dry-down event was the most severe and widespread. Abundance of several fishes decreased significantly through time, and the number of days post-dry-down covaried significantly with abundance for several species. Processes operating at the regional scale appear to play important roles in regulating large fishes. The most pronounced patterns in abundance and community structure occurred at the regional scale, and the effect size for region was greater than the effect size for sites nested within region for abundance of all species combined, all predators combined, and each of the seven most abundant species. Non-metric multi-dimensional scaling revealed distinct groupings of sites corresponding to the three regions. We also found significant variation in community structure through time that correlated with the number of days post-dry-down. Our results suggest that hydroperiod and water management at the regional scale influence large fish communities of Everglades marshes.

Relating precipitation and water management to nutrient concentrations in the oligotrophic ‘‘upside-down’’ estuaries of the Florida Everglades

We present 8 yr of long-term water quality, climatological, and water management data for 17 locations in Everglades National Park, Florida. Total phosphorus (P) concentration data from freshwater sites (typically ,0.25 mmol L21, or 8 mg L21) indicate the oligotrophic, P-limited nature of this large freshwater–estuarine landscape. Total P concentrations at estuarine sites near the Gulf of Mexico (average ø0.5 m mol L21) demonstrate the marine source for this limiting nutrient. This ‘‘upside down’’ phenomenon, with the limiting nutrient supplied by the ocean and not the land, is a defining characteristic of the Everglade landscape. We present a conceptual model of how the seasonality of precipitation and the management of canal water inputs control the marine P supply, and we hypothesize that seasonal variability in water residence time controls water quality through internal biogeochemical processing. Low freshwater inflows during the dry season increase estuarine residence times, enabling local processes to control nutrient availability and water quality. El Nin˜o–Southern Oscillation (ENSO) events tend to mute the seasonality of rainfall without altering total annual precipitation inputs. The Nin˜o3 ENSO index (which indicates an ENSO event when positive and a La Nin˜a event when negative) was positively correlated with both annual rainfall and the ratio of dry season to wet season precipitation. This ENSO-driven disruption in seasonal rainfall patterns affected salinity patterns and tended to reduce marine inputs of P to Everglades estuaries. ENSO events also decreased dry season residence times, reducing the importance of estuarine nutrient processing. The combination of variable water management activities and interannual differences in precipitation patterns has a strong influence on nutrient and salinity patterns in Everglades estuaries.

Importance of storm events in controlling ecosystem structure and function in a Florida gulf coast estuary

From 8/95 to 2/01, we investigated the ecological effects of intra- and inter-annual variability in freshwater flow through Taylor Creek in southeastern Everglades National Park. Continuous monitoring and intensive sampling studies overlapped with an array of pulsed weather events that impacted physical, chemical, and biological attributes of this region. We quantified the effects of three events representing a range of characteristics (duration, amount of precipitation, storm intensity, wind direction) on the hydraulic connectivity, nutrient and sediment dynamics, and vegetation structure of the SE Everglades estuarine ecotone. These events included a strong winter storm in November 1996, Tropical Storm Harvey in September 1999, and Hurricane Irene in October 1999. Continuous hydrologic and daily water sample data were used to examine the effects of these events on the physical forcing and quality of water in Taylor Creek. A high resolution, flow-through sampling and mapping approach was used to characterize water quality in the adjacent bay. To understand the effects of these events on vegetation communities, we measured mangrove litter production and estimated seagrass cover in the bay at monthly intervals. We also quantified sediment deposition associated with Hurricane Irene's flood surge along the Buttonwood Ridge. These three events resulted in dramatic changes in surface water movement and chemistry in Taylor Creek and adjacent regions of Florida Bay as well as increased mangrove litterfall and flood surge scouring of seagrass beds. Up to 5 cm of bay-derived mud was deposited along the ridge adjacent to the creek in this single pulsed event. These short-term events can account for a substantial proportion of the annual flux of freshwater and materials between the mangrove zone and Florida Bay. Our findings shed light on the capacity of these storm events, especially when in succession, to have far reaching and long lasting effects on coastal ecosystems such as the estuarine ecotone of the SE Everglades.

Population structure of spotted sunfish (Lepomis punctatus) in the Florida Everglades as revealed by DNA microsatellite analysis

Genetic diversity can be used to describe patterns of gene flow within and between local and regional populations. The Florida Everglades experiences seasonal fluctuations in water level that can influence local population extinction and recolonization dynamics. In addition, this expansive wetland has been divided into water management regions by canals and levees. These combined factors can affect genetic diversity and population structure of aquatic organisms in the Everglades. We analyzed allelic variation at six DNA microsatellite loci to examine the population structure of spotted sunfish (Lepomis punctatus) from the Everglades. We tested the hypothesis that recurrent local extinction and recent regional divisions have had an effect on patterns of genetic diversity. No marked differences were observed in comparisons of the heterozygosity values of sites within and among water management units. No evidence of isolation by distance was detected in a gene flow and distance correlation between subpopulations. Confidence intervals for the estimated F-statistic values crossed zero, indicating that there was no significant genetic difference between subpopulations within a region or between regions. Notably, the genetic variation among subpopulations in a water conservation area was greater than variation among regions (Fsp>FPT). These data indicate that the spatial scale of recolonization following local extinction appears to be most important within water management units.

Spatio-temporal patterns and nutrient status of macroalgae in a heavily managed region of Biscayne Bay, Florida, USA

The coastal bays of South Florida are located downstream of the Florida Everglades, where a comprehensive restoration plan will strongly impact the hydrology of the region. Submerged aquatic vegetation communities are common components of benthic habitats of Biscayne Bay, and will be directly affected by changes in water quality. This study explores community structure, spatio-temporal dynamics, and tissue nutrient content of macroalgae to detect and describe relationships with water quality. The macroalgal community responded to strong variability in salinity; three distinctive macroalgal assemblages were correlated with salinity as follows: (1) low-salinity, dominated by Chara hornemannii and a mix of filamentous algae; (2) brackish, dominated by Penicillus capitatus, Batophora oerstedii, and Acetabularia schenckii; and (3) marine, dominated by Halimeda incrassata and Anadyomene stellata. Tissue-nutrient content was variable in space and time but tissues at all sites had high nitrogen and N:P values, demonstrating high nitrogen availability and phosphorus limitation in this region. This study clearly shows that distinct macroalgal assemblages are related to specific water quality conditions, and that macroalgal assemblages can be used as community-level indicators within an adaptive management framework to evaluate performance and restoration impacts in Biscayne Bay and other regions where both freshwater and nutrient inputs are modified by water management decisions.

Groundwater-surface water interactions on tree islands in the Everglades, south Florida

The marked decline in tree island cover across the Everglades over the last century, has been attributed to landscape-scale hydrologic degradation. To preserve and restore Everglades tree islands, a clear understanding of tree island groundwater-surface water interactions is needed, as these interactions strongly influence the chemistry of shallow groundwater and the location and patterns of vegetation in many wetlands. The goal of this work was to define the relationship between groundwater-surface water interactions, plant-water uptake, and the groundwater geochemical condition of tree islands. Groundwater and surface water levels, temperature, and chemistry were monitored on eight constructed and one natural tree island in the Everglades from 2007–2010. Sap flow, diurnal water table fluctuations and stable oxygen isotopes of stem, ground and soil water were used to determine the effect of plant-water uptake on groundwater-surface water interactions. Hydrologic and geochemical modeling was used to further explore the effect of plant-groundwater-surface water interactions on ion concentrations and potential mineral formation.^