In vitro studies of the submerged angiospermRuppia maritima: Auxin and cytokinin effects on plant growth and development

Axenic tissue cultures ofRuppia maritima L. were established and propagated clonally in vitro from terminal rhizome segments collected from Tampa Bay, Florida, USA. Cultures were maintained in a base medium consisting of synthetic seawater supplemented with half-strength Murashige and Skoog salts and 1% sucrose at pH 5.6. The effects of five cytokins [6-furfurylaminopurine (kinetin), 6-benzylaminopurine (BAP), 2-isopentyladenine (2iP), 6-(4-hydroxy-3-methyl-but-2-enylamino) purine (zeatin), andn-phenyl-n-1,2,3-thidiazol-5yl urea (thidiazuron)] and one auxin [napthalene acetic acid (NAA)] on explant growth and development were investigated. Cytokinin additions resulted in a 3- to 4-fold increase in nodal production, branching, and biomass ofR. maritima after 12 wk in culture. Cultures responded in a dose-dependent manner to 2iP but exhibited broad dose-response curves to kinetin, BAP, zeatin, and thidiazuron. NAA addition resulted in increased leaf and internodal lengths, but reduced the number of leaves per node and the rhizome biomass. The addition of NAA almost completely suppressed root growth in media without cytokinins and had an antagonistic effect on nodal production and branching in cytokinin-supplemented media.     

The effect of pulsed versus gradual salinity reduction on the physiology and survival of Halophila johnsonii Eiseman

Plants of Halophila johnsonii Eiseman were exposed, in mesocosms, to either pulsed hyposalinity treatments of 30, 15, 10, and 8 or gradual salinity reductions of two every 2?days. When salinity was pulsed, survivorship (>80?%) and maximum quantum yields (>0.7) were high in the 30 and 15 salinity treatments, but both declined in the 10 and 8 salinity treatments. Leaf osmolality declined with respect to salinity treatment, but the difference between leaf and media osmolality remained relatively constant (675?±?177?mmol?kg^?1). In contrast, when salinity was gradually reduced, survivorship remained high from salinities of 30 to 4, and maximum quantum yields remained high from salinities of 30 to 6. Leaf osmolality declined linearly with respect to media osmolality and, similar to the pulsed treatments, the difference between leaf and media osmolality remained relatively constant from salinities of 30 to 2 (638?±?161?mmol?kg^?1). Trolox equivalent antioxidant capacity declined over time in both pulsed and gradual salinity reduction. The results indicate that H. johnsonii is more tolerant of hyposalinity than has previously been reported and that gradually reducing salinity extended its low-salinity tolerance threshold by approximately a salinity of 10.     

Photosynthetic utilization of CO2(aq) and HCO 3 - in Thalassia testudinum (Hydrocharitaceae)

The effects of total dissolved inorganic carbon (DIC), free carbon dioxide [CO_2(aq)], and bicarbonate (HCO ₃ ^- ) concentrations on net photosynthetic oxygen evolution of the marine angiosperm Thalassia testudinum Banks ex König collected from Biscayne Bay (1988) and from Tampa Bay (1990), Florida, USA, were examined. Rates of photosynthesis declined by 85% from pH 7.25 to 8.75 in buffered seawater media with constant DIC concentration (2.20 mM), suggesting a strong influence of CO_2(aq) concentration. A plateau in the pH-response curve between pH 7.75 and 8.50 indicated possible utilization of HCO ₃ ^- . Responses of photosynthesis measured in buffered seawater media of varying DIC concentrations (0.75 to 13.17 mM) and pH (7.8 to 8.61) demonstrated that photosynthesis is rate-limited at ambient DIC levels. Photosynthesis increased in media with increasing HCO ₃ ^- concentrations but near-constant CO_2(aq) levels, confirming HCO ₃ ^- assimilation. Calculated half-saturation constants (K _s )for CO_2(aq) and HCO ₃ ^- indicated a high affinity for the former [K _s (CO₂)=3 to 18 M] and a much lower affinity for the latter [K _s (HCO ₃ ^- )=1.22 to 8.88 mM]. Calculated V _max values for HCO ₃ ^- were generally higher than those for CO_2(aq), suggesting relatively efficient HCO ₃ ^- utilization, despite the apparent low affinity for this carbon form.     

Effect of hyposalinity on the photophysiology of <Emphasis Type="Italic">Siderastrea radians</Emphasis>

Tolerance to hyposalinity of the scleractinian coral S. radians was examined in a mesocosm study. Colonies of S. radians were collected from five basins in Florida Bay, USA, which occur along a northeast-to-southwest salinity gradient. Salinity treatments were based on historical salinity records for these basins. Photophysiology of the endosymbiont Symbiodinium spp. (maximum quantum yield; F _v/F _m) was measured as an indicator of holobiont stress to hyposalinity. Colonies from each basin were assigned four salinity treatments [The Practical Salinity Scale (PSS) was used to determine salinity. Units are not assigned to salinity values because it is a ratio and has no unit as defined by UNESCO (UNESCO Technical papers no. 45, IAPSO Pub. Sci. No. 32, Paris, France, 1985)] (30, 20, 15, and 10) and salinities were reduced 2 per day from ambient (30) to simulate a natural salinity decrease. Colonies treated with salinities of 20 and 15 showed no decrease in F _v/F _m versus controls (i.e. 30), up to 5 days after reaching their target salinity. This indicates a greater ability to withstand reduced salinity for relatively extended periods of time in S. radians compared to other reef species. Within 1 day after salinity of 10 was reached, there was a significant reduction in F _v/F _m, indicating a critical threshold for hyposaline tolerance. At the lowest treatment salinity (10), F _v/F _m for the more estuarine, northeast-basin colonies were significantly higher than the most marine southwest-basin colonies (Twin Key Basin). Our results suggest that historical salinity ranges within basins determine coral population salinity tolerances.     

Diurnal variation in chlorophyll fluorescence of Thalassia testudinum seedlings in response to controlled salinity and light conditions

Diurnal variability in chlorophyll fluorescence caused by dynamic irradiance conditions is an important issue when using pulse amplitude modulation fluorometry to measure physiological conditions of plants at the landscape scale. We examined the use of slopes and y-intercepts of diurnal effective photochemical efficiency of photosystem II (PSII) (ΔF/F _m′) versus photosynthetically active radiation (PAR) regressions in addition to direct measurements of maximum photochemical efficiencies of PSII (F _v/F _m) values to assess physiological status of Thalassia testudinum seedlings in a controlled mesocosm study. Seedlings were exposed to two light treatments (full sun and 50–70 % light reduction) and three salinity treatments (20, 35, and 50). Measurements were taken at 0600, 0900, 1200, 1500, 1800, and 2100 hours in order to assess the diurnal variation in photochemical efficiency of PSII and PAR, with measurements at 2100 providing F _v/F _m. Results indicated significant effects of light and salinity on regression y-intercepts and measured F _v/F _m values. Shaded seedlings had higher values for both parameters, suggesting low-light acclimation. The highest salinity treatment resulted in significant reductions for both parameters, suggesting stress. Stress was also indicated by significant reductions in both seedling leaf growth and mean differences between seedling leaves and media osmolalities in the hypersaline treatments (152.0 ± 26.4 vs. 630 ± 40.2 mmol kg^?1 for the control treatments). Slopes of ΔF/F _m′ versus PAR significantly differed with varying light treatments, with full sun seedlings exhibiting shallower slopes than shaded seedlings, indicating higher efficiency of dissipation of excess energy (photoprotection). These experimental results confirm field data suggesting that diurnal ΔF/F _m′ versus PAR regressions are responsive to changes in the physiological status of T. testudinum and that the y-intercepts of diurnal regressions may be used as a proxy for F _v/F _m.     

A comparative seasonal study of two populations of Hypnea musciformis from the East and West Coasts of Florida,USA I. Growth and chemistry

Dry weight levels of the red alga Hypnea musciformis (Wulfen) Lamouroux from Atlantic and Gulf of Mexico coastal sites in florida, USA were lowest in the late winter and early spring, increasing through the summer and highest in the fall. There was a two-month lag in the Gulf coast population's dry weight pattern, indicating differing growth patterns. Chlorophyll a, phycoerythrin and phycoerythrin/chlorophyll a ratios were highest in the winter and lowest in the summer for both populations. Total pigment levels for H. musciformis from the Atlantic coast site were significantly greater than those of the Gulf coast. Protein and carbohydrate percentages were inversely related in both populations, with carbohydrate levels highest in summer and protein levels highest in winter. The Gulf coast population contained significantly more protein than the Atlantic coast plants. Carrageenan levels were highest in spring and lowest in fall, the Atlantic coast population generally had higher levels than the Gulf coast population. The differences in seasonal patterns and levels of the chemical constituents were reflected by distinct morphological characteristics for each population. The Atlantic coast population was larger, darker, more coarse in texture and possessed more crozier branch tips than the Gulf coast plants. These distinctions represent acclimitization responses that relate to habitat differences.     

Effects of salinity and possible interactions with temperature and pH on growth and photosynthesis of <Emphasis Type="Italic">Halophila johnsonii</Emphasis> Eiseman

The effects of salinity, temperature, and pH variations on growth, survival, and photosynthetic rates of the seagrass Halophila johnsonii Eiseman were examined. Growth and survival responses to salinity were characterized by aquarium experiments in which plants were exposed to seven different salinity treatments (0, 10, 20, 30, 40, 50, and 60 psu) during 15 days. Photosynthetic behavior was assessed for short-term salinity exposures (1 or 20 h) by incubation experiments in biological oxygen demand (BOD) bottles and by measuring photosynthesis versus irradiance (PI) responses in an oxygen electrode chamber. In the bottle experiments the possible effects of interactions between salinity and temperature (15, 25, and 35°C) or pH (5, 6, 7, and 8.2) were also examined. Growth and survival of H. johnsonii were significantly affected by salinity, with maximum rates obtained at 30 psu. Salinity also altered the parameters of the PI curves. Light-saturated photosynthesis (P _max) and the photosynthetic efficiency at subsaturating light (α) increased significantly up to an optimum of 40 psu, decreasing again at the highest salinities. Dark respiration rates and compensating irradiance (I _c) showed minimum values at 40 and 50 psu, while light-saturation point (I _k) was maximum at 30–50 psu. An interaction between salinity and temperature was not found although an increase of temperature alone produced an increase in α, P _max, respiration rates, and I _k. An interaction between salinity and pH was only found in the P _max response: P _max increased with pH=5 at 30 psu. In addition, reducing the pH increased α significantly. In the BOD bottles experiment a significant reduction in the dark respiration with decreasing pH was observed, but the opposite trend was observed in the photosynthetic rate. These results suggest that the endemic seagrass H. johnsonii could be negatively affected by hypo- or hypersalinity conditions, although salinity changes did not seem to alter the tolerance of this species to other environmental factors, such as temperature or pH.     

A comparative seasonal study of two populations of Hypnea musciformis from the East and West Coasts of Florida,USA II. Photosynthetic and respiratory rates

Respiration rates of Hypnea musciformis (Wulfen) Lamouroux in Florida, USA, generally increased with increased temperature. Gulf coast H. musciformis respired at significantly higher rates than the Atlantic coast population, which exhibited a region of temperature independence between 24°–32°C. Respiration rates were highest in the fall and winter, during the periods of rapid growth. Respiration rates were lowest in the summer indicating a period of storage and low metabolism. Photosynthetic responses to various levels of light and temperature indicated that the Gulf coast population was more tolerant to high light intensities than the Atlantic coast population. Maximum photosynthetic responses for both populations occurred between 24° and 32°C which corresponds to the shallow slope region of the respiration-temperature curves. The results indicate that water temperature rather than light intensity is a significant factor in modifying seasonal photosynthetic capacities. The greatest seasonal variation in photosynthetic responses occurred at the light-temperature levels of highest responses while little seasonal variation was demonstrated at tolerance limits.     

Effect of El Nĩno on demographic,morphological, and chemical parameters in turtle-grass (Thalassia testudinum): an unexpected test of indicators

We examined the response of demographic, morphological, and chemical parameters of turtle grass (Thalassia testudinum), to much-higher-than-normal rainfall associated with an El Niño event in the winter of 1997–1998. Up to 20 inches of added rain fell between December 1997 and March 1998, triggering widespread and persistent phytoplankton blooms along the west coast of Florida. Water-column chlorophyll concentrations estimated from serial SeaWiFS imagery were much higher during the El Niño event than in the previous or following years, although the timing and magnitude of phytoplankton blooms varied among sites. Seagrass samples collected in 1997, 1998, and 1999 provided an excellent opportunity to test the responsiveness of Thalassia to decline and subsequent improvement of water quality and clarity in four estuaries. Using a scoring technique based on temporal responsiveness, spatial consistency, and statistical strength of indicators, we found that several morphological parameters (Thalassia shoot density, blade width, blade number, and shoot-specific leaf area) were responsive and consistent measures of light stress. Some morphological parameters, such as rhizome apex density, responded to declines and subsequent improvement in water clarity, but lacked the statistical discriminating power necessary to be useful indicators. However, rhizome sugar, starch, and the total carbohydrate concentrations also exhibited spatially and temporally consistent variation as well as statistical strength. Because changes in shoot density, as well as water clarity, affect rhizome carbohydrate levels, a composite metric based on Thalassia shoot density and rhizome carbohydrate levels together is probably more useful than either parameter alone as an indicator of seagrass health.