Physiological and morphological effects of long-term ammonium or nitrate deposition on the green and red (shade and open grown) Sphagnum capillifolium

Sphagna are vulnerable to enhanced nitrogen (N) deposition. This article reports how the green (shade, under Calluna) and red (open grown) Sphagnum capillifolium respond to ammonium and nitrate additions of 56 kg N ha⁻¹ y⁻¹ over the background of 8-10 kg N ha⁻¹ y⁻¹ on an ombrotrophic bog in the Scottish Borders after seven years. Samples and measurements were made during a range of hydrated and desiccated conditions in the summer of 2009. Both ammonium and nitrate increased moss N concentration, but while ammonium decreased cross-sectional area of leaf hyaline cells and the leaf hyaline/chlorophyllose cell area ratio, nitrate increased both of them and capitulum pH. The changes in leaf morphology have not previously been reported to our knowledge. Especially the red S. capillifolium was affected by ammonium with significant changes in shoot N concentration (+71%) and the cross-sectional area of leaf chlorophyllose cells (+67%), and reductions in shoot dry weight (−30%) and fresh weight (−42%), the cross-sectional area of leaf hyaline cells (−24%), the leaf hyaline/chlorophyllose cell area ratio (−54%), as well as in chlorophyll fluorescence (measured as F_v/F_m) of desiccated capitulum (−65%) (all p < 0.05). These observations show that N deposition may affect moss physiology also through changes in leaf anatomy and morphology. The results also highlight potential sampling issues and causes of variability in N responses when collecting variably pigmented Sphagna.     

Contrasting responses of seagrass transplants (Posidonia australis) to nitrogen, phosphorus and iron addition in an estuary and a coastal embayment

Addition of nutrients to sediments has been proposed as a means of enhancing transplantation success in seagrasses. The effects of nutrient and iron additions to natural sediments on the growth and morphology of Posidonia australis transplants were evaluated in underwater plots in two contrasting environments: a coastal embayment (Princess Royal Harbour) with sandy sediments and little riverine input, and an estuary (Oyster Harbour) with organic-rich sediments and subject to seasonal river flow from a large rural catchment. Sixty six planting units spaced 1 m apart were transplanted in situ in each location. Nitrogen (N) and phosphorus (P) were added in a randomized factorial design using slow release fertilizer granules at the start of the experiment and repeated every 4-5 months for 2 years. In a concurrent experiment, chelated iron Fe EDTA was added to modify the sediment sulphur cycle.In Oyster Harbour, the addition of N significantly increased leaf N concentrations but reduced total biomass and biomass of leaves. Addition of P significantly increased leaf P concentrations and number of living leaves per transplant, leaf area, leaf length, length of longest rhizome axis and total rhizome length. Combined N + P addition resulted in a significant increase in leaf P concentrations and leaf area per plant only. In Princess Royal Harbour, addition of N produced significant increases in leaf variables (total and leaf biomass, number of shoots and living leaves, leaf area, and leaf length) but there were no significant differences observed in below ground plant parts (rhizomes). Addition of P had no significant effects on any growth measurements. Addition of N + P combined increased number of living leaves and leaf area significantly. δ¹⁵N in mature leaf tissue were significantly more negative for N and N + P treatments at both locations.Our results indicated that N limitation was occurring in the coastal embayment, Princess Royal Harbour whereas in the more estuarine Oyster Harbour, P was limiting plant growth. Addition of FeEDTA produced equivocal results at both sites and we suggest these results are confounded by the addition of N and C in the EDTA. We caution the use of nutrient addition to transplants of slow growing seagrasses such as P. australis without a thorough understanding of the nutrient status of the system, estuarine or coastal embayment, in which they are to be transplanted.     

Chemical profile of the North American native Myriophyllum sibiricum compared to the invasive M. spicatum

Myriophyllum spicatum L. is a nonindigenous invasive plant in North America that can displace the closely related native Myriophyllum sibiricum Komarov. We analyzed the chemical composition (including: C, N, P, polyphenols, lignin, nonpolar extractables, and sugars) of M. spicatum and M. sibiricum and determined how the chemistry of the two species varied by plant part with growing environment (lake versus tank), irradiance (full sun versus 50% shading), and season (July through September). M. spicatum had higher concentrations of carbon, polyphenols and lignin (C: 47%; polyphenols: 5.5%; lignin: 18%) than M. sibiricum (C: 42%; polyphenols: 3.7%; lignin: 9%) while M. sibiricum had a higher concentration of ash under all conditions (12% versus 8% for M. spicatum). Apical meristems of both species had the highest concentration of carbon, polyphenols, and tellimagrandin II, followed by leaves and stems. Tellimagrandin II was present in apical meristems of both M. spicatum (24.6 mg g⁻¹ dm) and M. sibiricum (11.1 mg g⁻¹ dm). Variation in irradiance from 490 (shade) to 940 (sun) μmol of photons m⁻² s⁻¹ had no effect on C, N, and polyphenol concentrations, suggesting that light levels above 490 μmol of photons m⁻² s⁻¹ do not alter chemical composition. The higher concentration of polyphenols and lignin in M. spicatum relative to M. sibiricum may provide advantages that facilitate invasion and displacement of native plants.     

δN and δC analysis of a Posidonia sinuosa seagrass bed

Potential food sources and dominant invertebrates and fishes were collected for the examination of variability in ¹³C/¹²C and ¹⁵N/¹⁴N to determine the sources of carbon available to consumers within a Western Australian Posidonia sinuosa-dominated seagrass bed. Autotrophs showed a wide distribution of δ¹³C values, with P. sinuosa at −11.3 ± 0.8‰ and macroalgae ranging from −16.6 to −31.7‰. This variation allowed us to successfully identify macroalgae as the main contributor of carbon to the trophic structure, although no distinction could be made between epiphytic macroalgae on seagrass, or allochthonous macroalgal sources. The range in δ¹⁵N ratios among potential food items at the trophic base was too small to make it useful as tracer of nitrogen flow pathways, but it consistently increased from macrophytes and detritus (4.1–6.8‰), to invertebrates (5.7–7.4‰) located near the middle of the food web, to fishes (8.3–11.9‰), with piscivorous species such as Leviprora inops generally having a higher ¹⁵N. δ¹³C of seston (−12.8‰) and sedimentary organic matter (−8.7‰) indicate that seagrass material is the main contributor to these two carbon pools, and that very little of it contributes to animal biomass.     

Decomposition of Phragmites australis rhizome in a shallow lake

Decomposition of Phragmites australis (Cav. Trin ex Steudel) rhizome was studied at Lake Fert?/Neusiedler See using the litter bag technique. Samples were analysed for rhizome dry mass, fibre (cellulose, hemicellulose, lignin) and nutrient content (C, N, P and S), litter-associated fungal biomass, potential microbial respiration (electron transport activity: ETS) and cellulolitic bacteria. The mass loss of decomposing rhizome was rapid in the initial period and only 13.6% of the dry mass remained at the end of the experiment during 953 days. Substantial quantities of C, N, S and P were lost during 99 days; only 18% C, 19% N, 14% S and 6.4% of the P remained after 953 days. Hemicellulose degraded more rapidly than the other fibres whilst the lignin had the slowest rate of decomposition. Bacteria were found to be the primary colonizers of plant detritus, which was followed by fungal growth. An antagonistic relationship was observed between bacteria and fungi. Fungal biomass as determined by ergosterol concentrations ranged between 4.1 and 420 μg g⁻¹ and peaked every year in September. The number of cellulolitic bacteria varied from 0 to 22 MPN g⁻¹ with higher values in summer. The ETS-activity ranged between 0.1 and 1.6 mg O₂ g⁻¹ h⁻¹. The changes in ETS-activity varied almost in parallel with the in situ temperature of the lake water.     

Few long-term effects of simulated climate change on volatile organic compound emissions and leaf chemistry of three subarctic dwarf shrubs

Climate change is exposing arctic ecosystems to higher temperature, increased nutrient availability and shading due to the increasing cloud cover and the expanding forests. In this work, we assessed how these factors affect the emissions of biogenic volatile organic compounds (BVOCs) from three subarctic dwarf shrub species in a field experiment after 18 treatment years. Of the studied species the willow Salix phylicifolia L. was the only isoprene-emitter with an emission potential of 16.1 ± 8.4 μg g⁻¹ dw h⁻¹ (at 30 °C and photosynthetic photon flux density of 1000 μmol m⁻² s⁻¹). The dwarf birch Betula nana L. had significant emissions of various reactive BVOCs, including monoterpenes and sesquiterpenes. The evergreen Cassiope tetragona (L.) D. Don emitted high amounts of mono- and sesquiterpenes. Due to chance, the temperature in the warming treatment (employing open-top plastic tents) and the unwarmed treatments was similar at the time of the measurements, and therefore long-term indirect effects of warming could be assessed without interference of temperature differences at the time of measurement. The long-term warming had not altered foliar N, P or condensed tannin concentrations, but it had led to other chemical changes detected in the near-infrared reflectance spectra of the leaves. Nevertheless, there were no significant differences in the BVOC emissions per unit leaf mass measured by the dynamic enclosure method and gas chromatography-mass spectrometry. Annual additions of NPK fertilizer, which mimicked increased nutrient availability, had accumulated P in the leaves of all species. In addition, fertilization marginally increased the leaf N concentration of B. nana. The only significant fertilization effect on BVOC emissions was a stimulation of emission of the sesquiterpene β-selinene from S. phylicifolia. The shading treatment obtained by dome-shaped hessian tents did not cause clear long-term changes in leaf chemistry or BVOC emissions. The only observed change was a marginally significant increase in sesquiterpene emissions from B. nana. When the treatment effects on long-term biomass changes in the different treatments were taken into account by proportioning the BVOC emissions to the biomass of each species in the field treatments, warming led to a significant increase and shading to a decrease in the total BVOC emissions per unit ground area from B. nana. These results highlight the importance of an integrated approach for realistic assessment of responses to climate change.     

Morphological and physiological adaptive traits of Mediterranean narrow endemic plants: The case of Centaurea gymnocarpa (Capraia Island,Italy)

A case study on Centaurea gymnocarpa Moris & De Not., a narrow endemic species, was carried out by analyzing its morphological, anatomical, and physiological traits in response to natural habitat stress factors under Mediterranean climate conditions. The results underline that the species is particularly adapted to the environment where it naturally grows. At the plant level, the above-ground/below-ground dry mass (1.73 ± 0.60) shows its investment predominately in the above-ground structure with a resulting total leaf area per plant of 1399 ± 94 cm². The senescent attached leaves at the base of the plant contribute to limit leaf transpiration by shading soil around the plant. Moreover, the dense C. gymnocarpa leaf pubescence, leaf rolling, the relatively high leaf mass area (LMA = 12.3 ± 1.3 mg cm⁻²) and leaf tissue density (LTD = 427 ± 44 mg cm⁻³) contribute to limit leaf transpiration, also postponing leaf death under dry conditions. At the physiological level, a relatively low respiration/photosynthesis ratio (R/P_N) in spring results from high R [2.26 ± 0.59 μmol (CO₂) m⁻² s⁻¹] and P_N [12.3 ± 1.5 μmol (CO₂) m⁻² s⁻¹]. The high photosynthetic nitrogen use efficiency [PNUE = 15.5 ± 0.4 μmol (CO₂) g⁻¹ (N) s⁻¹] shows the large amount of nitrogen (N) invested in the photosynthetic machinery of new leaves, associated to a high chlorophyll content (Chl = 35 ± 5 SPAD units). On the contrary, the highest R/P_N ratio (1.75 ± 0.19) in summer is due to a significant P_N decrease and increase of R in response to drought. The low PNUE [1.5 ± 0.2 μmol (CO₂) g⁻¹ (N) s⁻¹] in this season is indicative of a greater N investment in leaf cell walls which may contribute to limit transpiration. On the contrary, the low R/P_N ratio (0.05 ± 0.02) in winter is resulting from the limited enzyme activity of the respiratory apparatus [R = 0.23 ± 0.08 μmol (CO₂) m⁻² s⁻¹] while the low PNUE [3.5 ± 0.2 μmol (CO₂) g⁻¹ (N) s⁻¹] suggests that low temperatures additionally limit plant production. The experiment of the imposed water stress confirms that the C. gymnocarpa growth capability is in conformity with the severe conditions of its natural habitat, likewise as it may be the case with others narrow endemic species that have occupied niches with similar extreme conditions.     

Shade-induced response and recovery of the seagrass Posidonia sinuosa

The effect of shading on the seagrass Posidonia sinuosa Cambridge et Kuo was investigated to identify mechanisms that prolong its survival during periods of low light and permit its subsequent recovery. We also tested whether the responses were consistent in plants growing at different depths. Shade treatments were low (LS; 70 – 100% of ambient Photosynthetic Photon Flux Density), medium (MS; 12 – 39%) and heavy (HS; 5 – 4%) at the shallow (3 – 4 m) site, whilst the deep (7 – 8 m) site had no HS treatment. HS at the shallow and MS at the deep site were below minimum light requirements (MLR) for the long-term survival of P. sinuosa. Physiological, morphological and growth attributes were repeatedly measured during 198 d of shade treatments and a subsequent 384 d recovery period at ambient PPFD. Shoot density declined by 82% within 105 d under HS treatment, though 6% of shoots remained after 198 d. We estimate that complete shoot loss in HS would have taken 2 years. Rhizome sugar concentrations declined to 32 – 52% of the controls at the end of the most severe shading treatments but after shoot loss, sugar concentrations declined more slowly or increased, suggesting a return to positive carbon balance. In the treatments below MLR, shading induced changes in physiological, morphological and growth characteristics, including reduced leaf length and width, reduced δ¹³C and photosynthetic adaptation to low light (increased α, reduced E_k and ETR_max), though not consistently. After removal of shading, photosynthetic characteristics became more typical of high light adaptation, possibly induced by greater light penetration through the thinned canopy, including reversal of the changes in α, E_k and ETR_max and induction of non-photochemical quenching. Carbohydrate concentrations increased to ambient concentrations within 115 d at ambient PPFD. Recovery of shoot density was slow, remaining significantly reduced in the MS and HS treatments after 384 d recovery. Shoot density at the end of shading is an important determinant of the rate seagrass meadows will recover and we estimated that the moderately and heavily shaded meadows would require 3.5 to 5 years to recover.     

Characterization of the 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase multigene family of Malus domestica Borkh

Two 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase (ACO) genes have been cloned from RNA isolated from leaf tissue of apple (Malus domestica cv. Royal Gala). The genes, designated MD-ACO2 (with an ORF of 990 bp) and MD-ACO3 (966 bp) have been compared with a previously cloned gene of apple, MD-ACO1 (with an ORF of 942 bp). MD-ACO1 and MD-ACO2 share a close nucleotide sequence identity of 93.9% in the ORF but diverge in the 3′ untranslated regions (3′-UTR) (69.5%). In contrast, MD-ACO3 shares a lower sequence identity with both MD-ACO1 (78.5%) and MD-ACO2 (77.8%) in the ORF, and 68.4% (MD-ACO1) and 71% (MD-ACO2) in the 3′-UTR. Southern analysis confirmed that MD-ACO3 is encoded by a distinct gene, but the distinction between MD-ACO1 and MD-ACO2 is not as definitive. Gene expression analysis has shown that MD-ACO1 is restricted to fruit tissues, with optimal expression in ripening fruit, MD-ACO2 expression occurs more predominantly in younger fruit tissue, with some expression in young leaf tissue, while MD-ACO3 is expressed predominantly in young and mature leaf tissue, with less expression in young fruit tissue and least expression in ripening fruit. Protein accumulation studies using western analysis with specific antibodies raised to recombinant MD-ACO1 and MD-ACO3 produced in E. coli confirmed the accumulation of MD-ACO1 in mature fruit, and an absence of accumulation in leaf tissue. In contrast, MD-ACO3 accumulation occurred in younger leaf tissue, and in younger fruit tissue. Further, the expression of MD-ACO3 and accumulation of MD-ACO3 in leaf tissue is linked to fruit longevity. Analysis of the kinetic properties of the three apple ACOs using recombinant enzymes produced in E. coli revealed apparent Michaelis constants (K_m) of 89.39 μM (MD-ACO1), 401.03 μM (MD-ACO2) and 244.5 μM (MD-ACO3) for the substrate ACC, catalytic constants (K_cat) of 6.6 × 10⁻² (MD-ACO1), 3.44 × 10⁻² (Md-ACO2) and 9.14 × 10⁻² (MD-ACO3) and K_cat/K_m (μM s⁻¹) values of 7.38 × 10⁻⁴ μM s⁻¹ (MD-ACO1), 0.86 × 10⁻⁴ M s⁻¹ (MD-ACO2) and 3.8 × 10⁻⁴ μM s⁻¹ (MD-ACO3). These results show that MD-ACO1, MD-ACO2 and MD-ACO3 are differentially expressed in apple fruit and leaf tissue, an expression pattern that is supported by some variation in kinetic properties.     

Diurnal courses of net photosynthesis and photosystem II quantum efficiency of submerged Lagarosiphon major under natural light conditions

An optode device for net-photosynthesis measurements, based on oxygen-depending quenching of fluorescence from O₂-specific sensors, and PAM fluorometry have been used to study diurnal courses of net-photosynthesis and the F_v/F_m ratio of the submerged plant Lagarosiphon major. Plants were pre-cultivated and studied in large mesocosm flow-through outdoor tanks under 50% and 80% shade cloth, respectively. Growth under the different shade cloths resulted in similar light compensation points (∼20 μmol photons m⁻² s⁻¹), but strongly different light saturation levels, with about 150 μmol m⁻² s⁻¹ for plants grown under 80% shade cloth and about 350 μmol m⁻² s⁻¹ for plants grown under 50% shade cloth. Plants under both growth conditions showed a transient reduction of the maximum F_v/F_m value in the afternoon (down to 70% of the morning control values under 80% shade cloth and down to 85% under 50% shade cloth), which was not accompanied by a reduction of the net photosynthetic rate. This indicated that the fluorescence parameter F_v/F_m must not be a reliable indicator of the rate of photosynthesis under all conditions. The new photo-optical device became evidenced as a valuable tool not only for laboratory experiments, but also for field studies of gas exchange of submerged plants.     

Synergistic effects of high temperature and sulfide on tropical seagrass

To examine the synergism of high temperature and sulfide on two dominant tropical seagrass species, a large-scale mesocosm experiment was conducted in which sulfide accumulation rates (SAR) were increased by adding labile carbon (glucose) to intact seagrass sediment cores across a range of temperatures. During the initial 10 d of the 38 d experiment, porewater SAR in cores increased 2- to 3-fold from 44 and 136 μmol L^− 1 d^− 1 at 28-29 °C to 80 and 308 μmol L^− 1 d^− 1 at 34-35 °C in Halodule wrightii and Thalassia testudinum cores, respectively. Labile C additions to the sediment resulted in SAR of 443 and 601 μmol L^− 1 d^− 1 at 28-29 °C and 758 to 1,557 μmol L^− 1 d^− 1 at 34-35 °C in H. wrightii and T. testudinum cores, respectively. Both T. testudinum and H. wrightii were highly thermal tolerant, demonstrating their tropical affinities and potential to adapt to high temperatures. While plants survived the 38 d temperature treatments, there was a clear thermal threshold above 33 °C where T. testudinum growth declined and leaf quantum efficiencies (Fv/Fm) fell below 0.7. At this threshold temperature, H. wrightii maintained shoot densities and leaf quantum efficiencies. Although H. wrightii showed a greater tolerance to high temperature, T. testudinum had a greater capacity to sustain biomass and short shoots under thermal stress with labile C enrichment, regardless of the fact that sulfide levels in the T. testudinum cores were 2 times higher than in the H. wrightii cores. Tropical seagrass tolerance to elevated temperatures, predicted in the future with global warming, should be considered in the context of the sediment-plant complex which incorporates the synergism of plant physiological responses and shifts in sulfur biogeochemistry leading to increased plant exposure to sulfides, a known toxin.     

Cacao Cultivation under Diverse Shade Tree Cover Allows High Carbon Storage and Sequestration without Yield Losses

One of the main drivers of tropical forest loss is their conversion to oil palm, soy or cacao plantations with low biodiversity and greatly reduced carbon storage. Southeast Asian cacao plantations are often established under shade tree cover, but are later converted to non-shaded monocultures to avoid resource competition. We compared three co-occurring cacao cultivation systems (3 replicate stands each) with different shade intensity (non-shaded monoculture, cacao with the legume Gliricidia sepium shade trees, and cacao with several shade tree species) in Sulawesi (Indonesia) with respect to above- and belowground biomass and productivity, and cacao bean yield. Total biomass C stocks (above- and belowground) increased fivefold from the monoculture to the multi-shade tree system (from 11 to 57 Mg ha^-1), total net primary production rose twofold (from 9 to 18 Mg C ha^-1 yr^-1). This increase was associated with a 6fold increase in aboveground biomass, but only a 3.5fold increase in root biomass, indicating a clear shift in C allocation to aboveground tree organs with increasing shade for both cacao and shade trees. Despite a canopy cover increase from 50 to 93%, cacao bean yield remained invariant across the systems (variation: 1.1–1.2 Mg C ha^-1 yr^-1). The monocultures had a twice as rapid leaf turnover suggesting that shading reduces the exposure of cacao to atmospheric drought, probably resulting in greater leaf longevity. Thus, contrary to general belief, cacao bean yield does not necessarily decrease under shading which seems to reduce physical stress. If planned properly, cacao plantations under a shade tree cover allow combining high yield with benefits for carbon sequestration and storage, production system stability under stress, and higher levels of animal and plant diversity.     

Photosynthetic and morphological responses of eelgrass (Zostera marina L.) to a gradient of light conditions

Zostera marina L. (eelgrass) from Great Bay Estuary, New Hampshire and Maine (USA), was transplanted in outdoor mesocosms and subjected to four light treatments (100, 58, 34 and 11% surface irradiance, SI) between May and September 2003 to investigate the relationship between light availability and the growth and survival of eelgrass. Evaluating eelgrass seedlings and adult mature plants demonstrated no differences in photosynthetic response after 22 days of acclimation. During at least the first 19 days of shading, maximum electron transport rate (ETR_max) rate of eelgrass did not differ significantly between light treatments. After 40 days, a significant reduction in ETR_max and minimum saturating light was observed in plants growing at 34% SI and below. Morphological responses exhibited a linear increasing trend with greater light. 34% SI exhibited drastic reductions (to less than 25% of control) in rhizome growth, shoot density, shoot production, number of nodes per plant and plant weight at the end of the study (81 days). Shoot to root ratio at 34% SI increased by > 50%. Plants shaded to 58% SI showed no significant difference from the control in plant parameters except an increased rate of rhizome elongation. Our results link the lower shoot densities with shading to the slow growth rate of horizontal rhizomes and a total lack of lateral expansion at 11% SI. ETR_max declined over time in plants at 11% SI resulting in 81% mortality, no lateral branching and no morphological development, indicating that the minimum light required for long-term eelgrass growth and survival is greater than the previously suggested 11% SI. We demonstrate that eelgrass plants at these latitudes can persist at light levels of 58% SI and above, and are light-limited at 34% SI and below.     

Biomass and primary production of a 8–11 m depth meadow versus <3 m depth meadows of the seagrass Cymodocea nodosa (Ucria) Ascherson

Current knowledge about the abundance, growth, and primary production of the seagrass Cymodocea nodosa (Ucria) Ascherson is biased towards shallow (depth <3 m) meadows although this species also forms extensive meadows at larger depths along the coastlines. The biomass and primary production of a C. nodosa meadow located at a depth of 8–11 m was estimated at the time of maximum annual vegetative development (summer) using reconstruction techniques, and compared with those available from shallow meadows of this species. A depth-referenced data base of values at the time of maximum annual development was compiled to that end. The vegetative development of C. nodosa at 8–11 m depth was not different from that achieved by shallow (depth <3 m) meadows of this species. Only shoot density, which decreased from 1637 to 605 shoots m⁻², and the annual rate of elongation of the horizontal rhizome, which increased from 23 to 71 cm apex⁻¹ year⁻¹, were different as depth increased from <3 to 8–11 m. Depth was a poor predictor of the vegetative development and primary production of C. nodosa. The biomass of rhizomes and roots decreased with depth (g DW m⁻² = 480 (±53, S.E.) − 32 (±15, S.E.) depth (in m); R² = 0.12, F = 4.65, d.f. = 35, P = 0.0381) which made total biomass of the meadow to show a trend of decrease with depth but the variance of biomass data explained by depth was low. The annual rate of elongation of the horizontal rhizome showed a significant positive relationship with depth (cm apex⁻¹ year⁻¹ = 18 (±5.1, S.E.) + 5.0 (±1.33, S.E.) depth (in m); R² = 0.50, F = 14.07, d.f. = 14, P = 0.0021). As shoot size and growth did not change significantly with depth, the reduction of shoot density should drive any changes of biomass and productivity of C. nodosa as depth increases. The processes by which this reduction of C. nodosa abundance with depth occur remain to be elucidated.     

Treatment of poultry slaughterhouse wastewater in hybrid upflow anaerobic sludge blanket reactor packed with pleated poly vinyl chloride rings

In this study, the performance of 5.4 L hybrid upflow anaerobic sludge blanket (HUASB) reactor for treating poultry slaughterhouse wastewater under mesophilic conditions (29-35 °C), was investigated. After starting-up, the reactor was loaded up to an OLR of 19 kg COD/m³ d and achieved varied TCOD and SCOD removal efficiencies of 70-86% and 80-92%, respectively. The biogas was varied between 1.1 and 5.2 m³/m³ d with the maximum methane content of 72%. The maximum methane yield was 0.32 m³/kg COD_removed at an OLR of 9.27 kg COD/m³ d. Black matured granules of size between 2.5 and 5 mm were observed at the end of 225 d operation. RTD study showed the flow behavior was in mixed regime at the end of performance study. Step wise polynomial regression analysis was fitted well. Methanobacterium and Methanosaeta bacteria were dominant at the end of start-up whereas Methanosarcina, Cocci and rods were predominant at the end of performance studies.     

Shoot age as a confounding factor on detecting the effect of human-induced disturbance on Posidonia oceanica growth performance

The response of orthotropic rhizome elongation and primary production of Posidonia oceanica to anthropogenic perturbations and potential confounding effects of shoot age were assessed using a Linear Multilevel Model (LMM). This model examined the confounding effect of age by comparing the estimates of impact and variance components obtained by excluding and including Age as an explanatory variable. Age had a negative effect on rhizome elongation and primary production with an annual decrease of 0.6 mm y^− 1 and 7 mg dw y^− 1 respectively. According to the LMM when age effect was omitted, the differences between disturbed and control locations in rhizome elongation and primary production were 2.62 mm y^− 1 and 0.044 g dw y^− 1 respectively. These effects were statistically not significant. On the contrary, when age effect was included in the statistical model, impacts became evident for both variables, with significant differences between disturbed and control locations of 5.85 mm y^− 1 and 0.081 g dw y^− 1 for rhizome elongation and primary production, respectively. Thus, particular attention should be paid to the potential confounding effect of shoots age in analyses of impacts on growth performance of P. oceanica.     

Seasonal variations in bulk tissue, fatty acid and monosaccharide δC values of leaves from mesotrophic grassland plant communities under different grazing managements

Leaves of 26 grass, herb, shrub and tree species were collected from mesotrophic grasslands to assess natural variability in bulk, fatty acid and monosaccharide δ¹³C values under different grazing management (cattle- or deer-grazed) on three sample dates (May, July and October) such that interspecific and spatiotemporal variations in whole leaf tissues and compound-specific δ¹³C values could be determined. The total mean leaf bulk δ¹³C value for plants was −28.9‰ with a range of values spanning 7.5‰. Significant interspecific variation between bulk leaf δ¹³C values was only determined in October (P = <0.001) when δ¹³C values of the leaf tissues from both sites was on average 1.5‰ depleted compared to during July and May. Samples from May were significantly different between fields (P = 0.03) indicating an effect from deer- or cattle-grazing in young leaves. The average individual monosaccharide δ¹³C value was 0.8‰ higher compared with whole leaf tissues. Monosaccharides were the most abundant components of leaf biomass, i.e. arabinose, xylose, mannose, galactose and glucose, and therefore, fluctuations in their individual δ¹³C values had a major influence on bulk δ¹³C values. An average depletion of ca. 1‰ in the bulk δ¹³C values of leaves from the deer-grazed field compared to the cattle-grazed field could be explained by a general depletion of 1.1‰ in glucose δ¹³C values, as glucose constituted >50% total leaf monosaccharides. In October, δ¹³C values of all monosaccharides varied between species, with significant variation in δ¹³C values of mannose and glucose in July, and mannose in May. This provided an explanation for the noted variability in the tissue bulk δ¹³C values observed in October 1999. The fatty acids C_16:0, C_18:2 and C_18:3 were highly abundant in all plant species. Fatty acid δ¹³C values were lower than those of bulk leaf tissues; average values of −37.4‰ (C_16:0), −37.0‰ (C_18:2) and −36.5‰ (C_18:3) were determined. There was significant interspecific variation in the δ¹³C values of all individual fatty acids during October and July, but only for C_18:2 in May (P = <0.05). This indicated that seasonal trends observed in the δ¹³C values of individual fatty acids were inherited from the isotopic composition of primary photosynthate. However, although wide diversity in δ¹³C values of grassland plants ascribed to grazing management, interspecific and spatiotemporal influences was revealed, significant trends (P = <0.0001) for fatty acid and monosaccharide δ¹³C values: δ¹³C_16:0 < δ¹³C_18:2 < δ¹³C_18:3 and δ¹³C_arabinose > δ¹³C_xylose > δ¹³C_glucose > δ¹³C_galactose, respectively, previously described, appear consistent across a wide range of species at different times of the year in fields under different grazing regimes.     

Leachate treatment and greenhouse gas emission in subsurface horizontal flow constructed wetland

Organic and nitrogen removal efficiencies in subsurface horizontal flow wetland system (HSF) with cattail (Typha augustifolia) treating young and partially stabilized solid waste leachate were investigated. Hydraulic loading rate (HLR) in the system was varied at 0.01, 0.028 and 0.056 m³/m² d which is equivalent to hydraulic retention time (HRT) of 28, 10 and 5 d. Average BOD removals in the system were 98% and 71% when applied to young and partially stabilized leachate at HLR of 0.01 m³/m² d. In term of total kjeldahl nitrogen, average removal efficiencies were 43% and 46%. High nitrogen in the stabilized leachate adversely affected the treatment performance and vegetation in the system. Nitrogen transforming bacteria were found varied along the treatment pathway. Methane emission rate was found to be highest at the inlet zone during young leachate treatment at 79–712 mg/m² d whereas CO₂ emission ranged from 26–3266 mg/m² d. The emission of N₂O was not detected.     

Phospholipid synthesis and exchange in castor bean endosperm homogenates

Leaves on a bush of Hyptis emoryi Torr. varied in length from less than 1 cm when development occurred in full sunlight (e.g. 40 Mjoules m⁻²) to over 7 cm when the total daily solar irradiance was less than 3 Mjoules m⁻². The 1-cm sun leaves were 3-fold higher than the 7-cm shade leaves in chlorophyll per unit area, mesophyll thickness, and the internal to external leaf area ratio (A^mes/A). The higher A^mes/A caused a 1.2-cm leaf to have a 3-fold lower CO₂ liquid phase resistance than did a 7.1-cm leaf. Large thin shade leaves captured photosynthetically active radiation effectively (less than 7% passed through), but were not adapted to full sunlight. Specifically, when a 6.9-cm leaf was placed at 910 w m⁻² for 30 min, its temperature exceeded that of the air by nearly 8 C. For the common daytime air temperatures above 30 C for this desert shrub, large shade leaves would have temperatures far in excess of that optimum for photosynthesis for H. emoryi, 29 to 32 C.     

Physiological,morphological and anatomical trait variations between winter and summer leaves of Cistus species

Morphological, anatomical and physiological summer and winter leaf traits of Cistus incanus subsp. incanus, C. salvifolius and C. monspeliensis growing at the Botanical garden of Rome were analyzed. With regard to differences between summer and winter leaves of the considered species, leaf thickness (L) was 21% higher in summer than in winter leaves (mean of the considered species) and this increase was mostly the result of the increased palisade parenchyma thickness over the spongy parenchyma one (24 and 16% higher in summer than in winter leaves, respectively). Leaf mass area (LMA) and leaf tissue density (LTD) were 38% and 17% higher in summer than in winter leaves, respectively (mean of the considered species). The photosynthetic rate (P_N), stomatal conductance (g_s) and chlorophyll content (Chl) of summer leaves were 54%, 17% and 14% lower, respectively, than in winter leaves. C. monspeliensis summer leaves had the highest LMA, LTD, adaxial cuticle thickness (14.6 ± 1.8 mg cm⁻², 1091 ± 94 mg cm⁻³, and 5.8 ± 1.7 μm, respectively) and the lowest mesophyll intercellular spaces (f_ias 38 ± 3%). Moreover, C. monspeliensis had the highest P_N in summer (2.6 ± 0.1 μmol m⁻² s⁻¹) and C. incanus the highest P_N and WUE (84% and 59% higher than the other species) in the favorable period, associated to a higher f_ias (42 ± 2%). C. salvifolius had the highest P_N (54% higher than the other species) in winter. The plasticity index could allow a better interpretation of the habitat preference of the considered species. The physiological plasticity (PI_p = 0.39, mean value of the considered species) was higher than the morphological (PI_m = 0.22, mean value) and anatomical (PI_a = 0.13, mean value) plasticity. Moreover, among the considered species, C. salvifolius and C. incanus are characterized by a larger PI_a (0.14, mean value) which seems to be correlated with their wider ecological distribution and the more favorable conditions of the environments where they naturally occur. The highest PI_m (0.29) of C. monspeliensis indicates that it can play a high adaptive role in highly stressed environments, like fire degraded Mediterranean areas in which it occurs.