Annual nitrogen budget of a temperate coastal barrier salt-marsh system along a productivity gradient at low and high marsh elevation

An annual nitrogen budget was established for a temperate back barrier salt-marsh system along a productivity gradient at low and high marsh elevation. We measured plant biomass and nitrogen content in three plant compartments to deduce plant N-allocation patterns. Measurements were done along a successional sequence in a salt-marsh system. In addition, N-mineralization, wet and dry atmospheric N-deposition and sediment N-deposition were measured.

Plant-species dominance changed along the successional sequence. In early stages, Elymus farctus and Spergularia media formed a large part of total plant biomass. Festuca rubra and Puccinellia maritima were dominant at intermediate stages, whereas Elymus pycnanthus and Limonium vulgare were dominant at late stages of succession. Shoot biomass was highest in June, whereas litter biomass was highest in September and December. Root biomass formed by far the largest fraction of total plant biomass, especially at a low-marsh elevation.

Wet deposition of nitrate and ammonium was 1.7 g N m⁻² yr⁻¹, whereas throughfall deposition (dry and wet deposition) amounted to 2.1–3.6 g N m⁻² yr⁻¹, and was positively related to the height of an artificial plant canopy. Sediment organic nitrogen deposition rate was 0.3–5.4 g N m⁻² yr⁻¹, and negatively related to marsh elevation. Nitrogen mineralization rate increased from 2.5–2.8 g N m⁻² yr⁻¹ in young marshes towards 8.0–12.7 g N m⁻² yr⁻¹ at older marshes, depending on marsh elevation.

At a low-marsh elevation, plant N-availability depended equally on tidal N, atmospheric N and mineralized N, especially in young marshes, whereas the decomposition pathway became more important in older marshes. Tidal N contributed most to ecoystem N-accumulation rate at early successional stages, whereas atmospheric N was more important at later stages. Tidal influence was low at high-marsh elevation sites. Here, atmospheric deposition was the dominant exogenous nitrogen source both in young and old marshes.     

Annual biomass and production of epiphytes in three monospecific seagrass communities of Thalassia hemprichii (Ehrenb.) Aschers

The biomass of epiphytes and seagrasses has been measured in relation to leaf age in three monospecific seagrass stands of Thalassia hemprichii (Ehrenb.) Aschers. in Papua New Guinea. From June 1981 through August 1982, biomass values for epiphytes at the three sites ranged from 5 to 70 g ADW m⁻² sediment surface at site 1, from 5 to 14 g ADW m⁻² at site 2, and from 3.5 to 7.0 g ADW m⁻² at the site 3. Annual mean epiphyte biomass values for the different sites were 1.3 g ADW m⁻² leaf surface at site 1, 1.7 g ADW m⁻² leaf surface at site 2, and 1.5 g ADW m⁻² leaf surface at site 3.

The annual mean standing crop of T. hemprichii leaves was highest at site 1 (103 g ADW m⁻². Values for site 2 and site 3 were 60 g ADW m⁻² and 41 g ADW m⁻², respectively.

Production of epiphytes was calculated in three different ways: firstly, by using biomass values for each specific leaf-age group, with corrections for colonization; secondly, by fitting the biomass values with a specific growth curve; and thirdly, by estimated the rate of biomass accumulation. On an area basis, production of epiphytes on leaves of T. hemprichii ranged from 0.55 to 3.97 g ADW m⁻² day⁻¹ at site 1, from 0.17 to 0.73 g ADW m⁻² day⁻¹ at site 2, and from 0.24 to 0.68 g ADW m⁻² day⁻¹ at site 3.     

Growth, nodulation and nitrate reductase activity in the aquatic legume Neptunia plena (L.) benth. At different external nitrate concentrations

The effects of different external nitrate concentrations (0 (control), 1, 50, 100, 500, 1000 and 20 000 mmol m⁻³) on growth, nodulation and nitrate-reductase activity (NRA) of inoculated Neptunia plena (L.) Benth. were examined.

Plants given 500 and 1000 mmol m⁻³ nitrate had greater (P < 0.05) shoot length, leaf, stem and root dry mass, and carbon and nitrogen contents than the controls and plants given 20 000 mmol m⁻³ nitrate. Nodule number was not significantly affected by nitrate concentration up to 50 mmol m⁻³, but 100 mmol m⁻³ nitrate reduced nodulation by 68% and concentrations above 100 mmol m⁻³ completely inhibited nodule development. Plants given 100–20000 mmol m⁻³ nitrate had a greater nitrate content per g leaf, stem and root dry mass (DM) than controls. Nitrate per g root DM did not increase with external nitrate concentration above 500 mmol m⁻³, but levels in leaf and stem were greater at 20 000 mmol m⁻³ nitrate than at all other concentrations. NRA per g leaf, stem and root fresh mass (FM) was greater for plants given 500–20000 mmol m⁻³ than for controls, but there was no significant increase with nitrate concentration above 500 mmol m⁻³. Substantial proportions of total plant nitrate and NRA were found in both root and shoot over the entire range of external nitrate concentrations given.

Findings for N. plena are compared with data obtained previously for terrestrial legumes.     

Transpiration rates of the understory annual Impatiens capensis (Balsaminaceae) in response to the autumnal changes in canopy leaf area

Leaf transpiration rates of Impatiens capensis were measured beneath a broadleaved deciduous forest canopy over successive growing seasons using a steady-state porometer. The transpiration measurements, which continued into early autumn, provided a framework for assessing whether I. capensis exhibits stomatal opening in response to the autumnal increase in available direct-beam radiation reaching the forest floor. The deciduous canopy LAI (leaf area index) decreased from a growing season maximum of 3.94 m² m⁻², while the understory I. capensis population located along a stream channel maintained LAI values ranging from 0.58 to 1.05 m² m⁻² late into the growing season. Late morning and early afternoon leaf transpiration rates during the months of June and July averaged about 8 μg cm⁻² s⁻¹, with a mean stomatal conductance of 0.5 cm s⁻¹. In August, leaf transpiration averaged almost 12 μg cm⁻² s⁻¹, with stomatal conductance exceeding 1.5 cm s⁻¹. However, beginning in early to mid-September, before canopy leaf-fall, the persistent green leaves of I. capensis exhibited a sharp decline in transpiration, possibly a result of decreasing vapor pressure deficits or non-lethal physiological damage induced by cold stress. This physiological decline offsets any advantage that could have been gained by the increased exposure to direct-beam radiation after canopy leaf-fall in mid-October. Although green leaf area and seed-bearing capsules may persist until the first frost in October or early November, there is no evidence of stomatal opening suggestive of carbon assimilation for enhanced seed development during this early autumn period. We conclude that the persistent green leaf area of I. capensis fails to exploit the increase in available direct-beam radiation in the final stage of its life cycle.     

The response of an emergent sedge Bolboschoenus medianus to salinity and nutrients

The potential for nutrient load (30, 100 and 350 g N m⁻² per year) to alter plant performance under saline conditions (control, 4.5, 9 and 13 dS m⁻¹) was examined in the sedge Bolboschoenus medianus. Relative growth rates (RGR) across nutrient loadings ranged from 30.2 to 41.8 mg g⁻¹ per day in controls and were reduced to 20.9–28.5 mg g⁻¹ per day by salinities of 13 dS m⁻¹. Whilst higher nutrient loads generally increased RGR, the response was smaller at higher salinities. Responses to salinity and nutrient load were specific. Nutrient load increased the RGR via increases in the leaf area ratio (LAR). The LAR ranged from 1.9 to 2.1 m² kg⁻¹ across salinity treatments at 30 g N m⁻² per year, and increased to 2.5–2.8 m² kg⁻¹ at 350 g N m⁻² per year. Salinity reduced the RGR via a reduction in the net assimilation rate (NAR). The NAR in control plants ranged from 14.7 to 16 g m⁻² per day across nutrient loadings and decreased to 11–12 g m⁻² per day at 13 dS m⁻¹. Carbon isotope discrimination of leaves decreased by 2–3‰ in response to 13 dS m⁻¹ at the lower nutrient loadings. A prominent response of B. medianus to salinity was a change in biomass allocation from culms to tubers. In contrast, the response to nutrient load was characterised by a shift in biomass allocation from roots to leaves.     

Solar irradiance level alters the growth of basil (Ocimum basilicum L.) and its content of volatile oils

The experiments were commenced in March 2003 and repeated in June 2003 at Sutton Bonington Campus, the University of Nottingham, UK, to investigate the effect of irradiance on plant growth and volatile oil content and composition in plants of basil. Four levels of irradiance were provided in the glasshouse, i.e. no shade (control), 25, 50 and 75% glasshouse irradiance. It suggested that basil grows well in full sun, however it can tolerate light shade. Heavy shading (75%) to provide a light integral of 5.3 moles m⁻² d⁻¹ resulted in shorter plants, lower weight, smaller leaf area, less shoots and higher specific leaf area, and also strongly reduced the rate of photosynthesis. There was no difference in CO₂ assimilation rate between 24.9 moles m⁻² d⁻¹ light integrals (no shading) and 13.5 moles m⁻² d⁻¹ light integrals (25% shading). Shading effectively reduced leaf temperature when air temperature was less than 30 °C, but heavy shading (75%) could not reduce leaf temperature when air temperature was above 36 °C due to a limitation of free air convection. Consequently, leaf temperature increased. Heavy shading strongly reduced total volatile oil content in fresh leaves, especially in older plants (shading treatment applied at the 3 leaf-pair growth stage). There were three chemical compounds in basil leaves, namely linalool, eugenol and methyl eugenol, influenced by the shading treatments. Linalool and eugenol, which contribute to the characteristic taste of basil, were significantly increased by high daily light integrals, whereas methyleugenol was increased by lower daily light integrals. No differences in the relative content of 1,8-cineole, one of the key aromatic compounds of Ocimum species, were observed.     

Effect of light-path length in outdoor flat plate reactors on output rate of cell mass and of EPA in Nannochloropsis sp.

The effect of light-path length (i.e. reactor width or thickness) of flat plate glass reactors on outdoor production of eicosapentaenoic acid (EPA) and cell mass of Nannochloropsis sp. was tested, using a range of light-paths from 1.3 to 17.0 cm. Volumetric productivity of cell mass and optimal, as well as maximal cell density which represents the highest sustainable cell density under the experimental conditions, decreased with increase in light-path. Daily areal output rate (g dry weight m⁻² day⁻¹) increased with increased light-path, in contrast with results obtained in similar reactors with Spirulina cultures, in which areal output rates increased when the light-path was reduced. Maximal areal productivity of Nannochloropsis sp. (12.8 and 22.4 g ash-free dry weight per day per m² of irradiated reactor surfaces, in winter and summer, respectively), reflecting maximal efficiency in light utilization, was obtained with the long light-paths, i.e. 10.4 and 17.0 cm. Increasing the light-path from 1.3 to 17.0 cm resulted in an increase in areal EPA productivity, from 66.7 to 278.2 mg m⁻² day⁻¹ in winter and from 232.1 to 515.7 mg m⁻² day⁻¹ in summer. This enhancement in areal productivity of EPA stems from increased productivity of cell mass which was associated with the increase in light-path. We concluded that the optimal light-path, which must be defined for each algal species, represents an important parameter which determines optimal culture density (i.e. resulting in the highest output rate of cell mass per irradiated reactor surface), as well as productivity of cell mass and cell products. Under our conditions the optimal light-path for culturing Nannochloropsis in vertical reactors was ca 10 cm.     

Sulfate reduction in lake sediments inhabited by the isoetid macrophytes Littorella uniflora and Isoetes lacustris

Sulfur cycling was examined in sediments inhabited with the isoetids Littorella uniflora and Isoetes lacustris in the oligotrophic soft-water Lake Kalgaard, Denmark. Based on short-term tracer incubations sulfate reduction was measured along a transect from the shore (0.6 m) to profundal sediments (4.6 m). The sulfate reduction rates were low (0.008–0.8 mmol m⁻² d⁻¹) in the sandy shallow sediments with low organic content (<1.3 mmol C g⁻¹ sed DW) and high redox potentials (>100 mV), whereas sulfate reduction was higher at the deeper sites (2.7–4.6 mmol m⁻² d⁻¹) with high organic content (max. 11.5 mmol C g⁻¹ sed DW) and lower redox potentials (<100 mV). High concentrations of dissolved organic carbon (DOC) were found in the low particulate organic sediments (up to 18.4 mM), and most of the DOC pool consisted of acetate (40–77%). Reoxidation of sulfides due to root oxygen release was probably important at all sites and a positive efflux of sulfate across the sediment–water interface was measured, attaining rates (up to 4.8 mmol m⁻² d⁻¹) similar to the sulfate reduction rates. Reoxidation of sulfides was also manifested by high fraction (>80%) of reduced sulfides being accumulated as elemental sulfur or pyrite (chromium reducible sulfur, CRS). The largest pools of CRS were found in high organic sediment with vertical distributions resembling those of the sulfate reduction rates. The overall effect of isoetid growth on sulfur cycling in the rhizosphere is a suppression of sulfate reduction in low organic sediments and the governing of sulfide reoxidation in sediments with higher organic content.     

Fungi on Leaf Blades of <Emphasis Type="Italic">Phragmites australis</Emphasis> in a Brackish Tidal Marsh: Diversity,Succession, and Leaf Decomposition

Although fungi are known to colonize and decompose plant tissues in various environments, there is scanty information on fungal communities on wetland plants, their relation to microhabitat conditions, and their link to plant litter decomposition. We examined fungal diversity and succession on Phragmites australis leaves both attached to standing shoots and decaying in the litter layer of a brackish tidal marsh. Additionally, we followed changes in fungal biomass (ergosterol), leaf nitrogen dynamics, and litter mass loss on the sediment surface of the marsh. Thirty-five fungal taxa were recorded by direct observation of sporulation structures. Detrended correspondence analysis and cluster analysis revealed distinct communities of fungi sporulating in the three microhabitats examined (middle canopy, top canopy, and litter layer), and indicator species analysis identified a total of seven taxa characteristic of the identified subcommunities. High fungal biomass developed in decaying leaf blades attached to standing shoots, with a maximum ergosterol concentration of 548 ± 83 μg g^–1 ash-free dry mass (AFDM; mean ± SD). When dead leaves were incorporated in the litter layer on the marsh surface, fungi experienced a sharp decline in biomass (to 191 ± 60 μg ergosterol g^–1 AFDM) and in the number of sporulation structures. Following a lag phase, species not previously detected began to sporulate. Leaves placed in litter bags on the sediment surface lost 50% of their initial AFDM within 7 months (k = −0.0035 day^–1) and only 21% of the original AFDM was left after 11 months. Fungal biomass accounted for up to 34 ± 7% of the total N in dead leaf blades on standing shoots, but to only 10 ± 4% in the litter layer. These data suggest that fungi are instrumental in N retention and leaf mass loss during leaf senescence and early aerial decay. However, during decomposition on the marsh surface, the importance of living fungal mass appears to diminish, particularly in N retention, although a significant fraction of total detrital N may remain associated with dead hyphae.     

Protein-based complex medium design for recombinant serine alkaline protease production

This work reports on the design of a complex medium based on simple and complex carbon sources, i.e. glucose, sucrose, molasses, and defatted-soybean, and simple and complex nitrogen sources, i.e. (NH₄)₂HPO₄, casein, and defatted-soybean, for serine alkaline protease (SAP) production by recombinant Bacillus subtilis carrying pHV1431::subC gene. SAP activity was obtained as 3050 U cm⁻³ with the initial defatted-soybean concentration C_soybean^o=20 kg m⁻³ and initial glucose concentration C_G^o=8 kg m⁻³; whereas, addition of the inorganic nitrogen source (NH₄)₂HPO₄ decreased SAP production considerably. Further increase in SAP production (3850 U cm⁻³) was obtained when sucrose was replaced with glucose at C_sucrose^o=15 kg m⁻³ and C_soybean^o=20 kg m⁻³. Nevertheless, when molasses was replaced with sucrose, the maximum activity was obtained with molasses having 10 kg m⁻³ initial sucrose concentration and C_soybean^o=15 kg m⁻³as 2130 U cm⁻³; moreover, when casein was replaced with defatted-soybean SAP production decreased considerably (ca. 250 U cm⁻³). Thereafter, the effects of inorganic ionic compounds were investigated; and except phosphate, inorganic compounds supplied from defatted-soybean were found to be sufficient for the bioprocess. The highest SAP activity was obtained as 5350 U cm⁻³ in the medium that contained (kg m⁻³): C_soybean^o=20, C_sucrose^o=15, C_Na2HPO4^o=0.021, and C_NaH2PO4^o=2.82, that was 6.5-fold higher than that of the SAP produced in the defined medium. By using the designed complex medium, oxygen transfer characteristics of the bioprocess were investigated; and, Damköhler number that is the oxygen transfer limitation increases with the cultivation time until t=14 h; and, at t>20 h both mass transfer and biochemical reaction resistances were effective. Overall oxygen transfer coefficient varied between 0.010 and 0.044 s⁻¹; volumetric oxygen uptake rate varied between 0.001 and 0.006 mol m⁻³ s⁻¹; and specific oxygen uptake rate varied between 0.0001 and 0.0022 mol kg⁻¹ DW s⁻¹ throughout the bioprocess.