Contribution of several nitrogen sources to growth of Alexandrium catenella during blooms in Thau lagoon,southern France

A monitoring program with a weekly sampling frequency over a 15-month period indicates that urea concentrations above a certain threshold level may trigger the blooms of Alexandrium catenella in Thau lagoon. However, urea concentrations were also sometimes related to ammonium and dissolved organic nitrogen concentrations, indicating that the role of urea may not be a direct one. An original approach is used to assess the relative contribution of several nitrogen sources (nitrate, nitrite, ammonium, urea) to growth of A. catenella by comparing nitrogen uptake rates to nitrogen-based growth rates estimated from dilution experiments during four blooms over a 4-year period (2001–2004) in Thau lagoon. Nitrate and nitrite contributed 0.1–14% and 0.1–5% respectively of growth requirements. Ammonium and urea were the main N sources fueling growth of A. catenella (30–100% and 2–59%, respectively). Indirect estimates indicated that an unidentified N source could also contribute significantly to growth at specific times. Concerning ammonium and urea uptake kinetics, half-saturation constants varied between 0.2 and 20 μgat N L⁻¹ for ammonium and between 0.1 and 44 μgat N L⁻¹ over the 4-year period, indicating that A. catenella can have a competitive advantage over other members of the phytoplankton even under low concentrations of ammonium and urea. However, the observed large changes in ammonium and urea uptake kinetics on a short time scale (days) during blooms preclude more precise estimates of those contributions to growth and require further investigation.     

Alexandrium catenella and Alexandrium minutum blooms in the Mediterranean Sea: Toward the identification of ecological niches

Annual recurrent blooms of the toxic dinoflagellates Alexandrium catenella and Alexandrium minutum were detected from 2000 to 2003 in harbours along the Catalan coast. The interrelation study between the occurrence of the blooms and specific external conditions at the study sites demonstrated that different factors are required for the bloom of each Alexandrium species. Concentrations higher than 10⁵ cells l⁻¹ of A. catenella were only detected in Tarragona harbour. These blooms were associated with water surface temperature between 21 and 25 °C and salinities of around 34 psu or higher than 37 psu. A. minutum appeared widely spread along the Catalan coast, though the most intensive and recurrent blooms of this species were observed in Arenys de Mar harbour. Concentrations of millions of cells per litre of A. minutum were associated with water temperatures below 14 °C and salinities of around 34–36 psu. A. minutum cell densities showed a positive significant correlation with NO₃ but a negative correlation with NH₄. On the other hand, A. catenella blooms dominated when both NO₃ and NH₄ levels were high. The prevailing inorganic nitrogen form (NO₃ vs. NH₄) could explain why these two species rarely coincide in the same harbours. Accumulation of cysts in the sediment was found to be an important potential factor for the recurrence of these species. The 4.3 × 10³ A. catenella cysts cm⁻³ of wet sediment in Tarragona harbour and the 3.02 × 10³ A. minutum cysts cm⁻³ of wet sediment in Vilanova harbour were the highest concentrations observed from the cyst study. Confined waters such as harbours play an important role as reservoirs for the accumulation of cysts and vegetative cells, which contributes to the expansion of these dinoflagellates in the region. However, the particular environmental conditions are also decisive factors of bloom intensity.     

NITROGENOUS NUTRITION OF ALEXANDRIUM CATENELLA (DINOPHYCEAE) IN CULTURES AND IN THAU LAGOON,SOUTHERN FRANCE1

Alexandrium catenella (Whedon et Kofoid) Balech was isolated from Thau lagoon (northern Mediterranean) and its growth and uptake characteristics measured for nitrate, ammonium, and urea. Although affinity constants did not indicate a preference for ammonium over nitrate, there was a strong inhibition of nitrate uptake by ammonium when both nitrogen (N) sources were present. Nitrogen budgets during growth in cultures revealed major imbalances between decreases in dissolved N and increases in particulate N, indicating excretion of dissolved organic N during the early part of the growth phase and uptake during the later part. A quasi‐unialgal bloom in November 2001 (4×10⁶ cells·L^?1) allowed measurements of uptake of nitrate, nitrite, ammonium, and urea; net and gross growth rate of A. catenella; and grazing rates on this organism. The affinity constants indicate that it is not a strong competitor for the N nutrients tested when these are in low concentrations (<10 μgat N·L^?1), compared with other members of the phytoplankton community. Indirect evidence from cultures indicate that dissolved organic N compounds could be important in triggering those blooms. Finally, the strongly unbalanced growth observed in the field indicates that A. catenella exhibits a storage rather than a growth response to a nutrient pulse and is adapted to low frequency events such as the passage of frontal disturbances. The disappearance of A. catenella was due to grazing that balanced growth at the peak of the bloom.     

Towards an optimal sampling strategy for Alexandrium catenella (Dinophyceae) benthic resting cysts

The study proposes methodological developments to optimize sampling strategy of resting cysts of Alexandrium catenella to estimate their abundance with a predefined error. This work also aims to provide information on spatial distribution of resting cysts in sediments. The distribution mode of A. catenella resting cysts related to the abundance variability was studied through sediment cores sampling on four different spatial scales and using Ludox CLX gradient density method. The quantification method underestimates by a factor of 2 the resting cysts abundance in one gram of sediment. Application of Taylor's power law allowed us to define a compromise between sampling effort and abundance estimation error. In the case of A. catenella resting cysts from Thau lagoon, the optimal sampling strategy consists of sampling 10 stations on a surface of 2 km² for a given coefficient of variability (C) of 15%, sampling 3 sediment cores at each station (C = 30%) and counting only one replicate by core (C = 18%). Results related to the application of Taylor's power law are closely dependent on resting cyst density and aggregation in a given sediment. In our area, A. catenella resting cysts are mainly observed in the upper 3 cm of sediment. Horizontally, their heterogeneity is lower on 10 cm² surface and tends to stabilize itself beyond a surface of 10 m². Each author has to carry out this pre-sampling effort for his own resting cysts-forming species, in his own area, in order to increase accuracy of resting cyst mapping.     

Comparing diatom and Alexandrium catenella/tamarense blooms in Thau lagoon: Importance of dissolved organic nitrogen in seasonally N-limited systems

Diatom blooms in Thau lagoon are always related to rain events leading to inputs of inorganic nutrients such as phosphate, ammonium and nitrate through the watershed with time lags of about 1 week. In contrast, blooms of Alexandrium catenella/tamarense can occur following periods of 3 weeks without precipitation and no significant input of conventional nutrients such as nitrate and phosphate. Field results also indicate a significant drop (from 22–25 to 15–16 μM over 3 days) in dissolved organic nitrogen (DON) at the bloom peak, as well as a significant inverse relationship between A. catenella/tamarense cell density and DON concentrations that is not apparent for diatom blooms. Such dinoflagellate blooms are also associated with elevated (6–9 μM) ammonium concentrations, a curious feature also observed by other investigators, possibly the results of ammonium excretion by this organism during urea or other organic nitrogen assimilation.The potential use of DON by this organism represents short cuts in the nitrogen cycle between plants and nutrients and requires a new model for phytoplankton growth that is different from the classical diatom bloom model. In contrast to such diatom blooms that are due to conventional (nitrate, phosphate) nutrient pulses, Alexandrium catenella/tamarense blooms on the monthly time scale are due to organic nutrient enrichment, a feature that allows net growth rates of about 1.3 d⁻¹, a value higher than that generally attributed to such organisms.     

Temperature as a factor regulating growth and toxin content in the dinoflagellate Alexandrium catenella

Controlled laboratory culture of Alexandrium catenella was used to determine the effects of a range of temperatures between 10 and 16 °C on the growth and saxitoxin content of this dinoflagellate, using strain ACC02 isolated from seawater at Aysen, XI Region, Southern Chile. Cell cultures were made using L1 culture medium at 30‰ salinity, and a photon flux density of 59.53 μmol m² s⁻¹. The results showed that the duration of the exponential growth phase was determined by the experimental temperature, with maximum cell concentrations obtained at 12 °C; significantly lower cell concentrations and growth rates were obtained at 16 °C. Cell dry weight and chlorophyll a values followed cell growth trends. The toxicity of A. catenella was variable at all the experimental temperatures, with a tendency towards having an inverse relation to temperature, with the highest values occurring at 10 °C and the lowest at 16 °C. The optimal range of temperature for the growth of the Chilean strain of A. catenella differed from rates reported for this species isolated at other latitudes, and was correlated with natural temperature conditions predominant in the environment from which it was isolated. The inverse relation of toxicity with temperature in the laboratory was broadly reflected in observations on the toxicity of this dinoflagellate in the field, and coincided with results from the literature.     

Comparison of the functional and numerical responses of resistant versus non-resistant populations of the copepod Acartia hudsonica fed the toxic dinoflagellate Alexandrium tamarense

The functional and numerical responses of grazers are key pieces of information in predicting and modeling predator–prey interactions. It has been demonstrated that exposure to toxic algae can lead to evolved resistance in grazer populations. However, the influence of resistance on the functional and numerical response of grazers has not been studied to date. Here, we compared the functional and numerical responses of populations of the copepod Acartia hudsonica that vary in their degree of resistance to the toxic dinoflagellate Alexandrium tamarense. In common environment experiments carried out after populations had been grown under identical conditions for several generations, female copepods were offered solutions containing different concentrations of either toxic A. tamarense or the non-toxic green flagellate Tetraselmis sp. ranging from 25 to 500 μgC L⁻¹, and ingestion and egg production rates were measured. Throughout most of the range of concentrations of the toxic diet, copepod populations that had been historically exposed to toxic blooms of Alexandrium exhibited significantly higher ingestion and egg production rates than populations that had little or no exposure to these blooms. In contrast, there were no significant differences between populations in ingestion or egg production for the non-toxic diet. Hence, the between population differences in functional and numerical response to A. tamarense were indeed related to resistance. We suggest that the effect of grazer toxin resistance should be incorporated in models of predator and toxic prey interactions. The potential effects of grazer toxin resistance in the development and control of Alexandrium blooms are illustrated here with a simple simulation exercise.     

Relative Uptake of Urea and Ammonium by Dinoflagellates or Cyanobacteria in Shrimp Mesocosms

The relative role of the organic nitrogen source, urea, versus ammonium as a nitrogen source for two species of dinoflagellates was compared with one species of cyanobacteria. Experiments were conducted opportunistically in nutrient-rich marine water during blooms of 34either cyanobacteria or dinoflagellates in outdoor mesocosms. These replicate mesocosms, which were stocked with shrimp fed high-protein formulated feeds, contained high biomasses of phytoplankton (mean chlorophyll a concentrations, 439.2–811.2 μg l⁻¹). ¹⁵N-urea and ammonium uptake rates for dinoflagellate-dominated blooms (Gymnodinium pulchellum-complex (Larsen), Karlodinium micrum (Larsen) (Dinophyceae)) were compared with blooms of the cyanobacterium, Romeria sp. (Cyanophyceae) in mesocosms with mean urea and ammonium concentrations ranging from 2.32 to 3.24 μM, and 7.39 to 64.85 μM, respectively. Urea uptake rates were significantly (p < 0.005) lower than ammonium uptake rates irrespective of which algal species dominated the bloom. Additionally urea uptake rates were not significantly higher in G. pulchellum-complex or K. micrum-dominated blooms than in Romeria sp. blooms. These results suggest that G pulchellum complex and K. micrum may not be gaining a competitive advantage in waters high in dissolved organic matter simply by preferentially utilizing urea. The periodic dominance of these species in highly organic environments, such as shrimp ponds, is likely to have a more complex explanation.     

The dinoflagellate Alexandrium minutum in Cape Town harbour (South Africa): Bloom characteristics, phylogenetic analysis and toxin composition

A novel bloom of Alexandrium minutum occurred in an inner basin of the Cape Town harbour from November 2003 to February 2004. Cellular concentrations reached a maximum of 1.4 × 10⁸ cells l⁻¹ during the mid-December period with corresponding chlorophyll a concentrations of 243 mg m⁻³. Primary productivity measurements conducted during the latter part of the bloom revealed a maximum assimilation number of 11.17 mg C mg Chl a⁻¹ h⁻¹ during the middle of the day. Productivity during this post-peak period was sustained largely by the reduced nitrogen species NH₄ and urea (96%) as measured using ¹⁵N tracer techniques. The large subunit ribosomal DNA sequence of A. minutum isolates from Cape Town harbour was identical to conspecifics collected in Western Europe and in Australia. The composition of tetrahydropurine neurotoxins associated with paralytic shellfish poisoning (PSP) was limited to gonyautoxins (GTX1-GTX4). This profile combined with evidence of a low toxin cell quota (1.5 fmol GTX cell⁻¹) supports a close association of this taxon with other members of the A. minutum species complex, particularly from Europe. Toxin analysis from black mussels collected during this bloom indicated that the accumulated PSP toxins originated from A. minutum and not from Alexandrium catenella as is most often the case along the South African coast.     

Nutrition of the giant clam Tridacna gigas (L.). II. Relative contributions of filter-feeding and the ammonium-nitrogen acquired and recycled by symbiotic alga towards total nitrogen requirements for tissue growth and metabolism

We compare the relative contributions of filter-feeding and ammonium-nitrogen translocated from algal symbionts (zooxanthellae) towards the total nitrogen requirements for tissue growth and metabolism in the giant clam Tridacna gigas. Isotope enrichments suggest that zooxanthellae effected most if not all primary assimilation from high concentrations ( 15 μM) of ¹⁵N-ammonium. There was a net daily uptake of ammonium from natural seawater, which was modulated according to nutritional history, light history, ammonium concentration, and possible biological rhythmicity. Zooxanthellae also assimilated nitrogen excreted by host tissues, including ¹⁵N absorbed from ingested alga. Nearly 100% of the ¹⁵N subsequently released from zooxanthellae was incorporated in host tissues, with no significant loss from the clam over at least 10 days. Zooxanthellae therefore conserve and recycle essentially all nitrogenous end-products within T. gigas, affording giant clams a nutritional advantage over non-symbiotic bivalves. In further contrast with heterotrophic bivalves from particle-rich environments, T. gigas show longer gut passage times and high net and gross efficiencies (78.1 ± 9.4% and 99.2 ± 0.4%, respectively) with which organic nitrogen is absorbed from ingested particles. Filter-feeding and/or other non-autotrophic processes were essential as a source of nitrogen to maintain observed rates of juvenile growth, but declined in significance with increasing clam size. Even under cloudy conditions of limited irradiance, the total contribution of nitrogen acquired as ammonium by zooxanthellae exceeded the contribution from ingested particles in T. gigas larger than about 0.1 g, increasing to at least 45% of total nitrogen requirements in clams of 10 g. Ammonium-nitrogen translocated from zooxanthellae also satisfied all basal requirements for nitrogen in particle-starved clams larger than 0.1 g. Irrespective of size, ingested particles and ammonium-nitrogen acquired by zooxanthellae together supplied about 70% of the total observed nitrogen requirements. We stress the need to determine effects of irradiance on uptake of dissolved inorganic nitrogen, and to quantify dissolved organic matter as a potential additional significant source of nitrogen, that might further explain the high rates of growth in this clam species.     

Bench-scale fermentation of Trichoderma viride on wastewater sludge: Rheology, lytic enzymes and biocontrol activity

Conidiation and lytic enzyme production by Trichoderma viride at different solids concentration of pre-treated municipal wastewater sludge was examined in a 15-L fermenter. The maximum conidia concentration (5.94 × 10⁷ CFU mL⁻¹ at 96 h) was obtained at 30 g L⁻¹ suspended solids. The maximum lytic enzyme activities were achieved around 12–30 h of fermentation. Bioassay against a fungal phytopathogen, Fusarium sp. showed maximum activity in the sample drawn around 96 h of fermentation at 30 g L⁻¹ suspended solids concentration. Entomotoxicity against spruce budworm larvae showed maximum value ≈17290 SBU μL⁻¹ at 30 g L⁻¹ suspended solids concentration at the end of fermentation (96 h). Plant bioassay showed dual action of T. viride, i.e., disease prevention and growth promotion. The rheological analyses of fermentation sludges showed the pseudoplastic behaviour. In order to maintain required dissolved oxygen concentration ≥30%, the agitation and aeration requirements significantly increased at 35 g L⁻¹ compared to 30 and 25 g L⁻¹. The oxygen uptake rate and volumetric oxygen mass transfer coefficient, k_La at 35 g L⁻¹ did not increase in comparison to 30 g L⁻¹ due to rheological complexity of the broth during fermentation. Thus, the successful fermentation operation of the biocontrol fungus T. viride is a rational indication of its potential for mass-scale production for agriculture and forest sector as a biocontrol agent.     

The chemical nature of host hatching factors in the monogenean skin parasites Entobdella soleae and Acanthocotyle lobianchi

Fully-developed eggs of the monogenean Entobdella soleae from the skin of the common sole (Solea solea) hatch when treated with dilute solutions of urea or ammonium chloride in sea water. There is some evidence that arginine may stimulate hatching but the eggs do not respond when treated with sea water solutions containing trimethylamine oxide or glutamine. Sole skin mucus contains sufficient urea to stimulate hatching but insufficient ammonia. Solutions of urea in sea water stimulate hatching in the monogenean parasite Acanthocotyle lobianchi found on ray skin. Sea water solutions containing ammonium chloride and trimethylamine oxide failed to hatch the eggs of A. lobianchi and the eggs were also insensitive to various amino acids made up at concentrations found in host mucus. Experiments with urease confirmed that urea in ray ventral skin mucus is the host hatching factor for A. lobianchi. Skin mucus from the common sole failed to stimulate hatching in A. lobianchi. The role as hatching factors of excretory products in host gill effluent, skin mucus and urine is discussed.     

A comparative analysis of Alexandrium catenella/tamarense blooms in Annaba Bay (Algeria) and Thau lagoon (France); phosphorus limitation as a trigger

Environmental conditions ultimately leading to blooms of the toxic dinoflagellate Alexandrium catenella/tamarense were investigated at two Mediterranean sites (Annaba Bay, Algeria and Thau lagoon, France). Three years were examined in details: 1992 (a pre-Alexandrium period), 2002 (a year with the first bloom in Annaba) and 2010 (a year with a major bloom in Annaba). Most conditions were similar, but ammonium concentrations were much higher in Annaba (up to 100 μM) than in Thau (up to 10 μM). First records of A. catenella/tamarense were in 1995 for Thau and 2002 for Annaba, and coincided with soluble reactive phosphorus (SRP) decreasing below a concentration of about 1 μM. No other environmental variable could be related to those blooms. Thus, it is likely that the large reductions in SRP at both sites led to phosphorus limitation of a certain number of phytoplankton species and favored the development of A. catenella/tamarense.     

Effects of food nitrogen content and concentration on the forms of nitrogen excreted by the calanoid copepod, Acartia tonsa

Nitrogen excreted as ammonium, urea, and dissolved primary amines (DPA), and nitrogen ingested by the planktonic calanoid copepod, Acartia tonsa, were measured while fed 4 foods with different N/C ratios in high (500 μg C l^− 1) and low (50 μg C l^− 1) concentrations. Adult copepods were fed the ciliate, Uronema marinum (N/C = 0.26), the diatom, Thalassiosira weissflogii, in log-phase growth (N/C = 0.20), and in senescent-phase growth (N/C = 0.12), and detritus derived from the saltmarsh grass, Spartina alterniflora, (N/C = 0.04). Total nitrogen excreted ranged from 0.06 to 0.18 μg N copepod^− 1 d^− 1 whereas nitrogen ingested exhibited considerably more variation (0.01 to 0.39 μg N copepod ^− 1d ^− 1). Ammonium was the dominant form of nitrogen excreted and was influenced by both food concentration and N/C ratio. Copepods fed foods with N/C ratios resembling their own body composition (log-phase diatoms and ciliates) excreted more ammonium when fed higher concentrations of food. In contrast, copepods fed foods with lower N/C ratios than their own body composition excreted more ammonium when fed lower concentrations of food, suggesting that they were catabolizing body protein for survival. Excretion of urea varied with food N/C ratio, with more urea excreted when the copepods were fed higher N/C foods. The excretion of DPA did not vary with either food concentration or food N/C ratio. Homeostasis serves to conserve the N/C ratio of copepods. Thus nitrogen excretion by healthy copepods should be expected to increase with ingestion only when copepods have high quantities of nitrogen-rich foods relative to the body composition of the copepods.     

紫云英配施化肥对水稻氮素吸收利用和紫云英氮在水稻-土壤体系分配、残留的影响

明确紫云英配施化肥条件下水稻对氮素吸收利用和紫云英氮在水稻-土壤体系的吸收利用、分配及残留规律,能够为豫南稻区合理施肥提供依据。本研究利用原状土柱模拟和¹⁵N示踪技术,研究等氮条件下不施肥(CK)、化肥+22500 kg·hm^-2紫云英(FM1)、化肥+30000 kg·hm^-2紫云英(FM2)、化肥+37500 kg·hm^-2紫云英(FM3)、化肥+22500 kg·hm^-2紫云英+石灰(FM1+CaO)、化肥+30000 kg·hm^-2紫云英+石灰(FM2+CaO)、化肥+37500 kg·hm^-2紫云英+石灰(FM3+CaO)对水稻氮素吸收利用、水稻-土壤体系氮素养分平衡和紫云英矿化分解的氮在水稻各部位吸收利用、分配及残留的影响。结果表明: 与CK相比,施肥显著提高了稻谷氮吸收量、稻秆氮吸收量和氮素表观损失量、氮素盈余量。稻谷氮吸收量、稻秆氮吸收量和水稻氮利用率随紫云英翻压量增加呈先升高后降低趋势,氮素表观损失量和氮素盈余量随紫云英翻压量增加呈先降低后上升趋势,均以翻压30000 kg·hm^-2紫云英配施化肥处理效果较好。增施石灰可提高水稻稻谷氮吸收量、稻秆氮吸收量和水稻氮利用率,降低氮素表观损失量和氮素盈余量,以FM2+CaO处理效果最好。各施肥处理水稻吸收的氮来源于紫云英的比例为6.3%～13.2%,来源于土壤和肥料的比例为86.8%～93.7%;水稻对紫云英氮的当季利用率为23.8%～33.6%,水稻各部位对紫云英氮的利用率表现为籽粒>茎叶>根;紫云英氮在土壤中的残留率为37.6%～62.4%,损失率为7.8%～38.6%。综合考虑水稻氮素吸收利用、水稻-土壤体系氮素养分平衡和紫云英氮在水稻中的分配状况,该研究区以FM2+CaO处理为最优。     

Screening of chemical composition, antimicrobial and antioxidant activities of Artemisia essential oils

The chemical composition of essential oils isolated from aerial parts of seven wild sages from Western Canada – Artemisia absinthium L., Artemisia biennis Willd., Artemisia cana Pursh, Artemisia dracunculus L., Artemisia frigida Willd., Artemisia longifolia Nutt. and Artemisia ludoviciana Nutt., was investigated by GC–MS. A total of 110 components were identified accounting for 71.0–98.8% of the oil composition. High contents of 1,8-cineole (21.5–27.6%) and camphor (15.9–37.3%) were found in Artemisia cana, A. frigida, A. longifolia and A. ludoviciana oils. The oil of A. ludoviciana was also characterized by a high content of oxygenated sesquiterpenes with a 5-ethenyltetrahydro-5-methyl-2-furanyl moiety, of which davanone (11.5%) was the main component identified. A. absinthium oil was characterized by high amounts of myrcene (10.8%), trans-thujone (10.1%) and trans-sabinyl acetate (26.4%). A. biennis yielded an oil rich in (Z)-beta-ocimene (34.7%), (E)-beta-farnesene (40.0%) and the acetylenes (11.0%) (Z)- and (E)-en-yn-dicycloethers. A. dracunculus oil contained predominantly phenylpropanoids such as methyl chavicol (16.2%) and methyl eugenol (35.8%). Artemisia oils had inhibitory effects on the growth of bacteria (Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis), yeasts (Candida albicans, Cryptococcus neoformans), dermatophytes (Trichophyton rubrum, Microsporum canis, and Microsporum gypseum), Fonsecaea pedrosoi and Aspergillus niger. A. biennis oil was the most active against dermatophytes, Cryptococcus neoformans, Fonsecaea pedrosoi and Aspergillus niger, and A. absinthium oil the most active against Staphylococcus strains. In addition, antioxidant (beta-carotene/linoleate model) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities were determined, and weak activities were found for these oils.     

Inorganic and organic nitrogen uptake by the toxigenic diatom Pseudo-nitzschia australis (Bacillariophyceae)

The nitrogen uptake capabilities of the toxigenic diatom Pseudo-nitzschia australis (Frenguelli), freshly isolated from Monterey Bay California, were examined in unialgal laboratory cultures at saturating photosynthetic photon flux densities (100 μmol photons m⁻² s⁻¹) and 15 °C. The kinetics of nitrogen (nitrate, ammonium, urea and glutamine) uptake as a function of substrate concentration were estimated from short (20.5 min) incubations using the ¹⁵N-tracer technique. Based on the estimated maximum specific uptake rates and measures of N affinity (the initial slope of the uptake versus nutrient concentration curve), nitrate is the preferred nitrogen substrate, followed by glutamine and ammonium, which are equivalent. Rates of urea uptake by P. australis did not saturate at concentrations as high as 36 μg-at N L⁻¹, and urea uptake as a function of concentration could not be described by Michaelis–Menten kinetics over the concentration gradient tested. Although there is a clear preference for nitrate at equivalent concentrations (compared to ammonium, urea, and glutamine), these laboratory results demonstrate the capability of this pennate diatom to utilize both inorganic and organic forms of nitrogen, supporting field observations that P. australis blooms during both upwelling and non-upwelling conditions off the west coast of North America. Substantial differences in the nitrogenous nutrition of P. australis can be expected in these environments, and anthropogenic inputs of N substrates such as ammonium and urea can support its growth, and may contribute significantly to both harmful diatom blooms and the maintenance of seed populations at non-bloom abundances, particularly during periods of reduced or absent upwelling.     

Growth promotion and yield enhancement of peanut (Arachis hypogaea L.) by application of plant growth-promoting rhizobacteria

Although plant growth-promoting rhizobacteria (PGPR) have been reported to influence plant growth, yield and nutrient uptake by an array of mechanisms, the specific traits by which PGPR promote plant growth, yield and nutrient uptake were limited to the expression of one or more of the traits expressed at a given environment of plant–microbe interaction. We selected nine different isolates of PGPR from a pool of 233 rhizobacterial isolates obtained from the peanut rhizosphere on the basis of ACC-deaminase activity. The nine isolates were selected, initially, on the basis of germinating seed bioassay in which the root length of the seedling was enhanced significantly over the untreated control. All the nine isolates were identified as Pseudomonas spp. Four of these isolates, viz. PGPR1, PGPR2, PGPR4 and PGPR7 (all fluorescent pseudomonads), were the best in producing siderophore and indole acetic acid (IAA). In addition to IAA and siderophore-producing attributes, Pseudomonas fluorescens PGPR1 also possessed the characters like tri-calcium phosphate solubilization, ammonification and inhibited Aspergillus niger and A. flavus in vitro. P. fluorescens PGPR2 differed from PGPR1 in the sense that it did not show ammonification. In addition to the traits exhibited by PGPR1, PGPR4 showed strong in vitro inhibition to Sclerotium rolfsii. The performances of these selected plant growth-promoting rhizobacterial isolates were repeatedly evaluated for 3 years in pot and field trials. Seed inoculation of these three isolates, viz. PGPR1, PGPR2 and PGPR4, resulted in a significantly higher pod yield than the control, in pots, during rainy and post-rainy seasons. The contents of nitrogen and phosphorus in soil, shoot and kernel were also enhanced significantly in treatments inoculated with these rhizobacterial isolates in pots during both the seasons. In the field trials, however, there was wide variation in the performance of the PGPR isolates in enhancing the growth and yield of peanut in different years. Plant growth-promoting fluorescent pseudomonad isolates, viz. PGPR1, PGPR2 and PGPR4, significantly enhanced pod yield (23–26%, 24–28% and 18–24%, respectively), haulm yield and nodule dry weight over the control in 3 years. Other attributes like root length, pod number, 100-kernel mass, shelling out-turn and nodule number were also enhanced. Seed bacterization with plant growth-promoting P. fluorescens isolates, viz. PGPR1, PGPR2 and PGPR4, suppressed the soil-borne fungal diseases like collar rot of peanut caused by A. niger and PGPR4 also suppressed stem rot caused by S. rolfsii. Studies on the growth patterns of PGPR isolates utilizing the seed leachate as the sole source of C and N indicated that PGPR4 isolate was the best in utilizing the seed leachate of peanut, cultivar JL24. Studies on the rhizosphere competence of the PGPR isolates, evaluated on the basis of spontaneous rifampicin resistance, indicated that PGPR7 was the best rhizoplane colonizer and PGPR1 was the best rhizosphere colonizer. Although the presence of growth-promoting traits in vitro does not guarantee that an isolate will be plant growth promoting in nature, results suggested that besides ACC-deaminase activity of the PGPR isolates, expression of one or more of the traits like suppression of phytopathogens, solubilization of tri-calcium phosphate, production of siderophore and/or nodulation promotion might have contributed to the enhancement of growth, yield and nutrient uptake of peanut.