GROWTH AND NITROGEN FIXATION OF THE DIAZOTROPHIC FILAMENTOUS NONHETEROCYSTOUS CYANOBACTERIUM TRICHODESMIUM SP. IMS 101 IN DEFINED MEDIA: EVIDENCE FOR A CIRCADIAN RHYTHM1

Trichodesmium sp. IMS 101, originally isolated from coastal western Atlantic waters by Prufert-Bebout and colleagues and maintained in seawater-based media, was successfully cultivated in two artificial media. Its characteristics of growth, nitrogen fixation, and regulation of nitrogen fixation were compared to those of natural populations and Trichodesmium sp. NIBB 1067. Results indicate that the culture grown in artificial media had nitrogen fixation characteristics similar to those when the culture is grown in seawater-based medium and to those of Trichodesmium sp. in the natural habitat. The study provides practical artificial media to facilitate the physiological studies of these important diazotrophic cyanobacteria, as well as the cultivation of other Trichodesmium species in future studies. Manipulations of the light/dark cycle were performed to determine whether or not the daily cycle of nitrogen fixation is a circadian rhythm. Cultures grown under continuous light maintained the cycle for up to 6 days. We demonstrated that the daily cycle of nitrogen fixation in Trichodesmium sp. IMS 101 was at least partially under the control of a circardian rhythm.     

Enhancement of the Gibberella zeae growth inhibitory lipopeptides from a Bacillus subtilis mutant by ion beam implantation

Bacillus subtilis JA antagonized the growth of Gibberella zeae. In order to reduce growth of this fungi pathogen to a greater extent, low-energy ion beam implantation was applied in mutant breeding. We studied the effects of different energies and different doses of nitrogen ion implantation. The mutant strain designated as JA026 was obtained showing higher inhibition activity in the screening plate. Its inhibition zone against indicator organism increased by 14.3% compared to the original strain. The electrospray ionization mass spectrometry (ESI/MS) analysis indicated that the antifungal lipopeptides produced by the mutant were identical to those produced by the wild-type strain. The mutant strain exhibited favorable properties including the high yield of antifungal lipopeptides production and faster growth over the parent strain, which suggested that this strain would be a promising biocontrol candidate in agriculture.     

Pyrimidine ribonucleoside catabolic enzyme activities ofPseudomonas pickettii

Pyrimidine ribonucleoside catabolic enzyme activities of the opportunistic pathogenPseudomonas pickettii were examined. Of the pyrimidine and related compounds tested, only dihydrouracil (nitrogen source) and ribose (carbon source) supported growth. Thin-layer chromatographic separation of the uridine and cytidine catabolities produced byP. pickettii extracts indicated that this pseudomonad contained nucleoside hydrolase activity. Its presence was confirmed by enzyme assay. Hydrolase activity was elevated in both glucose- and ribose-grown cells relative to succinate-grown cells. Nucleoside hydrolase activity was depressed when dihydrouracil served as a nitrogen source. Cytosine deaminase activity was present in extracts prepared from succinate-, glucose- or ribose-grown cells when (NH₄)₂SO₄ served as the nitrogen source although cells grown on glucose or ribose exhibited a higher enzyme activity. Cytosine deaminase activity was not detected in extracts prepared from cells grown on dihydrouracil as a nitrogen source. Both dihydropyrimidine dehydrogenase and dihydropyrimidinase activities were measurable inP. pickettii. The dehydrogenase activity was higher with NADH than with NADPH as its nicotinamide cofactor when uracil served as its substrate. Carbon source did not affect dehydrogenase or dihydropyrimidinase activity greatly but both activities were diminished in cells grown on the nitrogen source dihydrouracil.     

A preliminary study on the suitability of gum arabic as medium for fungal growth

Summary AlthoughRhizopus stolonifer, Aspergillus niger andRhizoctonia solani could be grown on gum arabic solutions of 1–4% concentrations, yet their growth was poor presumably due to lack of sufficient nitrogen. Gum arabic was found to be a good carbon source when it replaced sucrose in Czapek-Dox medium. The sporulation ofRhizopus was not affected by the presence of gum in the medium, while that ofAspergillus was appreciably affected. The number of sporulating heads was reduced, irregularity in the shape of the head due to variation in the length of spore chains was also noticed. A new medium of pH 5.6 was found satisfactory for fungal growth and could be recommended for soil isolations. Its composition is: 4% gum arabic, 1% peptone, 0.025% sodium chloride and 0.001% ferrous sulphate.     

Some nutritional factors influencing mevinolin production byAspergillus terreus strain

Aspergillus terreus produces the hypocholesterolemic compound mevinolin. Its growth and mevinolin production were affected by the composition of the culture medium. Both were at a maximum with glucose (6%) as the sole carbon source and in the presence of a mixture of yeast extract and sodium nitrate as nitrogen source. Influence of the concentration of some inorganic salts are also discussed.     

Nitrogen amendments reduce the growth of extramatrical ectomycorrhizal mycelium

The effect of three different nitrogen sources on the growth of external ectomycorrhizal mycelium was studied in Perspex micorocosms. Nonsterile peat was used as substrate. Five different fungal isolates growing in symbiosis with pine seedlings were investigated: two isolates of Paxillus involutus, one of Suillus bovinus and two unidentified ectomycorrhizal fungi isolated from ectomycorrhizal root tips. Three different nitrogen sources were used: ammonium as (NH₄)₂SO₄, nitrate as NaNO₃ and a complete nutrient solution (Ingestad 1979), and three different nitrogen concentrations, 1, 2 or 4 mg N/g dry wt. of peat. The mycelial growth of all fungi was found to be negatively affected by the nitrogen amendments, although the sensitivity to nitrogen varied between the isolates. One of the unidentified isolates was extremely sensitive and growth was completely inhibited by all nitrogen treatments. In contrast, the growth of one of the P. involutus isolates was only slightly reduced by the nitrogen amendments. The different nitrogen sources all reduced growth, and since no significant difference was found between the nitrogen sources or between the different nitrogen concentrations the results were pooled to give one value that summarized the effect of nitrogen on mycelial growth. Thus, the mycelial growth of one of the two P. involutus isolates was reduced to approximately 80% of the growth in the control, the other P. involutus and one of the unidentified fungi, vgk 2 89.10, were reduced to 40–50% of the control growth, S. bovinus to 30% of the control and the most sensitive fungus, the unidentified isolate vg 1 87.10, was reduced to 3% of the growth in the control treatment. In all experiments, the shoot to root ratio generally increased, mainly as a result of increased shoot growth.     

Effect of <Emphasis Type="Italic">Nostoc</Emphasis> sp. on soil characteristics,plant growth and nutrient uptake

The arid desert area is expanding by 6 million ha year⁻¹. Nostoc, a terrestrial cyanobacterium, is known as a pioneer organism which can photosynthesize, fix atmospheric nitrogen, and secrete polysaccharides. In order to explore the potential of Nostoc as a countermeasure to soil desertification, we have investigated the effects of Nostoc on the chemical and physical properties of soil and on plant growth in outdoor and laboratory experiments. The effect of Nostoc on soil properties, when applied on the surface of soil in plastic containers, was determined after cultivation for 90 days outdoors. Its effect on plant growth and nutrient uptake was determined by growing Nostoc with Komatsuna (Brassica rapa var. peruviridis) on a soil-less culture in a plastic petri dish incubated inside a growth chamber for 14 days. Two colonies of Nostoc with different shapes were also examined for their tolerance to dryness and salinity. Nostoc application resulted in an increase in the organic carbon and nitrogen content of the surface soil and enhanced plant growth and plant iron uptake, but soil temperature and moisture were maintained. Spherical-shaped Nostoc showed a higher tolerance to dryness and salinity than irregular-shaped Nostoc, and the former was able to rectify problematic soil better than the latter. The results indicate that the application of Nostoc to soil has a potential for increasing soil organic matter and reclaiming degraded soil ecosystems. Presented at the 6th Meeting of the Asian Pacific Society of Applied Phycology, Manila, Philippines.     

Gamma-aminobutyric acid (GABA) increases in vitro germ-tube formation and phospholipase B1 mRNA expression in Candida albicans

Candida albicans is a commensal yeast in humans that disseminates in immunocompromised persons. Its spreading is modulated by melanin, hormones, or some neurotransmitters, among other factors. The neurotransmitter gamma-aminobutyric acid (GABA) is used by bacteria, plants, and fungi as a carbon and nitrogen source. In this article, the in vitro effect of different doses of GABA on germ-tube formation and expression of phospholipase B1 (PLB1) mRNA in two Candida albicans strains was investigated. Results demonstrated that GABA increases both germ-tube formation and PLB1 mRNA expression in the two Candida strains in a dose-dependent manner, which suggests that GABA promotes the growth of C. albicans.     

The capability of the algaeChlorella vulgaris andScenedesmus obliquus of utilizing amino acids as the sole nitrogen source

The capability of utilizing 20 amino acids and 2 amides as the sole nitrogen source for growth was studied in two green algae (Chlorophyceae). A comparison was made of the growth rate of algae in a mineral nutrient solution containing nitrate as the nitrogen source, with that in the same solution in which nitrogen in the form of nitrate was substituted by an equivalent nitrogen amount in the form of various amino acids. In addition to this, another series of experiments was carried out in whioh both culture media were supplied with glucose. The results show that both algae utilize a series of amino acids in dependence of their structure (mostly 3-carbon amino acids). The growth rate ofChlorella in the presence of these sources is the same as in nitrate, that ofScenedesmus even much higher. In the cultures containing glucose both algal species exhibit a higher growth rate in the media with the nitrate nitrogen source than in those with amino acids (with the exception of glycine inScenedesmus).     

Auxin biosynthetic gene TAR2 is involved in low nitrogen‐mediated reprogramming of root architecture in Arabidopsis

In plants, the plasticity of root architecture in response to nitrogen availability largely determines nitrogen acquisition efficiency. One poorly understood root growth response to low nitrogen availability is an observed increase in the number and length of lateral roots (LRs). Here, we show that low nitrogen‐induced Arabidopsis LR growth depends on the function of the auxin biosynthesis gene TAR2 (tryptophan aminotransferase related 2). TAR2 was expressed in the pericycle and the vasculature of the mature root zone near the root tip, and was induced under low nitrogen conditions. In wild type plants, low nitrogen stimulated auxin accumulation in the non‐emerged LR primordia with more than three cell layers and LR emergence. Conversely, these low nitrogen‐mediated auxin accumulation and root growth responses were impaired in the tar2‐c null mutant. Overexpression of TAR2 increased LR numbers under both high and low nitrogen conditions. Our results suggested that TAR2 is required for reprogramming root architecture in response to low nitrogen conditions. This finding suggests a new strategy for improving nitrogen use efficiency through the engineering of TAR2 expression in roots.     

Internal Glutamine and Glutamate Pools in Klebsiella pneumoniae Grown Under Different Conditions of Nitrogen Availability

Internal pool sizes of glutamine and glutamate in Klebsiella pneumoniae grown under nitrogen limitation or nitrogen sufficiency were measured to study the signal transduction of external nitrogen limitation. K. pneumoniae cells were grown in an anaerobic, ammonium-limited chemostat culture. At a growth rate of 0.217 h⁻¹, the steady state ammonium concentration in the culture was 55 μm, correlating with repression of the nitrogen fixation (nif) genes. At growth rates below 0.138 h⁻¹, the ammonium concentration in the culture dropped below 0.5 μm and the nif genes became derepressed. During the transition from nitrogen sufficiency to nitrogen limitation, the internal glutamine pool in K. pneumoniae decreased by a factor of approximately 6. The glutamate pool, however, remained stable. Similarly, in anaerobic batch cultures with different limiting nitrogen sources, the glutamine pool generally decreased by a factor of 7 to 9 when nif gene derepression was achieved. All the limiting nitrogen sources used resulted in decreased growth rates compared with growth under nitrogen excess, suggesting an inverse relationship between glutamine pool size and doubling time. These studies indicate that K. pneumoniae perceives external nitrogen limitation as internal glutamine limitation. Received: 3 April 2000 / Accepted: 5 June 2000     

Tillage effect on seasonal nitrogen availability in corn supplied with legume green manures

Colonization of sorghum by Macrophomina phaseolina in field plots was determined at nitrogen fertilization rates of 0, 56, 112, and 168 kg ha^-1 in 1988 and 0, 84, 168, and 256 kg ha^-1 in 1989. Above ground plant tissue and roots were sampled monthly to determine total nitrogen and percent colonization of root segments by natural inoculum. Root infection was not affected by nitrogen treatment, but was affected by growth stage and environment. High root infection occurred before reproductive development (growth stage 3) in 1988 and was associated with hot, dry weather early in the growing season. In 1989, when the weather was cool and wet, root infection began after reproductive development (growth stage 4). The effect of nitrogen treatments on lesion length was determined in sorghum stalks artificially inoculated with M. phaseolina. Lesion lengths were significantly affected by both nitrogen treatments and growth stage. Lesions were significantly longer with all nitrogen treatments at growth stage 9 than with the no-nitrogen treatment, and lesions tended to increase with increased levels of nitrogen fertilization. Significant increases in lesion length occurred between growth stages 5, 7, and 9 in 1988 and between 7 and 9 in 1989. This study demonstrates that nitrogen fertilization affects colonization of sorghum stalks but not root infection by M. phaseolina.     

Nodulation and growth response ofAllocasuarina andCasuarina species to phosphorus fertilization

To examine how soil phosphorus status affects nitrogen fixation by the Casuarinaceae —Frankia symbiosis,Casuarina equisetifolia and two species ofAllocasuarina (A. torulosa andA. littoralis) inoculated or fertilized with KNO₃ were grown in pots in an acid soil at 4 soil phosphate levels. InoculatedC. equisetifolia nodulated well by 12 weeks after planting and the numbers and weight of nodules increased markedly with phosphorus addition. Growth ofC. equisetifolia dependent on symbiotically fixed nitrogen was more sensitive to low levels of phosphorus (30 mg kg^–1 soil) than was growth of seedings supplied with combined nitrogen; at higher levels of phosphorus, the growth response curves were similar for both nitrogen fertilized and inoculated plants. The interaction between phosphorus and nitrogen treatments (inoculated and nitrogen fertilized) demonstrated that there was a greater requirement of phosphorus for symbiotic nitrogen fixation than for plant growth when soil phosphorus was low.WithAllocasuarina species, large plant to plant variation in nodulation occurred both within pots and between replicates. This result suggests genetic variation in nodulation withinAllocasuarina species. Nodulation ofAllocasuarina species did not start until 16 weeks after planting and no growth response due toFrankia inoculation was obtained at the time of harvest. Addition of nitrogen starter is suggested to boost plant growth before the establishment of the symbiosis. Growth ofAllocasuarina species fertilized with nitrogen responded to increasing levels of phosphorus up to 90 mg P/kg soil after which it declined by 69% forA. littoralis. The decrease in shoot weight ofA. littoralis, A. torulosa, C. equisetifolia andC. cunninghamiana at high phosphorus was confirmed in a sand culture experiment, and may be atributable to phosphorus toxicity.     

Effects of nitrogen supply and irradiance on growth and nitrogen status in the moss Dicranum majus from differently polluted areas

Abstract

A growth experiment was undertaken to study the effects of nitrogen supply and irradiance on growth and nitrogen status in the moss Dicranum majus Sm. from two areas receiving different amounts of atmospheric nitrogen deposition. Intact samples of D. majus carpets were taken from two Picea abies forests, one located in southern Norway (high-N site) and the other in central Norway (low-N site). The moss carpets were grown for 120 days at three irradiance levels (PPFD: 20,40 or 80 μmol m^?2 s^?l) and sprayed daily with equal amounts of a nutrient solution containing 30, 180 or 330 μM nitrogen as NO₃ - and NH₄ +. Concentrations and total amounts of nitrogen, soluble proteins and chlorophyll were highest in moss plants from the high-N site, both at the start and the end of the experiment. The elongation growth was highest at the lowest irradiance level. As total biomass production did not differ between nitrogen and light treatments, moss growth was presumably limited by other factors, even at the lowest supply rates. Concentrations and total amounts of nitrogen increased with increasing nitrogen supply in moss plants from both sites. Accumulated nitrogen was partly stored as protein and chlorophyll. Recycling of nitrogen from old to young tissues is discussed as a possible explanation for the rather low nitrogen demand in D. majus and the persistently higher nitrogen contents in moss plants from the high-N site.     

Influence of nitrogen stress on Isochrysis galbana strain U4, a candidate for biodiesel production

Lipid accumulation has been investigated in numerous microalgal species to assess their potential with respect to biodiesel production. The present work determines the effect of nitrogen stress on physiological and ultrastructural changes in Isochrysis galbana U4. This study is unique in showing the correlations between growth, lipid production, pigmentation and ultrastructural changes in Isochrysis cells undergoing nitrogen starvation. The continuation of algal growth after the complete depletion of external nitrogen was shown to be supported by internal nitrogen stores, possibly in the pyrenoid. Cell growth ceased and lipid accumulation was initiated after the internal store of nitrogen had become exhausted. The depletion of intracellular nitrogen reservoirs to critical thresholds initiated the onset of the stationary phase, a decline in chlorophyll content and the initiation of lipid and carotenoid accumulation. The most notable ultrastructural changes, upon nitrogen stress, were the accumulation of plastidial and cytoplasmic lipid bodies and the dismantling of the chloroplast. The size of the pyrenoid when external nitrogen became depleted was found to decrease significantly, up to four‐fold. This was attributed to the remobilization of nitrogen from Rubisco. The level of expression of heterochromatin was found to increase when cells were nitrogen starved. This is thought to favor long‐term dormancy in this species because aging cells have been noted to recover rapidly when returned to conditions favorable for growth. The observations of this study are consistent with the hypothesis that the responses of Isochrysis cells to nitrogen starvation are regulated by the internal reserves of nitrogen, and the depletion of these reserves is an important trigger for lipid accumulation in this species. The findings of this study also indicate that Isochrysis galbana U4 is a promising candidate for biodiesel lipid production.     

Mobilization of minerals and moisture loss during senescence of the energy crops Miscanthus×giganteus,Arundo donax and Phalaris arundinacea in Wales,UK

Miscanthus×giganteus, Arundo donax and Phalaris arundinacea were planted at three sites in Wales, UK, to study the timing of mineral and moisture decline during autumn and winter. The mineral and moisture content at several harvest dates throughout the period were compared with power station threshold levels. All crops showed a highly significant decrease in moisture content in all years with the exception of Arundo which did not lose moisture in the final year. The Miscanthus and Arundo crops remained above the moisture content threshold in all years and would require further drying before combustion. The Phalaris fell below the moisture threshold in January. The Miscanthus leaves fell below the nitrogen threshold before leaf abscission at one site but not at another, although the cane remained mostly within the guideline range. Both its leaves and cane remained above the potassium threshold level. Its leaf sulphur fell within acceptable levels during November and its cane remained below the lowest threshold from November onwards. The Arundo leaves remained above the nitrogen, potassium and sulphur thresholds throughout the period and it did not lose its leaves. The cane nitrogen increased above the threshold during autumn whereas its potassium remained higher than its threshold, with the exception of one sampling date at one site. Its sulphur remained within the guidelines. The Phalaris crop fell to within nitrogen guidelines at one site but increased at the other. Its potassium decreased to below the threshold at both sites in January, and its sulphur content was consistently within the guidelines.     

Does nitrogen limitation promote intraguild predation in an aphidophagous ladybird?

Reciprocal intraguild predation occurs between the two aphidophagous ladybird beetles Coccinella septempunctata L. and Harmonia axyridis Pallas (Coleoptera: Coccinellidae). However, its direction is asymmetrical; H. axyridis generally acts as an intraguild predator, and C. septempunctata as an intraguild prey. According to Denno and Fagan's prediction that nitrogen shortages in predators may promote intraguild predation, it was hypothesized that growth of intraguild predator H. axyridis is more limited by nitrogen than that of intraguild prey C. septempunctata, and that H. axyridis growth is enhanced by feeding on C. septempunctata compared to feeding on aphids. To determine nitrogen‐limited growth in H. axyridis, the following two predictions were examined. First, it was predicted that the nitrogen content of H. axyridis would be higher than that of C. septempunctata when both feed on aphids. However, nitrogen content did not differ between the two ladybirds. Second, it was predicted that nitrogen‐use efficiency of H. axyridis would be lower than that of C. septempunctata. However, there was no significant difference between species. These results did not support the hypothesis that growth of H. axyridis is more limited by nitrogen than that of C. septempunctata. In addition, the present study showed that dry mass and nitrogen growth of H. axyridis were not enhanced, but rather decreased, by eating high‐nitrogen C. septempunctata, compared to eating low‐nitrogen aphids. Overall, the present study did not support the hypothesis that nitrogen shortages in predators may promote intraguild predation.     

Inorganic nitrogen source for autotrophic growth of Euglena mutabilis Schmitz

The effect of inorganic nitrogen source on population growth of Euglena mutabilis, an acidophillic benthic protozoa colonizing on the sediment of acid mine drainage, was investigated. Sodium nitrate, ammonium chloride, and ammonium sulfate were tested as nitrogen sources. The population density of E. mutabilis at equilibrium density cultivated in ammonium chloride‐ and ammonium sulfate‐containing media was 9–11 times higher than that in sodium nitrate‐containing medium at the optimal salt medium concentration. The population growth of E. mutabilis in ammonium sulfate‐containing medium was rapid and reached half of the equilibrium density after ca. 228 h, which was ca. 77 h earlier than that in ammonium chloride‐containing medium. Culture medium with ammonium sulfate as the nitrogen source achieved the highest maximum population density and the fastest growth rate among the three nitrogen salts used as nitrogen sources.     

On the physiology of the red alga Asterocytis ramosa in axenic culture

The red alga Asterocytis ramosa has been cultivated aseptically in artificial seawater ASP6 F. The alga shows optimal growth at 22·5–25° C. Optimal light conditions as well as phosphorus and nitrogen requirements were investigated. Nitrate and ammonium salts as well as organic nitrogen can be used as a nitrogen source. Arginine was an extremely good source of nitrogen, presumably because it splits off urea. Asterocytis is vitamin B₁₂ heterotrophic and growth is obtained with cyanocobalamin, Factor III and Factor Z₁. A mixture of other vitamins gives further stimulation. In the newly isolated culture, hexoses and pentoses enhanced growth. Acetate increased growth in an old axenic culture. In darkness no growth was obtained.