Effect of temperature and irradiance on the uptake of ammonium and nitrate by <Emphasis Type="Italic">Laurencia brongniartii</Emphasis> (Rhodophyta,Ceramiales)

The effects of temperature (20, 24 and 28 °C) and irradiance (15 and 40 μmol photon m⁻² s⁻¹) on the nitrate and ammonium uptake rates of the subtropical red alga, Laurencia brongniartii, were investigated to prepare for tank cultivation. Nitrate uptake followed saturation kinetics and was faster at higher irradiances and temperatures. In contrast, ammonium uptake was linear over the experimental range and was not affected by an increase in temperature. A parameter, β, was calculated to compare substrate uptake rates of nitrate along the linear portion of the uptake curve with that of ammonium. For nitrate, β was lower at low irradiance and higher at high irradiance (β = 0.007 ± 0.003 and 0.030 ± 0.002 [μmol N L⁻¹ (μmol N g_ww⁻¹ d⁻)⁻¹], respectively). However, β was 0.023 ± 0.002 and 0.034 ± 0.002 [μmol N L⁻¹ (μmol N g_ww⁻¹ d⁻¹)⁻¹] for ammonium, suggesting a preference for ammonium over nitrate.     

The vitamin content of microalgae used in aquaculture

The vitamin content in four Australian microalgae, a Nannochloropsis-like sp., Pavlova pinguis, Stichococcus sp. and Tetraselmis sp., were examined. These were grown under a 12:12 h light:dark regimen (100 μmol photon m⁻²s⁻¹) and harvested during late-logarithmic phase. Typically, the content showed a two- to three fold range between the species. When expressed on a dry weight basis, the content of ascorbate ranged from 1.3 to 3.0 mg g⁻¹, β-carotene from 0.37 to 1.05 mg g⁻¹, α-tocopherol from 0.07 to 0.29 mg g⁻¹, thiamine from 29 to 109 μg g⁻¹, riboflavin from 25 to 50 μg g⁻¹, total folates from 17 to 24 μg g⁻¹, pyridoxine from 3.6 to 17 μg g⁻¹, cobalamin from 1.70 to 1.95 μg g⁻¹ and biotin from 1.1 to 1.9 μg g⁻¹. Retinol was detected only in Tetraselmis sp. (2.2 μg g⁻¹); any vitamins D₂ or D₃ were below the detection limit (≤0.45 μg g⁻¹). Nannochloropsis sp. was also grown under a 24:0 h light:dark light cycle and harvested at stationary phase. The content of most vitamins in Nannochloropsis sp. cultures differed significantly, and the degree of variation was similar to that observed between the four species grown under 12:12 h light:dark regimen (100 μmol photon m⁻²s⁻¹) and harvested during late-logarithmic phase. Thiamine content was also examined in six non-Australian strains commonly used in aquaculture, Chaetoceros muelleri, Thalassiosira pseudonana, Nannochloris atomus, Nannochloropsis oculata, Isochrysis sp. (T.ISO) and Pavlova lutheri. Values (average 61 μg g⁻¹; range 40 to 82) were similar to those in the Australian strains (average 61 μg g⁻¹; range 29 to 109) and increased during stationary phase (average 94 μg g⁻¹; 38 to 131). Comparison of the data with the known nutritional requirements for marine fish species and prawns suggests that the microalgae should provide excess or adequate levels of the vitamins for aquaculture food chains. The data may be used to guide the content of vitamins included in micro-diets developed as replacements for live diets. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of temperature and ammonium on growth,pigment production and nitrogen uptake by four species of <Emphasis Type="BoldItalic">Porphyra</Emphasis> (Bangiales,Rhodophyta) native to the New England coast

Porphyra is one of the world’s most valued maricultured seaweeds and has been cultivated for several hundred years in Asia. The objective of this study was to produce critical information as a guide for the selection of an appropriate Porphyra species from coastal New England for the development of a land-based aquaculture system. Four Northwest Atlantic Porphyra species: P. leucosticta, P. amplissima, P. linearis and P. umbilicalis, were cultivated for 1 and 2 weeks at saturated light intensities (100–150 μmol photons m⁻²s⁻¹) and six combinations of ammonium (25 and 250 μmoles L⁻¹) and temperature (10, 15 and 20°C). Specific growth rate (SGR) increased with decreasing temperature in P. leucosticta, P. linearis and P. umbilicalis and increased with increasing temperature in P. amplissima. The SGR of all species was greater at the higher ammonium concentration. Porphyra linearis had the highest SGR, increasing in biomass by approximately 16% day⁻¹. Phycoerythrin (PE) content was higher at 10°C and 250 μmoles L⁻¹ in all species except P. amplissima. The PE content, measured as fresh weight (FW), of P. linearis (29 mg g⁻¹ FW⁻¹) and P. umbilicalis (26 mg g⁻¹ FW⁻¹) was significantly higher than the other two species. Tissue nitrogen content of all species measured in dry weight was on average 1.45% higher at 250 μmoles L⁻¹ than at 25 μmoles L⁻¹ ammonium concentration. Porphyra umbilicalis had the highest tissue nitrogen contents (6.76%) at 10°C and 250 μmoles L⁻¹ ammonium. Based on these results, P. linearis and P. umbilicalis should be considered as potential candidates for bioremediation with finfish and shellfish mariculture.     

Effect of light and temperature on the cyanobacterium <Emphasis Type="Italic">Arthronema africanum</Emphasis> - a prospective phycobiliprotein-producing strain

The effect of light intensity (50–300 μmol photons m⁻² s⁻¹) and temperature (15–50°C) on chlorophyll a, carotenoid and phycobiliprotein content in Arthronema africanum biomass was studied. Maximum growth rate was measured at 300 μmol photons m⁻² s⁻¹ and 36°C after 96 h of cultivation. The chlorophyll a content increased along with the increase in light intensity and temperature and reached 2.4% of dry weight at 150 μmol photons m⁻² s⁻¹ and 36°C, but it decreased at higher temperatures. The level of carotenoids did not change significantly under temperature changes at illumination of 50 and 100 μmol photons m⁻² s⁻¹. Carotenoids were about 1% of the dry weight at higher light intensities: 150 and 300 μmol photons m⁻² s⁻¹. Arthronema africanum contained C-phycocyanin and allophycocyanin but no phycoerythrin. The total phycobiliprotein content was extremely high, more than 30% of the dry algal biomass, thus the cyanobacterium could be deemed an alternative producer of C-phycocyanin. A highest total of phycobiliproteins was reached at light intensity of 150 μmol photons m⁻² s⁻¹ and temperature of 36°C, C-phycocyanin and allophycocyanin amounting, respectively, to 23% and 12% of the dry algal biomass. Extremely low (<15°C) and high temperatures (>47°C) decreased phycobiliprotein content regardless of light intensity.     

Enhanced extracellular production of trans-resveratrol in Vitis vinifera suspension cultured cells by using cyclodextrins and methyljasmonate

In this work, the effect of different inducing factors on trans-resveratrol extracellular production in Monastrell grapevine suspension cultured cells is evaluated. A detailed analysis provides the optimal concentrations of cyclodextrins, methyljasmonate and UV irradiation dosage, optimal cell density, elicitation time and sucrose content in the culture media. The results indicate that trans-resveratrol production decreases as the initial cell density increases for a constant elicitor concentration in Monastrell suspension cultured cells treated with cyclodextrins individually or in combination with methyljasmonate; the decrease observed in cell cultures elicited with cyclodextrins alone is far more drastic than those observed in the combined treatment. trans-Resveratrol extracellular production observed by the joint use of cyclodextrins and methyljasmonate (1,447.8 ± 60.4 μmol trans-resveratrol g⁻¹ dry weight) is lower when these chemical compounds are combined with UV light short exposure (669.9 ± 45.2 μmol trans-resveratrol g⁻¹ dry weight). Likewise, trans-resveratrol production is dependent on levels of sucrose in the elicitation medium with the maximal levels observed with 20 g l⁻¹ sucrose and the joint action of cyclodextrins and 100 μM methyljasmonate. The sucrose concentration did not seem to limit the process although it affects significantly the specific productivity since the lowest sucrose concentration is 10 g l⁻¹, the highest productivity is reached (100.7 ± 5.8 μmol trans-resveratrol g⁻¹ dry weight g⁻¹ sucrose) using cyclodextrins and 25 μM methyljasmonate.     

Partial quantification of pigments extracted from the zooxanthellate octocoral <Emphasis Type="Italic">Sinularia flexibilis</Emphasis> at varying irradiances

Chlorophyll-a (chl-a) and carotenoid pigments of the zooxanthellate octocoral Sinularia flexibilis were analyzed using high performance liquid chromatography following exposure to three light intensities for over 30 days. From the coral fragments located at different light intensities, a total carotenoid of >41 μg g⁻¹ dry weight, including peridinin, xanthophylls (likely diadinoxanthin + diatoxanthin), and chl-a as the most abundant pigments, with minor contents of astaxantin and β-carotene were detected. The whole content of chl-a weighed 5 μg g⁻¹ dry weight in all coral colonies. Chl-a and carotenoids contributed 11.2% and 88.2%, respectively, to all pigments detected, and together accounted for 99.4% of the total pigments present. The highest contents of carotenoids and chl-a was observed in the coral grafts placed in an irradiance of 100 μmol quanta m⁻² s⁻¹; they showed lower ratios of total carotenoids: chl-a compared to those exposed to 400 μmol quanta m⁻² s⁻¹ after >30 days of incubation. The ratios of peridinin and xanthophylls with respect to chl-a from the colonies at 400 μmol quanta m⁻² s⁻¹ were approximately double those observed at irradiances of 100 and 200 μmol quanta m⁻² s⁻¹. Partial quantification of pigments in this study showed that the carotenoids of S. flexibilis showed a decrease at irradiances above 100 μmol quanta m⁻² s⁻¹, with the exception of an increase in β-carotene at 200 μmol quanta m⁻² s⁻¹.     

Kinetics of nitrate uptake by New Zealand marine macroalgae and evidence for two nitrate transporters in <Emphasis Type="Italic">Ulva intestinalis</Emphasis> L.

Kinetic constants were determined for nitrate uptake in three species, Pterocladiella capillacea (S.G. Gmelin) Santelices et Hommersand (Rhodophyceae, Gelidiales), Ulva intestinalis L. (Chlorophyceae, Ulvales) and Xiphophora chondrophylla (Turner) Montagne ex Harvey (Phaeophyceae, Fucales), of New Zealand macroalgae, with K _m values ranging from 10 to 17 μM and V _max values from 3 to 65 μmole g⁻¹ dry weight h⁻¹. There was no effect of ammonium on nitrate uptake by Pterocladiella capillacea or Xiphophora chondrophylla. Ammonium inhibited nitrate uptake by 40% in Ulva intestinalis from a site with relatively low seawater ammonium concentrations. In contrast, U. intestinalis from an ammonium-enriched site had lower rates of nitrate uptake that were insensitive to inhibition by ammonium. It is suggested that there are (at least) two transport systems for nitrate in U. intestinalis; a constitutive transporter, which is insensitive to ammonium, and a transporter that is sensitive to ammonium inhibition and down-regulation by ammonium; the implications of this for our understanding of macroalgal blooms is discussed. Handling editor: K. Martens     

Effect of irradiation intensity on cell growth and kalata B1 accumulation in <Emphasis Type="Italic">Oldenlandia affinis</Emphasis> cultures

Light irradiation had remarkable effects on callus growth of Oldenlandia affinis with an optimum intensity of 35 μmol m⁻² s⁻¹. Biosynthesis of kalata B1, the main cyclic peptide in O. affinis, was induced and triggered with rising irradiation intensities. The highest concentration of kalata B1, 0.49 mg g⁻¹ DW characterised by the maximum productivity of 3.88 μg per litre and day was analysed at 120 μmol m⁻² s⁻¹, although callus growth was repressed. The light saturation point was established to be 35 μmol m⁻² s⁻¹, where kalata B1 productivity was in a similar order (3.41 μg per day) due to the higher growth index. O. affinis suspension cultures were shown to accumulate comparable specific kalata B1 concentrations in a delayed growth associated production pattern. These were dependent on irradiation intensity (0.16 mg g⁻¹ at 2 μmol m⁻² s⁻¹; 0.28 mg g⁻¹ at 35 μmol m⁻² s⁻¹). The batch cultivation process resulted in a maximum productivity of 27.30 μg per litre and day with culture doubling times of 1.16 d⁻¹. Submers operation represented a 8-fold product enhancement compared to callus cultivation.     

Accumulation of Heavy Metals by Ectomycorrhizal Fungi Colonizing Birch Trees Growing in an Industrial Desert Soil

The results of analyses conducted to determine contents of Cd(II) and Pb(II) in mushrooms of mycorrhizal fungi and selected parts of birches growing in an industrial desert surrounding a nonferrous works are presented in this study. The fruiting bodies of fungi accumulated several times higher contents (up to 80 μg g⁻¹ dry weight) of Cd(II) compared to those found in the soil (20 μg g⁻¹ dry weight). In contrast, the mushroom contents of Pb(II) were only slightly increased (up to 895 μg g⁻¹ dry weight) than those present in the soil (500 μg g⁻¹ dry weight). However, fivefold higher concentrations of the metals were found in the mycorrhizal roots. Comparing the distribution of the metals analysed, the protective role of the ectomycorrhizae in relation to the host tree was indicated. Mycorrhizal fungi persistently fixed heavy metals, forming an efficient biological barrier that reduced movement of the metals in birch tissues.     

Regulation of the Expression of the Photosynthetic Apparatus of Rhodobacter capsulatus Grown in Nitrogen-Limited Chemostat Cultures

Rhodobacter capsulatus was grown in chemostat cultures under different dilution rates and with ammonium ions as the limiting nutrient. The maximal growth rate (μmax) and the Monod cell growth saturation coefficient (K_s), were calculated from batch cultures grown at different concentrations of NH₄ ⁺. The experiments in chemostat were carried out at 0.25 mM (NH₄)₂SO₄, and the dilution rates were varied between 38% and 75% of μmax. The results indicated that under continuous culture conditions the cell yield coefficient (Y) (mg dry weight × μmol consumed ammonium sulfate⁻¹) decreased with increasing dilution rate (D). On the contrary, the cell yield was constant when expressed as mg cellular protein ×μmol consumed ammonium sulfate⁻¹. This occurred as a consequence of both an increase in the consumed ammonium sulfate and a simultaneous decrease in the cell biomass production at increasing growth rates. The cells produced at higher growth rates had a higher protein content per cell. The specific content of bacteriochlorophyll (Bchl) decreased (between 3 and 4 times) with increasing growth rates measured in either cells or chromatophores. However, the absorption spectra of the cells indicated that the ratio LHI (light-harvesting complex I) to LHII (light-harvesting complex II) Bchl complexes did not change. The reaction center (RC) complex content varied in parallel with the total Bchl content, yielding a constant photosynthetic unit of 65 mol Bchl × mol RC⁻¹ at different Ds. On the other hand, the uncoupled ATPase-specific activity measured in chromatophores was usually between 30% and 40% higher at the highest growth rates reached in these experiments. Received: 22 January 1996 / Accepted: 9 March 1996     

Secondary metabolites during ontogenetic phase of reproductive structures in Hypericum maculatum

The distribution patterns of flavonoids hyperoside, isoquercitrin, quercitrin, quercetin, I3,II8-biapigenin and naphtodianthrones hypericin and pseudohypericin were studied in reproductive structures during ontogenetic phase of flowering in Hypericum maculatum Crantz. Considerable differences in the content of these secondary metabolites, in the particular flower parts were found. The content of all the metabolites studied is stable during the whole period of flowering in green flower parts (sepals). In petals, stamens and pistils their content undergoes considerable change associated with the biological functions of particular metabolites. The most conspicuous changes during ontogenetic phase of flowering were the decrease of hyperoside and isoquercitrin content in petals (average content in buds 1.589 mg g⁻¹ dry weight, average content in overblown flowers 0.891 mg g⁻¹ dry weight), the decrease of the I3,II8-biapigenin content in stamens (in buds 1.189 mg g⁻¹ dry weight, in overblown flowers 0.319 mg g⁻¹ dry weight), and the increase of hypericin and pseudohypericin content in both petals (total average content of hypericins in the buds 0.547 mg g⁻¹ dry weight; in overblown flowers 0.792 mg g⁻¹ dry weight) and stamens (in buds 0.189 mg g⁻¹ dry weight; in overblown flowers 0.431 mg g⁻¹ dry weight). Hypericins are absent in the pistil. The flavonoids hyperoside and isoquercitrin, the content of which decreased during ontogenetic phase of flowering, reach the highest contents in the pistil.     

The distribution of dimethylsulfoniopropionate in tropical Pacific coral reef invertebrates

Dimethylsulfoniopropionate (DMSP) is an important component of the global sulfur cycle and may be involved, via its cleavage product dimethylsulfide, in climate regulation. Although it is common in many algae, reports of DMSP in animals, particularly tropical invertebrates, are limited. This study examined the distribution of DMSP in a diverse group of coral reef invertebrates. DMSP was present in all 22 species of cnidarians and ranged from 9 to 723 μmol g⁻¹ of dry mass (DM) with a mean (± 1SD) of 110 ± 180 μmol g⁻¹ DM. It was not detected in a flatworm and an ascidian or in two of five sponges. Concentrations in sponges ranged from undetectable to 16 μmol g⁻¹ DM with a mean of 4 ± 7 μmol g⁻¹ DM. Within the cnidarians, DMSP concentrations did not differ among orders. Among cnidarian species, DMSP concentrations were correlated with symbiotic zooxanthellae densities. Within cnidarian species, DMSP concentrations of individuals were positively correlated with zooxanthellae densities in three of the four species examined. We speculate that DMSP is dietarily derived in sponges and derived from zooxanthellae in the cnidarians. The functions of DMSP in coral reef invertebrates are not known.     

Nitrate regulation of nitrate uptake and nitrate reductase expression in barley grown at different nitrate:ammonium ratios at constant relative nitrogen addition rate

Barley (Hordeum vulgare L. cv. Golf) was cultured using the relative addition rate technique, where nitrogen is added in a fixed relation to the nitrogen already bound in biomass. The relative rate of total nitrogen addition was 0.09 day^?1 (growth limiting by 35%), while the nitrate addition was varied by means of different nitrate: ammonium ratios. In 3- to 4-week-old plants, these ratios of nitrate to ammonium supported nitrate fluxes ranging from 0 to 22 μmol g^?1 root dry weight h^?1, whereas the total N flux was 21.8 ± 0.25 μmol g^?1 root dry weight h^?1 for all treatments. The external nitrate concentrations varied between 0.18 and 1.5 μM. The relative growth rate, root to total biomass dry weight ratios, as well as Kjeldahl nitrogen in roots and shoots were unaffected by the nitrate:ammonium ratio. Tissue nitrate concentration in roots were comparable in all treatments. Shoot nitrate concentration increased with increasing nitrate supply, indicating increased translocation of nitrate to the shoot. The apparent V_max for net nitrate uptake increased with increased nitrate fluxes. Uptake activity was recorded also after growth at zero nitrate addition. This activity may have been induced by the small, but detectable, nitrate concentration in the medium under these conditions. In contrast, nitrate reductase (NR) activity in roots was unaffected by different nitrate fluxes, whereas NR activity in the shoot increased with increased nitrate supply. NR-mRNA was detected in roots from all cultures and showed no significant response to the nitrate flux, corroborating the data for NR activity. The data show that an extremely low amount of nitrate is required to elicit expression of NR and uptake activity. However, the uptake system and root NR respond differentially to increased nitrate flux at constant total N nutrition. It appears that root NR expression under these conditions is additionally controlled by factors related to the total N flux or the internal N status of the root and/or plant. The method used in this study may facilitate separation of nitrate-specific responses from the nutritional effect of nitrate.     

Effects of low nitrogen medium on endogenous changes in ethylene, auxins, and cytokinins in in vitro shoot formation from rhizomes of Cymbidium kanran

Summary Shoot formation from rhizome explants of Cymbidium kanran was promoted on Murashige and Skoog (MS) medium: (1) with 1 mgl⁻¹ (4.4μM) 6-benzyladenine (BA) and 0.1 mgl⁻¹ (0.54μM) α-naphthaleneacetic acid (NAA); (2) with ethylene inhibitor (silver nitrate, AgNO₃); or (3) by reducing ammonium nitrate (NH₄NO₃) and potassium nitrate (KNO₃) to 25 and 50%, respectively, of their original concentrations. Shoot formation by BA and NAA was strongly inhibited with the application of ethephon, an ethylene releaser. The ethylene production from the rhizome explants was reduced 30–55% on low nitrogen medium after 1–3 mo. of culture and 52% on BA and NAA medium after 1 mo. of culture compared with explants on standard MS medium. No difference in endogenous auxin (indole-3-acetic acid, IAA) and cytokinin (isopentenyl adenosine, iPA) contents in the rhizomes was found between the treatments. Low ethylene levels were correlated with higher frequency of shoot formation from the rhizomes.     

Modeling nitrogen cycling in a coastal fresh water sediment

Increased nitrogen (N) loading to coastal marine and freshwater systems is occurring worldwide as a result of human activities. Diagenetic processes in sediments can change the N availability in these systems, by supporting removal through denitrification and burial of organic N (N_org) or by enhancing N recycling. In this study, we use a reactive transport model (RTM) to examine N transformations in a coastal fresh water sediment and quantify N removal rates. We also assess the response of the sediment N cycle to environmental changes that may result from increased salinity which is planned to occur at the site as a result of an estuarine restoration project. Field results show that much of the N_org deposited on the sediment is currently remineralized to ammonium. A rapid removal of nitrate is observed in the sediment pore water, with the resulting nitrate reduction rate estimated to be 130 μmol N cm⁻² yr⁻¹. A model sensitivity study was conducted altering the distribution of nitrate reduction between dissimilatory nitrate reduction to ammonium (DNRA) and denitrification. These results show a 40% decline in sediment N removal as NO ₃ ⁻ reduction shifts from denitrification to DNRA. This decreased N removal leads to a shift in sediment-water exchange flux of dissolved inorganic nitrogen (DIN) from near zero with denitrification to 133 μmol N cm⁻² yr⁻¹ if DNRA is the dominant pathway. The response to salinization includes a short-term release of adsorbed ammonium. Additional changes expected to result from the estuarine restoration include: lower NO ₃ ⁻ concentrations and greater SO ₄ ²⁻ concentrations in the bottom water, decreased nitrification rates, and increased sediment mixing. The effect of these changes on net DIN flux and N removal vary based on the distribution of DNRA versus denitrification, illustrating the need for a better understanding of factors controlling this competition.     

Microbial degradation of tetraethyl lead in soil monitored by microcalorimetry

Sieved agricultural soil samples were treated with the anti-knock agent tetraethyl lead (Et₄Pb), and the resulting effects were analyzed by microcalorimetry. Et₄Pb additions resulted in an increase of the heat production rate, provided that oxygen was present and that the soil was not autoclaved. The increased heat production rate was accompanied by degradation of Et₄Pb, as verified by speciation analysis (GC-MS) of the remaining Et₄Pb and its ionic degradation products (triethyl lead and diethyl lead cations). Conclusive evidence was obtained that these transformations were mediated mainly by microbes. At an initial Et₄Pb concentration of 2 g Pb/kg dry weight the biodegradation rate was about 780 μmol day⁻¹ kg dry weight⁻¹, whilst the chemical decomposition was only 50 μmol day⁻¹ kg dry weight⁻¹. A fivefold rise of the initial Et₄Pb concentration resulted in a decrease of the biodegradation rate to 600 μmol day⁻¹ kg dry weight⁻¹ and an increase of the chemical decomposition to 200 μmol day⁻¹ kg dry weight⁻¹. The biodegradation rate was not influenced by the addition of glucose, which means that no indication for a cometabolic attack of Et₄Pb was found. Received: 25 February 1997 / Received revision: 22 April 1997 / Accepted: 27 April 1997     

Nucleotide-Dependent Isomerization of Glutamate Dehydrogenase in Relation to Total RNA Contents of Peanut

The physiological function of glutamate dehydrogenase (GDH) was investigated by treating germinating peanut (Arachis hypogaea L.) seeds with nucleoside triphosphate (NTP) solutions in order to alter the isoenzyme distribution patterns. The free nucleosides and nucleotides of the GTP-treated peanut were the highest [8.7 μmol g⁻¹(f.m.)], and they decreased through the ATP-treated peanut [5.8 μmol g⁻¹(f.m.)], and CTP-treated peanut [5.5 μmol g⁻¹(f.m.)], to the UTP-treated peanut [4.1 μmol g⁻¹(f.m.)]. The combination of 4 NTPs induced 20 % higher content of Pi [173 nmol g⁻¹(f.m.)] than in the control, but the combined ATP+UTP treatment induced the lowest (93.0 nmol g⁻¹(f.m.)] Pi. The 4 NTP treatment also induced the highest number of GDH isoenzymes (28) followed by the purine NTP treatments (15 to 20), but the pyrimidine NTP treatments and the combined purine + pyrimidine NTP treatments induced the lowest numbers (<15) of isoenzymes. The deamination/amination ratios were generally higher in the UTP (0.11), and CTP (0.06) treated peanuts than in the GTP (0.04), and ATP (0.07) treated peanuts. There were mutual relationships between higher numbers of GDH isoenzymes present in the GTP-, and ATP-treated peanuts and higher RNA (236.5 and 239.4 μg g⁻¹, respectively) contents on one hand, and between the lower numbers of isoenzymes in the CTP-, and UTP-treated peanuts and lower RNA (162.0 and 152.5 μg g⁻¹, respectively) contents. The recurrent relationships of the effects of the NTP treatments of peanut were UTP > ATP > CTP > GTP. This revised version was published online in July 2006 with corrections to the Cover Date.     

Improvement of P450(BM-3) whole-cell biocatalysis by integrating heterologous cofactor regeneration combining glucose facilitator and dehydrogenase in E. coli

Escherichia coli BL21, expressing a quintuple mutant of P450_BM-3, oxyfunctionalizes α-pinene in an NADPH-dependent reaction to α-pinene oxide, verbenol, and myrtenol. We optimized the whole-cell biocatalyst by integrating a recombinant intracellular NADPH regeneration system through co-expression of a glucose facilitator from Zymomonas mobilis for uptake of unphosphorylated glucose and a NADP⁺-dependent glucose dehydrogenase from Bacillus megaterium that oxidizes glucose to gluconolactone. The engineered strain showed a nine times higher initial α-pinene oxide formation rate corresponding to a sixfold higher yield of 20 mg g⁻¹ cell dry weight after 1.5 h. The initial total product formation rate was 1,000 μmol h⁻¹ μmol⁻¹ P450 leading to a total of 32 mg oxidized products per gram cell of dry weight after 1.5 h. The physiological functioning of the heterologous cofactor regeneration system was illustrated by a sevenfold increased α-pinene oxide yield in the presence of glucose compared to glucose-free conditions.     

Critical Levels of Selenium in Different Crops Grown in an Alkaline Silty Loam Soil Treated with Selenite-Se

Critical levels of selenium in raya (Brassica juncea Czern L.), maize (Zea mays L.), wheat (Triticum aestivum L.) and rice (Oryza sativa L.) were worked out by growing these crops in an alkaline silty loam soil treated with different levels of selenite-Se ranging from 1 to 25 μg g⁻¹ soil. Significant decrease in dry matter yield was observed above a level of 5 μg Se g⁻¹ soil in raya and maize; 4 μg Se g⁻¹ soil in wheat and 10 μg Se g⁻¹ soil in rice shoots. The critical level of Se in plants above which significant decrease in yield would occur was found to be 104.8 μg g⁻¹ in raya, 76.9 μg g⁻¹ in maize, 41.5 μg g⁻¹ in rice and 18.9 μg g⁻¹ in wheat shoots. Significant coefficients of correlation were observed between Se content above the critical level and dry matter yield of raya as well as rice (r = −0.99, P ≤ 0.01), wheat (r = −0.97, P ≤ 0.01) and maize ((r = −0.96, P ≤ 0.01). A synergistic relationship was observed between S and Se content of raya (r = 0.96, P ≤ 0.01), wheat (r = 0.89, P ≤ 0.01), rice (r = 0.85, P ≤ 0.01) and maize (r = 0.84, P ≤ 0.01). Raya, maize and rice absorbed Se in levels toxic for animal consumption (i.e. > 5 mg Se kg⁻¹) when the soil was treated with more than 1.5 μg Se g⁻¹. In case of wheat, application of Se more than 3 μg g⁻¹ soil resulted in production of toxic plants.     

Growth, water use efficiency, and proline content of hydroponically grown tomato plants as affected by nitrogen source and nutrient concentration

Tomato plants (Lycopersicon esculentum Mill. cv. Counter) were grown in 12-L polyethylene containers in aerated and CaCO₃-buffered nutrient solutions containing different concentrations of complete stock solutions with either nitrate (stock solution N) or ammonium (stock solution A) as the only nitrogen source (X1 = standard concentration with 5 mM NO₃ ^–-N or NH₄ ⁺-N, and X3, X5.5, X8 and X11 = 3, 5.5, 8, 11 times the standard concentration), or a mixture of both stock solutions (N:A ratio = 100:0, 75:25, 50:50, 25:75, 0:100) at moderate nutrient concentration (X3). Total dry matter production and fruit dry weight were only slightly affected by increasing nutrient concentration if nitrate was supplied as the sole nitrogen source. Compared to nitrate, ammonium nitrogen caused a decrease in total dry weight (32–86% between X1 and X11), but led to an increase in both total dry weight and fruit dry weight (11% and 30%) at low concentration if supplied in addition to nitrate nitrogen (N:A ratio = 75:25). Dry matter partitioning in plants was affected by the strength of the nutrient solution, but even more by ammonium nitrogen. Fruits accumulated relatively less dry matter than did the vegetative parts of tomato plants when supplied with nutrient solutions containing ammonium as the only nitrogen source (fruit dry weight to total dry weight ratio 0.37 and 0.15 at low and high nutrient concentration), while nitrate nitrogen rather supported an increase in dry matter accumulation in the reproductive organs (fruit dry weight to total dry weight ratio 0.39–0.46). The water use efficiency (WUE) was only slightly affected by the strength of the nutrient solutions containing nitrate nitrogen (2.9–3.4 g DW (kg H₂O)^–1), while ammonium nitrogen led to a decrease in WUE from 2.4 to 1.3 g DW (kg H₂O)^–1at low (X1) and high (X11) nutrient concentration, respectively. The proline content of leaves fluctuated (0.1–5.0 mol (g fresh weight)^–1) according to nutrient concentration and global radiation, and reflected enhanced sensitivity of plants to these potential stress factors if ammonium was the predominant N source supplied. It was concluded, that proline is a reliable indicator of the environmental stress imposed on hydroponically grown tomato plants.