Nutrients can impact and regulate cellular metabolism and cell function which is particularly important for the activation and function of diverse immune subsets. Among the critical nutrients for immune cell function and fate, glutamine is possibly the most widely recognised immunonutrient, playing key roles in TCA cycle, heat shock protein responses and antioxidant systems. In addition, glutamine is also involved with inter-organ ammonia transport, and this is particularly important for not only immune cells, but also to the brain, especially in catabolic situations such as critical care and extenuating exercise. The well characterised fall in blood glutamine availability has been the main reason for studies to investigate the possible effects of glutamine replacement via supplementation but many of the results are in poor agreement. At the same time, a range of complex pathways involved in glutamine metabolism have been revealed via supplementation studies. This article will briefly review the function of glutamine in the immune system, with emphasis on metabolic mechanisms, and the emerging role of glutamine in the brain glutamate/gamma-amino butyric acid cycle. In addition, relevant aspects of glutamine supplementation are discussed.

Glufosinate resistance in Palmer amaranth (Amaranthus palmeri S. Watson) was recently detected in three accessions from Arkansas, USA. Amaranthus palmeri is the first and only broadleaf weed species resistant to this herbicide, and the resistance mechanism is still unclear. A previous study characterized the glufosinate resistance level in the accessions from Arkansas. A highly glufosinate-resistant accession was further used to investigate the mechanism conferring glufosinate resistance in A. palmeri. Experiments were designed to sequence the herbicide target enzyme cytosolic and chloroplastic glutamine synthetase isoforms (GS1 and GS2, respectively) and quantify copy number and expression. Absorption, translocation, and metabolism of glufosinate using the 14C-labeled herbicide were also evaluated in the resistant and susceptible accessions. The glufosinate-resistant accession had an increase in copy number and expression of GS2 compared with susceptible plants. All accessions showed only one GS1 copy and no differences in expression. No mutations were identified in GS1 or GS2. Absorption (54% to 60%) and metabolism (13% to 21%) were not different between the glufosinate-resistant and glufosinate-susceptible accessions. Most residues of glufosinate (94% to 98%) were present in the treated leaf. Glufosinate translocation to tissues above the treated leaf and in the roots was not different among accessions. However, glufosinate translocation to tissues below the treated leaf (not including roots) was greater in the resistant A. palmeri (2%) compared with the susceptible (less than 1%) accessions. The findings of this paper strongly indicate that gene amplification and increased expression of the chloroplastic glutamine synthetase enzyme are the mechanisms conferring glufosinate resistance in the A. palmeri accession investigated. Thus far, no additional resistance mechanism was observed, but further investigations are ongoing.

Glutamine synthetase (GS) and glutamate synthase (GOGAT) play a central role in plant nitrogen (N) metabolism. In order to study the effect of powdery mildew (Blumeria graminis f. sp. tritici, Bgt) on N metabolism, field experiments were carried out to evaluate GS and GOGAT activity, GS expression and grain protein content (GPC) in susceptible (Xi'nong 979) and resistant (Zhengmai 103) wheat cultivars under three treatments. The three treatments were no inoculation (CK), inoculated once with Bgt (MP) and inoculated nine times with Bgt (HP). For Xi'nong 979, the activities of GS and GOGAT in grains as well as GS activity in flag leaves increased at 10–15 days after anthesis (DAA), and decreased significantly at 15 or 20–30 DAA in HP and MP. However, GS activity in grains decreased from 20 DAA, which was later than that of flag leaves (15 DAA). At the same time, GS expression in grains was up-regulated at early stage, with GS1 at 10 DAA and GS2 at 15 DAA, followed by a continuous down-regulation. This result indicated that GS and GOGAT activity as well as GS expression were inhibited by powdery mildew, indicating that N metabolism in grains was inhibited at 20–30 DAA. The current study also found out that the yield of the susceptible cultivar decreased significantly, while its GPC increased obviously in HP. It was shown that the increase of GPC was not due to the enhancement of N metabolism, but due to the passive increase caused by yield reduction.

The transgenic Liberty Link® (LL) soybean is tolerant to glufosinate, conferred by the enzyme phosphinothricin acetyltransferase (PAT), which is encoded by the pat gene from Streptomyces viridochromogenes. Because symptoms of injury can be observed in soybean [Glycine max (L.) Merr.] plants in some situations, this study evaluated the effects of rates of glufosinate on agronomic performance; quality of LL soybean seeds; and the ammonia, glufosinate, and N-acetyl-l-glufosinate concentration (NAG) in soybeans with and without the pat gene after application of increasing glufosinate rates. Field and greenhouse experiments were conducted; the first evaluated the selectivity of glufosinate in LL soybeans, and the second evaluated the metabolic changes in soybeans with (LL) and without (RR2) the pat gene, after application of glufosinate. For fieldwork, application of glufosinate at rates up to four times the maximum recommended caused initial injury symptoms (up to 38.5%) in LL soybean plants. However, no negative effect was found on seed quality and agronomic performance of LL plants, including yield. This shows the selectivity of glufosinate promoted by pat gene insertion for application in POST (V4), in LL soybean. For the greenhouse experiment, it was concluded that the LL soybean plants presented high glufosinate metabolism, lower ammonia concentration, and no reduction in dry matter, in comparison with RR2 soybean, after application of high rates of glufosinate.

Tolerance to glufosinate has been bioengineered into cotton through the expression of a gene encoding the enzyme phosphinothricin acetyl transferase (PAT). Studies were conducted to determine thermal limitations on herbicide efficacy in bioengineered cotton. The 50% inhibition (I50) of glufosinate of the target-site enzyme glutamine synthetase was thermally dependent with the lowest values between 25 and 35 C. Larger values of I50 were measured above and below the 25 to 35 C range. The apparent Michaelis constant KM of the enzyme PAT was relatively stable from 15 to 30 C and increased more rapidly from 30 to 45 C. The two components in combination suggest the aggregate tolerance to glufosinate would not be thermally limited between 15 and 45 C. The thermal dependence of the aggregate tolerance in cotton suggests that glufosinate would not damage the crop over a range of temperatures. This prediction is in agreement with the results of field studies carried out over a number of years, which showed the glufosinate-tolerant cotton to be undamaged by glufosinate over a wide range of temperatures.

Greenhouse experiments were conducted to determine the effects of glufosinate on ammonium accumulation and photosynthetic inhibition in Palmer amaranth. Glufosinate applied at 410 g ha−1 reduced glutamine synthetase activity and enhanced ammonium content 30 min after treatment. Glufosinate application resulted in rapid inhibitions of the photosynthetic rate and stomatal conductance during the first 2 h after treatment (HAT), whereas the ammonium concentration increased over the same time period. Ammonium content 6 HAT was 22 times higher in treated plants than in untreated plants, whereas the photosynthetic rate of treated plants decreased by 63%. At 24 HAT, the ammonium content was 53 times higher in treated plants than in untreated plants; however, no further inhibition of photosynthesis occurred. Photosynthetic inhibition in Palmer amaranth coincided with the rapid accumulation of ammonium and decrease in stomatal conductance shortly after glufosinate application.

Glufosinate efficacy was examined in two major grass weed species, rigid ryegrass and sterile oat. Dose–response pot experiments under controlled environmental conditions showed that sterile oat was more successfully controlled by glufosinate than was rigid ryegrass. Glutamine synthetase was extracted from both species and assayed in vitro. Glufosinate readily inhibited glutamine synthetase activity in both species, indicating no differential sensitivity to the target enzyme. Thin-layer chromatography analysis of glufosinate showed no significant metabolism of glufosinate in either species. Absorption and translocation studies with 14C-glufosinate showed that the radiolabel was rapidly absorbed into the leaves of both species. However, translocation of radiolabeled glufosinate from the treated leaf to the meristematic regions was significantly greater in sterile oat, whereas translocation to the tip of the leaf was significantly greater in rigid ryegrass. This indicates that there is a difference in glufosinate distribution between the two species. It is likely that this difference in the distribution of glufosinate results in sterile oat being more easily controlled by glufosinate than is rigid ryegrass.

Resistance to glufosinate has been confirmed in glyphosate-resistant Italian ryegrass populations collected in hazelnut orchards in Oregon. Dose–response, ammonia accumulation, and enzyme activity studies were conducted to test the sensitivity of three glyphosate-resistant and three susceptible Italian ryegrass populations to glufosinate. The glufosinate rates required to reduce the growth by 50% (GR50) were 0.15, 0.18, and 0.21 for the control populations C1, C2, and C3, respectively, whereas for the resistant populations OR1, OR2, and OR3, the GR50 values were 0.49, 0.42, and 0.40 kg ai ha−1, respectively, exhibiting an average resistance index of 2.4. The same trend was observed in ammonia accumulation studies between 48 and 96 h after glufosinate treatment where the susceptible populations accumulated on average two times more ammonia than the resistant populations. The glufosinate concentration required to reduce the glutamine synthetase enzyme activity by 50% (I50) was not different for the resistant and susceptible populations. The I50s ranged from 3.1 to 3.6 µM for the resistant populations and from 3.7 to 4.3 µM for the susceptible populations; therefore, an insensitive target site is not responsible for the glufosinate resistance.

The objective of this study was to evaluate the response of hairy beggarticks plants to different doses of glufosinate ammonium and the range in sensitivities of the plants and their progenies to the herbicide. Three studies were conducted, all in a greenhouse and repeated at different times. In the first study, two experiments were conducted to examine the dose–response curve, and the treatments were seven different doses of the herbicide glufosinate ammonium (0, 50, 100, 200, 400, 800, and 1,600 g ai ha−1), with four replications each. In the second study, which examined the range in sensitivity of hairy beggarticks to glufosinate ammonium, 44 plants were sprayed with a dose of 200 g ai ha−1 of the herbicide. Finally, in the third study, the range in sensitivity of the progeny of hairy beggarticks to glufosinate ammonium was investigated; in this experiment, the progenies of seven of the previous plants were sprayed with 200 g ai ha−1 of herbicide. The ammonium contents in the tissues were measured and percent injury wase visually assessed. Ammoniun content in hairy beggarticks leaves was increased more than seven times by glufosinate application and the maximum ammonium content was observed for the highest dose of the herbicide. Variability existed in the ammonium content among the individuals of the population of hairy beggarticks; however, the behavior was not replicated in the same way in the progenies. The survival of the plants after application of the herbicide allows the production of progenies with wide variability in their sensitivity to the product, independent of the behavior for the progenitor plants.

Satellite glial cells (SGCs) are specialized cells that form a tight sheath around neurons in sensory ganglia. In recent years, there is increasing interest in SGCs and they have been studied in both intact ganglia and in tissue culture. Here we studied phenotypic changes in SGCs in cultured trigeminal ganglia from adult mice, containing both neurons and SGCs, using phase optics, immunohistochemistry and time-lapse photography. Cultures were followed for up to 14 days. After isolation virtually every sensory neuron is ensheathed by SGCs, as in the intact ganglia. After one day in culture, SGCs begin to migrate away from their parent neurons, but in most cases the neurons still retain an intact glial cover. At later times in culture, there is a massive migration of SGCs away from the neurons and they undergo clear morphological changes, and at 7 days they become spindle-shaped. At one day in culture SGCs express the glial marker glutamine synthetase, and also the purinergic receptor P2X7. From day 2 in culture the glutamine synthetase expression is greatly diminished, whereas that of P2X7 is largely unchanged. We conclude that SGCs retain most of their characteristics for about 24 h after culturing, but undergo major phenotypic changes at later times.

We recently proposed a model of cytogenesis which assumes that primitive ancestral mammals and premammalian vertebrates had a retinal composition that consisted of about seven neurons per Müller cell, comprising 1–2 cone photoreceptors, 1–2 rod photoreceptors, 2–3 bipolar cells, 1–2 amacrine cells, less than 1 ganglion cell, and less than 1 horizontal cell (Reichenbach & Robinson, 1995). The Australian lungfish (Neoceratodus forsten) closely resembles the lobe-finned ancestors of land vertebrates, and has an extremely plesiomorphic nervous system. The present study, therefore, has examined the relative frequencies of retinal neurons and Müller cells (identified by immunolabelling for glutamine synthetase) in the lungfish retina. It was found that for each Müller cell there is an average of 1.9 cone photoreceptors, 1.7 rod photoreceptors, 3.1 amacrine/bipolar/horizontal cells, and 0.6 ganglion cells; amounting to a ratio of 7.3 neurons per Müller cell. These results support our conjecture that the sequence of cytogenesis in mammals is constrained by a developmental program that predates the evolution of mammals. The study also provides the first detailed morphological descriptions of lungfish Müller cells and their relationship with adjacent neurons. It was found that individual Müller cells in lungfish have a volume (more than 12,000 μm3) that is an order of magnitude higher than in mammals, yet the proportion of total retinal volume occupied by these cells (20%) is very similar.

Biomphalaria glabrata embryonic (Bge) cells have been shown to be a valuable in vitro cellular model for the study of snail host–parasite interactions. They both promote the growth and differentiation of various trematode species including Schistosoma mansoni, and Echinostoma caproni and share some morphological and functional features with circulating haemocytes. As an approach to investigate snail genes potentially regulated following exposure to trematode excretory–secretory (ES) products, we compared gene expression profiles of Bge cells exposed to saline solution, or saline solution containing ES products from S. mansoni or E. caproni, two trematode species parasitizing B. glabrata. Following differential display RT-PCR analysis we characterized 23 differentially displayed cDNAs and we focussed on the 5 cDNAs showing sequence similarity to known genes for expression validation. Using RT-PCR, we confirmed that ES products from S. mansoni and E. caproni differentially affect the expression levels of 4 out of the 5 transcripts. These partial transcripts corresponded to novel B. glabrata sequences, and showed significant sequence similarity to genes coding for (i) cytochrome C, (ii) methyl-binding proteins, (iii) glutamine synthetases, and (iv) protease inhibitors from the Kunitz family. The possible significance of these gene expression changes in host–parasite molecular interactions is discussed.

The activities of nitrate reductase and glutamine synthetase were evaluated in young plants of Faidherbia albida, a tropical woody legume, fed with different N sources under hydroponic conditions. Results showed that assimilation of both NO3− and NH4+ preferentially took place in shoots. A basal amount of nitrate reductase activity was detected in shoots of plants grown with an NO3−-free solution or placed under N2-fixing conditions, and also in nodules of N2-fixing plants. This strongly suggests that constitutive nitrate reductase activity is present in these organs. Analyses of the soluble nitrogenous content showed that the major form of N in the different organs was α-amino acids (particularly amides), irrespective of the N status of the culture conditions. The same result was obtained for nodulated plants grown in local sandy soil. In this case, amide-N generally accounted for more than 40% of the total soluble N. This was especially true in nodules. Ureide-N never exceeded 9% of the total soluble N and did not appear to increase with increasing nodule nitrogenase activity. Amides were also predominant in three N2-fixing Sahelian acacias (Acacia seyal, A. nilotica and A. tortilis), showing that F. albida does not differ from Sahelian Acacia in terms of the metabolism of fixed N. However, like another Sahelian acacia growing preferentially near water (A. nilotica), F. albida can be distinguished from acacias growing strictly in arid zones (A. seyal and A. tortilis) in terms of initial growth, water and nitrate management.

The purpose of the present experiments was to evaluate the contribution of the glutamate-glutamine cycle in retinal glial (Müller) cells to photoreceptor cell synaptic transmission. Dark-adapted isolated rat retinas were superfused with oxygenated bicarbonate-buffered media. Recordings were made of the b-wave of the electroretinogram as a measure of light-induced photoreceptor to ON-bipolar neuron transmission. L-methionine sulfoximine (1–10 mM) was added to superfusion media to inhibit glutamine synthetase, a Müller cell specific enzyme, by more than 99% within 5–10 min, thereby disrupting the conversion of glutamate to glutamine in the Müller cells. Threo-hydroxyaspartic acid and D-aspartate were used to block glutamate transporters. The amplitude of the b-wave was well maintained for 1–2 h provided 0.25 mM glutamate or 0.25 mM glutamine was included in the media. Without exogenous glutamate or glutamine the amplitude of the b-wave declined by about 70% within 1 h. Inhibition of glutamate transporters led to a rapid (2–5 min) reversible loss of the b-wave in the presence and absence of the amino acids. In contrast, inhibition of glutamine synthetase did not alter significantly either the amplitude of the b-wave in the presence of glutamate or glutamine or the rate of decline of the b-wave found in the absence of these amino acids. Excellent recovery of the b-wave was found when 0.25 mM glutamate was resupplied to L-methionine sulfoximine–treated retinas. The results suggest that in the isolated rat retina uptake of released glutamate into photoreceptors plays a more important role in transmitter recycling than does uptake of glutamate into Müller cells and its subsequent conversion to glutamine.

The physiological potential for acquisition of atmospheric ammonia (NH3) was investigated in three European meadow grasses (Arrhenatherum elatius, Bromus erectus and Brachypodium pinnatum) competing in chalk grasslands. Experiments were carried out with plants cultivated for about three months on a soil–sand mixture at high root nitrogen supply or in nutrient solutions at low root nitrogen supply. Two different root nitrogen regimes were applied to the solution-grown plants: 130 μmol NO3 plant−1 wk−1 (approx. 50 kg nitrogen ha−1 in three months); or 130 μmol NO3− plus 130 μmol NH4+ plant−1 wk−1. Each regime was combined with two levels of NH3 fumigation (0 and 70 nmol mol−1 air for 24 d). Uptake of gaseous NH3 in the shoots was investigated under controlled environmental conditions including NH3 concentrations ranging from 0 to 30 nmol mol−1 air. Concurrently, photosynthesis, glutamine synthetase activity, nitrogen allocation, biomass allocation and apoplastic cation composition were measured. For A. elatius, the influence of photorespiration on NH3 acquisition was also assessed. Independently of plant nitrogen status, ammonia compensation points in A. elatius and B. erectus plants were <0.5 nmol mol−1. The total leaf conductance to NH3 absorption remained constant at increasing NH3 concentrations, showing that the capacity for assimilation was unaltered. Whereas internal factors in the leaves did not cause differences in the potential for NH3 acquisition between the species, other factors of NH3 acquisition were quite different: B. erectus had higher stomatal conductance and, thus, higher NH3 uptake rates per unit leaf area compared to A. elatius and B. pinnatum; higher stomatal conductances of B. erectus were to a large extent offset by a lower leaf area per plant, resulting from a lower growth rate and thicker leaves than in the two other species. The rate of photorespiration in Arrhenatherum constituted at least 15% of the net photosynthetic rate. Surprisingly, suppression of photorespiration indicated that NH3 uptake was supported by photorespiration. Bromus responded to fumigation with 70 nmol NH3 mol−1 air for 24 d by lowering the root[ratio ]shoot ratio and increasing the nitrogen concentration in the stem dry matter. The total leaf conductance to NH3 uptake decreased in all three species upon exposure to NH3, while the stomatal conductance was unaffected. The NH3 exposure caused lower apoplastic concentrations of H+, Mg2+ and Ca2+ in A. elatius and B. erectus.

Dry deposition of NH3 and NOx (NO and NO2) can affect plant metabolism at the cellular and whole-plant level. Gaseous pollutants enter the plant mainly through the stomata, and once in the apoplast NH3 dissolves to form NH4+, whereas NO2 dissolves to form NO3− and NO2−. The latter compound can also be formed after exposure to NO. There is evidence that NH3-N and NOx-N can be reversibly stored in the apoplast. Temporary storage might affect processes such as absorption rate, assimilation and re-emission. Once formed, NO3− and NO2− can be reduced, and NH4+ can be assimilated via the normal enzymatic pathways, nitrate reductase (NR), nitrite reductase and the glutamine synthetase/glutamate synthase (GS/GOGAT) cycle. Fumigation with low concentrations of atmospheric NH3 increases in vitro glutamine synthetase activity, but whether this involves both or only one of the GS isoforms is still an open question. There seems to be no correlation between fumigation with low concentrations of NH3 and in vitro GDH activity. The contribution of atmospheric NH3 and NO2 deposition to the N budget of the whole plant has been calculated for various atmospheric pollutant concentrations and relative growth rates (RGRs). It is concluded that at current ambient atmospheric N concentrations the direct impact of gaseous N uptake by foliage on plant growth is generally small.

The activities of the two key enzymes involved in glutamine metabolism, glutaminase (EC 3.5.1.2) and glutamine synthetase (EC 6.3.1.2), have been measured in the various tissues of the gastrointestinal (GI) tract of the rat, from the mouth to the rectum. Glutaminase activity was particularly high in the mucosa of the small intestine, where its activity accounted for more than 80% of the total activity of the GI tract. In contrast, the mouth and oesophagus had very low activities, accounting for less than 2% of the total. Glutamine synthetase was mainly confined to the lower part of the stomach, which accounted for almost 90% of the total activity of the GI tract. Activity in the small intestine was very low, accounting for less than 2% of the total, and similarly low levels were found in the mouth and oesophagus. The data provide the most complete information on the distribution of these enzymes in the GI tract of the rat and suggest: (a) that the mucosa of the small intestine has the highest capacity for glutamine breakdown but the lowest capacity for its synthesis, and so requires an external source of this amino acid; (b) that there is little potential for glutamine synthesis or breakdown in the mouth and oesophagus; and (c) that the lower stomach has a substantial capacity to synthesize glutamine, in contrast to the rest of the GI tract. The results of the investigation are relevant to sites of glutamine metabolism in therapeutic studies involving glutamine administration discussed with reference to reports of the effects of glutamine administration on GI tract injury.

Using field plots, three species (Mercurialis perennis L., Rubus fruticosus L., and Trientalis europaea L.) were tested for their potential to emit gaseous ammonia to the atmosphere. Canopies were misted with 5 mM methionine sulphoximine (MSO) to inhibit glutamine synthetase (GS), the enzyme of ammonium assimilation. Leaf tissue NH4+ concentration of control plants was 0·03–0·1 μmol g−1 f. wt. Although NH4+ accumulated in the leaf tissue of MSO-treated plants of all three species to similar concentrations (6-10 μmol g−1 f. wt after 4 d), emissions were only detected from the leaves of M. perennis, with potential rates of 2·5 nmol m−2 leaf s−1. Experiments carried out in a controlled environment confirmed this rate of emission over 9 d, during which time leaf tissue ammonium increased to 66 μmol g−1 f. wt. Comparisons with Hordeum vulgare grown under the same conditions showed that tissue NH4+ concentration reached a plateau of about 40 μmol g −1f. wt after 2 d. Emissions of NH3 during the 5 d of treatment reached a maximum rate of 10 nmol m−2 s−1 by the third day.

Apoplastic pH of the plants was determined, and it is suggested that this is an important factor explaining the differences in NH3 emission between species. The higher the apoplastic pH, the greater the likelihood of loss of NH3 from sub-stomatal spaces to the atmosphere. T. europaea (non-emitter) had an apoplastic pH of 5, R. fruticosus (non-emitter) a pH of c. 5·6, whereas that of M. perennis (emitter) was c. pH 6·3. The apoplastic pH is thought to be dictated in part by the N nutrition of a species, nitrophilous species tending to have high pH. Without NO3− fertilization, H. vulgare had an apoplastic pH of 6·8 but this increased to 7·3, 3 d after feeding with NO3−.

Short-term fumigation (2 h) of shoots of H. vulgare with 60 μg (≈32 mg NH3 m−3) of labelled gaseous 15N-NH3 (in the absence of MSO) showed that a substantial proportion (60%) of the applied label was found in the leaves, as well as in stems and roots (3%). There was also a change in amino acid pools, with an increase in shoot amino acids and a decrease in those in the root, while tissue NH4+ was very low in both shoots and roots. This provided indirect evidence that some of the applied label had been incorporated into an organic form. Following the fumigation treatment, emissions of NH3 were collected for 3 h, then c. 6·5 μg of N was recovered, of which c. 17% was 15N-labelled. Some of this label could have resulted from desorption of NH3 from leaf surfaces, but it was more likely that the remaining 14N isotope was from sub-stomatal emissions of NH3.

It is argued that non-nitrophilous plants tend to rely on mixed sources of N (NO3−, NH4+ or organic-N) and are more likely to favour root rather than shoot assimilation. Under these circumstances, their apoplastic pH is relatively low (compared with that of nitrophiles, which tend to assimilate NO3− mainly in their shoots), and at atmospheric concentrations most wild species are likely to be net assimilators, rather than emitters, of atmospheric ammonia.