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PubMed1994

Identification of chromosomal genes located downstream of dctD that affect the requirement for calcium and the lipopolysaccharide layer of Rhizobium leguminosarum.

In Rhizobium leguminosarum both the C4-dicarboxylate transport system and wild-type lipopolysaccharide layer (LPS) are essential for nitrogen fixation. A Tn5 mutant (RU301) of R. leguminosarum bv. viciae 3841, was isolated that is only able to synthesize LPS II, which lacks the O-antigen. Strain RU301 exhibits a rough colony morphology, flocculates in culture and is unable to swarm in TY agar. It also fails to grow on organic acids, sugars or TY unless the concentration of calcium or magnesium is elevated above that normally required for growth. The defects in the LPS and growth in strain RU301 were complemented by a series of cosmids from a strain 3841 cosmid library (pRU3020-pRU3022) and a cosmid from R. leguminosarum bv. phaseoli 8002 (pIJ1848). The transposon insertion in strain RU301 was shown to be located in a 3 kb EcoRI fragment by Southern blotting and cloning from the chromosome. Sub-cloning of pIJ1848 demonstrated that the gene disrupted by the transposon in strain RU301 is located on a 2.4 kb EcoRI-PstI fragment (pRU74). R. leguminosarum bv. viciae VF39-C86, which is one of four LPS mutants previously isolated by U. B. Priefer (1989, J Bacteriol 171, 6161-6168), was also complemented by sub-clones of pIJ1848 but not by pRU74, suggesting the mutation is in a gene adjacent to that disrupted in strain RU301. Complementation and Southern analysis indicate that the region contained in pIJ1848 does not correspond to any other cloned Ips genes. Two dctA mutants, RU436 and RU437, were also complemented by pIJ1848 and pRU3020. Mapping of pIJ1848 and Southern blotting of plasmid-deleted strains of R. leguminosarum revealed that dctD and the region mutated in strain RU301 are located approximately 10 kb apart on the chromosome. Analysis of homogenotes demonstrated that there is not a large region important in calcium utilization, organic acid metabolism or LPS biosynthesis located between the gene disrupted in strain RU301 and dctD. Strain VF39C-86 also required an elevated concentration of calcium for growth on succinate, while strains mutated in the alpha-chromosomal or beta-plasmid group of Ips genes grew at the same calcium concentrations as the wild type, demonstrating that the additional calcium requirement is not a property of all LPS rough mutants. Strain RU301 nodulates peas, but does not reduce acetylene, demonstrating that the gene mutated in this strain is essential for nitrogen fixation.

Bacterial Proteins↗
PubMed2006

Fixation of nitrogen in an electrospray mass spectrometer.

The formation of an iron-nitrogen compound in an electrospray ionization instrument is reported. The iron was released from the stainless steel electrospray needle by acetic acid added to displace trifluoroacetic acid and the nitrogen was present as the drying gas. The product is an iron nitride ion, [N(2)FeOH](1+), m/z 100.9438, and is capable of addition to peptides and proteins.

Acetic Acid↗
PubMed2005

Effects of enhanced ultraviolet-B radiation on algae and cyanobacteria.

This article provides an overview of existing literature on the ultraviolet-B (UV-B) radiation effects on algae and cyanobacteria. We report on the effects of UV-B radiation to the growth and development, biomass, sensitivity, photosynthetic pigments, UV-B absorbing compounds, photosynthesis, protein and DNA damage, enzyme activity, nitrogen fixation and assimilation of nitrogen, protective mechanisms of algae and cyanobacteria, the accommodation of algae and cyanobacteria to environmental stress, and the effects to ecology system. Many of the studies show the dramatic effects of UV-B radiation; but typically these studies were conducted under conditions with supplemental UV-B irradiance that was higher than would ever occur outside experimental conditions or natural condition. A few of the studies reviewed used experimental conditions and supplemental UV-B irradiance that approached realism. Enhanced UV-B generally decreased chlorophyll content, whereas it increased UV-B absorbing compounds in many algae. Decrease in photosynthesis, particularly at higher UV-B doses, was due to both direct (effect on photosystem) and indirect (decrease in pigments) effects. The decreases in chlorophyll pigments and photosynthesis resulted in lower biomass. However, algae and cyanobacteria have evolved various avoidance and repair mechanisms to protect themselves against the damaging effects of UV radiation to acclimate to enhanced UV-B radiation. The review points to areas where further studies on the relationships among nitrogenase, Rubisco, antioxidase activity, signal, antioxidants, and free radicals under enhanced UV-B are needed to quantify the effects of UV-B radiation on algae and cyanobacteria. These studies are needed in order to develop dose response functions that can facilitate development of dynamic simulation models for use in UV-B and other environmental impact assessments.

Cyanobacteria↗
PubMed2001

Role of Escherichia coli nitrogen regulatory genes in the nitrogen response of the Azotobacter vinelandii NifL-NifA complex.

The redox-sensing flavoprotein NifL inhibits the activity of the nitrogen fixation (nif)-specific transcriptional activator NifA in Azotobacter vinelandii in response to molecular oxygen and fixed nitrogen. Although the mechanism whereby the A. vinelandii NifL-NifA system responds to fixed nitrogen in vivo is unknown, the glnK gene, which encodes a PII-like signal transduction protein, has been implicated in nitrogen control. However, the precise function of A. vinelandii glnK in this response is difficult to establish because of the essential nature of this gene. We have shown previously that A. vinelandii NifL is able to respond to fixed nitrogen to control NifA activity when expressed in Escherichia coli. In this study, we investigated the role of the E. coli PII-like signal transduction proteins in nitrogen control of the A. vinelandii NifL-NifA regulatory system in vivo. In contrast to recent findings with Klebsiella pneumoniae NifL, our results indicate that neither the E. coli PII nor GlnK protein is required to relieve inhibition by A. vinelandii NifL under nitrogen-limiting conditions. Moreover, disruption of both the E. coli glnB and ntrC genes resulted in a complete loss of nitrogen regulation of NifA activity by NifL. We observe that glnB ntrC and glnB glnK ntrC mutant strains accumulate high levels of intracellular 2-oxoglutarate under conditions of nitrogen excess. These findings are in accord with our recent in vitro observations (R. Little, F. Reyes-Ramirez, Y. Zhang, W. Van Heeswijk, and R. Dixon, EMBO J. 19:6041-6050, 2000) and suggest a model in which nitrogen control of the A. vinelandii NifL-NifA system is achieved through the response to the level of 2-oxoglutarate and an interaction with PII-like proteins under conditions of nitrogen excess.

Azotobacter vinelandii↗
PubMed1977

[The effect of biocides on the microflora of soils and their degradation. 4. The influence of substances inhibiting the straw decomposition on the nitrogen transformation (author's transl)].

1. Active ingredients retarding the carbon-transformation of straw (TCP, Thiuram, TBTO) also retard the nitrification. Their active period depends on the concentration and on the presence of organic matter. High doses of urea can inhibit the nitrification as well. 2. The activity of urease and the ammonification could not be influenced by adequate doses of the tested active ingredients. 3. The species of bacterium Azotobacter fixing the nitrogen from the air were inhibited by TCP, urea and D-chloramphenicol and promoted by Thiuram and WTZ II 277 after a short period of inhibition. 4. The fungi are very important in the biological nitrogen fixation after manuring straw and in prevention of nitrogen leaching in the autumn. 5. The sufficient concentration of the active ingredients to inhibit the nitrification are lower than the fungicidal concentrations. The soil bacteria could even not be restrained by doses of 5000 ppm. The bacilli were more retarded in presence of organic matters than in a not manured soil.

Ammonia↗
PubMed1996

[Agricultural and nutritional importance of legumes].

The main ecophysiologic, agronomic and economic feature of legume plants is the development of tubercles and nodules in their apical system. Nodule formation occurs in most legume species provided a compatible type of Rhizobium bacteria is present in the soil. Nitrogen fixation in nodules renders these plants independent of nitrogen fertilizers, the most expensive of all goods in modern cereal agriculture. Considering that soils may get enriched in nitrogen through fixation in nodules and the decomposition of foliage when the aerial parts of legume plants are used as green fertilizers, only through the inclusion of legume crops within planned harvest schemes, it would be possible to achieve success in large scale production strategies. Legume crops are extensively produced in temperate climates areas in which, in addition to their use in animal nutrition, yields of 18 kg per person per year are obtained. In contrast, in the Third World countries located in tropical areas, legume production is scarce, with annual yields of 9 kg per person per year. Currently, it is proposed that the energy and protein intake should match that of the developed countries 40 years ago (i.e. 3000 Kcal and 70 g protein per day); for this, it would be necessary to have an average availability of 60 g of legume seeds per person per day. Therefore, the production of legume seeds should be increased. In addition, research aimed to study and exploit the agronomic potential of this rich botanical family should be strengthened through the formation of interdisciplinary groups.

Agriculture↗