Possible evidence for novel metabolism of nitrogen gas by a green alga

We report the isolation of a cukaryotic green alga ( Chlorella , strain WPI-2) which accumulates large stores of nitrogen (N) during growth in N-free medium and seems to incorporate¹⁴N₂, yet does not reduce acetylene to ethylene. Total N accumulation during growth on N-free medium and in gases free of combined N was measured by three methods: Kjeldahl, oxidative pyrolysis via chemiluminescence (Antek N analyzer), and Dumas (Coleman N analyzer). Increases in N ranging from 22–64%± 1% were observed. Isotope dilution studies using cells labelled with ¹⁵NO ₃- and then shifted to ¹⁴N₂ in N-free medium showed dilution of the ¹⁵N isotope by ¹⁴N from 5.67 to 5.32%± 0.05%. Using a variety of conditions, we were unable to demonstrate the reduction of acctylene to ethylene by WPI-2, although diazotrophic cyanobacteria gave positive results. Although the data on WPI-2 are not conclusive in establishing this alga as a diazotroph, the data do suggest that within the Chlorophyceae there may exist a novel form of nitrogen gas metabolism.     

The effect of oxygen on nitrate and nitrite assimilation in leaves of Zea mays L. under dark conditions

The assimilation of nitrate under dark-N₂ and dark-O₂ conditions in Zea mays leaf tissue was investigated using colourimetric and ¹⁵N techniques for the determination of organic and inorganic nitrogen. Studies using ¹⁵N indicated that nitrate was assimilated under dark conditions. However, the rate of nitrate assimilation in the dark was only 28% of the rate under non-saturating light conditions. No nitrite accumulated under dark aerobiosis, even though nitrate reduction occurred under these conditions. The pattern of nitrite accumulation in leaf tissue in response to dark-N₂ conditions consisted of three phases: an initial lag phase, followed by a period of rapid nitrite accumulation and finally a phase during which the rate of nitrite accumulation declined. After a 1-h period of dark-anaerobiosis, both nitrate reduction and nitrite accumulation declined considerably. However, when O₂ was supplied, nitrate reduction was stimulated and the accumulated nitrite was rapidly reduced. Anaerobic conditions stimulated nitrate reduction in leaf tissue after a period of dark-aerobic pretreatment.     

Seasonal variation of nitrogenous compounds in the xylem sap of Citrus

Samples of tracheal sap of Citrus sinensis (L.) Osbeck cv. Washington Navel were taken from field trees throughout the year and the nitrogen composition of the sap was determined. The nitrogenous fraction of the sap was composed mainly of free amino acids (92–97% of total nitrogen) and nitrates throughout the year. Proline was the most abundant amino acid during almost the entire cycle, and its concentration was especially high during the autumn and winter period. Nevertheless, a significant part (40–60%) of the total organic nitrogen was transported as arginine. Total nitrogen as well as amino acids and nitrates were maximal at spring flush. At spring flush and summer flush there was also a diversification of α-amino nitrogen among different amino acids. During the spring flush, nitrates, asparagine and γ-aminobutyric acid in the xylem sap seemed to have a radicular origin, whereas glutamic acid and arginine were released from the surrounding parenchyma. The results suggest a metabolic transformation in the wood parenchyma of nitrogenous compounds coming from the roots (including reduction of nitrates) and a turnover of different nitrogen metabolites between the xylem and surrounding cells.     

Nutrition of winter wheat during the life cycle. I. Yield and accumulation of dry matter and minerals

Mineral uptake by winter wheat (Trilicum aestivum L. cv. Martonvasari 8) was studied throughout the life cycle. Accumulation of macronutrients (i.e. total nitrogen, phosphorus, potassium, sodium, magnesium and calcium) and the water content of roots and shoots of plants grown in complete nutrient solution were higher than those of plants grown in two types of soils. The supply of macronutrients was in some cases limiting for soil-grown plants as revealed by a comparison of available and accumulated amounts of these nutrients. Their supply was abundant, however, for solution-grown plants. This led to a doubling of grain yield for the latter plants with a three fold increase in accumulation of dry matter and a five-fold increase in fresh weight. The efficiency ratios of solution-grown plants to soil-grown plants were approximately 1 for N and Na, 0.5 for Mg and Ca, and 0.3 for P and K.     

The influence of a deep-storage reservoir on the physicochemical limnology of a Central Texas River

A study was conducted to determine a physicochemical profile of a new deep-storage reservoir and to determine the influence of impoundment and thermal stratification in the reservoir on the physicochemical limnology of the parent river. The presence of thermal stratification from May through November caused the most significant change in water conditions. Of 23 parameters studied, 12 remained unchanged, 10 improved, and 2 deteriorated (Table VI).The greatest downstream changes in water conditions from those upstream from the reservoir were a decrease in temperature, an increase of ammonia, and the presence of hydrogen sulfide during the period of thermal stratification. Ammonia did not increase to a level considered to be toxic to aquatic species. It could, however, serve as a nutrient for certain species of plants and result in a change in community structure.Water tempeature downstream from the reservoir was always within the annual temperature range of the river upstream from the reservoir; however, the summer maximum in the tailrace was decreased to a temperature that could interfere wih the normal life cycle of many species.     

Electron paramagnetic resonance of single crystal deoxycobaltohemoglobin

Single crystals of horse ^CoHb were obtained by reduction of ^CoHb⁺ crystals with dithionite. Epr measurements showed that the g? and ^Coà tensors are both axial and share the same principal axis systems. Of the four subunits, the “heme” normals of C? and d? subunits ãb?plane 29 ± 1° from b?; they have the same orientation as the hemes in methemoglobin. The normals of “hemes” à and B? are 47 above the ãb? plane as compared to 16° in methemoglobin.     

The Effect of Light and Exogenously Supplied Ammonium Ions on Glutamate Dehydrogenase Activity and Isoforms in Young Mustard (Sinapis alba L.) Seedlings

Glutamate dehydrogenase (GDH, E.C. 1.4.1.3) of mustard cotyledons was investigated during the first 4 days of seedling development. The enzyme was found to be composed of seven catalytically active isoforms (each with a molecular mass of 270 kDa) which exhibited a charge heterogeneity when investigated by isoelectric focusing. Antibodies against the purified isoform 7, raised in rabbits, cross-reacted with each of the isoforms in Western blotting experiments. In addition, each of the isoforms was composed of four immunopositive reacting polypeptides with 19, 21, 23 and 25 kDa. During development of the seedlings, a shift in the isoform pattern towards the more acidic forms was found which was more pronounced when the seedlings were supplied with 15 mM NH₄Cl. The time course of changes in total GDH level can be correlated with the time course of disappearance of storage proteins. Both parameters are negatively regulated by light possibly via the photoreceptor, phytochrome. There are some indications that GDH in young mustard cotyledons mainly acts in the deaminating direction.     

Assessment of genetic variability for N2 fixation between and within provenances of Leucaena leucocephala and Acacia albida estimated by 15N labelling techniques

Nitrogen fixed in 13 provenances of Acacia albida and 11 isolines of Leucaena leucocephala inoculated with effective Rhizobium strains was measured by ¹⁵N techniques and the total N difference method. In the test soil, on the average, L. leucocephala derived about 65% of its total N from atmospheric N₂ fixation compared to about 20% by A. albida. Significant differences in the percentage of N derived from atmospheric N₂ (% Ndfa) occurred, between provenances or isolines within species. The % Ndfa ranged from 37 to 74% within L. leucocephala and from 6 to 37 within A. albida; (equivalent to 20–50 mg N plant^–1 and 4–37 mg N plant^–1 for the two species over three months, respectively) and was correlated with the nodule mass (r=0.91). The time course of N₂ fixation of three selected provenances (low, intermediate and good fixers) was followed at 12 weekly intervals over a 36 week period. The % Ndfa of all provenances and isolines increased with time; and except for one of the L. leucocephala provenances, % Ndfa was similar within species at the 36 weeks harvest. There was a significant correlation between % Ndfa and the amount of N₂ fixed (r=0.96). Significant interactions occurred between provenances and N treatments and often growth of uninoculated but N fertilized plants was less variable than for inoculated unfertilized plants.     

Growth and carbon partitioning in compatible and incompatible peach/plum grafts

The shoot growth of compatible ( Prunus persica L. Batsch cv. Springtime grafted on Prunus cerasifera L. Ehrh cv. Myrobolan P2032) and incompatible ( Prunus persica L. Batsch cv. Springtime grafted on Prunus cerasifera L. Ehrh cv. Myrobolan P18) peach/plum grafts was observed over a period of 100 days after grafting under controlled conditions. Leaf and root activities were determined by studying carbon assimilation and partitioning, leaf mineral contents and water relations. Shoot length and leaf number were not significantly affected in the incompatible combination during the first 55 days after grafting, but then, shoot growth rate was significantly reduced. Final total dry weights of the shoot were similar in both graft combinations. The incompatible combination did not show any water stress. Soluble sugar and starch contents increased in the leaves of the incompatible combination, accounting for about 36% of the increase of leaf dry weight per unit area. Photosynthesis was affected by the compatibility of the grafts. Leaf nitrogen content (% dry weight) fell in the incompatible graft combination 65 days after grafting. However, nitrogen content on an area basis was not affected. The possibility of nitrogen stress is discussed.     

Enhancement of leaf nitrogen,phosphorus and potassium and seed protein in Vigna radiata by pyridoxine application

Soaking the seeds of mungbean (Vigna radiata L. Wilczek cv. K-851) in pyridoxine solution significantly enhanced leaf N, P and K concentrations at different growth stages, and seed protein concentration at harvest. Leaf N, P and K were significantly correlated with root length and seed protein. Thus, pyridoxine application not only enhanced the availability of nutrients to plants but also was responsible for the maintenance of a favourable source-sink relationship, thus ensuring more nutritious seeds of mungbean.