Denitrification in undisturbed cores from a solodized solonetz B horizon

Summary An electrolytic respirometer was used to study the effects of oxygen, moisture and glucose on denitrification in undisturbed cores from a solodized solonetz B horizon. Under the experimental conditions, nitrous oxide present after 144 h was a reasonable index of total denitrification. Nitrous oxide evolution was positively correlated with moisture, quantity of added glucose, and negatively correlated with per cent oxygen. With glucose present, denitrification proceeded at high oxygen and moderate moisture levels, but was most rapid at low oxygen and high moisture. At low oxygen content, denitrification increased with increasing water content. In horizons with high potential denitrifying activity, denitrification could occur under field conditions at moderate aeration and moderately high moisture contents.This work was supported by the Australian Wheat Industry Research Council as part of a programme of investigation of nitrogen conservation in Australian wheat soils.Formerly holder of an Australian Wheat Industry Research Council Studentship, and Associate Professor, respectively, Department of Agronomy, University of New England, Armidale, New South Wales, Australia. The senior author is now with CSIRO, P.O. Box 21, Katherine, N.T. 5780, Australia.     

Heterotrophic Growth and Production of Xanthophylls by Chlorella pyrenoidosa

The growth and level of xanthophylls of several representative species of green algae were investigated as a possible source of pigmentation for the egg yolk and broiler markets. Chlorella pyrenoidosa 7-11-05 was selected for fermentation studies because of its high level of xanthophylls and wide temperature range for growth. The heterotrophic metabolism was preferred because of the ease of adaptability to present fermentation equipment. When used as the sole carbon source, glucose was the only sugar, among many tested, that gave appreciable growth in illuminated shaken flasks. A dry cell weight of 90 g per liter and total xanthophylls of 450 mg per liter were obtained from 190 g per liter of glucose monohydrate in 168-hr illuminated shaken flasks. Higher levels of glucose decreased yields. In combination with glucose, monosaccharides, such as fructose and galactose, were readily assimilated. The 7-11-05 strain was adapted to galactose as the sole carbon source after six vegetative passages. Light of the proper intensity and duration stimulated total xanthophylls approximately 35%. The effect on dry cell weight and total xanthophylls of seven antibiotics added at various levels in shaken flasks was studied. Erythromycin was essentially stable throughout the fermentation and nontoxic up to 25 μg/ml, with only slight toxicity at higher levels. Both erythromycin and ristocetin were effective in controlling a high incidence of bacterial contamination in 30-liter fermentors. With the higher agitation and aeration rates possible in 30-liter fermentors, dry cell weights in excess of 100 g per liter and total xanthophylls of 467 to 512 mg per liter were readily obtained from 230 to 260 g per liter of glucose in 162-hr illuminated batch-type fermentations. Continuous-feed runs yielded a dry cell weight of 302 g per liter and total xanthophylls of 650 mg per liter from 520 g per liter of glucose. The type of Chlorella cell produced was an important consideration with respect to the availability of the xanthophylls in pigmenting egg yolks and broilers.     

Effect of Shaking on the Growth of Diluted Cultures of Tetrahymena

ABSTRACT. Cultures of Tetrahymena pyriformis, T. thermophila and T. pigmentosa have been studied with regard to growth rates in shaken and unshaken flasks. In the standard medium, a minimum doubling time of 170 min was obtained for T. pyriformis at 28° C in the unshaken cultures. If the depth of the medium was less than 1 cm, the gyratoric shaking increased the doubling time to 340 min. The effect of shaking could be reduced by the addition of dextrane. Cells subjected to shaking were observed in different media and at different growth temperatures. If cultures were inoculated with 10⁴ cell·ml⁻¹ or more, the effect of shaking was absent. However, with inoculates of 10³ or 10² cell·ml⁻¹, the doubling times for T. pyriformis increased to 240 and 275 min, respectively. Periods of 2 min shaking followed by rest for 60 min could not induce an effect.     

RESPIRATORY ACTIVITY OF GUINEA PIG BRAIN NUCLEI

Abstract— Preparations of guinea pig brain nuclei, obtained by discontinuous gradient centrifugation in sucrose solutions of pH 6.7–6.8, containing 3 mM-MgCl₂ and phosphate exhibited steady and reproducible oxygen uptake. Oxygen uptake was stimulated 60–70 per cent by glucose, pyruvate, oxalacetate or α-ketoglutarate and 267 per cent by succinate. This respiratory activity was unaffected by the relative sodium or potassium ion content of the medium and by variations in the concentration of inorganic phosphate. Agents known to inhibit citric acid cycle oxidation, oxidative phosphorylation and glycolysis diminished oxygen uptake, but antibiotics inhibiting nucleic acid or protein synthesis did not. Treatment of the nuclear preparation with DNase decreased respiratory capacity, which was partially restored by the addition of polyacrylic acid.     

Wheat Viability at High Moisture Content under Hermetic and Aerobic Storage Conditions

Wheat seeds (Triticum durum) were stored under both hermeticand aerobic conditions at 25 °C with moisture contents from15 to 33 per cent. Under hermetic storage, seeds lost viabilitymore rapidly the higher the moisture content, whereas in aerobicstorage, seed longevity was enhanced as the moisture contentwas increased from 24 to 31 per cent, and over this range ofmoisture content the seeds survived longer under aerobic thanhermetic storage. On the contrary, an apparent reversal of thistrend occurred when moisture content was increased above 31per cent. The possibility that the changes in longevity occurring at highermoisture contents might be due to the activation of seed metabolismwas supported by the enhanced incorporation of [³H]leucine intoTCA insoluble material (indicating increased protein synthesis)and the reduced leakage of glucose (indicating increased membranerestitution) when seeds were allowed to achieve higher moisturecontents during the prehydration period. The highest level ofseed activation was found in seeds preconditioned to about 31per cent moisture content. Moreover, these seeds, when subsequentlystored under aerobic conditions, maintained a higher rate ofprotein synthesis and lower membrane permeability during thestorage period than seeds at lower water contents. It is suggested that seeds stored at a sufficient hydrationlevel in the presence of oxygen can sustain an effective metabolismduring extended storage, thus permitting the repair of cellulardamage. However, it might be possible that at about 33 per centmoisture content seed could suffer from an excessive advancementof metabolism. Triticum durum, seed storage, effects of high moisture content and oxygen     

The Influence of Temperature, Moisture, and Oxygen on Period of Seed Viability in Barley, Broad Beans, and Peas

The viability of seeds of barley, broad beans, and peas hasbeen examined in hermetic storage over a range of temperaturesfrom 25? to 45? C and over a range of moisture contents fromabout 12 to 18 per cent. It was found that the survival curvesunder nearly all conditions can be described as negative cumulativenormal distributions. Under very severe storage conditions,however, when the mean viability period is of the order of aweek or less, the survival curves may become slightly skew.The spread of the distribution is linearly proportional to themean viability period. There is a negative linear relationshipbetween log mean viability period and both temperature and moisturecontent. Because of these relationships it is possible to predictthe percentage germination of these species after any givenperiod under a wide range of storage conditions. This patternof loss of viability in barley, broad beans, and peas is consistentwith that previously shown for wheat and rice. Oxygen has been shown to have a deleterious effect on the viabilityof barley, beans, and peas. Most of the deleterious effect isproduced by increasing the oxygen level from 0 per cent (nominal)to 21 per cent at atmospheric pressure; a further increase to100 per cent has comparatively little effect. The deleteriouseffect of oxygen is more pronounced at high moisture contents.Experiments at a low moisture content (12 per cent) have demonstratedthat the effect of oxygen is independent of the activity ofmicro-organisms. There is also some indication that the effectof oxygen is independent of the rate of seed respiration. The gas-exchange of pea seeds has been investigated in hermeticstorage at 25? C and 18.4 per cent moisture content. The seedsshowed a constant rate of gas-exchange and a constant R.Q. (0.63)over a period of time during which the concentration of oxygendecreased from 21 to I.4 per cent and the carbon dioxide concentrationrose from 0.03 to 12 per cent.     

Different Arginine Transfer Ribonucleic Acid Species Prevalent in Shaken and Unshaken Cultures of Neurospora

When the arginyl-transfer ribonucleic acid (tRNA) species isolated from unshaken and from shaken cultures of Neurospora were compared by co-chromatography, a marked change in the relative abundance of the two main tRNA(arg) species was found. The two arginine tRNA species had different codon responses in ribosome binding assays. The tRNA(arg) eluting first (prevalent in shaken cultures) bound strongly to polyadenylic-guanylic acid [poly(A,G)] and to a lesser extent to polycytidylic-guanylic-adenylic acid [poly(C,G,A)]. The second tRNA(arg) species (prevalent in unshaken cultures) bound to poly(C,G,A) but not to poly(A, G). The possible significance of these observations is briefly discussed. Several modifications that improve the yield of tRNA from Neurospora were introduced in a standard isolation procedure.     

Effect of Shaking Speed and Type of Closure on Shake Flask Cultures

Growth of microorganisms and biosynthesis of microbial products in shake flasks may be limited by operating conditions which provide inadequate supplies of oxygen. Methods are described for meeting the oxygen requirements of test organisms by using standard baffled flasks with pad-type closures and shaking at relatively high speeds. Growth of Escherichia coli B in a rich medium and production of candidin by Streptomyces viridoflavus were the test systems used. Flasks shaken at 230 to 385 rev/min gave sulfite oxidation rates of 1 to 8 mmoles of oxygen per liter per min over a useful working volume range (40 to 150 ml in 300-ml flasks). These rates are as high as those obtained in agitated fermentors under usual operating conditions.     

Dormancy of Paddy Seeds of a 'Non-dormant' Variety

Seeds of a non-dormant rice variety, Satika, were studied underdifferent natural and controlled humid conditions of storageat room temperature. Just after harvest the germination andpercentages of grain moisture were 9.2 and 30.0 respectively.Under humid conditions of storage (94 per cent) there was onlypartial break of dormancy after 42 d (32.8 per cent ger mination)with high grain moisture (23.8 per cent); in all other treatmentswhere the grain moisture varied from 4.4 to 16.4 per cent thegermination percentages were over 96 per cent after 42 d. This delaying effect of high moisture contents on the breakingof dormancy became very pronounced at about 26 per cent. Itis suggested that high moisture content may inhibit dormancy-breakingreactions possibly by interrupting air-diffusion inside theseeds.     

Physiology and microbial community structure in soil at extreme water content

A sandy loam soil was brought to 6 water contents (13-100% WHC) to study the effects of extreme soil moistures on the physiological status of microbiota (represented by biomass characteristics, specific respiration, bacterial growth, and phospholipid fatty acid, PLFA, stress indicators) and microbial community structure (assessed using PLFA fingerprints). In dry soils, microbial biomass and activity declined as a consequence of water and/or nutrient deficiency (indicated by PLFA stress indicators). These microbial communities were dominated by G+ bacteria and actinomycetes. Oxygen deficits in water-saturated soils did not eliminate microbial activity but the enormous accumulation of poly-3-hydroxybutyrate by bacteria showed the unbalanced growth in excess carbon conditions. High soil water content favored G bacteria.     

Microclimate and forest management alter fungal-to-bacterial ratio and N<Subscript>2</Subscript>O-emission during rewetting in the forest floor and mineral soil of mountainous beech forests

The effects of site exposure (microclimate) and forest management (thinning) on fungal-to-bacterial (F:B) respiratory ratio and N₂O emission from forest floor and Ah layer samples were studied at untreated and thinned beech forests. Microclimate effects were studied by selecting sites facing north-east (NE) or south-west (SW). The F:B respiratory ratio was estimated using substrate-induced respiration in combination with inhibitors either affecting fungi or bacteria. N₂O production was evaluated after moistening samples initially pre-incubated at different moisture levels to 100% of the water holding capacity (WHC). F:B respiratory ratios were significantly affected by microclimate and thinning, with site exposure having the strongest effect on fungal-to-bacterial ratio and N₂O production both for the forest floor and the Ah layer. Significantly more N₂O was produced from soils pre-incubated under low (15% WHC) moisture conditions as compared to soils pre-incubated under air dry (5% WHC) or wet conditions (30–60% WHC). A positive correlation between N₂O emission and F:B respiratory ratio for Ah layer samples and a negative correlation between bacterial substrate induced respiration (SIR) and N₂O emission for both Ah layer and forest floor samples indicated that net N₂O production was the result of predominantly fungal N₂O production and predominantly bacterial N₂O consumption. The latter hypothesis was further supported by increased N₂O emission from samples treated with bacterial inhibitor.     

Nitrogen and phosphorus removal through laboratory batch cultures of microalga Chlorella vulgaris and cyanobacterium Planktothrix isothrix grown as monoalgal and as co-cultures

Batch cultures of the green microalga Chlorella vulgaris and cyanobacterium Planktothrix isothrix and their corresponding co-cultures were grown in municipal wastewater in order to study their growth as well as the nitrogen (NH₄–N) and phosphorus (PO₄³⁻–P) removal. The cultures were grown under two irradiances of 20 and 60 μmol photons m⁻² s⁻¹ in shaken and unshaken conditions. The co-culture of unshaken Chlorella and Planktothrix showed the greatest growth under both irradiances. The monoalgal Planktotrix cultures showed better growth when unshaken than when shaken, whereas Chlorella cultures grew better when mixed, but only at the higher irradiance. The highest percentage of nitrogen removal (up to 80%) was attained by the unshaken co-cultures of Chlorella and Planktothrix. The amount of nitrogen recycled in the biomass reached up to 85% of that removed. Shaken monoalgal cultures of Chlorella showed phosphorus removal under both irradiances. They completely removed the initial phosphorus concentration (7.47 ± 0.17 mg L⁻¹) within 96 and 48 h under 20 and 60 μmol photons m⁻² s⁻¹, respectively.     

Production of a New Acidic Polysaccharide,Succinoglucan by Alcaligenes faecalis var. myxogenes

A slimy non-spore-forming bacterium strain 10C3 isolated from soil was motile with peritrichous flagella and named Alcaligenes faecalis var. myxogenes. Studies were made on the conditions necessary for maximal production of a new acidic succinoglucan polysaccharide by this strain in shaken cultures. Much production was observed with sucrose, glucose, xylose, galactose, cellobiose, maltose, fructose, mannose and rhamnose. The yield was greatest with sucrose and decreased in order with the above sugars from about 36 to 23 per cent. The most suitable medium contained 4 per cent sugar, 0.5 per cent yeast extract and one per cent calcium carbonate in tap water. The optimum temperature was 28°C.     

水分对林地和草地土壤氮初级转化速率的影响

水分含量是与土壤氮转化相关微生物活性的重要影响因素。本研究以黑龙江省北安市的草地和林地土壤为对象,通过室内培养试验,利用¹⁵N同位素标记技术和FLUAZ数值优化模型研究60%和100%田间持水量(WHC)条件下土壤氮初级矿化速率、初级固定速率、初级硝化速率和初级反硝化速率,以探讨土壤氮初级转化速率对水分含量变化的响应,阐明不同水分条件下土壤中氮的产生、消耗、保存机制及其生态环境效应。结果表明: 土壤水分变化不影响草地和林地土壤氮初级矿化速率和铵态氮固定速率,水分含量由60% WHC增加至100% WHC后显著增加了林地土壤的初级硝化速率,但对草地土壤的初级硝化速率没有显著影响。60% WHC条件下草地和林地土壤的初级反硝化速率可以忽略不计,水分含量增加至100% WHC后土壤初级反硝化速率显著提高,且草地土壤的初级反硝化速率显著低于林地土壤。100% WHC条件下林地土壤初级硝化速率与铵态氮固定速率比值(gn/ia)和N₂O排放量均显著高于60% WHC;100% WHC条件下草地土壤的N₂O排放量显著高于60% WHC,但两个水分条件下的gn/ia值无显著差异。表明短期内水分含量的增加可能会增加草地和林地土壤氮转化的负面环境效应,且对林地土壤的影响尤为显著。     

The production of bacteriophage μ2

In three series of experiments, 3-l., 20-l., and 150-l. bacterial cultures were grown in stirred, deep culture vessels to average bacterial cell densities of 71 × 10⁸, 63 × 10⁸, and 43 × 10⁸ viable organisms per milliliter, respectively, and then infected with phage. The average yield of progeny phage in each case was ca. 3000 mpfu (minimum plaque-forming units) per cell. Thus, the average mass of phage obtained in the 3-l. experiments was not less than 124 mg./l., calculated from the plaque counts, assuming a particle size of 3.6 × 10⁶ Daltons for the μ2 phage. This is about twentyfold higher than is obtainable by conventional methods in aerated, shaken culture flasks. The actual phage yields are probably much higher than the minimum values calculated from plaque counts. For example, in the case of one of our culture lysates which was purified at King's College, the efficiency of plating was shown to be only 19%. The carbon dioxide evolution rate of cultures was measured and used as a guide to the time at which phage should be added. In this way, greater control of cultural conditions was obtained than is possible in shaken flasks. For the best yield of phage per milliliter of culture, the optimum time for phage infection was such that bacterial lysis just prevented the carbon dioxide evolution rate from reaching its potential maximum. The major factor influencing the phage yield per milliliter of culture was the aeration capacity of the culture vessel used. All had maximum aeration capacities much higher than those obtainable in shaken culture flasks. Cultures grown and infected in 3-l. Vessel operated under conditions of low aeration gave poor yields of phage. The reason for this are discussed.     

Effect of small scale culture vessel type on hyphal fragment size and erythromycin production in Saccharopolyspora erythraea

Erythromycin production dynamics in stirred, baffled shaken and non-baffled shaken flasks was strongly correlated with the different distributions of hyphal particle diameters observed. Production only took place when hyphal fragments with diameters greater than 88 m were observed. Results are consistent with significant hyphal breakage rates, even in non-baffled shaken flasks.     

Loss of Viability in Lettuce Seeds and the Accumulation of Chromosome Damage under Different Storage Conditions

Loss of seed viability in lettuce (Lactuca sativa L.) duringstorage is associated with an increase in the frequency of cellsin the surviving seeds showing chromosome damage during firstmitoses. The relation is linear when probit of the frequencyof aberrant cells is plotted as a function of probit percentagenormal germination. The slope of the relation, however, variesaccording to moisture content so that the proportion of aberrantcells for any given loss of germination increases with decreasein moisture content over the range 13.0–5.5 per cent.At 3.3 per cent moisture content, however, the proportion ofaberrations was no greater than at 5.5 per cent moisture content;and at 18.1 per cent moisture content the proportion was noless than at 13.0 per cent moisture content. Despite these differences,the increase in chromosomal aberrations per unit time for agiven temperature was always less the lower the moisture content.Diplontic selection markedly reduced the frequency of chromosomalaberrations and eliminated the differences in these frequenciesbetween the different storage treatments. But even after fiveweeks' growth, root tips from aged seed still contained abouttwice as many aberrant cells as compared with similar root tipsderived from the original seed stock. Studies on the frequencyof recessive mutations indicated that excessive amounts of heritablemutations were not present in the progenies of aged seed, evenwhen stored at moisture contents as low as 5.5 per cent. Allthis and other evidence reinforces the view that orthodox seedsfor genetic conservation should be stored at not more than about5 per cent moisture content, and that even lower moisture contentsare worth considering. The results also emphasise the need formaintaining a high regeneration standard, i.e. the percentageto which seed viability is allowed to fall during storage beforethe seed stock is regenerated. Lactuca sativa, lettuce, seed storage, seed viability, chromosomal aberrations, phenotypic mutations     

Enhanced production of recombinant galactose oxidase from <Emphasis Type="Italic">Fusarium graminearum</Emphasis> in <Emphasis Type="Italic">E. coli</Emphasis>

The gene gaoA encoding the copper-dependent enzyme galactose oxidase (GAO) from Fusarium graminearum PH-1 was cloned and successfully overexpressed in E. coli. Culture conditions for cultivations in shaken flasks were optimized, and optimal conditions were found to be double-strength LB medium, 0.5% lactose as inducer, and induction at the reduced temperature of 25°C. When using these cultivation conditions ~24 mg of active GAO could be produced in shaken flasks per litre medium. Addition of copper to the fermentation medium decreased the enzyme production significantly. The His-tagged recombinant enzyme could be purified conveniently with a single affinity chromatography step. The purified enzyme showed a single band on SDS–PAGE with an apparent molecular mass of 66 kDa and had kinetic properties similar to those of the fungal wild-type enzyme.     

甲硫氨基酸对亚热带森林土壤硝化作用和N2O排放的影响

为探讨甲硫氨基酸对亚热带红壤硝化作用和N₂O排放的影响,选择福建省建瓯市万木林保护区的山地红壤为研究对象,在土壤饱和持水量(WHC)60%和90%的条件下,开展室内培养试验.试验分为对照(CK)、添加甲硫氨基酸(M)、甲硫氨基酸和硫酸铵(MA)、甲硫氨基酸和亚硝酸钠(MN)、甲硫氨基酸和葡萄糖(MC)5个处理.结果表明: 与对照相比,M处理使土壤NH₄⁺-N平均含量显著提高0.8%～61.3%,而NO₃^--N含量显著降低13.2%～40.7%;60%WHC条件下,MC处理土壤NO₂^--N含量高于M处理,MA、MN处理NO₃^--N含量高于M处理,且MN处理高于MA处理,M处理于试验后期最低,表明甲硫氨基酸抑制了硝化作用的亚硝化过程.碳添加处理使甲硫氨基酸在一定程度上降低NH₄⁺-N含量,抑制了土壤自养硝化,并且甲硫氨基酸和碳源共同作用下NO₃^--N含量变化与土壤水分条件有关,在90%WHC条件下,碳加入后反硝化作用更明显;而NO₃^--N含量降低不足以表明是异养硝化受到抑制所致.甲硫氨基酸在一定程度上促进土壤N₂O的释放,90%WHC条件下较60%WHC条件下释放量更大,且葡萄糖添加的促进效果更明显.     

甲硫氨基酸对亚热带森林土壤硝化作用和N2O排放的影响

为探讨甲硫氨基酸对亚热带红壤硝化作用和N₂O排放的影响,选择福建省建瓯市万木林保护区的山地红壤为研究对象,在土壤饱和持水量(WHC)60%和90%的条件下,开展室内培养试验.试验分为对照(CK)、添加甲硫氨基酸(M)、甲硫氨基酸和硫酸铵(MA)、甲硫氨基酸和亚硝酸钠(MN)、甲硫氨基酸和葡萄糖(MC)5个处理.结果表明: 与对照相比,M处理使土壤NH₄⁺-N平均含量显著提高0.8%～61.3%,而NO₃^--N含量显著降低13.2%～40.7%;60%WHC条件下,MC处理土壤NO₂^--N含量高于M处理,MA、MN处理NO₃^--N含量高于M处理,且MN处理高于MA处理,M处理于试验后期最低,表明甲硫氨基酸抑制了硝化作用的亚硝化过程.碳添加处理使甲硫氨基酸在一定程度上降低NH₄⁺-N含量,抑制了土壤自养硝化,并且甲硫氨基酸和碳源共同作用下NO₃^--N含量变化与土壤水分条件有关,在90%WHC条件下,碳加入后反硝化作用更明显;而NO₃^--N含量降低不足以表明是异养硝化受到抑制所致.甲硫氨基酸在一定程度上促进土壤N₂O的释放,90%WHC条件下较60%WHC条件下释放量更大,且葡萄糖添加的促进效果更明显.