Pyrolysis methylation—mass spectrometry of whole soils

In-source pyrolysis field ionization mass spectrometry (Py-FIMS) and Curie–point pyrolysis gas chromatography/mass spectrometry (Py-GC/MS) have been applied to the characterization of whole soils methylated off–line and on–line by two methylation reagents. Samples of a gleysolic Ap–horizon and a podzolic Bh horizon (C_org concentrations 2–3%) were investigated by conventional on–line and developed off–line methylation with tetramethylammonium hydroxide (TMAH) and by off–line methylation with diazomethane. For the first method, the soils were dried, milled and pretreated with TMAH for 10 min at 250°C at ambient pressure outside the pyrolyzers. For the second method, the dried and milled soils were methylated with an ether solution of diazomethane for 12 h at room temperature (～20°C). Pyrolysis methylation with TMAH enabled aliphatic C₂–C39 monocarboxylic acid methyl esters, C4–C₃₀ dicarboxylic acid dimethyl esters and benzenecarboxylic acid methyl esters to be detected. Methoxybenzenes from phenols, benzenediols and benzenetriols, methoxybenzenecarboxylic acid methyl esters from phenolic acids and furancarboxylic acid methyl esters from carbohydrates were also identified. Nitrogen–containing compounds in soil organic matter were obtained as N,N–dimethylamides. Using diazomethane as methylation reagent, distinct Py-FIMS signals were observed for aliphatic C₂–C₃₂ monocarboxylic acid methyl esters and C₃–C₂4 dicarboxylic acid dimethyl esters. Additionally, methoxybenzenes originating from lignins, methoxybenzenecarboxylic acid methyl esters from phenolic acids and N,N–dimethylamides from amides were detected. The more acid podzolic Bh horizon showed higher relative intensities for dicarboxylic acid dimethyl esters and methylated phenolic acids compared to the gleysolic Ap horizon. Similarly, benzenecarboxylic acids are connected mainly by ester linkages to the macromolecular network of soil organic matter. Both methylation procedures support conventional Py-FIMS and Py-GC/MS and give valuable additional information on the occurrence of aliphatic and aromatic carboxylic acids, substituted phenols, benzenediols, benzenetriols, phenolic acids and amides in soil organic matter.     

Nature and Origin of Lipids in Clay Fractions from a Fluvisol in a Sewage Sludge Deposition Field

Few studies on free lipids in total solvent extracts from soil clay fractions directly measured by gas chromatography–mass spectrometry (GC/MS) have been reported so far. In this study, we aimed to examine the free lipids in the clay fraction separated from a Fluvisol profile on which sewage sludge was deposited 6 years ago and provide information on the sources, diagenetic processes and organic pollutants derived from the sludge. Clay fractions were separated from the four horizons of a Fluvisol and analysed for the biochemically stable lipid pool. The GC/MS analysis of the lipid fraction showed that lipid signatures were dominated by alkanes (C₁₇–C₃₃), alkanoic acids (C₁₂–C₁₈), alkanols (C₁₄–C₃₀), aromatic acids and phthalate esters. Sources of lipids show predominant bacterial contribution as shown by the alkane, fatty acids and n-alkanol distributions. The preservation of lipids of microbial origin in the clay fraction was revealed by the presence of even number, branched alkanes and short-chain and branched fatty acids. These results imply that similar pedogenic processes took place in this soil profile affected by hydromorphic conditions with some quantitative changes in the clay lipid compositions between different soil horizons. Some aromatic acids and xenobiotics such as phthalates were detected in the lipid extracts of the clay fractions in depth (0–85 cm) of the soil profile, which has implications for pollution of soils and ground waters in situations of sewage sludge deposition.     

Changes in Concentrations of Malodorous Compounds During Controlled Aeration Composting

Effects of composting on odorous chemicals in dairy manure were investigated in replicated pilot-scale studies. Three 16-day composting trials were conducted, using 205 L vessels containing 83 kg mass. Fresh or 12-day-aged manure, from lactating cows, was mixed with sawdust (3man:1saw w/w). Vessels were either aerated continuously with high (2.3 kg/hr)/low (0.8 kg/hr) air flow controlled by thermostats or intermittently on a 5 min high air flow/55 min off clock controlled cycle. Six vessel replicates were conducted on each manure/air flow treatment combination. Temperatures, air flow, O₂ consumption, and CO₂ production were recorded every 10 minutes. Trapped NH₃ emissions were determined daily. Exhaust air was passed through water-cooled condensers to analyze emitted volatiles, and condensate volume, pH, and volatile fatty acids (VFAs) were quantified at 12 or 24 hour intervals. Solids were collected from each vessel initially, at remix at the end of day 7, and at the end of each trial (day 16) for analysis of moisture, pH, ash, C/N, and odorants. Phenolics and indolics were extracted with ethyl ether. VFAs were recovered with pH 2.0 water. Analysis was by flame or mass selective detection gas chromatography. Temperature increased most rapidly in continuously aerated vessels yet maintained a lower mean temperature (49 vs 58°C) than intermittent aeration. Both returned to near ambient temperature by day 16. Continuous aeration nearly doubled (11 vs 18 L) the amount of condensate released over 16 days. Fresh manure/sawdust mixes contained 6553, 795, 77, 51, 19, and 17μg/g of acetate, propionate, isobutyrate, isovalerate, phenol, and p-cresol. Aged manure mixes contained 9350, 3397, 2810, 445, 285, 441, 34, 176, and 18 μg/g acetate, propionate, butyrate, valerate, isobutyrate, isovalerate, phenol, p-cresol, and skatole, as well as a number of C₁₁ to C₁₇ fatty acids. Both aeration methods maintained conditions that resulted in the destruction of most of the odorous chemicals studied in the composting mass in 7 days with only small quantities of acetate, isobutyrate, and skatole present by the end of day 16. Continuous aeration, as opposed to intermittent, more than doubled (115 vs 55 g) the emissions of NH3-N and increased the emissions of VFAs in condensate four-fold. Whereas, limited aeration did not destroy the odorants as rapidly, they remained in the compost until destruction.     

Microbial diversity in three floodplain soils at the Elbe River (Germany)

Microbial communities in floodplain soils are exposed to periodical flooding. A long-term submerged Eutric Gleysol (GLe), an intermediate flooded Eutric Fluvisol (FLe), and a short-time flooded Mollic Fluvisol (FLm) at the Elbe River (Germany) with similar organic carbon contents (C_org) between 8.1% and 8.9% were selected to test the quality of phospholipid fatty acids (PLFA), soil microbial carbon (C_mic), basal respiration (BR), metabolic quotient (qCO₂), and C_mic/C_org ratio to characterize and discriminate these soils with microbial parameters.The three floodplain soils can be differentiated by C_mic and by total PLFA-biomass. Due to the different flooding durations and the time since the soils were last flooded C_mic and PLFA-biomass increase in the order GLe<FLe<FLm. Both parameters correlate significantly (r=0.999;p<0.05). The C_mic/C_org ratios are low in comparison to terrestrial soils and revealed the same ranking over the three soils like C_mic. Contrary, qCO₂ and BR are highest in GLe and lowest in FLm according to inundation regime. The diminished C_mic, high BR, and high qCO₂ values in GLe seem to be an unspecific response of aerobic soil microorganisms on the long flooding period and the resulting short time for developing after last flooding as well as the low pH value. Different plant communities and their residues may influence the microbial diversity additionally.The PLFA profiles were dominated by the group of saturated fatty acids that together constituted almost 62-72% of the total fatty acids identified in the soils. In GLe all groups of PLFA, inclusive monounsaturated fatty acids, are lowest and in FLm highest, while in FLe the PLFA fractions show an intermediary amount of the three soils. The FLm had most of the time aerobic conditions and revealed therefore the highest C_mic, PLFA-biomass, especially monounsaturated fatty acids, C_mic/C_org ratio as well as relatively low BR and qCO₂ value. These indicate that microorganisms in FLm are more efficiently in using carbon sources than those in GLe and FLe.All 26 identified PLFA were found in FLe and FLm, while the polyunsaturated fungi biomarker 18:2ω6,9c could not be detected in GLe. In this long-time submerged soil the environmental conditions which microorganisms are exposed might be disadvantageous for fungi.     

Accessibility of Starch Granules to Fatty Acyl Amides

Microscopic techniques were used to determine accessible regions in various starches under different swelling conditions. Fatty acyl chlorides of increasing chain length (C₆–C₁₆) were reacted with 2‐aminopyridine to produce the corresponding fatty acyl amides. Starch granules (common corn, waxy maize, potato) were treated with the series of fluorescent amides under a variety of conditions: anhydrous (hot pyridine), aqueous (no heat), and aqueous alkali (49°C, with and without swelling‐inhibiting salts). Isolated granules were then viewed by confocal laser scanning microscopy to determine reagent analog penetration. All populations of granules were heterogeneous with respect to fatty acyl amide penetration, but general patterns could be discerned. Observations also indicated that the area surrounding the hilum was more easily penetrated than was the rest of the granule matrix. No substantial differences in penetration of the fluorescent fatty acyl amides as the chain length increased (C₆–C₁₆ was observed in hot pyridine‐swollen common corn starch granules. Common corn and potato starch granules swollen in room temperature water showed cutoffs for granular exclusion at C₁₄ and C₁₂, respectively. Common corn, waxy maize, and potato starch granules treated under industrial etherification conditions (heat, pH ≈ 11, swelling‐inhibiting salts) were less accessible to C₆, C₈, and C₁₀ fluorescent amides when sodium citrate was present than when sodium sulfate was used, and less accessible in either case than in water alone or in hot anhydrous pyridine. However, appreciable differences between inhibition by sodium sulfate and sodium citrate were not observed in every case.     

Permanganate oxidation of methylated and non-methylated aquatic humus in Norway

The chemical composition of aquatic humus was investigated by permanganate oxidation. Both methylated and non-methylated samples were investigated and the results compared with those of different soil humic fractions investigated earlier.The total amount of oxidation products identified from the methylated sample was 2%, and from the non-methylated sample 0.9%. The composition of the oxidation products from methylated aquatic humus was 42% benzenecarboxylic acid methyl esters (8 different compounds), 43% methoxy-benzenecarboxylic acid methyl esters (12 compounds), 10% dimethoxy-benzenecarboxylic acid methyl esters (4 compounds), and 5% of 1, 2, 3-propanetricarboxylic acid trimethyl ester. The unmethylated aquatic humus yielded 84% benzenecarboxylic acid methyl ester (7 compounds), 7% methoxy-benzenecarboxylic acid methyl esters (2 compounds), and 9% of 1, 2, 3-propanetricarboxylic acid trimethyl ester. Three diazines isolated from methylated material were believed to be artefacts from diazomethane treatment. Two of the diazines have earlier been found by oxidation of methylated soil samples, the third, C₁₀H₁₂N₂O₆, is an oxidation product of methylated aquatic humus only.Oxidation of aquatic humus yielded more benzenecarboxylic acids and methoxy-benzenecarboxylic acids than soil humic fractions, and less dimethoxy-benzenecarboxylic acids. No aliphatic dicarboxylic acids were detected among the oxidation products of the aquatic humus.The compounds identified are mainly the same as those found by oxidation of different soil humic fractions, although their yields clearly demonstrated that the aquatic humus differed in composition from the soil fractions.     

Organic-geochemical Differentiation of Petroleum-type Pollutants and Study of Their Fate in Danube Alluvial Sediments and Corresponding Water (Pančevo Oil Refinery, Serbia)

A review is given in this paper of the up-to-date results observed in differentiation and transformation studies on petroleum-type pollutants in underground and surface waters. Water and particulate matter derived from the locality of Pan?evo Petroleum Refinery, Serbia (River Danube alluvial formations). It was shown that distributions of n-alkanes, steranes and triterpanes, and δ¹³C_PDB values of n-alkanes may successfully be used for qualitatively differentiating the petroleum-type pollutants from native organic matter in recent sedimentary formations. In underground waters, a petroleum-type pollutant is exposed to microbiological degradation which is manifested through relatively fast degradation of n-alkanes. Following an almost complete degradation of crude oil n-alkanes in underground water, the biosynthesis of novel, even carbon-number C₁₆–C₃₀ n-alkanes may be observed. It is shown that the n-alkane distribution observed in a petroleum-type pollutant may depend on the intensity of its previous interaction with water. The fate of petroleum-type pollutants in environmental waters may be predicted through laboratory simulative microbiological degradation experiments by using microorganism consortiums similar to those observed under relevant natural conditions, as well as on corresponding nutrient base.     

Quantification of long‐chain fatty acids in dissolved organic matter and soils

The objective was to develop and adapt a versatile analytical method for the quantification of solvent extractable, saturated long‐chain fatty acids in aquatic and terrestrial environments. Fulvic (FA) and humic (HA) acids, dissolved organic matter (DOM) in water, as well as organic matter in whole soils (SOM) of different horizons were investigated. The proposed methodology comprised extraction by dichloromethane/acetone and derivatization with tetramethylammonium hydroxide (TMAH) followed by gas chromatography/mass spectrometry (GC/MS) and library searches. The C_10:0 to C_34:0 methyl esters of n‐alkyl fatty acids were used as external standards for calibration. The total concentrations of C_14:0 to C_28:0 n‐alkyl fatty acids were determined in DOM obtained by reverse‐osmosis of Suwannee river water (309.3 μg g^—1), in freeze‐dried brown lake water (180.6 μg g^—1), its DOM concentrate (93.0 μg g^—1), humic acid (43.1 μg g^—1), and fulvic acid (42.5 μg g^—1). The concentrations of the methylated fatty acids (n‐C_16:0 to n‐C_28:0) were significantly (r² = 0.9999) correlated with the proportions of marker signals (% total ion intensity (TII), m/z 256 to m/z 508) in the corresponding pyrolysis‐field ionization (FI) mass spectra. The concentrations of terrestrial C_10:0 to C_34:0 n‐alkyl fatty acids from four soil samples ranged from 0.02 μg g^—1 to 11 μg g^—1. The total concentrations of the extractable fatty acids were quantified from a Podzol Bh horizon (26.2 μg g^—1), Phaeozem Ap unfertilized (48.1 μg g^—1), Phaeozem Ap fertilized (57.7 μg g^—1), and Gleysol Ap (66.7 μg g^—1). Our results demonstrate that the method is well suited to investigate the role of long‐chain fatty acids in humic fractions, whole soils and their particle‐size fractions and can be serve for the differentiation of plant growth and soil management.     

Hydrogen Sulfide and Odor Control in İzmir Bay

The city of ?zmir on the Aegean Sea shoreline is suffering from rotten odors emitted by anoxic river mouths. Anaerobic conditions in the shallowest portion of ?zmir Bay due to industrial and domestic wastewaters as well as eutrophication products in this very calm part of the Bay are responsible for this. The inner section of the Bay is becoming shallower with sediments rich in organic matter. Aerobic digestion of organic pollutants is limited by the oxygen input and the warm climate leads to an optimal medium for anaerobic processes when anoxic conditions take over. Anaerobic digestion products are odorous gases among which H₂S with a characteristic pungent odor is most effective in this case. Sulfur containing gases are formed from sulfides and sulfates in the sediment-water interface and are released into the air. Airborne H₂S concentrations are variable as they depend on factors such as high atmospheric diffusion coefficients under changing wind direction and speed, as well as variable such as water depths, organic loadings from rivers, air and water temperatures, sulfate concentrations in sediment and water phases, pH, and Eh. This study aimed to control noxious smells by inhibiting the anaerobic sulfate-reducing bacteria at the sediment surface. To achieve this aim, pH of the sediment-water interface was increased by spreading hydrated lime. Successful lime doses were investigated in laboratory models first and were then tested and applied in the estuary and downstream segment of one of the creeks. Rates of H₂S gas emissions and aqueous H₂S levels were measured. It was noted that waterborne H₂S levels at river mouths were reduced. Odor control efficiencies of 80–96% even after 10 days of lime addition in the field experiments were obtained. Therefore, the results of this study indicated that lime application to the sediment surface is a recommended method in odor control programs.     

Changes in soil organic compound composition associated with heat‐induced increases in soil water repellency

Soil heating, as for example experienced during vegetation fires, often increases soil water repellency; however, no detailed analysis of the soil chemical changes associated with this increase has been conducted to date. Here we characterize the changes in organic compound composition associated with heat‐induced increases in water repellency for three Australian eucalypt‐forest soils (one sandy loam, two sands). Laboratory heating (300°C) strongly increased water drop penetration times (WDPTs) in all soils. Soils were extracted by accelerated solvent extraction (ASE) with an iso‐propanol/ammonia mixture (IPA/NH₃ 95:5) and pure iso‐propanol (IPA). Extracts were fractionated into less and more polar fractions and analysed by GC‐MS. Water repellency was eliminated in unheated and heated soils by IPA/NH₃, but not by pure IPA. Before heating, total solvent extracts were dominated by n‐alkanols, terpenoids, C₁₆ acid, C₂₉ alkane, β‐sitosterol and polar compounds. After heating, dominant compounds were aromatic acids, aldehydes, levoglucosan, simple sugars and glycosides. Heating resulted in a sharp absolute decrease of homologous aliphatic series of alkanols and alkanes, a shift of fatty acid signature to members <C₂₀ and an increase in total content of aromatic compounds. Heating also caused the formation of complex high‐molecular‐weight compounds detected in the more polar fractionated extracts and low‐molecular‐weight oxo‐ and hydroxyacids and aromatics in the IPA/NH₃ solvent. We speculate that these compounds in conjunction with fatty acids of <C₁₂ interact with organic and mineral soil surfaces and cause the observed strong increases in soil water repellency following heating.     

Structure and biological activities of lipochitooligosaccharide nodulation signals produced by Bradyrhizobium japonicum USDA 138 under saline and osmotic stress

The establishment of a symbiotic interaction involves a signal exchange between the host legume (flavonoids) and the nitrogen-fixing rhizobia (nodulation factors (NFs)). Likewise, abiotic stress conditions, such as salinity and drought, strongly reduce the nodulation process, possibly affecting also the signal exchange. In this work we characterized the structure and biological activity of NFs produced by Bradyrhizobium japonicum USDA 138 under control, salt, and osmotic stress conditions. This strain is the most widely used in Argentine soybean culture; under control conditions, it produces a mixture of four types of NFs (V(C_16:0,MeFuc), V(C_18:1,MeFuc), IV(C_18:1), and V(C_18:1,Ac,MeFuc)). Interestingly, under stress conditions, this strain produces new types of NFs, one common for both stress conditions (V(C_16:1,MeFuc)) and another one only present under salt stress (IV(C_18:1,MeFuc)). All mixtures of NFs, extracted from control, salt, and osmotic stress conditions, showed biological activity in soybean plants, such as root hair deformation, and the radical application of purified NFs induced systemic differences in dry matter accumulation. The inoculation of soybean with genistein-induced bacteria cultured under both control and stress conditions had a positive effect on the number of nodules formed and in some cases on dry matter accumulation. These responses are not related to changes in chlorophyll fluorescence or greenness index.     

Comparison of two alkaline treatments in the extraction of organic compounds associated with water repellency in soil under Pinus taeda

Isopropanol:NH₃ and Methanol:KOH (saponification) alkaline treatments are usually applied in the study of soil organic matter. The first is used in studies of soil water repellency, and the latter in the extraction of ester-bound lipids from soil. In this study, isopropanol:NH₃ and methanol:KOH treatments were applied separately in a solvent extracted repellent soil, in order to compare their efficiency in the extraction of water repellent compounds. The soil sample was taken from a site under a 16 year old Pinus taeda stand. The amount and class of organic compounds released by each treatment were compared using gas chromatography-mass spectrometry (GC-MS). Both treatments resulted in wettable soil after alkaline extraction. In general, alkaline treatments yielded extracts with the same class of organic compounds. Alkanoic acids, α,ω-alkanedioic acids, hydroxyalkanoic acids, aromatic compounds, and alkanols were identified, indicating the preservation of suberin and cutin biopolyester in the soil. Large differences were observed in the amounts of ω-hydroxyalkanoic acids, as well as in the quantity and distribution of dihydroxyalkanoic and trihydroxyalkanoic acids. In contrast to methanol:KOH, isopropanol:NH₃ was not efficient in the extraction of whole aliphatic biopolyesters, mainly pine cutin-related products. Methanol:KOH was more effective in hydrolysis. The presence of biopolyesters in water repellent soil under the P. taeda stand seems to play an important role in water repellency.     

Statistical evaluation of the relationships between spatial variability in the organic carbon content in gray forest soils, soil density, concentrations of heavy metals, and topography

The spatial variability in the organic carbon content (C_org) in the gray forest soils was studied in relation to topography, soil density (D); and concentrations of Al, Fe, K, Ca, Mg, Mn, Cu, and Zn measured at 47 points in the upper (0–10 cm) and lower (10–20 cm) layers by the X-ray fluorescent method. The study area (48 by 104 m) under meadow vegetation included the break of slope of a river valley with strongly eroded gray forest soils and active development of erosional processes. Methods of geomorphometry were used for the quantitative characterization of topographic conditions. Statistical relationships between the studied characteristics were investigated by multiple regression methods with verification of the models according to specially developed criteria. The obtained statistical relationships were used to develop 3D cartographic models of the C_org and D distribution in the two soil layers. It was shown that the content of C_org in the upper layer increased on south-facing slopes, whereas the content of C_org in the lower layer gained its maximum of southwestern slopes, and the reasons for this distribution were determined. The major characteristics of topography affecting the distribution of C_org in the different soil layers were identified. The C_org content in any soil layer was most tightly correlated with the D values; a less tight correlation was observed between the C_org and Mg contents. The Zn and Cu contents correlated with the C_org in the lower (10–20 cm) layer, whereas the Ca and Fe contents correlated with the C_org in the upper (0–10 cm) layer. The interpretation of the observed regularities involved data on the stability of metal complexes with humic acids under different conditions of the soil acidity; the effect of the erosional processes was also taken into account.     

Organic compounds of different extractability in total solvent extracts from soils of contrasting water repellency

Previous studies examining organic compounds that may cause water‐repellent behaviour of soils have typically focussed on analysing only the lipophilic fraction of extracted material. This study aimed to provide a more comprehensive examination by applying single‐ and sequential‐accelerated solvent extraction (ASE), separation and analysis by GC/MS of the total solvent extracts of three soils taken from under eucalypt vegetation with different degrees of water repellency. Water repellency increased in all the soils after extraction with DCM/MeOH (95:5), but was eliminated with iso‐propanol/ammonia (95:5). Quantities of major lipid compound classes varied between solvents and soils. Iso‐propanol/ammonia (95:5) solvent released saccharides, glycerol, aromatic acids and other polar organic compounds, which were more abundant in fractionated extracts from the single extraction and the third step sequential ASE extraction, than in the extracts from the DCM/MeOH ASE solvent. Dominant compounds extracted from all soils were long‐chain alkanols (>C₂₂), palmitic acid, C₂₉ alkane, β‐sitosterol, terpenes, terpenoids and other polar compounds. The soil with the lowest repellency lacked >C₁₈ fatty acids and had the lowest concentrations of alkanols (C_26,C₂₈ and C₃₀) and alkanes (C₂₉, C₃₁), but a greater abundance of more complex polar compounds than the more repellent soils. We therefore speculate that the above compounds play an important role in determining the water repellency of the soils tested. The results suggest that one‐stage and sequential ASE extractions with iso‐propanol/ammonia and subsequent fractionation of extracts are a useful approach in providing a comprehensive assessment of the potential compounds involved in causing soil water repellency.     

THE ORIGIN AND DISTRIBUTION OF HYDROCARBONS IN AN UPLAND MOORLAND SOIL AND UNDERLYING SHALE

Investigations of plant and soil lipids at an upland moorland site revealed certain similarities in composition. In all lipid samples the polar compound fraction was the largest and the paraffinic the smallest. The range (C_16–36) and pattern of alkanes in the soil were also similar to those of the surface vegetation, both demonstrating a definite predominance of C_odd components. A slight peak in the alkanes identified was seen in the region C_27–C₃₀. The pattern of alkanes in the water running into the soil was quite different, and was not seen to influence the spectrum detected in the soil. Incubation of soil samples with and without added succinate, although causing a drop in lipid content, did not produce many significant changes in the soil alkanes. Plant-derived alkanes in the soil appeared to be resistant to microbial alteration under the experimental conditions employed. The origin of alkanes in an ancient sediment, the underlying shale, was not so clear since the carbon preference index was around unity. It seemed possible that microbial activity might have altered the spectrum of alkanes in the sediment during ageing.     

Leaf gas exchange and water-use efficiency of dry-land wheat genotypes under water stressed and non-stressed conditions

The objective of this study was to determine drought tolerance characteristics of dryland wheat genotypes based on leaf gas exchange and water-use efficiency in order to identify promising genotypes for drought tolerance breeding. Physiological responses of ten genetically diverse wheat genotypes were studied under non-stressed (NS) and water stressed (WS) conditions using a 2?×?10 factorial experiment replicated 3 times. A highly significant water condition?×?genotype interaction (P?<?0.001) was observed for photosynthetic rate (A), ratio of photosynthetic rate and internal CO₂ concentration (A/C_i), ratio of internal and atmospheric CO₂ (C_i/C_a), intrinsic (WUE_i) and instantaneous (WUE_inst) water-use efficiencies suggesting genotypic variability among wheat genotypes under both test conditions. Principal component analysis (PCA) identified three principal components (PC’s) under both test conditions accounting for 84% and 89% of total variation, respectively. Bi-plot analysis identified G339 and G344 as drought tolerant genotypes with higher values of A, T, gs, A/C_i, WUE_i and WUE_inst under WS condition. The current study detected significant genetic variation for drought tolerance among the tested wheat genotypes using physiological parameters. Genotypes G339 and G344 were identified to be drought tolerant with efficient A, T, gs, A/Ci and water-use under water stressed condition.     

Anaerobic Eury- and Crenarchaeota inhabit ectomycorrhizas of boreal forest Scots pine

Members of the euryarchaeotal genera Methanolobus and Halobacterium as well as group 1.1c Crenarchaeota were enriched from ectomycorrhizal samples and cultured under anaerobic conditions. 16S rRNA gene sequences of Methanolobus were obtained in a H₂ + CO₂ atmosphere and autofluorescent putatively methanogenic microbial cells were detected by epifluorescence microscopy of the anaerobic methane-producing enrichment cultures. Halobacterium and group 1.1c Crenarchaeota grew anaerobically when either H₂ or CH₄ was added to the atmosphere. Group 1.1c Crenarchaeota were also enriched under aerobic conditions on mineral media, but only when methane or methanol was added as carbon sources. The 16S rRNA gene sequences of 1.1c Crenarchaeota grown under both anaerobic and aerobic conditions were highly similar. Our study demonstrates the growth of group 1.1c Crenarchaeota and Halobacteria derived from non-extreme soil environment in non-saline enrichments under anaerobic conditions. The results suggest that 1.1c Crenarchaeota may play a role in the cycling of C-1 substrates in the boreal forest soil ecosystem.     

Solvent-extractable lipids in an acid andic forest soil; variations with depth and season

Total lipid extracts from an acid andic soil profile located on Madeira Island (Portugal) were analysed using gas chromatography (GC) and GC-mass spectrometry (GC/MS). The profile was covered mainly by grass. Bulk soil characteristics determined included soil pH (H₂O) ranging from 4.5 to 4.0 and TOC, ranging from 84 to 30 g kg⁻¹. A decrease of the contribution of lipids per TOC with depth was observed. The absence of typical bacterial markers might be an indication for reduced bacterial activity, most likely related to the low soil pH and the presence of Al and Fe (oxides). The distribution observed in the top layer with a dominant C₂₆n-alkanol, steroids and triterpenoids, reflected mainly an input by grass leaves. A strong decrease in both relative and absolute concentration of these leaf-derived compounds was observed when comparing the litter layer with the mineral soil. The presence of C₂₂-C₃₂n-alkanoic acids, C₂₂-C₂₆ ω-hydroxy acids, C₃₁n-alkane and C₂₂-C₃₂n-alkanols observed in the sub-soil is indicative of an important contribution by (grass) roots. In summer, a signal most likely reflecting the leaching of microbially derived products from the litter and/or aerial vegetation at the surface was observed.     

Phosphate-solubilizing microorganisms associated with the rhizosphere of oil palm (Elaeis guineensis Jacq.) in Colombia

This study determined the cell density in the field and the biological activity of culturable phosphate-solubilizing microorganisms (PSMs) present in the Elaeis guineensis Jacq. rhizosphere at two locations in a commercial plantation. Promising isolates found in two soils under different agronomic management conditions were selected. We first calculated the cell density of cultivable PSMs grown in SRS medium (Sundara-Rao and Sinha, 1963) supplemented with the insoluble phosphate sources Ca₃(PO₄)₂, AlPO₄, and FePO₄. Twenty-two bacteria, ten filamentous fungi, and eight yeast isolates were found. The 16 isolates with the clearest P solubilization halo in Petri dishes were selected to estimate their P solubilization potential in SRS medium with Ca₃(PO₄)₂. No solubilization activity was registered using AlPO₄ or FePO₄ as the P sources. Ten of the isolates presented solubilization efficiencies between 20 and 82%. Some of these isolates showed high percentages of identity with the 16S and ITS rDNA sequences of the genera Aspergillus, Penicillium, Klebsiella, Burkholderia, and Staphylococcus according to the NCBI and EzTaxon-e databases. The solubilization activity of the isolates was associated with a decrease in the pH and the release of organic acids, such as gluconic, citric, succinic, and acetic acids. Gluconic acid was mainly released by the genera Aspergillus and Penicillium, and these isolates also showed the highest solubilization activities (82 and 80%, respectively). Therefore, these isolates were selected as the most promising isolates present in the oil palm rhizosphere for phosphate solubilization.     

Effect of introduction of clays,mineral fertilizers,and soil ameliorants on decomposition of organic pollutants in oil-polluted sand under conditions of model experiment

The results of a model laboratory experiment in which the acidic oil-polluted sand was incubated for three months in the presence of palygorskite or bentonite clays supplemented with mineral fertilizers and lime under conditions of periodic wetting and drying are presented. A reliable decrease in the total content of nonpolar and weakly polar CCl₄ extractable organic compounds and of the weakly polar compounds was established. The content of the nonpolar fraction, the amount of C₁₄–C₃₆ alkanes in its composition, and the (C₁₇ + C₁₈)/(C _i–15 + C _i–16) ratio, which was indicative of microbiological destruction of hydrocarbons, decreased significantly in the experiment with the palygorskite clay.