Effects of ozone on organic acids in needles of Norway spruce and Scots pine

Summary The effect of ozone, needle age, and season on the pH of homogenate and acid contents of Scots pine and Norway spruce needles is presented. In addition enzyme activities of cytochrome C-oxidase (cyt. C-ox), phosphoenolpyruvate-carboxylase (PEPC), shikimic acid-dehydrogenase (SHDH) and malate-dehydrogenase (MDH) were measured in Scots pine needles. In freshly sprouted spruce needles the level of quinic acid is high and the pH of the needle homogenate is low. Shikimic acid starts at low levels, increases with increasing needle age and becomes dominant, whereas the quinic acid content decreases. Malic acid has a marked seasonal trend; no trend was found in citric acid. Ozone (200 g/m³) decreased shikimic acid and quinic acid, whereas pH, malic acid and citric acid increased. Ozone (100 g/m³) had a similar effect, except in the current-year spruce needles. In Scots pine needles ozone led to increased enzymatic activities of cyt. C-ox, PEPC and SHDH, and a decrease in the activity of MDH. This effect was more pronounced in summer than in autumn, but the visible damage was greater in autumn. These effects can be found with other stresses and are not specific for ozone.     

Decomposition of organic matter by fungi in saline soils

The object of the present investigation was to study the fungal decomposition of organic matter from different sources, added to the saline soils. The organic matter added was in the form of fresh cuttings of two salt tolerant harbaceous plants namelyDiplachne fusca (Kallar grass) andSesbania aculeata (Dhancha). In addition to these treatments, farm yard manure and press mud (waste of sugar industry) were also used as a source of organic matter. Twelve combinations of these treatments were studied. A number of fungi were isolated from differently treated saline soils incubated at 30 °C. Among the fungi isolatedAspergillus spp. andFusarium solani had the highest frequency of occurrence. The relative cellulolytic ability of 10 fungal species was also estimated.Alternaria humicola andNigrospora sp. were found to be the most cellulolytic among the fungi isolated. The decrease in organic matter due to microbial decomposition was found to be most rapid during first two weeks of incubation. Among the treatments with one organic matter source, press mud produced maximum humus equal to 0.28 %. The pH and electrical conductivity increased slightly due to the closed system where no leaching took place.     

Low-molecular-weight organic acids in rhizosphere soils of durum wheat and their effect on cadmium bioaccumulation

Cadmium (Cd) accumulation has been found to vary between cultivars of durum wheat (Triticum turgidum var. durum), and it is hypothesized that low-molecular-weight organic acids (LMWOAs) produced at the soil-root interface (rhizosphere) may play an important role in the availability and uptake of Cd by these plants. The objective of this study, therefore, was to (1) investigate the nature and quantity of LMWOAs present in the rhizosphere of durum wheat cultivars Arcola (low Cd accumulator) and Kyle (high Cd accumulator) grown in three different soils: Yorkton, Sutherland and Waitville, and (2) determine the relationship between Cd accumulation in these plants and LMWOAs present in the rhizosphere. Plants were grown for two weeks in pot-cultures under growth chamber conditions. Oxalic, fumaric, succinic, L-malic, tartaric, citric, acetic, propionic and butyric acids were found and quantified in the water extracts of rhizosphere soil, with acetic and succinic acids being predominant. No water extractable LMWOAs were identified in the bulk soil. Total amount of LMWOAs in the rhizosphere soil of the high Cd accumulator (Kyle) was significantly higher than that for the low Cd accumulator (Arcola) in all three soils. Furthermore, large differences in amounts of LMWOAs were found in the rhizosphere soil for the same cultivars grown in different soils and followed the pattern: Sutherland > Waitville > Yorkton. Extractable soil Cd (M NH4Cl) and Cd accumulation in the plants also followed the same soil sequence as LMWOA production. Cadmium accumulation by the high and low Cd accumulating cultivars was proportional to the levels of LMWOAs found in the rhizosphere soil of each cultivar. These results suggest that the differing levels of LMWOAs present in the rhizosphere soil played an important role in the solubilization of particulate-bound Cd into soil solution and its subsequent phytoaccumulation by the high and low Cd accumulating cultivars.     

Habitat models of wood-inhabiting fungi along a decay gradient of Norway spruce logs

Information on the habitat requirements of wood-inhabiting fungi is needed to understand the factors that affect their diversity. We applied culture-free DNA extraction and 454-pyrosequencing to study the mycobiota of decaying Norway spruce (Picea abies) logs in five unmanaged boreal forests. Fungal habitat preferences in respect of wood density gradient were then estimated with generalized additive mixed models. Fungal diversity and wood density were inversely related, i.e., OTU richness generally increased as the log became increasingly decomposed. White-rot fungi (e.g., Phellinus nigrolimitatus) and members of Hyphodontia did not show a clear response to the wood-density gradient, whereas abundance of Phellinus viticola and brown-rot fungi (e.g., Fomitopsis pinicola, Antrodia serialis, Coniophora olivaceae) peaked during intermediate decay and mycorrhizal fungi (e.g., Piloderma, Tylospora, Russula) increased in the later stages. This information on fungal habitat requirements facilitates the development of management practices that preserve fungal diversity in managed forests.     

Diversity of endophytic fungi of single Norway spruce needles and their role as pioneer decomposers

The diversity of endophytic fungi within single symptomless Norway spruce needles is described and their possible role as pioneer decomposers after needle detachment is investigated. The majority (90%) of all 182 isolates from green intact needles were identified as Lophodermium piceae. Up to 34 isolates were obtained from single needles. Generally, all isolates within single needles had distinct randomly amplified microsatellite (RAMS) patterns. Single trees may thus contain a higher number of L. piceae individuals than the number of their needles. To investigate the ability of needle endophytes to act as pioneer decomposers, surface-sterilized needles were incubated on sterile sand inoculated with autoclaved or live spruce forest humus layer. The dry weight loss of 13-17% found in needles after a 20-week incubation did not significantly differ between the sterilized and live treatments. Hence, fungi surviving the surface sterilization of needles can act as pioneer decomposers. A considerable portion of the needles remained green during the incubation. Brown and black needles, in which the weight loss had presumably taken place, were invaded throughout by single haplotypes different from L. piceae. Instead, Tiarasporella parca, a less common needle endophyte, occurred among these invaders of brown needles. Needle endophytes of Norway spruce seem thus to have different abilities to decompose host tissues after needle cast. L. piceae is obviously not an important pioneer decomposer of Norway spruce needles. The diversity of fungal individuals drops sharply when needles start to decompose. Thus, in single needles the decomposing mycota is considerably less diverse than the endophytic mycota.     

Non-structural carbohydrate status in Norway spruce buds in the context of annual bud structural development as affected by acidic pollution

The present study focused on changes in the annual dynamics of the contents of non-structural saccharides (NSS) of Norway spruce vegetative buds related to their structural development under the effect of acidic pollution during the year 1995. Two types of material were analysed: (1) 4-year-old trees treated for 2 years by simulated acid rain (SAR; pH 2.9 and 3.9), and (2) 40-60-year-old trees growing in natural mountain stands exhibiting different degrees of macroscopic damage. Our study revealed that the dynamics of the NSS content reflected the major morphogenetic and developmental changes occurring during the annual bud developmental cycle. No systematic changes in the annual dynamics of NSS content were observed in buds from both mountain sites, or as a consequence of the SAR. The total sugar content of bud tissues was composed of a combination of five main sugar components: sucrose, glucose, fructose, raffinose family oligosaccharides (RFO; combination of raffinose and stachyose), and a pinitol fraction (PF) probably of cyclitols with pinitol as a main member. The dynamics of individual sugar components also reflected possible carbohydrate mediated bud frost protection. Interesting results were obtained from buds in dormant state. In dormant buds of the SAR experiment the higher value of the ratio PF:RFO of the pinitol fraction and raffinose family oligosaccharides followed the higher dose of SAR treatment. When evaluating the ratio from both types of material we assumed that changes in PF:RFO ratio corresponded to early stages of damage or acute metabolic reaction. Thus, we suggest the ratio PF:RFO as a possible non-specific metabolic marker of early bud stress reaction which is, among other stress factors, sensitive to increasing load of acidic pollutants.     

RNA reveals a succession of active fungi during the decay of Norway spruce logs

Current knowledge of the succession of fungi in decaying wood is mostly based on fruit bodies and in vitro culture. Here, we investigated the changing community of metabolically active fungi during the decomposition of fallen Picea abies logs by directly extracting and barcode sequencing precursor rRNA. We also compared rRNA-derived amplicons of the 18S and ITS regions in 21 isolates and discuss the use of RNA as a marker of metabolically active fungi. The richness of active fungi, revealed as separated bands in DGGE, peaked in logs at an advanced stage of decay. Soft-rot fungi were common in the early stages but white- and brown-rot fungi became dominant as decay progressed. Ectomycorrhizal fungi were detected at an early stage, and they became the most abundant group in the late stages of succession. A comparison of rRNA-derived amplicons revealed that although ITS was detected in the form of precursor rRNA, introns within 18S rDNA were already spliced. As such, rRNA- and rDNA-derived amplicons would yield different profiles of active and total communities if profiling method is affected by amplicon length.     

Biological activity of humus acids isolated from spruce wood decomposed by fungi

Biological activity of humic acid, hymatomelanic acid, lignofulvic acid, and fulvic acids isolated from peat, from spruce wood decomposed by the fungusFomes marginatus (Fr.) Gillet, and from spruce wood decomposed by the fungusPoria cinerascens Bres. was tested on cress seedlings (Lepidium sativum L.). The results evaluated by the analysis of variance indicate that humus acids show a high degree of affinity in their biological activity which is a condition for the applicability of the decomposition of wood by fungi as a model of the humus formation. Humic and hymatomelanic acids, products of an advanced step of humification, have a predominantly stimulatory effect especially on the hypocotyl. Lignofulvic acids and fulvic acids exert a rather inhibitory effect with the root being more affected.     

Measured and modelled differences in nutrient concentrations between rhizosphere and bulk soil in a Norway spruce stand

The gradient in soil characteristics from the bulk soil to the root surface is important to roots and to the organisms that live in the rhizosphere. Our ability to measure ion concentrations at the root surface is extremely limited, and models are largely untested. We used data from a well studied Norway spruce stand in SW Sweden to compare the measured difference in nutrient concentrations between rhizosphere and bulk soil with the difference predicted by a steady-state simulation model based on ecosystem budgets of nutrient uptake. The simulation model predicted depletion of NH₄, Ca, Mg, K in the rhizosphere, which shows that budgeted uptake rates were greater than the mass flow of bulk solution towards the root. In plots treated with ammonium sulphate, the model predicted an accumulation of S in the rhizosphere. In contrast, the observed rhizosphere concentrations were generally enriched in nutrients, relative to bulk soil. Collecting rhizosphere soil adhering to root surfaces may not be an appropriate method for describing the concentration gradient around the root. In addition, the simulation model omits some processes affecting conditions in the rhizosphere that are important to explaining nutrient uptake.     

Production of organic acids by different white-rot fungi as detected using capillary zone electrophoresis

Capillary zone electrophoresis was used as an analytical method for the first time to determine organic acids accumulated by 15 white-rot fungi both in liquid and solid lignocellulose medium. Oxalic acid accumulation by Phanerochaete chrysosporium F1767 (ATCC 24725), Phlebia radiata 79 (ATCC 64658) and Ceriporiopsis sub-vermispora FP-90031-sp in the solid medium was maximal during the first and second weeks of growth. © Rapid Science Ltd. 1998     

Aluminium tolerance in plants and the complexing role of organic acids

The aluminium cation Al(3+) is toxic to many plants at micromolar concentrations. A range of plant species has evolved mechanisms that enable them to grow on acid soils where toxic concentrations of Al(3+) can limit plant growth. Organic acids play a central role in these aluminium tolerance mechanisms. Some plants detoxify aluminium in the rhizosphere by releasing organic acids that chelate aluminium. In at least two species, wheat and maize, the transport of organic acid anions out of the root cells is mediated by aluminium-activated anion channels in the plasma membrane. Other plants, including species that accumulate aluminium in their leaves, detoxify aluminium internally by forming complexes with organic acids.     

Dissolved organic carbon concentrations in four Norway spruce stands of different ages

Boreal forests are increasing in age partly due to reduced logging and efficient wildfire control. As a result, they also stock more carbon. Whether increased forest C stock causes greater production of dissolved organic carbon (DOC) is uncertain. DOC in bulk precipitation, throughfall and soil water was studied in 10-, 30-, 60- and 120-year-old stands of Norway spruce (Picea abies (L.) Karst.) DOC concentrations in throughfall and O horizon soil water followed the order 10 < 30 < 60 = 120 and 10 = 30 < 120 < 60, respectively. DOC fluxes followed the order 10 = 30 < 60 = 120 in throughfall, while no significant difference between stands was found for O horizon soil water. Above-ground tree litter varied according to 10 < 30 < 60 = 120, a pattern identical to that for DOC concentrations in throughfall and resembling but not identical to that for DOC concentrations in O horizon soil water. This indicates additional sources for DOC in soil water. Seasonality in DOC concentrations was observed at the base of the O horizon, and seasonality in DOC fluxes in both throughfall and O horizon soil water. Our results suggest differences in the polarity of DOC between the 10-year stand and the others, which we interpret as reflecting the lack of grown trees and possibly the different vegetation on the 10-year stand.     

Exudate components exert different influences on microbially mediated C losses in simulated rhizosphere soils of a spruce plantation

Aims

An emerging shoot experiences mechanical impedance (MI) prior to initiating photosynthesis, when it needs to break through soil that has a surface crust. This is the one of the first physical stresses that the shoot experiences. Surprisingly, few measurements have been made to understand the impact of this stress upon post-emergent shoot growth.

Methods

A system employed wax layers of different strengths to investigate shoot responses to MI of the soil surface. Experiments tested the responses of plants to MI using wax layers with different strengths, and tested different seed sizes, nitrogen and phosphorus nutrition and different wheat genotypes. Detailed leaf and root morphological responses and photosynthetic gas exchange and fluorescence were measured.

Results

MI produced permanent impairment to limit plant size, leaf growth rate and leaf photosynthetic function. Large seed sizes and N and P fertilization were able to overcome MI, especially for moderate levels of impedance. There was strong genotypic variation in the response to MI among 14 diverse wheat cultivars, and breeding for varieties suitable to no-tillage cropping systems appears to have facilitated selection in the ability to overcome MI of the soil surface.

Conclusions

This study has highlighted the importance of MI stress of the soil surface in limiting shoot growth and has broad implications for plant genotype selection and agricultural systems management, particularly with regard to nutrition and tillage systems.

     

Nitrogen cycling in a Norway spruce plantation in Denmark--a SOILN model application including organic N uptake

A dynamic carbon (C) and nitrogen (N) circulation model, SOILN, was applied and tested on 7 years of control data and 3 years of manipulation data from an experiment involving monthly N addition in a Norway spruce ( Picea abies, L. Karst) forest in Denmark. The model includes two pathways for N uptake: (1) as mineral N after mineralisation of organic N, or (2) directly from soil organic matter as amino acids proposed to mimic N uptake by mycorrhiza. The model was parameterised and applied to the data from the control plot both with and without the organic N uptake included. After calibration, the models performance was tested against data from the N-addition experiment by comparing model output with measurements. The model reproduced well the overall trends in C and N pools and the N concentrations in soil solutions in the top soil layers whereas discrepancies in soil-solution concentrations in the deeper soil layers are seen. In the control data, the needle-N concentration was well reproduced except for small underestimations in some years because of drought effects not included in the model. In the N-addition experiment, SOILN reproduces the observed changes; in particular, the changes in needle-N concentrations and the overall distribution within the ecosystem of the extra added 3.5 g N m(-2) year(-1) parallel the observations. When organic N uptake is included, the simulations indicate that in the control plot receiving c. 1.9 g N m(-2) year(-1), the organic N uptake in average supplies 35% of the total plant N uptake. By addition of an extra 35 kg N ha(-1) year(-1), the organic N uptake is reduced to 16% of the total N uptake. Generally, inclusion of the pathway for organic N uptake improves model performance compared with observations for both C and N. This is because mineral N uptake alone implies a larger mineralisation rate, leading to bigger concentrations of N in the soil and soil water, bigger N losses, and net loss of c. 100 kg C ha(-1) year(-1), thereby causing depletion of the organic soil layer.     

Chemical changes in organic matter after fungal colonization in a nitrogen fertilized and unfertilized Norway spruce forest

Background and aims

Mineralization of soil organic matter (SOM) constitutes a major carbon flux to the atmosphere. The carbon use efficiency (CUE) of the saprotrophic microorganisms mineralizing SOM is integral for soil carbon dynamics. Here we investigate how the CUE is affected by temperature, metabolic conditions, and the molecular complexity of the substrate.

Methods

We incubated O-horizon soil samples (with either ¹³C–glucose or ¹³C–cellulose) from a boreal coniferous forest at 4, 9, 14, and 19 °C, and calculated CUEs based on the amount of ¹³C–CO₂ and ¹³C–labelled microbial biomass produced. The effects of substrate, temperature, and metabolic conditions (representing unlimited substrate supply and substrate limitation) on CUE were evaluated.

Results

CUE from metabolizing glucose was higher as compared to cellulose. A slight decrease in CUE with increasing temperature was observed in glucose amended samples (but only in the range 9–19 °C), but not in cellulose amended samples. CUE differed significantly with metabolic conditions, i.e. CUE was higher during unlimited growth conditions as compared to conditions with substrate limitation.

Conclusions

We conclude that it is integral to account for both differences in CUE during different metabolic phases, as well as complexity of substrate, when interpreting temperature dependence on CUE in incubation studies.

     

Slow response of soil organic matter to the reduction in atmospheric nitrogen deposition in a Norway spruce forest

Global nitrogen (N) deposition rates in terrestrial environments have quadrupled since preindustrial times, causing structural and functional changes of ecosystems. Different emission reduction policies were therefore devised. The aim of our study was to investigate if, and over what timescale, processes of soil organic matter (OM) transformation respond to a decline in atmospheric N deposition. A N‐saturated spruce forest (current N deposition: 34 kg ha^?1 yr^?1; critical N load: 14 kg ha^?1 yr^?1), where N deposition has been reduced to 11.5 kg ha^?1 yr^?1 since 1991, was studied. Besides organic C and organic and inorganic N, noncellulosic carbohydrates, amino sugars and amino acids were determined. A decline in organic N in litter indicated initial effects at plant level. However, there were no changes in biomarkers upon the reduction in N deposition. In addition, inorganic N was not affected by reduced N deposition. The results showed that OM cycling and transformation processes have not responded so far. It was concluded that no direct N deposition effects have occurred due to the large amount of stored organic N, which seems to compensate for the reduction in deposited N. Obviously, the time span of atmospheric N reduction (about 14.5 years) is too short compared with the mean turnover time of litter to cause indirect effects on the composition of organic C and N compounds. It is assumed that ecological processes, such as microbial decomposition or recycling of organic N and C, react slowly, but may start within the next decade with the incorporation of the new litter.     

Determination of low molecular weight dicarboxylic acids and organic functional groups in rhizosphere and bulk soils of Tsuga and Yushania in a temperate rain forest

Low molecular weight organic acids (LMWOAs) derived from root exudates, decomposing organic matter, and other sources are important ligands. The species of these LMWOAs in the Tsuga rhizosphere soil (TRS), and Yushania rhizosphere soil (YRS), and bulk soil (BS) from an alpine forest region were identified. LMWOA and organic functional groups were used to those fresh twigs and leaves, litters, and roots as comparison. The objectives of this study were to (i) develop a method that could be used to determine LMWOAs in soil solution by gas chromatography (GC), (ii) assess methods for processing LMWOAs in soil samples, and (iii) determine the relative proportions of organic carbon functional groups in the TRS, YRS and BS, and fresh plant materials with¹³C nuclear magnetic resonance (¹³C NMR) analysis. The proportion of organic acid contents followed the order of YRS > TRS > BS, and also showed significant differences (P < 0.05) from GC analysis. The amounts of malonic, fumaric and succinic acids in the YRS samples were greater than in the TRS and BS. Samples analyzed after 1 month of deep freeze storage (–24°C) showed no signs of decomposition. The proportion of organic functional groups in the rhizosphere and bulk soils quantified by ¹³C NMR analyses followed the general order: alkyl-C > O-alkyl-C > N-alkyl-C > acetal-C > aromatic-C > carboxylic-C > phenolic-C.