In situ hybridization to study the origin and fate of identified neurons

Egg-laying behavior in Aplysia is mediated by a set of peptides, including egg-laying hormone (ELH), which are released by a cluster of identified neurons, the bag cells. A family of neuropeptide genes which includes the gene encoding ELH along with two additional genes encoding the A and B peptides thought to initiate the egg-laying process has been isolated and their nucleotide sequence has been determined. In situ hybridization and immunofluorescence was used to explore the origin and distribution of the neurons that express this family of genes. The ELH genes are expressed, not only in the bag cells, but in an extensive system of neurons distributed in four of the five ganglia of the central nervous system. The genes for ELH are expressed in these cells early in the animal's life cycle. As a result, it was possible to use in situ hybridization to trace the cells expressing ELH to their site of origin. The cells originate outside the central nervous system in the ectoderm of the body wall and appear to migrate to their final locations within the central nervous system by crawling along strands of connective tissue.     

Wind dispersal of oribatid mites as a mode of migration

Oribatid mites are important colonizers of young soils, but little is known about their immigration pathways. In this study, one often-stated hypothesis was tested quantitatively: that wind is an important dispersal pathway. The aim was (1) to detect wind dispersal in oribatid mites (using sticky traps at different heights above ground level) and to determine factors influencing wind dispersal, (2) to investigate whether oribatids can survive wind dispersal and immigrate by wind into young soils (using mini-pitfall traps in test plots with oribatid-free substrate, active immigration being prohibited) and (3) to find out whether those oribatids are able to colonize young soils (using soil cores from the test plots). The results demonstrate (1) that mainly arboreal oribatid species were dispersed by wind - even at 160 m height - and can therefore be spread over large distances. Nevertheless, about 10% of the wind-dispersed oribatid mites belonged to species able to live in the soil and may therefore be potential colonizers of young soils. The number of specimen and species transported by wind was the highest close to the soil surface and the number of dispersed individuals was mainly influenced by seasonality and humidity. The results also suggest that the probability of a soil oribatid being dispersed by wind depends on its original microhabitat (tree habitats > soil surface > deeper soil layers) and its body weight. It was also shown that soil-dwelling oribatid mites survived wind dispersal and immigrated by wind into the test plots and that colonization of the test plots took at least 2 months longer than immigration. However, colonization success was low during the first 2 years of investigation and only Trichoribates incisellus was found several times in the nutrient-poor substrate. Therefore, wind dispersal is an important migration pathway, especially for arboreal oribatids. We suggest that immigration into young soils most likely occurs by repeated short-distance dispersal. Only some species are able to survive the hostile conditions of wind dispersal as well as of pioneer soils, but those that do are potentially the founders of new populations.     

Genetic diversity within a range of cultivars and landraces of flax (Linum usitatissimum L.) as revealed by RAPDs

Analysis of the extent and distribution of genetic diversity incrop plants is essential for optimizing sampling and breedingstrategies. We used random amplified polymorphic DNA (RAPD)markers to assess genetic diversity and relationships in 22 Canadiancultivars, 29 selected world cultivars and 10 landraces of flax(Linum usitatissimum L.). RAPDvariation was generally low and more variation was detected among,than within, the investigated flax accessions. Based on 53 variableRAPD loci observed for the 61 accessions, the landraces had a lowerproportion of fixed recessive RAPD loci (0.427) (i.e.,more genetic variation) than all of the flax cultivars examined(0.492). The linseed cultivars had a lower proportion ofrecessive loci (0.469) than the fiber flax cultivars(0.529). Canadian linseed cultivars had a lower proportionof recessive loci (0.465) than the selected world flaxcultivars (0.512). A trend was also observed that the rateof loss in genetic variation in Canadian flax breeding programs overthe last fifty years was approximately two variable loci per 100 lociper 10 years. Clustering analyses based on similarity estimatesshowed that the fiber cultivars were more related (or similar toeach other) and were classified as a homogeneous group. All ofthe linseed cultivars were clustered in diverse groups with the ninelandrace accessions. Implications of these findings for flax breedingand germplasm management are discussed.     

Soil‐ and plant‐based nitrogen‐fertilizer recommendations in arable farming

Under‐ as well as overfertilization with nitrogen (N) will result in economic loss for the farmer due to reduced yields and quality of the products. Also from an ecological perspective, it is important that the grower makes the correct decision on how much and when to apply N for a certain crop to minimize impacts on the environment. To aggravate the situation, N is a substance that is present in many compartments in different forms (nitrate, ammonium, organic N, etc.) in the soil‐plant environment and takes part in various processes (e.g., mineralization, immobilization, leaching, denitrification, etc.). Today, many N‐recommendation systems are mainly based on yield expectation. However, yields are not stable from year to year for a given field. Also the processes that determine the N supply from other sources than fertilizer are not predictable at the start of the growing season. Different methodological approaches are reviewed that have been introduced to improve N‐fertilizer recommendations for arable crops. Many soil‐based methods have been developed to measure soil mineral N (SMN) that is available for plants at a given sampling date. Soil sampling at the start of the growing period and analyzing for the amount of NO ‐N (and NH ‐N) is a widespread approach in Europe and North America. Based on data from field calibrations, the SMN pool is filled up with fertilizer N to a recommended amount. Depending on pre‐crop, use of organic manure, or soil characteristics, the recommendation might be modified (±10–50 kg N ha^–1). Another set of soil methods has been established to estimate the amount of N that is mineralized from soil organic matter, plant residues, and/or organic manure. From the huge range of methods proposed so far, simple mild extraction procedures have gained most interest, but introduction into practical recommendation schemes has been rather limited. Plant‐analytical procedures cover the whole range from quantitative laboratory analysis to semiquantitative “quick” tests carried out in the field. The main idea is that the plant itself is the best indicator for the N supply from any source within the growth period. In‐field methods like the nitrate plant sap/petiole test and chlorophyll measurements with hand‐held devices or via remote sensing are regarded as most promising, because with these methods an adequate adjustment of the N‐fertilizer application strategy within the season is feasible. Prerequisite is a fertilization strategy that is based on several N applications and not on a one‐go approach.     

Growth and interactions of arbuscular mycorrhizal fungi in soils from limestone and acid rock habitats

Arbuscular mycorrhizal (AM) development in different soil types, and the influence of AM fungal hyphae on their original soil were investigated. Plantago lanceolata, which can grow in soils of a very wide pH range, was grown in two closely related limestone soils and an acid soil from rock habitats. Plants were colonised by the indigenous AM fungal community. The use of compartmented systems allowed us to compare soil with and without mycorrhizal hyphae. Root colonisation of P. lanceolata was markedly higher in the limestone soils (30-60%) than in the acid soil (5-20%), both in the original habitat and in the experimental study. Growth of extraradical AM fungal hyphae was detected in the limestone soils, but not in the acid soil, using the signature fatty acid 16:1ω5 as biomass indicator. Analysis of signature fatty acids demonstrated an increased microbial biomass in the presence of AM fungal hyphae as judged for example from an increased amount of NLFA 16:0 with 30 nmol g⁻¹ in one of the limestone soils. Bacterial activity, but not soil phosphatase activity, was increased by around 25% in the presence of mycorrhizal hyphae in the first harvest of limestone soils. AM fungal hyphae can thus stimulate microorganisms. However, no effect of AM hyphae was observed on the soil pH or organic matter content in the limestone soils and the available P was not depleted.     

Root colonisation by arbuscular mycorrhizal, fine endophytic and dark septate fungi across a pH gradient in acid beech forests

Colonisation by root endophytes can be beneficial to plants growing on acid, nutrient-poor soils. Arbuscular mycorrhizal (AM) fungi can supply herbs with nutrients and may give protection against aluminium toxicity. Two other root colonising fungi, fine endophytes (FE) and dark septate fungi (DSE), are less well known but are potentially of benefit to their host plant. AM fungi are the most prevalent symbionts in herbs at neutral to acidic soil pH. At extremely low pH, fungal growth can be limited and AM colonisation is usually rare. Fine and dark septate endophytes, on the other hand, have been observed more often under these conditions. In order to relate endophyte colonisation to a gradient in soil pH, we investigated root colonisation by AM, FE and DSE in Maianthemum bifolium, Galium odoratum, Mercurialis perennis and Stellaria nemorum, from a range of acidic beech forests. With decreasing pH, colonisation by AM decreased, whereas the other two endophytes increased. AM and FE colonisation were inversely correlated in Maianthemum bifolium. We compared changes in root colonisation with those in chemical composition of soil and leaf samples and found a positive correlation between leaf magnesium concentrations and the presence of DSE in Galium odoratum. Aluminium concentration in Maianthemum bifolium tended to be lower when FE colonisation was high, suggesting a possible role for the fungi in plant protection against Al. We suggest that FE and DSE may replace AM fungi in herbaceous vegetation at extremely low pH, counteracting some of the negative effects of high soil acidity on plants.     

The river elbe

Industrial activities in the river basin of the Elbe have a very long tradition, and have been resulting in the contamination of sediments for centuries. Contamination lasted until the fall of the iron curtain; since then, the situation has improved significantly. In the transition zone between freshwater systems and the marine environment, ports like Hamburg still have to bear this burden of history. An overall (contaminated) management strategy should be developed in the context of the European Water Framework Directive with the emphasis on source control. Axel Netzband gave a keynote lecture during the SedNet Inaugural Conference, April 22–24, 2002, which forms the basis of this article.     

Amounts and degradability of dissolved organic carbon from foliar litter at different decomposition stages

Litter is one of the main sources of dissolved organic carbon (DOC) in forest soils and litter decomposition is an important control of carbon storage and DOC dynamics. The aim of our study was to evaluate (i) effects of tree species on DOC production and (ii) relationships between litter decomposition and the amount and quality of DOC. Five different types of leaves and needles were exposed in litterbags at two neighboring forest sites. Within 12 months we sampled the litterbags five times and leached aliquots of field moist litter in the laboratory. In the collected litter percolates we measured DOC concentrations and recorded UV and fluorescence spectra in order to estimate the aromaticity and complexity of the organic molecules. Furthermore, we investigated the biodegradability of DOC from fresh and decomposed litter during 6 weeks incubations. Fresh sycamore maple litter released the largest amounts of DOC reaching about 6.2% of litter C after applying precipitation of 94 mm. We leached 3.9, 1.6, 1.0 and 3.3% carbon from fresh mountain ash, beech, spruce and pine litter, respectively. In the initial phase of litter decomposition significantly decreasing DOC amounts were released with increasing litter mass loss. However, after mass loss exceeds 20% DOC production from needle litter tended to increase. UV and fluorescence spectra of percolates from pine and spruce litter indicated an increasing degree of aromaticity and complexity with increasing mass loss as often described for decomposing litter. However, for deciduous litter the relationship was less obvious. We assume that during litter decomposition the source of produced DOC in coniferous litter tended toward a larger contribution from lignin-derived compounds. Biodegradability of DOC from fresh litter was very high, ranging from 30 to 95% mineralized C. DOC from degraded litter was on average 34% less mineralizable than DOC from fresh litter. Taking into account the large DOC production from decomposed needles we can assume there is an important role for DOC in the accumulation of organic matter in soils during litter decomposition particularly in coniferous forests.     

Spatial changes of soil solution and mineral composition in the rhizosphere of Norway‐spruce seedlings colonized by Piloderma croceum

Ectomycorrhizae (ECM) or the root‐fungal association in forest ecosystems provide a unique soil microenvironment where soil properties and processes differ from the bulk soil. In this study, we would like to better understand the role of ECM systems in mineral weathering and its implications to soil formation and nutrient cycling in forest ecosystems. Specifically, we would like to document the spatial variations in the composition of soil solution and mineralogy of the rhizosphere as influenced by the ECM of Norway spruce + Piloderma croceum. Two‐month‐old seedlings of Norway spruce (control and colonized by P. croceum) were cultivated in special rhizotrons designed to allow spatial collection of soil solution. We used A and C horizons of a Dystric Cambisol collected from Höglwald forest near Munich. Micro suction cups (5 mm x 1mm) were installed in colonized and control rhizotrons, and soil solution was collected from September to November 2000. Our results show that the concentrations of NH , Ca²⁺, and Mg²⁺ in the soil solution were lower in <1.0 cm than in >3.0 cm distance from the roots of Norway spruce, due to the possible range of influence of Piloderma mycelium reaching about 2–3 cm from the surface of the mycorrhizal root. In the rhizotron with soil from the A horizon, a higher phosphorus content in Piloderma‐colonized seedlings was observed. X‐ray diffraction data indicate that chlorite and possibly mica are being transformed to 2:1‐expanding clay minerals (probably smectite) within <1.0 cm distance from roots. The spatial variations in soil solution composition and mineral transformation are likely to be due to Piloderma colonization and concentrated mycelial growth within <1.0 cm distance from the roots. This is also evident in more intricate growth of mycelia on surfaces of micaceous minerals as compared to quartz. We assume that Piloderma modifies soil solution and mineralogy through acquisition of essential elements for its own survival and/or for the uptake by plant roots. However, the presence of spontaneous infection with wildtype ECM in the control plots may have altered the influence of Piloderma and must be taken into consideration when interpreting our results.     

Synthesis and insect growth regulating activity of thiosemicarbazones of methyl 2-pyridyl ketones

Thirty-six thiosemicarbazones of methyl 2-pyridyl ketone and of some other methyl ketones were tested on Dysdercus cingulatus, Drosophila melanogaster, Locusta migratoria, Aedes aegypti, Tenebrio molitor and Musca domestica. Several derivatives were toxic, delaying or preventing ecdysis. The structure-activity relationship is discussed. The mode of action of these compounds appears to be different from that of juvenile hormones, anti-juvenile hormones, or chitin biosynthesis inhibitors.