Changes in crown leaching composition induced by a sudden increase in atmospheric deposition. A case study in South-Western Bulgaria

Rain and throughfall chemistry has been monitored for five years (1987–1991) in three adjacent stands (Norway spruce, Silver fir and Scots pine) situated at an altitude of 1500 m above sea level in the Rhodopes mountains (South-Western Bulgaria). Throughfall collectors have been set up near the stem, below the opening between the crowns, and halfway between the stem and the border of the tree crown. Pollutant concentration in bulk precipitation strongly increased during the period of investigation. Because base cation deposition increased together with sulfate deposition, the increase in proton deposition remained moderate. The increase in throughfall concentration was parallel for different tree species. The time course of mineral concentration in throughfall was approximately similar to that of rainfall but the inter-event variability in concentration was reduced. Net throughfall fluxes of Ca, Mg and K increased during the study period whereas net throughfall fluxes of Cl, Na and S remained more stable. Although the wet deposition of protons increased, the proton input in net throughfall decreased or remained stable in relation to the increase in base cation concentration. This suggests a low dry deposition in the study region and conversely an increased net leaching of base cations. However, the seasonal variations in net throughfall as well as the time course of the difference between the concentrations of throughfall samples collected at different distances from the stems indicates that most of the increase in the net throughfall of Ca and Mg was due to dry deposition. The difficulties associated with the use of ions such as Na as deposition tracers for other base cations is emphasized.     

The Use of Crown Ethers in Peptide Chemistry – V. Solid-phase Synthesis of Peptides by the Fragment Condensation Approach using Crown Ethers as Non-covalent Protecting Groups

We have previously described the conditions by which peptide synthesis by the solid-phase fragment condensation approach can be carried out using crown ethers as non-covalent protection for the N^α -amino group. Here we demonstrate that the procedure can be extended to large, partially protected peptide fragments possessing free Lys and/or Arg residues. The first step was to ensure that complex formation on the side chain of amino acids was not detrimental to the methodology and exhibited the same solubility and coupling properties as N^α -complexed peptides. Thus, a model hexapeptide was synthesized using Fmoc chemistry containing Lys and Arg residues, which, when complexed with 18-Crown-6, was readily soluble in DCM and coupled quantitatively to a resin-bound tetrapeptide. Two tripeptides were then prepared, one containing a free Ser residue, the other free Tyr, to examine the possible occurrence of side reactions. After coupling using standard conditions only the former tripeptide exhibited the formation of the O-acylation by-product (5%). Another model hexapeptide containing Lys, Tyr, Ser and Asp protected with a TFA-stable adamantyl group was complexed with 18-Crown-6 and coupled to the resin-bound tetrapeptide with near quantative yield. Extending the length of the peptide to 21 and 40 residues, which represent sequences Gly⁵² to Leu⁷² (21-mer) and Pro³³ to Leu⁷² (40-mer) from Rattus norvegicus chaperonin 10 protein, respectively, resulted in partially protected fragments that were readily soluble in water, thus enabling purification by RP-HPLC. Complexation with 18-Crown-6 gave two highly soluble products that coupled to resin-board tetramer with 68% and 50% coupling efficiencies for the 21-mer and 40-mer, respectively. Treatment with 1% DIEA solutions followed by acidolytic cleavage and purification of the major product confirmed that the correct product had been formed, when analysed by amino acid analysis and ESI-MS. These results served to extend the methodology of non-covalent protection of large partially protected peptide fragments for the stepwise fragment condensation of polypeptides.     

Presence of a plant cell wall is not required for transformation of Nicotiana by Agrobacterium tumefaciens

Agrobacterium has been used to transform zero to six-day-old cell wall nonregenerating (CWNR) and cell wall regenerating (CWR) leaf protoplasts of tobacco. Transformed cells were selected by phoytohormone autotrophic growth and were verified by detection of the presence of lysopine dehydrogenase. Transformation frequencies in CWNR protoplasts were at least as high as those in CWR protoplasts, indicating that a plant cell wall is not required for the process of crown gall tumorigenesis. Transformation frequencies were highest in two-day-old protoplasts. This age coincides with the onset of DNA synthesis and the first mitosis within the cell populations. We suggest that the initiation of cell cycle activity may be important for the transformation process.     

Zinc in teeth from children with and without cystic fibrosis

This study analyzes Zn concentration levels in teeth from children with and without cystic fibrosis with respect to different variables, namely: gender, age, type of teeth, area, fluoridation of water supply, term of pregnancy, maternal smoking habit, and maternal drinking habit. The method of analysis is proton-induced X-ray emission on crown samples. Atomic absorption spectrometry (AAS) studies on some of these samples show close correlation to the PIXE data. In general, the concentration level of zinc in teeth of children greater than 10 years old is less than the concentration level in teeth of children aged 10 years or younger for children who took nontetracycline antibiotics. Breast feeding-mixed feeding-bottle feeding show no significant difference in the zinc concentration in teeth.     

Calcium and phosphorous in teeth from children with and without cystic fibrosis

The objective of this study is to analyze the percentage of Ca and P in teeth from children with and without cystic fibrosis with respect to different variables, namely, gender, age, type of teeth, area, fluoridation of water supply, term of pregnancy, and maternal smoking and drinking habits. The method of analysis is proton-induced X-ray emission and proton-induced gamma emission on tooth-crown samples. T-test results show less Ca in the teeth of the population of CF + NT (cystic fibrosis+ nontetracycline antibiotics) than in that of NCF (noncystic fibrosis) for incisors and the total tooth population. It also shows greater Ca for incisors than for nonincisors for NCF. Teeth from urban CF + NT have less Ca than those from urban NCF. This holds for the total area population. Both Ca and P in teeth of NCF population living in fluoridated areas are greater than in those living in nonfluoridated area. Ca is depleted in the teeth of CF + NT children whose mothers smoke and P is depleted in the teeth of NCF children whose mothers drink. High and low Ca and P values for individual teeth are reported.     

The ultrastructure of the zygospore in Endogone flammicorona Trappe & Gerdemann

The ultrastructural organization of the spores of the sporocarp of Endogone flammicorona was studied. Two types of organization are described. Initially the spore possessed a vacuolate protoplasm and was bound by two cell wall layers. The spore was surrounded by a hyphal mantle formed of a sheet of vacuolized hyphae with uniformly thin walls. Secondly, although the ultrastructural features of the spore appeared the same, it was now surrounded by a hyphal mantle with unevenly thickened walls (i. e., the so-called flaming crown) due to the gradual and irregular deposition of granules and lamellae. This crown gives the spore its most commonly observed morphological feature and is the preminent character employed taxonomically to speciate Endogone flammicorona Trappe & Gerdemann.     

Development of a semi-nested PCR based method for sensitive detection of tumorigenic Agrobacterium in soil

AIMS: To develop a specific, sensitive and rapid PCR-based method for detection of tumorigenic Agrobacterium in soil. METHODS AND RESULTS: Three newly designed primers complementary to tms2 gene amplified DNA of only the tumor-inducing agrobacteria of 113 strains tested, resulting in 617 bp and 458 bp products in the first and second rounds of semi-nested PCR respectively. As optimized method of pre-incubation of soil suspensions on selective medium, DNA isolation and two-round semi-nested PCR enabled detection of 1-2 bacterial cells in 1 g of soil. Using this method tumour-inducing Agrobacterium was detected in 67 of 69 samples of naturally infested soil originating from the field, where plants with crown gall symptoms occurred. The pathogen was detected only in two samples of 15 tested, collected from a nursery where crown gall symptoms were not observed. CONCLUSIONS: The semi-nested PCR-based method allowed for sensitive and rapid detection of tumorigenic agrobacteria in soil. SIGNIFICANCE AND IMPACT OF THE STUDY: The method is proposed for testing of soil in fields intended for nursery production of fruit trees, roses or other plants susceptible to crown gall, as well as a tool for ecological studies.     

Regulation of cell adhesion using a signal-responsive membrane substrate

We have developed a novel cell culture material that regulates cell adhesion by changes in potassium ion concentration. The material is a polyethylene substrate grafted to a copolymer of the thermoresponsive polymer N-isopropylacrylamide (NIPAM) and benzo-18-crown- 6-acrylamide (BCAm), with a pendant crown ether as sensor. The crown ether recognizes potassium ion concentrations and NIPAM conformational changes lead to changes in the hydrophobicity/hydrophilicity balance of the entire polymer at constant cell culture temperatures. Although cells were successfully cultured on the ion recognition material in normal culture medium at 37 degrees C, the cells could be detached from the material surface by adding potassium ions alone, without proteolytic enzymes, because the surface to which the cells were attached altered its surface characteristics to a more hydrophilic state. Therefore, cell layers with intact cell-to-cell junctions and high activities were successfully recovered. Furthermore, by changing the target sensors, this material will be able to control cell adhesion through various cellular signals.     

Population continuity vs. discontinuity revisited: dental affinities among late Paleolithic through Christian-era Nubians

The present study revisits a subject that has been a source of long-standing bioarchaeological contention, namely, estimation of Nubian population origins and affinities. Using the Arizona State University dental anthropology system, frequencies of 36 crown, root, and intraoral osseous discrete traits in 12 late Pleistocene through early historic Nubian samples were recorded and analyzed. Specifically, intersample phenetic affinities, and an indication of which traits are most important in driving this variation, were determined through the application of correspondence analysis and the mean measure of divergence distance statistic. The results support previous work by the author and others indicating that population discontinuity, in the form of replacement or significant gene flow into an existing gene pool, occurred sometime after the Pleistocene. This analysis now suggests that the break occurred before the Final Neolithic. Samples from the latter through Christian periods exhibit relative homogeneity, which implies overall post-Pleistocene diachronic and regional population continuity. Yet there are several perceptible trends among these latter samples that: 1) are consistent with documented Nubian population history, 2) enable the testing of several existing peopling hypotheses, and 3) allow the formulation of new hypotheses, including a suggestion of two post-Pleistocene subgroups predicated on an age-based sample dichotomy.     

A high‐resolution genetic map of the cereal crown rot pathogen Fusarium pseudograminearum provides a near‐complete genome assembly

Fusarium pseudograminearum is an important pathogen of wheat and barley, particularly in semi‐arid environments. Previous genome assemblies for this organism were based entirely on short read data and are highly fragmented. In this work, a genetic map of F. pseudograminearum has been constructed for the first time based on a mapping population of 178 individuals. The genetic map, together with long read scaffolding of a short read‐based genome assembly, was used to give a near‐complete assembly of the four F. pseudograminearum chromosomes. Large regions of synteny between F. pseudograminearum and F. graminearum, the related pathogen that is the primary causal agent of cereal head blight disease, were previously proposed in the core conserved genome, but the construction of a genetic map to order and orient contigs is critical to the validation of synteny and the placing of species‐specific regions. Indeed, our comparative analyses of the genomes of these two related pathogens suggest that rearrangements in the F. pseudograminearum genome have occurred in the chromosome ends. One of these rearrangements includes the transposition of an entire gene cluster involved in the detoxification of the benzoxazolinone (BOA) class of plant phytoalexins. This work provides an important genomic and genetic resource for F. pseudograminearum, which is less well characterized than F. graminearum. In addition, this study provides new insights into a better understanding of the sexual reproduction process in F. pseudograminearum, which informs us of the potential of this pathogen to evolve.