Microbial characteristics of sandy soils exposed to diazinon under laboratory conditions

Changes in microbiological characteristics in response to diazinon, applied at three different dosages of 7, 35 and 700 mg kg^?1 soil, were studied in pots filled with sandy soils of different texture. The insecticide dosages corresponded to the maximum predicted environmental concentration (PEC) in field conditions and five or hundred times this rate, respectively. To ascertain these changes, activities of selected soil enzymes, numbers of some microbial populations and bacterial community structure (r/K-strategists) were determined. Acid phosphatase activity and concentrations of ammonium ions either increased or were unaffected by the diazinon dosages. In contrast, nitrate ions, alkaline phosphatase, urease and especially dehydrogenase activities decreased in both soils treated with the higher dosages of the insecticide. The diazinon treatments increased the numbers of culturable bacteria and fungi; however, N-fixing bacteria and nitrifiers (but not denitrifiers) were decreased in both soils. When the high concentration of diazinon (100-times PEC) was added to the loamy sand soil (LS), simulating an undiluted chemical spill, the bacterial community structure shifted towards domination of K-strategists over time. However, r-strategists dominated in the community exposed to the same dosage of the insecticide in the sandy loam soil (SL). Generally, SL soil characteristics were more suitable for microbial activity and growth than those of LS soil were. The results indicate that except a situation of undiluted spill diazinon seems to be a non-hazardous chemical for the culturable soil microorganisms when applied at recommended concentrations.     

Immunocytochemical investigations on the presence of neuron-specific antibodies in the CSF of Parkinson's disease cases

The CSF and sera of 7 patients with Parkinson's disease were investigated immunocytochemically, in order to see if antibodies were present which recognized DA-ergic cell bodies in glutaraldehyde fixed rat brain. In 2 patients a marked labeling of DA-ergic neurons in the substantia nigra was observed, identified by anti-DA antiserum and by 6-OHDA induced degeneration, but also other neurons in the ventral mesencephalon were recognized. The other patients were weakly positive or negative. Sera gave unspecific labelling of all neurons. In one patient the sub-classes of IgG were investigated and found to be of IgG₃ (labeling nerve terminals) and of IgG_1–2, low affinity type (recognizing perikarya). The epitopes recognized have not been identified, but are unlikely to be DA-like, since blocking experiments and ELISA-tests gave negative results. The possible clinical importance of the results are discussed.Special Issue dedicated to Prof. Holger Hydén.     

Thermal requirements for development of aphidophagous Coccinellidae (Coleoptera), Chrysopidae,Hemerobiidae (Neuroptera), and Syrphidae (Diptera): some general trends

Summary The sum of effective temperatures (SET) and lower development threshold (LDT) were established for eggs and/or pupae of central European populations of 20 species of chrysopid, coccinellid, hemerobiid, and syrphid predators of aphids. LDT ranged between 5.6° and 12.2°C, SET between 38.3 and 140.9 day degrees (dd), with broad overlap among stages and taxa. When LDT was plotted against SET, the data for both eggs and pupae were scattered along a single regression line which predicted a 0.47°C decrease in LDT per 10 dd increase in SET (r=-0.77, p<0.001). A regression calculated from published data from all over the world predicted a 0.24°C/10 dd decrease in LDT, and the data were more scattered (r=-0.38, p<0.01). This is perhaps the first report on the functional relationship between LDT and SET at the interspecific level. The species and stages differed in typical development length (VDL) and in the extent of its deceleration by low temperatures (DD). DD increased with increasing VDL, but the relative effect of low temperature on development length (DD/VDL ratio) reflected thermal adaptations consistent with the life history of the species. Polyvoltine species were less affected by low temperatures than monovoltine species, particularly the thermophilic ones.     

Parthenogenetic activation decreases the polyphosphoinositide content of frog eggs

Polyphosphoinositides were quantified in metaphase II-arrested eggs of the amphibian Xenopus laevis and 8-10 min later in eggs activated by pricking. The content of phosphatidylinositol 4,5-biphosphate (PIP2) was remarkably high in metaphase II-arrested eggs with respect to that of phosphatidylinositol 4-phosphate (PIP). It was found to drop dramatically at activation. In contrast PIP content did not change significantly.     

Two atavistic characters of some Lumbriculidae and their importance for the classification of Oligochaeta

The rudimentary atria, and the posterior sperm funnels and sperm ducts, which occur in some species of the Lumbriculidae, are discussed. It is shown that the posterior location of funnels and sperm ducts is the result of a forward shift of the atria, which refutes Stephenson's supposition that the Lumbriculidae is the most archaic family of the present-day Oligochaeta.     

Purification and characterization of wheat and pine small basic protein substrates for plant calcium-dependent protein kinase.

A wheat basic protein (WBP) was purified to homogeneity from wheat germ by a protocol involving extraction, centrifugation, batchwise elution from carboxymethylcellulose (CM-52), acidification with trifluoroacetic acid, neutralization and HPLC on a SP5PW cation exchange column. WBP is a 10 kDa protein and is phosphorylated on serine residues by wheat germ Ca(2+)-dependent protein kinase (CDPK). [32P]phosphoWBP exactly comigrates with WBP on SDS-PAGE. WBP does not inhibit either wheat germ CDPK or calmodulin-dependent myosin light chain kinase. Apart from histone H1, WBP is the best endogenous substrate yet found for wheat embryo CDPK. A 12 kDa pine basic protein (PBP) was purified to homogeneity from seeds of stone pine (Pinus pinea L.) by a simple procedure involving batchwise elution from carboxymethylcellulose and cation exchange HPLC. PBP is also a good substrate for CDPK and is phosphorylated on Ser residues. N-terminal sequencing of WBP and PBP revealed that these proteins are homologous to a family of small basic plant proteins having a phospholipid transfer function.     

Effect of temperature on development rate and intrinsic rate of increase ofAphis gossypii reared on greenhouse cucumbers

Thermal requirements for development and life table statistics ofAphis gossypii Glover (Homoptera, Aphididae) were determined over a range of constant temperatures from 10 to 30°C. The lower development threshold and the sum of effective temperatures were 6.9°C and 90.1°C, respectively, for preimaginal development, and 5.8°C and 113.6°C from birth to the onset of reproduction. Mean total fecundity ranged from 36 larvae per female at 10°C to 76 larvae at 30°C. On a time scale of days, net reproductive rate (R _o ) increased with increasing temperature while generation time (T) decreased causing the intrinsic rate of increase (r _m ) to increase linearly from 0.115 to 0.465. On a day-degree scaler _m only varied from 0.019 to 0.028 because the growth ofR _o was compensated by an increase inT with increase in temperature. The nearly constantr _m in terms of day-degrees, over a wide range of temperatures, greatly simplifies the prediction of future population numbers ofA. gossypii.     

Purification and characterization of (1→3, 1→4)-β-glucan endohydrolases from germinated wheat (Triticum aestivum)

A (13, 14)--glucan 4-glucanohydrolase [(13, 14)--glucanase, EC 3.2.1.73] was purified to homogeneity from extracts of germinated wheat grain. The enzyme, which was identified as an endohydrolase on the basis of oligosaccharide products released from a (13, 14)--glucan substrate, has an apparent pI of 8.2 and an apparent molecular mass of 30 kDa. Western blot analyses with specific monoclonal antibodies indicated that the enzyme is related to (13, 14)--glucanase isoenzyme EI from barley. The complete primary structure of the wheat (13, 14)--glucanase has been deduced from nucleotide sequence analysis of cDNAs isolated from a library prepared using poly(A)⁺ RNA from gibberellic acid-treated wheat aleurone layers. One cDNA, designated LW2, is 1426 nucleotide pairs in length and encodes a 306 amino acid enzyme, together with a NH₂-terminal signal peptide of 28 amino acid residues. The mature polypeptide encoded by this cDNA has a molecular mass of 32085 and a predicted pI of 8.1. The other cDNA, designated LW1, carries a 109 nucleotide pair sequence at its 5 end that is characteristic of plant introns and therefore appears to have been synthesized from an incompletely processed mRNA. Comparison of the coding and 3-untranslated regions of the two cDNAs reveals 31 nucleotide substitutions, but none of these result in amino acid substitutions. Thus, the cDNAs encode enzymes with identical primary structures, but their corresponding mRNAs may have originated from homeologous chromosomes in the hexaploid wheat genome.     

Molecular evolution of plant β-glucan endohydrolases

The evolutionary relationships of two classes of plant β-glucan endohydrolases have been examined by comparison of their substrate specificities, their three-dimensional conformations and the structural features of their corresponding genes. These comparative studies provide compelling evidence that the (1→3)-β-glucanases and (1→3,1→4)-β-glucanases from higher plants share a common ancestry and, in all likelihood, that the (1→3,1→4)-β-glucanases diverged from the (1→3)-β-glucanases during the appearance of the graminaceous monocotyledons. The evolution of (1→3,1→4)-β-glucanases from (1→3)-β-glucanases does not appear to have invoked ‘modular’ mechanisms of change, such as those caused by exon shuffling or recombination. Instead, the shift in specificity has been acquired through a limited number of point mutations that have resulted in amino acid substitutions along the substrate-binding cleft. This is consistent with current theories that the evolution of new enzymic activity is often achieved through duplication of the gene encoding an existing enzyme which is capable of performing the required chemistry, in this case the hydrolysis of a glycosidic linkage, followed by the mutational alteration and fine-tuning of substrate specificity. The evolution of a new specificity has enabled a dramatic shift in the functional capabilities of the enzymes. (1→3)-β-Glucanases that play a major role, inter alia, in the protection of the plant against pathogenic microorganisms through their ability to hydrolyse the (1→3)-β-glucans of fungal cell walls, appear to have been recruited to generate (1→3,1→4)-β-glucanases, which quite specifically hydrolyse plant cell wall (1→3,1→4)-β-glucans in the graminaecous monocotyledons during normal wall metabolism. Thus, one class of β-glucan endohydrolase can degrade β-glucans in fungal walls, while the other hydrolyses structurally distinct β-glucans of plant cell walls. Detailed information on the three-dimensional structures of the enzymes and the identification of catalytic amino acids now present opportunities to explore the precise molecular and atomic details of substrate-binding, catalytic mechanisms and the sequence of molecular events that resulted in the evolution of the substrate specificities of the two classes of enzyme.     

Separation and properties of α-mannosidase and mannanase from the basidiomycetePhellinus abietis

Proteins of a crude enzyme preparation obtained from the cultivation medium of the basidiomycetePhellinus abietis were separated by gel filtration and ion-exchange chromatography. The preparation contained a minimum of three enzymes capable of splitting α-d-mannosidic bonds: α-mannosidase, exomannanase, and endomannanase, which were separated. Some properties of the mannanase complex of the crude enzyme preparation, and of a partially purified α-mannosidase were examined. The mannanase complex exhibited two pH optima, its temperature optimum being at 46 °C The pH optimum of purified α-mannosidase was at pH 5.0, the temperature optimum was at 60 °C; the enzyme had a relatively high heat stability. The K_m of α-mannosidase forp-nitrophenyl α-d-mannopyranoside was 1.5 x 10⁻⁵ M. Pure α-mannosidase did not split mannan.