Microbial-feeding nematodes and protozoa in soil: Their effectson microbial activity and nitrogen mineralization in decomposition hotspots and the rhizosphere

Food web studies from a range of ecosystems have demonstrated that the fauna contributes about 30% of total net nitrogen mineralization. This results mainly from the activities of microbial-feeding microfauna (nematodes and protozoa). Microbial and microfaunal activity is concentrated at spatially discrete and heterogeneously distributed organic substrates, including the rhizosphere. The dynamics of microfauna and their effect on nutrient cycling and microbial processes at these sites is reviewed. The potential manipulation of microfauna, either as an experimental tool to further understand soil microbial ecology or as a practical means of managing nutrient flows in agroecosystems, is discussed.     

Artificial destratification of a small tropical reservoir: effects upon the phytoplankton

Seasonal changes in the phytoplankton community of a small tropical reservoir were monitored over a four year period comprising of an initial two seasonal cycles during which the water column stratified strongly for extended periods each year, and two further seasonal cycles after installation of a mechanical aeration system to induce artificial destratification. In the unmanaged reservoir, the concentration of chlorophyll a at 0.5 m reached maximum values (on one occasion > 90 mg m⁻³) when the water column was stratified and the epilimnion was very shallow (ca 2 m depth). The hypolimnion at this time was anoxic (less than 2% oxygen saturation) and had a high concentration of bacteriochlorophyll (100–200 mg m⁻³). The phytoplankton community of the unmanaged reservoir was generally dominated by cyanobacteria (Cylindrospermopsis raciborskii, Anabaena tenericaulis) during the warmer months of the year (November–March) (but replaced by chlorophyta, dinophyceae and euglenophyceae after periods of intense rain) and by bacillariophyceae (Synedra ulna var. chaseana, S. tenera) during the cooler, dry months. In the artificially destratified reservoir (8 h aeration day⁻¹), the phytoplankton community was largely dominated by diatoms except after depletion of the silica content of the water column which caused diatoms to be replaced by cyanobacteria (dominated by A. tenericaulis) and a range of chlorophytes. The changing pattern of stratification and circulation of the water column in the unmanaged reservoir caused repeated disruption of the established phytoplankton assemblage with peaks of high biomass associated with transient cyanobacterial blooms. Continuous aeration and the consequent increase in the ratio mixed: euphotic depth provided conditions suitable for dominance of the phytoplankton by diatoms, as long as silica was available, and resulted in average chlorophyll levels higher than in the unmanaged reservoir (120 ± 10 v. 64 ± 9 mg m⁻²). Hierarchical fusion analysis based on the biomass of species differentiated the phytoplankton samples into cluster groups that could be related primarily to stratification or mixing of the water column.     

Enzymological and mutational analysis of a complex primary hyperoxaluria type 1 phenotype involving alanine:Glyoxylate aminotransferase peroxisome-to-mitochondrion mistargeting and intraperoxisomal aggregation

Primary hyperoxaluria type 1 (PH1) is a rare autosomal recessive disease caused by a deficiency of the liver-specific peroxisomal enzyme alanine:glyoxylate aminotransferase (AGT). Three unrelated PH1 patients, who possess a novel complex phenotype, are described. At the enzymological level, this phenotype is characterized by a complete, or nearly complete, absence of AGT catalytic activity and reduced AGT immunoreactivity. Unlike normal individuals in whom the AGT is confined to the peroxisomal matrix, the immunoreactive AGT in these three patients was distributed approximately equally between the peroxisomes and mitochondria. The peroxisomal AGT appeared to be aggregated into amorphous core-like structures in which no other peroxisomal enzymes could be identified. Mutational analysis of the AGT gene showed that two of the three patients were compound heterozygotes for two previously unrecognized point mutations which caused Gly41→Arg and Phe152→Iso amino acid substitutions. The third patient was shown to be a compound heterozygote for the Gly41→Arg mutation and a previously recognized Gly170→Arg mutation. All three patients were homozygous for the Pro11→Leu polymorphism that had been found previously with a high allelic frequency in normal populations. It is suggested that the Phe152→Iso and Gly170→Arg substitutions, which are only eighteen residues apart and located in the same highly conserved internal region of 58 amino acids, might be involved in the inhibition of peroxisomal targeting and/or import of AGT and, in combination with the Pro11→Leu polymorphism, be responsible for its aberrant mitochondrial compartmentalization. On the other hand, the Gly41→Arg substitution, either in combination with the Pro11→Leu polymorphism or by itself, is predicted to be responsible for the intraperoxisomal aggregation of the AGT protein.     

Plasmid Suppressors Active in the Sexual Cycle of Neurospora Intermedia

We have discovered that, in certain crosses of natural isolates of Neurospora intermedia, linear and circular mitochondrial plasmids of the maternal parent are not transmitted to the progeny. This contrasts with the maternal transmission of organellar genetic elements generally observed in crosses between laboratory strains and between other natural isolates. Formally, failure of plasmid transmission is a type of plasmid suppression. The present cases represent the first report of plasmid suppressors in natural populations of fungi. Strains used as female parents can transmit or not transmit plasmids depending on the strain used as male parent. Males that act to suppress in one cross fail to suppress in others. Therefore, the suppression of plasmids depends on a strain-specific interaction and is not determined exclusively by the males. Since suppression is a specific interaction we inferred that it must be genetically based and tested this hypothesis by seeking segregation of suppressed and nonsuppressed phenotypes in octads. Segregation of the original full suppression of all plasmids was indeed observed in each of the three sets of testcrosses examined. The interaction type of suppression must be initiated in ascogenous tissue during the sexual cycle. It is a nonautonomous type of suppression, affecting all descendent cells. In any one case of suppression, either one, several, or all plasmids can be lost. Both linear and circular plasmids can be eliminated by the same suppressor genotype. In addition, several strains were found to contain suppressors that act after ascospore delineation. This autonomous type of suppression has been observed previously in laboratory strains, but not in natural isolates. All the cases of plasmid suppression identified in this study involved a range of apparently neutral circular and linear plasmids. Using one senescent Kalilo strain of N. intermedia, we did not detect any case of suppression of the senescence-determining linear plasmid kalDNA.     

Mycorrhizal associations in Hong Kong Fagaceae

Polyphenols histochemically detected in fresh uninfected roots of Quercus, Castanopsis and Lithocarpus growing in Hong Kong and shown to be condensed tannins were found mainly as intracellular material in the cells of the root cap, the epidermal layer and the endodermis. The cell walls of the outer cortex and the endodermis also contained suberin. Following invasion by compatible ectomycorrhizal symbionts, condensed tannins disappeared from cells of the root cap and the epidermal layer but hyphae were prevented from colonizing the cortex presumably due to suberin barriers. In vitro experiments indicated that a number of broad-host ectomycorrhizal fungi could utilise various polyphenolic compounds, including tannins found in the root exudates of the host trees, with different degrees of efficiency.     

Proteomics in Drosophila melanogaster.

To be able to understand cellular mechanisms, we require fully integrated data sets combining information about gene expression, protein expression, post-translational modification states, sub-cellular location and complex formation. Proteomics is a very powerful technique that can be applied to interrogate changes at the protein level. Studying this effectively requires specialised facilities within research institutes. Here, we describe the setting up and operation of such a facility, providing a resource for the Arabidopsis and Drosophila research communities.     

Secretory lysosomes - a special mechanism of regulated secretion in haemopoietic cells

The secretory granules o f cells derived from the haemopoietic lineage are 'secretory lysosomes' containing both lysosomal hydrolases and secretory proteins. Studies on cytotoxic T lymphocytes (CTLs) have elucidated several of the mechanisms that regulate protein sorting to, and secretion from, this unusual secretory organelle. In particular, recent findings from a CTL mutant have led to the hypothesis that CTLs, and other cells of the haemopoietic lineage, use specialized sorting and secretory mechanisms in which the lysosome functions as a regulated secretory granule.     

Assembly of vaccinia virus: incorporation of p14 and p32 into the membrane of the intracellular mature virus.

The cytoplasmic assembly of vaccinia virus begins with the transformation of a two-membraned cisterna derived from the intermediate compartment between the endoplasmic reticulum and the Golgi complex. This cisterna develops into a viral crescent which eventually forms a spherical immature virus (IV) that matures into the intracellular mature virus (IMV). Using immunoelectron microscopy, we determined the subcellular localization of p32 and p14, two membrane-associated proteins of vaccinia virus. p32 was associated with vaccinia virus membranes at all stages of virion assembly, starting with the viral crescents, as well as with the membranes which accumulated during the inhibition of assembly by rifampin. There was also low but significant labelling of membranes of some cellular compartments, especially those in the vicinity of the Golgi complex. In contrast, anti-p14 labelled neither the crescents nor the IV but gave strong labelling of an intermediate form between IV and IMV and was then associated with all later viral forms. This protein was also not significantly detected on identifiable cellular membranes. Both p32 and p14 were abundantly expressed on the surface of intact IMV. Our data are consistent with a model whereby p32 would become inserted into cellular membranes before being incorporated into the crescents whereas p14 would be posttranslationally associated with the viral outer membrane at a specific later stage of the viral life cycle.