Lipid changes during development of Blastocladiella emersonii

The lipid composition of swimming spores, cysts and five hour germlings was established. Spores utilized triglycerides first, then phospholipids. Upon encystment all glycolipid components decreased, while in germlings the phospholipids, monoglycerides and sterol esters exhibited a marked increase.     

Respiration of the hydrogenosome-containing fungus Neocallimastix patriciarum

Mass spectrometric determinations of O₂ affinities by the rumen fungus Neocallimastix patriciarum indicated a stable respiration under liquid phase O₂ concentrations up to 10 M, the apparent K _m for O₂ under these conditions was 4.0 M. Exposure to O₂ concentrations in excess of 10 M resulted in rapid inactivation of the observed respiration. Calculated H₂ evolution rates for the organism are 8.1 nmol min^-1 per mg of protein. Exposure to liquid-phase O₂ concentrations in excess of 1.4 M caused 50% inhibition of H₂ production. That superoxide and peroxide are amongst the products of respiration was shown by the use of ESR spectroscopy with the spin trapping agent 5,5-dimethyl-l-pyrroline-N-oxide. An active superoxide dismutase was present, but catalase could not be detected.Abbreviations ESR electron spin resonance - DMPO 5,5-dimethyl-l-pyrroline-N-oxide - DETAPAC diethylene-triamine pentaacetic acid     

Formation of Metallogenium-like structures by a manganese-oxiding fungus

Structures resembling Metallogenium spp. were observed in agar and in liquid cultures of a Mn-oxidizing basidiomycetous fungus only when Mn²⁺ was oxidized. Fungal viability was necessary for formation of the structures; Mn²⁺ concentration and the presence or absence of agar in the medium were important factors determining their morphology. Slide cultures revealed no identifiable cells in any stage of development. Fluorescent dyes that stained nucleic acids and polysaccharides in the fungal hyphae did not stain the Metallogenium-like structures. Likewise, Rhodamine 123, a fluorescent probe for membrane potential, stained fungal mitochondria, but did not stain the structures. Thin sections through the structures showed no biological membranes or other cellular features. Only the characteristic ultrastructure of biological Mn oxides were observed in serial thin sections. In agar, unfixed structures disappeared permanently during reduction of Mn oxides with hydroxylamine. Glutaraldehyde fixation stabilized these structures. Fixed structures lost most of their original phase density during reduction with hydroxylamine, but continuous microscopic observations showed that their phase density could be restored by staining with Coomassie blue. Structures that formed in liquid medium did not require stabilization with glutaraldehyde during reduction of Mn oxides. They, too, lost their original phase density during reduction with hydroxylamine; phase density could be restored by staining with cationic colloidal iron or Coomassie blue. The results suggest that the Metallogenium-like structures were formed as a result of Mn oxidation associated with exopolymers produced by the fungus.Non-standard abbreviations HEPES (N-hydroxyethylpiperazine-N-2-ethane sulfonic acid) - DAPI (4,6-diamidino-2-phenylindole) - PIPES (piperazine-N,N-bis[2-ethane sulfonic acid])     

Hyphal fusion during initial stages of trap formation in Arthrobotrys oligospora

Hyphal fusion during initial stages of trap formation by Arthrobotrys oligospora was studied by video-enhanced contrast and electron microscopy. Trap initials grew perpendicularly to the parent hypha, then curved around and anastomosed with a peg that developed on the hypha. Trap initials usually developed 40–140 m apart while the anastomosis occurred 20–25 m from the initial. Vigorous cytoplasmic movements in trap initials and developed traps corresponded to intense staining with fluorescein diacetate (FDA) of these cells. In addition, bundles of microfilaments were seen in developing loops of traps. On fusion organelle migration took place from the tip cell of the trap into the peg. Later on a septum was formed at the site of fusion.     

Methanogenic bacteria as a key factor involved in changes of town gas stored in an underground reservoir

Growth of Phanerochaete chrysosporium in a nitrogen-limited medium buffered with sodium acetate, instead of the commonly used 2,2-dimethylsuccinate (DMS), resulted in quantitative and qualitative differences in the production of various extracellular lignin peroxidases (LIPs) and manganese-dependent peroxidases (MNPs) involved in lignin degradation. The results indicate that production of LIPs and MNPs can be selectively enhanced by manipulation of culture conditions. Partial N-terminal analyses of the major LIPs and MNPs have made it possible to assign a specific protein to the specific genes and cDNAs that have been reported recently. The LIPs and MNPs differed widely in their ability to decolorize various dyes that are known to be degraded by the lignin degrading enzyme system of P. chrysosporium.     

Ethephon and auxin induce mycorrhiza-like changes in the morphology of root organ cultures of Mugo pine

The effects of ethylene and auxin on the morphology and anatomy of root organ cultures of Pinus mugo Turra var. mugo were investigated to test the hypothesis that changes in root morphology associated with formation of ectomycorrhizae may be related to ethylene produced by ectomycorrhizal fungi or by host plant roots in response to fungus-produced auxin. Morphological changes characteristic of mycorrhizal infection include dichotomous branching of lateral roots, inhibition of root hair formation and enlargement of cortical cells. Lateral roots on non-mycorrhizal root organ cultures, grown in a defined medium, underwent dichtotomous branching while root hair formation was inhibited in response to the ethylene released by 50 and 100 μ M ethephon (2-chloroethylphosphonic acid), but no effect on cortical cell dimensions was observed. The auxin, naphthaleneacetic acid (1 and 10 μ M ) also stimulated dichotomous branching and inhibited root hair formation, but to a lesser extent and with a greater lag time than ethephon. Auxin-stimulated ethylene production by root organ cultures was demonstrated. This appeared to be responsible, at least in part, for the auxin-induced dichotomous branching since the ethylene action inhibitor, silver thiosulfate (0.1 m M ) inhibited the response to auxin by 35%.     

In vivo development and pathogenicity of Ascosphaera proliperda (Ascosphaeraceae) to the alfalfa leafcutting bee, Megachile rotundata

First instar larvae of the leafcutting bee, Megachile rotundata, were fed on either artificial or natural provisions containing spores of Ascosphaera proliperda. Two isolates were used as a source of inocula: one originated from in vitro isolates obtained while culturing what was thought to be pure spores of A. aggregata, the second originated from in vitro cultures from Denmark. Histological and scanning electron microscopy studies revealed that the spores germinated in the gut lumen and the developing hyphae invaded all tissues, after which they penetrated through larval integument and began the sexual phase of the life cycle aerially. Virtually all fungus-exposed larvae developed symptoms of disease regardless of source of inoculum, type of provision, and spore dose (1.5 × 10³ to 3 × 10⁶) per insect. It was concluded that the fungus was pathogenic to the alfalfa leafcutting bee under laboratory conditions and future studies should be conducted to determine its etiology, cross infectivity, and natural distribution in other bee taxa.