Effect of carbon levels from three organic substrates on egyptian soil fungi

Summary The fungal composition of soils with an equivalent percentage of carbon levels 2%, 5%, 10% and 20% fromOlea europea,, Citrus sinensis andEucalyptus rostrato leaves was studied after 10, 45 and 120 days using the dilution-plate method. They all induced significant increases in the total count of fungi, but their effect on the individual fungi was selective. The best substrate for the total fungus flora wasCitrus sinensis and the poorest wasOlea europeae leaves. The best three colonisers (possessing the highest counts), at carbon levels 2%, 5%, 10% and 20% at the end of the experimental periods were, respectively, as follows: forOlea europeae leaves,Scopulariaopsis brevicaulis, Drechslera halodes andCylindrocarpon sp.;Aspergillus quadrilineatus, A. sydowi andCurvularia lunata; A. quadrilineatus, Alternaria alternata andC. lunata andA. quadrilineatus, A. egyptiacus andA. terreus; forCitrus sinensis leaves,A. quadrilineatus, A. fumigatus andS. brumptii; A. fumigatus, A. quadrilineatus andA. versicolor; A. quadrilineatus, A. terreus andS. brumptii; andA. quadrilineatus, A. flavus andA. sydowi; and forEucalyptus rostrata leaves,A. quadrilineatus, Rhizopus stolonifer andA. sydowi; A. quadrilineatus, A. niger andA. terreus; A. niger, A. quadrilineatus andS. brevicaulis; andA. terreus, A. niger andA. quadrilineatus.     

A rapid identification method for aflatoxin-producing strains ofAspergillus flavus andA. parasiticus by ammonia vapor

The colony reverse of aflatoxin (AF)-producing strains ofAspergillus flavus andA. parasiticus turned pink when their cultures were exposed to ammonia vapor. The color change was visible for colonies grown on media suitable for AF production such as potato dextrose, coconut, and yeast extract sucrose agars after 2 d incubation at 25°C. Of the 120 strains ofA. flavus, A. parasiticus, and two related species inA. flavus group:A. oryzae andA. sojae tested in this study, only the AF-producing strains ofA. flavus andA. parasiticus showed the pink pigmentation. The color change occurred immediately after the colony was contacted with ammonia vapor. This method was useful for rapid screening the AF-producing strains ofA. flavus andA. parasiticus.     

Arbuscular mycorrhizal fungi on adjacent semi-natural grasslands with different vegetation in Japan

Arbuscular mycorrhizal (AM) fungi on Japanese semi-natural grasslands were investigated at three adjacent sites with different vegetation. The predominant grasses at the three sites were 1)Pleioblastus chino, 2)Miscanthus sinensis andArundinella hirta (M. sinensis/A. hirta), and 3)Zoysia japonica, respectively. The degree of colonization was higher inM. sinensis/A. hirta than inP. chino andZ. japonica. AM fungi were recovered by spore extraction and by pot cultures started from soil inoculum or from transplanting of field plants. Total spore number obtained by the spore extraction method was highest in the rhizosphere ofM. sinensis/A. hirta and lowest in that ofP. chino. AGlomus sp. resemblingG. geosporum predominated in association withM. sinensis/A. hirta andP. chino. FromZ. japonica, three species,Acaulospora gerdemannii, Glomus leptotichum, and a species resemblingG. clarum, were isolated by pot culture from soil and two species,A. longula andScutellospora cerradensis, by pot culture from transplanting ofZ. japonica. FromM. sinensis/A. hirta, one species,A. longula, was found by pot culture from soil. FromP. chino, no AM fungus was detected by either method. Single-spore culture confirmed thatG. leptotichum andA. gerdemannii are conspecific.     

Immunocytochemical identification of laticifers

Summary sensitive immunocytochemical method for the identification of laticifers has been developed. Frozen sections of various laticifer-bearing plant material, mounted on slides, were first flooded with the IgG fraction of rabbit anti-latex antiserum, prepared using whole latex ofAsclepias syriaca, then flooded with fluorescein-conjugated IgG fraction goat anti-rabbit IgG to visualize laticifers. Positive fluorescence was observed for laticifers in shoots and embryos ofA. syriaca andStapelia bella and embryos ofA. tuberosa. Laticifers did not fluoresce in shoots ofA. tuberosa andEuphorbia tirucalli, in embryos ofE. marginata, or in petioles ofMusa paradisiaca andCichorium intybus. Controls prepared with uninjected rabbit serum were negative (no fluorescence).     

Nucleation of calcite on the sheaths of cyanobacteria using a simple diffusion cell

A method allowing observation of calcium carbonate nucleation on the sheaths of cyanobacteria and other microorganisms is described. Nucleation of calcite was obtained on the sheaths of Lyngbya aestuarii and Scy‐tonema myochrous cultures but not on Entophysalis major and Pseudanabaena minuta. Differences in the rate of nucleation were also noted in media of differing salinity.     

Restriction enzyme analysis of the chloroplast and nuclear 45s ribosomal DNA ofAllium sectionsCepa andPhyllodolon (Alliaceae)

Estimates of the phylogenetic relationships among cultivated and wildAllium species would benefit from identification of molecular characters. Restriction enzyme analysis of the chloroplast DNA (cpDNA) of the bulb onion (Allium cepa), Japanese bunching onion (A. fistulosum), wildAllium species in sect.Cepa andPhyllodolon, and the outgroupsA. ampeloprasum andA. tuberosum detected 39 polymorphisms.Allium cepa andA. vavilovii were identical for all characters. Cladistic analysis generated three most-parsimoniousWagner trees of 44 steps differing only in a zero-length branch.Allium fistulosum andA. altaicum (sect.Phyllodolon) comprised a monophyletic lineage separated from theA. cepa andA. vavilovii of sect.Cepa. The unresolved node was composed ofA. galanthum, A. roylei, and the lineage containingA. cepa, A. vavilovii, A. fistulosum, andA. altaicum. The clade containingA. altaicum, A. cepa, A. fistulosum, A. galanthum, A. roylei, andA. vavilovii remained resolved for strict consensus ofWagner trees of 48 steps or less.Allium pskemense andA. oschaninii were increasingly distant.Allium oschaninii has been proposed as the progenitor of the bulb onion, but was more closely related to the common progenitor of all species in sect.Cepa andPhyllodolon. Phylogenies estimated from cpDNA characters usingDollo parsimony resulted in a single most-parsimonious tree of 46 steps and agreed with phylogenies based onWagner parsimony. Polymorphic restriction enzyme sites in the 45s ribosomal DNA were not used to estimate phylogenies because of uncertain homologies, but are useful for identifying interspecific hybrids. The maternal phylogenies estimated in this study help to distinguish wildAllium species closely related to the bulb onion. Although not in agreement with classifications based on morphology, the phylogenies closely reflected crossability among species in sect.Cepa andPhyllodolon.     

Preparation and regeneration of mycelial protoplasts ofPleurotus florida andP. ostreatus

A method for isolation of higher frequency of regenerated protoplast fromPleurotus florida andP. ostreatus is reported.     

Home range and mating system of two sympatric field mouse species,Apodemus speciosus andApodemus argenteus

The size and spatial distribution of home ranges in two sympatric field mouse species,Apodemus speciosus andApodemus argenteus, were revealed by the capture-recapture method in a temperate deciduous forest from June 1987 to June 1990. InA. speciosus, the home ranges of males were significantly larger tha those of females, and overlapped with those of other males and females during the breeding season. InA. argenteus, the home range of each male overlapped only that of a single female throughout the year. These results suggested thatA. speciosus was promiscuous or polygynous andA. argenteus was monogamous.     

Based on biochemical and physiological behavior,where isAspergillus egyptiacus better placed?

Physiological and biochemical properties were tested in 45 isolates ofAspergillus egyptiacus (16 isolates),Emericella nidulans (16) andAspergillus versicolor (13). The three fungal species exhibited common and similar features. The big similarity betweenA. egyptiacus andE. nidulans was greater than betweenA. egyptiacus andA. versicolor. It included the inability to produce base either from sodium citrate or lactic acid media, growth at 45 °C (thermophilicity), and production of very similar pigmentations onAspergillus flavus andparasiticus agar.A. egyptiacus is therefore better placed in theAspergillus nidulans-Emericella assemblage.     

Toxicity of a number of pesticides to strains ofTyphlodromus pyri andAmblyseius andersoni (Acari: Phytoseiidae)

Side effects of ten pesticides used in orchards and vineyards were tested with a laboratory method on several Dutch and Italian strains of the predatory mitesTyphlodromus pyri Scheuten andAmblyseius andersoni (Chant). Resistant and susceptible strains of both species were studied. Results showed that a test which evaluates mortality of various developmental stages and fecundity of adult females is better than one that measures only survival of adult females. A definite resistance to certain pesticides was found in ItalianT. pyri andA. andersoni. The level of resistance to parathion, azinphos-methyl and carbaryl was particularly high in some strains ofA. andersoni. The high level of resistance to certain pesticides was often associated with a marked reduction in fecundity.      

Identification of pathogenic fungi in surgical and autopsy specimens by immunofluorescence

Summary A method is presented for the preparation of immune sera and detection by immunofluorescence ofC. immitis, S. schenckii, B. dermatitidis, C. neoformans, andC. albicans in surgical and autopsy material. Formalin fixation does not affect the antigens of the mycotic agents. There are no cross reactions except withC. immitis andC. neoformans, which can be differentiated by the site of the specific fluorescence in each organism.     

Method for distinguishingLimnodrilus hoffmeisteri andLimnodrilus claparedeianus (Oligochaeta,Tubificidae) and its applicability in Lake Suwa

It is still next to impossible to distinguish species using immature worms of theLimnodrilus genus. A method was developed to separate mixed immature populations ofLimnodrilus hoffmeisteri andLimnodrilus claparedeianus into each species. The ratio of setal upper tooth length to lower tooth length, inL. hoffmeisteri andL. claparedeianus, ranged from 0.9 to 1.6 and from 1.3 to 2.1, respectively. Even if the median value of the frequency overlapped, this indistinguishable portion did not exceed 6% of each population.     

A new method for the histochemical demonstration of O-acyl sugars in human colonic epithelial glycoproteins

Summary A new general method has been developed for the specific histochemical identification ofO-acyl sugars in any epithelial glycoprotein. These sugars include hexose, 6-deoxyhexose andN-acetylhexosamine with an ester substituenent(s) located on a potentialvicinal diol(s). In the procedure reported [the periodic acid-borohydride reduction-saponification-selective periodate oxidation-borohydride reduction-periodic acid-Schiff (PA-Bh-KOH-PA-Bh*-Bh-PAS) method] the initial PA-Bh treatment rendersvicinal diols located on either sialic acid or neutral sugars PAS unreactive. In the subsequent steps ester substituents are removed from bothO-acyl sugars andO-acyl sialic acids by saponification (KOH), sialic acidvicinal diols are selectively removed by the PA^*-Bh sequence andO-acyl sugars are stained with the PAS technique. This method has the advantage that the results are obtained with a single section and the results are either positive or negative. Consequently, it is superior to the three indirect methods investigated because it does not require an observer to compare the intensity or the shade of the staining obtained with serial sections.Using the PA-Bh-KOH-PA^*-Bh-PAS method we have demonstrated, for the first time, thatO-acyl sugars occur in the epithelial goblet cell glycoproteins of adult human colon. The effect of the presence ofO-acyl sugars on the interpretation of a number of other methods for the histochemical investigation of glycoproteins is discussed. It is recommended that the results obtained with the PA-Bh-KOH-PA^*-Bh-PAS method be evaluated before histochemical procedures for the investigation of neutral sugars andO-acyl sialic acids are selected.     

Experimental transmission and pathogenicity of some zoosporic fungi toTilapia fish

Seventeen fungal species belong to six genera were recovered from the four organs ofTilapia fish and the most common wereSaprolegnia ferax, S. diclina, Achlya dubia, A. americana, A. racemosa andA. flagellata, Dictyuchus sterile,Pythium undulatum andAphanomyces sp. Severe infection followed by death of all fish was incited byS. parasitica andS. ferax through experiment I. 30–70% ofT. nilotica andT. galileae were killed through experiment II byS. parasitica andS. ferax. T. galileae was more susceptible to fungal infection thanT. nilotica.     

A dynamic model of an environmental system withn interacting components andp degrees of freedom

The nonlinear differential equations of growth proposed by Volterra (1931) are used as the basis for a dynamic model of ann-element system withp specified, terminal conditions andp missing initial conditions. The resulting two-point boundary-value problem can be solved, ifp is not large, by the shooting method used previously by Huddleston (1967). A numerical example withn=4 andp=2 is solved on a digital computer, and some results are presented.     

Phylogeny of the GenusTrichodermaBased on Sequence Analysis of the Internal Transcribed Spacer Region 1 of the rDNA Cluster

Sequences of the internal transcribed spacer region 1 (ITS1) of the ribosomal DNA were used to determine the phylogenetic relationships of species ofTrichodermasect.Pachybasium.To this end, 85 strains—including all the availableex-type strains—were analyzed. Parsimony analysis demonstrated that the section is nonmonophyletic, distributing the 85 strains among three main groups that were supported by bootstrap values. Group A comprises two clades (A1 and A2), with A1 includingT. polysporum, T. piluliferum,andT. minutisporum,while A2 includedT. hamatum, T. pubescens,andT. strigosumin addition to species previously included in sect.Trichoderma(i.e.,T. viride, T. atroviride,andT. koningii). Theex-type strain ofT. fasciculatumformed a separate branch basal to clade A. Clade B contained the sect.PachybasiummembersT. harzianum, T. fertile, T. croceum, T. longipile, T. strictipile, T. tomentosum, T. oblongisporum, T. flavofuscum, T. spirale,and the anamorphs ofHypocrea semiorbisandH. cf. gelatinosa.Sequence differences among clades A1, A2, and B were in the same order of magnitude as between each of them andT. longibrachiatum,which was used as an outgroup in these analyses. Sequence differences within clades A1, A2, and B were considerably smaller: in some cases (i.e.,T. virensandT. flavofuscum; T. strictipileandH. cf. gelatinosa), the ITS1-sequences were identical, suggesting conspecifity. In other cases (e.g.,T. crassumandT. longipile; T. harzianum, T. inhamatum, T. croceum, T. fertile,andH. semiorbis; T. hamatumandT. pubescens;andT. viride, T. atroviride,andT. koningii) differences were in the range of 1–3 nt only, suggesting a very close phylogenetic relationship. The sequence of a previously described aggressive mushroom competitor group ofT. harzianumstrains (Th2) was strikingly different from that of theex-type strain ofT. harzianumand closely related species and is likely to be a separate species.     

Entwicklungsgeschichte und Ultrastruktur von Pollenkitt und Exine bei nahe verwandten entomophilen und anemophilen Sippen derOleaceae,Scrophulariaceae, Plantaginaceae undAsteraceae

Different members of the Angiosperm familiesOleaceae, Scrophulariaceae, Plantaginaceae andAsteraceae with variable modes of pollination were studied in relation to ultrastructure of pollenkitt and exine. In entomophilous representatives the mostly electron-dense and homogeneous pollenkitt is located on the outside layer of the exine, making the pollen sticky. In related anemophilous taxa the pollenkitt mostly is electrontransparent and inhomogeneous; it becomes inactive by remaining in the loculus or sinking to the bottom of the exine caves; this makes the pollen powdery. According to such criteria,Fraxinus ornus andRhinanthus serotinus are entomophilous, whileFraxinus excelsior is anemophilous. The pollen ofLathraea squamaria eventually becomes powdery and suited for wind dispersal. The broad transition field between entomo- and anemophily is demonstrated byPlantago andArtemisia. Plantago lanceolata is anemophilous, whilePl. serpentina andPl. media are amphiphilous.Artemisia nitida andA. mutellina are entomophilous,A. absinthium, A. pedemontana andA. gabriellae are amphiphilous, but tend towards entomo- (A. absinthium) or anemophily (A. gabriellae), whileA. chamaemelifolia and especiallyA. vulgaris are anemophilous.

     

Rapid detection ofRhizoctonia species, causal agents of rice sheath diseases, by PCR-RFLP analysis using an alkaline DNA extraction method

Restriction fragment length polymorphism (RFLP) analysis for DNA products amplified by the polymerase chain reaction (PCR) was used for the direct detection ofRhizoctonia solani AG 1 IA and AG 2-2 IIIB,R. oryzae, R. oryzae-sativae andR. fumigata from the diseased rice sheaths. A rapid DNA extraction method with a solution of sodium hydroxide was conducted to extract parasite DNA from diseased rice sheaths. 28S ribosomal DNA (rDNA) derived from fungal genomic DNA extracted by the alkaline method was specifically PCR-amplified. The results of PCR-RFLP analysis for DNA samples from artificially inoculated rice sheath tissues with eachRhizoctonia spp. and the corresponding culture on the medium using two restriction enzymes.HhaI andMspI, showed identical polymorphisms. PCR-RFLP analysis using DNA samples from naturally infected rice sheath tissues also revealed the possibility of direct diagnosis ofR. solani AG 1 IA,R. oryzae andR. oryzae-sativae.     

The 5S DNA units ofAcacia species (Mimosaceae)

The DNA sequence structure of 5S DNA units inAcacia species, including representatives from the three subgenera ofAcacia, have been determined. The data was interpreted to suggest that at least three lineages of 5S DNA sequences exist inAcacia and the proposal was made that the lineages be named5S Dna-1, 5S Dna-2, and5S Dna-3. The5S Dna-1 lineage was represented by units fromA. boliviana andA. bidwilli, the5S Dna-2 lineage by units fromA. melanoxylon, A. pycnantha, A. ulicifolia, A. boliviana, A. bidwillii, andA. albida, and the5S Dna-3 lineage by units fromA. bidwillii, A. boliviana, andA. senegal. Based on this interpretation of the sequence data, the Australian species of subg.Phyllodineae grouped together as a cluster, quite separate from the subgeneraAculeiferum andAcacia. As expected from the analyses of morphological characters, the 5S DNA units fromAcacia albida (syn.Faidherbia albida) were quite separate from the otherAcacia spp.     

Phylogenetic relationships of the hyphomycete generaChaetopsina andKionochaeta based on 18S rDNA sequences

A setiform dematiaceous hyphomycete,Kionochaeta spissa was newly collected and isolated from evergreen broad-leaved forests in the southern parts of Japan. Except for its dematiaceous nature, the species is morphologically similar to a nectriaceous hyphomycete,Chaetopsina fulva. The morphology and cultural properties of the Japanese isolates ofK. spissa were described, and the phylogenetic relationships betweenChaetopsina (C. fulva (type species)) andKionochaeta (K. ramifera (type species),K. spissa andK. ivoriensis) were inferred based on nuclear encoded small subunit (18S) rDNA sequences using the neighbor-joining method.Chaetopsina andKionochaeta were found to be phylogenetically related to the Hypocreales and Sordariales, respectively. Both should be maintained as separate genus for phylogenetic classification. The morphological resemblance especially betweenC. fulva andK. spissa is an example of the convergent evolution.