Recovery of growth of Hyphochytrium catenoides after exposure to environmental stress.

The survival of an isolate of Hyphochytrium catenoides collected from soil in the Blue Mountains in eastern New South Wales, Australia, was tested under extreme conditions in the laboratory. This isolate recovered growth after being subjected to drying on filter paper, to heat while desiccated, to hypersalinity, to strict anaerobic conditions, to freezing temperatures, and to a short period in solutions at pH 2.8-11.2. The capacity to survive under these conditions in the laboratory suggests adaptation to fluctuating conditions in the soil. The partial DNA sequence of the 28S ribosomal RNA gene in the isolate from New South Wales was 98% similar to that in an isolate from Arizona with a similar morphology.

Freeze tolerance of soil chytrids from temperate climates in Australia.

Very little is known about the capacity of soil chytrids to withstand freezing in the field. Tolerance to freezing was tested in 21 chytrids isolated from cropping and undisturbed soils in temperate Australia. Samples of thalli grown on peptone-yeast-glucose (PYG) agar were incubated for seven days at -15 degrees C. Recovery of growth after thawing and transferring to fresh medium at 20 degrees C indicated survival. All isolates in the Blastocladiales and Spizellomycetales survived freezing in all tests. All isolates in the Chytridiales also survived freezing in some tests. None of the isolates in the Rhizophydiales survived freezing in any of the tests. However, some isolates in the Rhizophydiales recovered growth after freezing if they were grown on PYG agar supplemented with either 1% sodium chloride or 1% glycerol prior to freezing. After freezing, the morphology of the thalli of all isolates was observed under LM. In those isolates that recovered growth after transfer to fresh media, mature zoosporangia were observed in the monocentric isolates and resistant sporangia or resting spores in the polycentric isolates. Encysted zoospores in some monocentric isolates also survived freezing. In some of the experiments the freezing and thawing process caused visible structural damage to the thalli. The production of zoospores after freezing and thawing was also used as an indicator of freeze tolerance. The chytrids in this study responded differently to freezing. These data add significantly to our limited knowledge of freeze tolerance in chytrids but leave many questions unanswered.

Novel soil lineages of Archaea are present in semi-arid soils of eastern Australia.

The diversity of Archaea was studied in vertisolic and loam soils of a semi-arid region in Australia. Sampling was undertaken at an agricultural site, two grassland environments, and a brigalow (Acacia harpophylla) woodland. Archaeal community structure was profiled using amplified ribosomal DNA restriction analysis (ARDRA) combined with rDNA sequencing of an example of each restriction fragment length polymorphism type. Sequence comparison and phylogenetic analysis demonstrated that both crenarchaeotal and euryarchaeotal Archaea were present at oxic depths in the soil at all field sites. Along with previously described soil archaeal lineages, novel soil lineages and the deeply divergent Pendant-33 group of Euryarchaeota were also detected. A novel statistical method for comparing ARDRA derived data was demonstrated and implemented using the archaeal communities from the four field sites. Archaeal diversity, as measured by this method, was significantly higher in the agricultural site than at either of the grassland sites or the brigalow woodland.

Molecular diversity of soil basidiomycete communities in northern-central New South Wales, Australia.

Basidiomycete communities were profiled using terminal RFLP (TRFLP) and amplified ribosomal DNA restriction analysis (ARDRA) approaches at seven field sites under differing land use in northern-central New South Wales (NSW), Australia. TRFLP data indicated greater basidiomycete species richness at sites with natural vegetation. Sixty-seven basidiomycete ARDRA-types were detected. Various putatively ectomycorrhizal fungi were detected at all sites with native vegetation. Most ectomycorrhizal taxa had affinities to the genus Tomentella, while two Pisolithus taxa and putatively ectomycorrhizal Cantharellales taxa were also detected. Although soils under woodland or grassland communities supported a range of putatively saprotrophic taxa, only members of the Ceratobasidiales were detected in soils under agricultural land use. This study is the first investigation of fungal communities in soils of northern-central NSW, Australia.

Can soil Chytridiomycota survive and grow in different osmotic potentials?

Twenty isolates from soil in the orders Spizellomycetales, Blastocladiales and Chytridiales (Chytridiomycota) grew on complex solid media supplemented with 10 gl(-1) sodium chloride. In a synthetic liquid medium, 4.4 gl(-1) sodium chloride strongly inhibited growth in three of the five isolates, possibly because of the effect of the ions or osmolarity of the solution. The maximum concentration for growth in synthetic liquid medium with different osmotic potentials using polyethylene glycol (PEG) varied considerably amongst the isolates. Three patterns of growth with increasing concentrations of PEG were evident among isolates within the genus Rhizophydium. Up to the concentration where growth ceased, the dry weight of each isolate either decreased, remained constant, or in one case, increased. Most of the fungi survived when incubated at room temperature for 7d in complex liquid media supplemented with 35 gl(-1) sodium chloride or 300 gl(-1) PEG. These data indicate that soil Chytridiomycota can survive various osmotic potentials that may occur during the wetting and drying phases in soils.

Access to organic and insoluble sources of phosphorus varies among soil Chytridiomycota.

The sources of minerals accessed by fungi in the Chytridiomycota (chytrid) in soil are largely unknown. The ability of ten species of soil chytrids to use various sources of phosphorus was examined in vitro. While all grew on orthophosphate, fifty per cent of isolates grew on phytic acid, and one isolate grew on DNA as the sole source of phosphorus. All isolates solubilised and utilised CaHPO(4). Most isolates utilised hydroxyapatite when NH (4) (+) was the nitrogen source. When ammonium was omitted, 50% of isolates solubilised hydroxyapatite. Many soil chytrids may utilise phosphomonoesters as the sole source of phosphorus, and access to DNA appears limited. We suggest that the capacity to use different sources of phosphorus may influence the diversity of chytrids found in Australian soils.

Trichocomaceae in bark survive high temperatures and fire.

Fifty-six species in the Trichocomaceae were recovered from bark of trees and shrubs from hot arid and temperate regions, and following one fire in a temperate region of Australia. Fungi were recovered from dry bark after incubation for up to 1 h at up to 105 degrees C. Fourteen species also regenerated on agar after their conidia were heated for 1 h at 105 degrees C. Anamorphic species were commonly recovered and widespread. Teleomorphic species were only recovered after heating the bark. In addition, anamorphic fungi were recovered from one plant species following a natural fire. The results support the view that both anamorphic and teleomorphic fungi may tolerate extreme temperatures in their environment while dry.

Utilisation of carbon substrates by orchid and ericoid mycorrhizal fungi from Australian dry sclerophyll forests.

The utilisation of a range of cell-wall-related and aromatic carbon substrates by multiple genotypes of three ericoid mycorrhizal fungal taxa was compared with two orchid mycorrhizal fungal taxa. Both groups of fungi catabolised most common substrates, though significant inter- and intraspecific variability was observed in the use of a few carbon substrates. Orchid mycorrhizal fungi had limited access to tannic acid as a carbon source and did not use phenylalanine, while the ericoid mycorrhizal fungi used both. Utilisation of tryptophan was limited to single genotypes of each of the orchid mycorrhizal fungi, and to only two of the three ericoid mycorrhizal fungi examined. Although broadly similar, some significant differences apparently exist in carbon catabolism of ericoid and orchid mycorrhizal fungi from the same habitat. Functional and ecological implications of these observations are discussed.

Diversity of the Trichocomaceae in the Katandra Nature Reserve, Central Coast, NSW, Australia.

The diversity of the family Trichocomaceae, which includes the major anamorph genera Aspergillus and Penicillium, was studied in the Katandra Nature Reserve, Central Coast, NSW, Australia. Soil, living leaves, leaf litter and detritus were examined by both direct and dilution plating techniques. Fungi were isolated on dichloran Rose Bengal chloramphenicol agar, and dichloran 18 % glycerol agar, media suitable for cultivation of many species within this family. Species of Trichocomaceae were isolated from all sites and all substrates examined. A high diversity was found, with more than 50 known species identified, and an equal number of undescribed species detected. More species of Penicillium were recovered than other genera, with Aspergillus species the next most common. Most of the species recovered were anamorphs, though 16 known and unknown ascosporic species were also isolated from heated and unheated soil. Soils, leaf litter, a scat from a native herbivore and leaves of living native plants yielded higher diversity than insects, worms or introduced plants. More species belonging to the family were isolated from soil in dry sclerophyll forest than in rainforest. Conversely, native rainforest plants harboured more diversity than the dry sclerophyll forest plants examined.

The growth response of some Chytridiomycota to temperatures commonly observed in the soil.

Chytridiomycota were isolated into pure culture from cool temperate and warm semi-arid soils of eastern Australia. In pure culture these fungi responded variably to the range of temperatures commonly recorded in their environment. All members of the Blastocladiales, Spizellomycetales and Chytridiales grew in culture at temperatures up to 30 degrees C. Some isolates from the Blastocladiales and Spizellomycetales continued to grow at or above 37 degrees. Some isolates of the Chytridiales grew up to but not beyond 35 degrees. All isolates in the Chytridiales were able to resume growth at 20 degrees after brief exposure to temperatures higher than the maximum growth temperature, but were killed by exposure to higher temperatures for 7 d. Because in the natural soil habitat temperature may exceed the maximum for growth it may be a limiting factor that determines the distribution of chytrids in the soil.

Some Chytridiomycota in soil recover from drying and high temperatures.

Rhizophlyctis rosea was found in 44% of 59 soil samples from national parks, urban reserves and gardens, and agricultural lands of eastern New South Wales, Australia. As some of the soils are periodically dry and hot, we examined possible mechanisms that enable survival in stressful environments such as agricultural lands. Air-dried thalli of R. rosea in soil and pure cultures of R. rosea, two isolates of Allomyces anomalus, one isolate of Catenaria sp., one of Catenophlyctis sp. and one of Spizellomyces sp. recovered following incubation at 90 degrees C for two days. Powellomyces sp. recovered following incubation at 80 degrees. Sporangia of all seven fungi shrank during air-drying, and immediately returned to turgidity when rehydrated. Some sporangia of R. rosea released zoospores immediately upon rehydration. These data indicate that some Chytridiomycota have resistant structures that enable survival through periodic drying and high summer temperatures typical of soils used for cropping. Eleven Chytridiomycota isolated from soil did not survive either drying or heat. Neither habitat of the fungus nor morphological type correlated with the capacity to tolerate drying and heat.

Some endophytic fungi reduce the density of pustules of Puccinia recondita f. sp. tritici in wheat.

The interaction between Puccinia recondita f. sp. tritici (now widely referred to as P. triticina) and endophytic fungi in wheat was examined in laboratory experiments to determine whether the presence of fungal endophytes suppresses leaf rust disease caused by this fungus. Endophytes and cell-free washings from culture plates of the endophytes reduced the density and size of pustules in a susceptible cultivar when inoculated 3, 7 and 14 d prior to the pathogen. Disease at 12 d was reduced when the endophytes were inoculated simultaneously up to 50 mm from the fungus. Interactions between endophytes and this Puccinia are most probably mediated by defence mechanisms induced in the host plant.