This issue of Mycologcal Research News features: Locust and grasshopper control with fungi.

Fifteen papers are included in this part. Nutrient use of mycorrhizal and saprotrophic fungi is compared; Spathulospora is shown by molecular methods to belong to the Sordariomycetes; and novel anamorphs belonging to two Tuber species have been discovered in soil.

Molecular phylogenetic studies around the species level consider: the Ophiostoma minus group; variation in Entoleuca mammata; the placement of Rhynchosporium alismatis in Plectosporium; the identity of Hygrophorus cossus based on sequences from 200 yr old specimens; the differentiation of two special forms in Fusarium oxysporum; and a new crop-associated species of Pythium in North America.

Volatile organic compounds produced by two wood-rotting fungi are characterised and detection methods compared; and a procedure for the more rapid production of gametangia and so the typing of races of Phytophthora ramorum is reported.

The infection processes in the plant pathogenic Helminthosporium solani and the potential biocontrol agent Plectosporium alismatis are documented in detail, and a new rust on Kalanchoe cultivars has been discovered.

The following new scientific names are introduced: Aecidium kalanchoe, and Pythium attrantheridium sp. nov.; and Plectosporium alismatis (syn. Septoria alismatis) comb. nov.

Stable isotopes in fruit bodies from field studies have been used to infer ectomycorrhizal or saprotrophic status and to understand carbon and nitrogen use, but few controlled culture studies have correlated source and fungal isotopic patterns. Here, we measured natural abundances of 15N and 13C in ten strains of ectomycorrhizal fungi and seven strains of saprotrophic fungi grown on agar with three different primary carbon sources: glucose, glucose plus malt extract, and potato dextrose agar. Eight fungal strains were also grown using position-specific, 13C-labelled glucose (C-1 through C-6 labelled). Most fungi resembled nitrogen sources in δ15N, suggesting that growth on agar media minimizes isotopic fractionation on uptake compared to growth on liquid media, and that in general saprotrophic and mycorrhizal fungi process nitrogen similarly. Saprotrophic fungi were more depleted in 13C than ectomycorrhizal fungi on all media, presumably because of assimilation of 13C-depleted, agar-derived carbon. Results on 13C-enriched glucose indicated that saprotrophic fungi obtained up to 45% of their carbon from the agar substrate. Fungi generally incorporated the individual carbon atoms of glucose in the order, C-4<C-1<C-2, C-3, C-5<C-6, ranging from a mean of 9% for the C-4 atom to 21% for the C-6 atom. Based on these incorporation patterns and intramolecular 13C patterns within glucose, differential incorporation of carbon atoms within glucose among fungal taxa contributed less than 1‰ to isotopic differences among taxa, whereas isotopic fractionation among taxa during metabolism varied up to 4‰. Parallel studies of 13C-enriched and natural abundance substrates were crucial to interpreting our results.

The phylogenetic position of the marine ascomycete genus Spathulospora was investigated using partial SSU and LSU DNA sequences obtained from dried herbarium specimens. Spathulospora was represented by the two species S. adelpha and S. antarctica. Phylogenetic analyses using Bayesian, parsimony, and neighbour-joining algorithms on SSU and LSU data sets agreed with the placement of Spathulospora. Both Spathulospora species are each others closest relatives, and group within the Lulworthiales (Sordariomycetes, Ascomycota) with support in all analyses. Members of the morphologically similar insect parasites in the Laboulbeniomycetes are not closely related to Spathulospora. Despite several striking morphological differences between Spathulospora and Lulworthiales, an important shared morphological character was found that until now had not been recognized. Ascospores of Spathulospora and some members of the Lulworthiales have apical chambers containing mucus believed to be involved in ascospore attachment. A closest relative to Spathulospora could not be determined.

This study reports novel terricolous mitosporic fungal morphs nested in the genus Tuber according to molecular phylogenetic analysis. Fungal DNA was amplified directly from field-collected anamorph samples. Nuclear ribosomal DNA (nrDNA) sequences including the ITS regions and the D1 and D2 domains of the LSU identify the anamorphs as mitosporic Tuber borchii and Tuber oligospermum. The link of the novel anamorphs to the genus Tuber is confirmed by the comparative analysis of five collections from four sampling sites.

Ectomycorrhizas with characteristic features of Tuber borchii ectomycorrhizas were found in the soil volume collected with one of the mitosporic T. borchii collections. A nrDNA sequence amplified from these ectomycorrhizae is identical with the corresponding anamorph sequence.

The possible role of the newly discovered anamorphs in the Tuber life-cycle and the potential significance of anamorphs for the propagation of ectomycorrhizal fungi are discussed.

In an attempt to clarify the relationship between fungi classified as Ophiostoma minus, but of different geographic origins and mating systems, sequencing of the 5.8S and ITS 2 rDNA, and β-tubulin gene was carried out. The β-tubulin gene was highly informative, supporting the sub-division of O. minus into two groups based on geographic origin. Furthermore, isolates previously classified as O. pseudotsugae were confirmed as being clearly distinct from O. minus. However, sequencing did not reveal any polymorphisms between isolates with homothallic as compared to heterothallic mating systems. This was supported by crosses using methylbenzamidazole-2-yl carbamate nuclear markers which showed that hybridisation between isolates of different mating systems was possible. However, we propose that different mating systems may still signal a divergence of isolates of O. minus.

Entoleuca mammata (syn. Hypoxylon mammatum) is a damaging pathogen of Populus tremuloides and P. grandidentata in North America and P. tremula in Europe, where the fungus occurs only sporadically in alpine regions and Scandinavia. It has been hypothesized that E. mammata was introduced to Europe from North America. In this study, E. mammata isolates collected from Europe and North America were compared by a sequence analysis of two DNA markers derived from DNA fingerprints. The objective of the study was to elucidate the relationship between North American and European E. mammata populations by testing two hypotheses: (1) North American and European isolates are conspecific; and (2) the fungus was introduced between continents causing both a founder effect and a genetic bottleneck. North American populations were found to be more polymorphic, but no major phylogenetic differences between fungal isolates collected from different continents were found. This result combined with the historical observations of the disease in Europe implies that E. mammata was introduced to Europe several centuries ago.

The phytopathogen Rhynchosporium alismatis, occurring on Alisma, Sagittaria and other genera in the Alismataceae, is under investigation as a mycoherbicide for alismataceous weeds in Australian rice crops. The type species of Rhynchosporium, R. secalis, belongs in the Helotiales while the phylogenetic relationships of R. alismatis were unknown. To identify fungi related to R. alismatis, the internal transcribed spacer (ITS) region of the ribosomal DNA (ITS1, 5.8S rRNA gene, ITS2) of 56 isolates was sequenced and compared to those available in databases. Analysis of ITS sequences revealed close relationships between R. alismatis and the teleomorph genus Plectosphaerella, as well as several anamorphic fungi which were primarily species of Verticillium. Rhynchosporium alismatis and Plectosphaerella cucumerina clustered together with 98% bootstrap support. Morphological comparisons supported this relationship indicating that R. alismatis and the anamorphic genus Plectosporium are congeneric. Rhynchosporium alismatis is transferred to Plectosporium, a name proposed for conservation.

Sowerby described Agaricus cossus in 1799. The fungus possessed a smell, resembling that of a wounded larva of Cossus cossus (Lepidoptera). The species belongs in Hygrophorus, and since more than one white Hygrophorus species has this distinctive smell the epithet cossus has been variously interpreted. The complete internal transcribed spacer (ITS) region of the original type collection made in 1794, preserved in the Royal Botanic Gardens Kew herbarium, was successfully sequenced. Comparison with the ITS sequences from four other white aromatic-acidulous smelling Hygrophorus species, including the type specimen of H. quercetorum, showed that H. cossus is a species associated with Quercus and an older name for H. quercetorum. The differences in basidiome colouration developing with age and host-tree association appear to be the most useful characters to discriminate between the four species with a Cossus cossus smell. A table of morphological and ecological characters is provided.

Two pathogenic special forms (f. sp.) of the Fusarium oxysporum species complex f. sp. lycopersici (Fol) and f. sp. radicis-lycopersici (Forl) are morphologically indistinguishable. Although they are pathogenic to the same host genus Lycopersicon (tomato), and infect the same tomato cultivar, they form distinct diseases; Fol causes wilt and Forl causes crown rot and root rot. These two special forms apparently exist as genetically isolated populations, based on vegetative compatibility and molecular variation at the DNA level. In seeking efficient diagnostic tools for differentiating Fol and Forl isolates, we examined three techniques: isozyme analysis, mitochondrial DNA (mtDNA) RFLP by HaeIII-digestion of total genomic DNA, and an osmotic method using high performance liquid chromatography (HPLC) to detect fungal pigments. The isolates were collected from geographically widespread locations. Distinct HPLC-profile differences were found between an endophytic non-pathogenic isolate and the other pathogenic isolates. However, the direct mtDNA RFLP technique proved to be an efficient diagnostic tool for routine differentiation of Fol and Forl isolates.

Pythium attrantheridium sp. nov. is a new species isolated from cavity spot lesions of carrots as well as apple and cherry seedlings from various locations widely distributed in Canada and the USA. This fungus is closely related to the heterothallic P. intermedium, but is distinguished by: (1) unique molecular characteristics; (2) unique morphological characteristics; and (3) mating incompatibility with P. intermedium. The ITS region of the nuclear rDNA of all strains of P. attrantheridium studied is different from that of all other known Pythium spp. The oogonia attract a large number of antheridia when compatible mating types contact each other. The positive mating type produces zoospores unlike those of P. intermedium. Thus, biological, morphological and molecular data support the recognition of a new species.

Volatile organic compounds (VOCs) emitted by two wood-rotting basidiomycete fungi, Serpula lacrymans (dry rot fungus) and Coniophora puteana (cellar fungus), and the timber of Pinus sylvestris (Scots pine), were identified. Several volatile collection techniques were employed including dichloromethane solvent extraction, solid-phase microextraction (SPME) and thermal desorption of VOCs entrained on Tenax GR. In addition, a new method of solid sample injection (SSI) is described which utilises a low injector temperature and an all-glass deactivated injector liner designed to minimise both the formation of pyrolysis products and analyte degradation. All the volatile compounds collected were analysed using electron impact capillary gas chromatography-mass spectrometry (GC-MS) on HP-5, HP-Innowax and β-cyclodextrin columns. SSI and Tenax thermal desorption were found to be the most effective extraction methods. A total of 19 VOCs were observed from S. lacrymans grown on glass slides and pine, 15 from C. puteana grown on glass slides and 12 from P. sylvestris timber. S. lacrymans was found to emit, in low abundance, six unique VOCs, of which 2-methylbutanal was the greatest. The major volatile compound emitted by S. lacrymans was 1-octen-3-ol, which was also found in lower abundance from C. puteana. Six VOCs, including diethylene glycol and 4-methyl methylbenzoate, were found to be unique to C. puteana, all in medium abundance. From P. sylvestris, the major volatiles identified were S-α-pinene and 3-carene.

Production of the carcinogenic mycotoxins fumonisins has been reported in several Fusarium species, most of which are members of the Gibberella fujikuroi (Gf) complex. In this study, we examined 15 Fusarium species in the Gf complex and 12 other species for fumonisin production and the presence of fumonisin biosynthetic genes (FUM). Among the species within the Gf complex, fumonisin production was detected only in F. fujikuroi, F. globosum, F. proliferatum, F. nygamai, F. oxysporum and F. verticillioides. These five species include members of two of the three major clades delineated in the Gf complex. The FUM genes were detected in these same five species and in F. anthophilum, a member of the third clade. Among the species outside the Gf complex, fumonisin production and FUM genes were detected only in F. oxysporum. Phylogenetic analyses of nucleotide sequences from two FUM gene fragments inferred relationships similar but not identical to those inferred from previous analyses of other genes. The results indicate the FUM genes are discontinuously distributed in the Gf complex and that this distribution gives rise to the differences in the abilities of closely related Fusarium species to produce fumonisins.

Until now gametangia have not been obtained between paired European A1 and American A2 isolates of Phytopthora ramorum in vitro. Their production in artificial culture relies on interspecific pairings. Using P. drechsleri and P. cambivora testers, 51 of 110 P. ramorum isolates from across Europe were all shown to be A1s; while 32 of 38 American isolates from across California and southwest Oregon were shown to be A2s. However, these interspecific pairings are complex, unusually slow and unpredictable. A range of culture media and conditions are described that were tested, unsuccessfully, with a view to enhancing the efficiency of the interspecific pairings. In further tests, gametangia were obtained between A1 and A2 isolates of P. ramorum when juvenile, pre-chlamydospore producing mycelia were mixed together on carrot agar. The gametangia formed in 3–10 d, sparsely to frequently, initially only within the boundaries of the mixed inocula but subsequently in the extended mycelial growth. Chlamydospores were also produced. This inoculum-mixing method, though again sometimes unpredictable, should enhance efficiency of testing for compatibility types and facilitate further studies on whether the sexual outcrossing system of P. ramorum is functional. Differences between sexual reproduction of P. ramorum and that of other heterothallic Phytophthora species are discussed.

Silver scurf is an important postharvest disease affecting potato tubers worldwide, caused by Helminthosporium solani. In the present study, key steps of infection of potato tubers (cv. ‘Dark Red Norland’) by H. solani were described using transmission (TEM) and scanning electron microscopy (SEM). The fungus entered potato tubers mainly via hyphae, although germ tubes were also able to directly penetrate the tubers. An extracellular sheath was observed around hyphae growing over the surface of tubers and the host cell wall appeared lyzed at the point of penetration. Observations suggested that both mechanical and enzymatic processes are involved in periderm penetration. Hyphae of H. solani, 9 h after tuber inoculation, were present intracellularly mostly in the periderm and in some cortical cells. Two days after inoculation, host cells were invaded and both infected and neighbouring host cells showed signs of necrosis (disrupted cytoplasm, absence of typical organelles or endomembrane systems, collapsed peridermal cells) that were not observed in healthy control tubers. Four days after inoculation, completing the infection cycle, conidiophores emerged from peridermal cells directly by erupting through the host cell walls.

In Australia, the endemic fungus Plectosporium alismatis (syn. Rhynchosporium alismatis) has potential use as a mycoherbicide for several species in the Alismataceae, a family of aquatic and semi-aquatic marsh herbs, which are considered to be important weeds in rice crops. Of five species identified in south-eastern Australia where rice is grown, two species, Sagittaria graminea and Sagittaria montevidensis are resistant (non-hosts), and no records of P. alismatis on these species have been reported. To better understand the interactions that lead to resistance in these pathosystems, the infection process of the fungus was studied on these species and also on the host Alisma plantago-aquatica, using light, fluorescent and scanning electron microscopy. On all three species both conidial germination and appressorium formation commenced within 6 h of inoculation with greater than 50% of conidia elongating to form germ tube structures and associated appressoria 12–18 h post inoculation. Germ tube elongation and appressorium formation occurred randomly over the leaf surface. Direct host penetration was facilitated by the production of penetration hyphae that emerged from beneath appressoria. Penetration sites were clearly identified by the presence of spherical holes 0.25–0.5 μm in diam, and were frequently accompanied by resistance reactions in non-host species. Visible symptoms of disease occurred 4–6 d after inoculation of susceptible (host) species.

A rust fungus found on cultivars of Kalanchoe blossfeldiana (Crassulaceae) is described as a new species, Aecidium kalanchoe sp. nov., and compared to the other described rusts on members of the Crassulaceae. Only one other rust is known to parasitize Kalanchoe spp. A DNA sequence of A. kalanchoe suggests that the teleomorph is related to Puccinia.

Mycological Research108 (2): 206–213 (February 2004)

Description and affinities of a new sequestrate fungus, Barcheria willisiana gen. et sp. nov. (Agaricales) from Australia

Teresa LEBEL, Deanne K. THOMPSON and Frank UDOVICIC

On p. 211 in this paper, Fig. 2 was inadvertently printed in black and white and not in full colour as intended. The correct full-colour version is therefore reproduced below. The publishers would like to apologise to the authors and readers for this error.

Mycological Research108 (5): 583–589 (May 2004)

Some Chytridiomycota in soil recover from drying and high temperatures

Frank H. GLEASON, Peter M. LETCHER and Peter A. McGEE

In this paper, on p. 584, left-hand column, line 14 up, ‘24 h’ should be corrected to ‘48 h’. In addition, on p. 586 the legend to Fig. 1 (b) should read ‘other thalli’ and not ‘another thallus’.

The authors and editors regret any confusion overlooking these points in the proof has caused.

Mycological Research108 (5): 533–570 (May 2004).

Taxonomy of the non-yellow species of Rhizocarpon (Rhizocarpaceae) in the Nordic countries, with hyaline and muriform ascospores

Per IHLEN

Owing to a change in the pagination of this paper after the original proofs were returned, the page references in the index to names treated were all two pages out. The correct page numbers are included in the replacement Table presented here