Karin Jacobs

Leptographium wingfieldii introduced into North America and found associated with exotic Tomicus piniperda and native bark beetles

Leptographium wingfieldii is a well-known fungal associate of the pine shoot beetle, Tomicus piniperda, in Europe. This fungus is pathogenic to pines and is an important cause of blue-stain in the sapwood of infested trees. Tomicus piniperda was first found in a Christmas tree plantation in Ohio, USA, 1992, but isolation of the fungi associated with these intercepted insects was not attempted. Fungal strains resembling L. wingfieldii were recently isolated from pines attacked by T. piniperda, Dendroctonus valens and Ips pini in the northeastern United States. These strains were morphologically similar to the ex-type and other reference strains of L. wingfieldii. Strains were also compared based on sequences of the partial ITS ribosomal DNA operon, beta-tubulin and elongation factor 1-alpha (EF-1alpha) genes. Based on these DNA sequence comparisons, reference strains of European L. wingfieldii were conspecific with North American strains from pines attacked by T. piniperda, D. valens and I. pini. A single strain from Canada, collected in 1993 near the Ontario border with the USA, shortly after the discovery of T. piniperda in that area and tentatively identified as L. wingfieldii, was also included in this study. Its identification was confirmed, suggesting that L. wingfieldii has been present in this region and probably over the whole range of the insects distribution for at least a decade. This represents the first record of L. wingfieldii associated with the introduced and damaging pine shoot beetle T. piniperda in North America. It shows that the fungus is well established and can become associated with other native bark beetles that attack stressed and/or dying trees. The occurrence and spread of this highly pathogenic fungus associated with North American bark beetles should be monitored.

Microbial diversity and community structure in Fynbos soil

The Fynbos biome in South Africa is renowned for its high plant diversity and the conservation of this area is particularly important for the region. This is especially true in the case of endangered vegetation types on the lowlands such as Sand Fynbos, of which only small fragments remain. The question is thus whether the diversity of the above‐ground flora is mirrored in the below‐ground microbial communities. In order to determine the relationship of the above‐ and below‐ground communities, the soil community composition of both fungal and bacterial groups in Sand Fynbos was characterized over space and time. A molecular approach was used based on the isolation of total soil genomic DNA and automated ribosomal intergenic spacer analysis of bacterial and fungal communities. Soil from four different sites was compared to resolve the microbial diversity of eubacterial and fungal groups on a local (alpha diversity) scale as well as a landscape scale (beta diversity). The community structures from different sites were compared and found to exhibit strong spatial patterns which remained stable over time. The plant community data were compared with the fungal and the bacterial communities. We concluded that the microbial communities in the Sand Fynbos are highly diverse and closely linked to the above‐ground floral communities.

Ophiostoma kryptum sp. nov. from Larix decidua and Picea abies in Europe, similar to O. minus

An unknown species of Ophiostoma was isolated from European larch (Larix decidua) infested by Tetropium gabrieli (Coleoptera: Cerambycidae) and Norway spruce (Picea abies) infested by Tetropium sp. in Austria. The fungus is similar to O. minus, but distinguished from it by the ecology, colony morphologies on OA and MEA, and phylogenetic analysis of aligned DNA sequences of the ITS region of the rDNA operon and the partial beta-tubulin gene. It is described here as O. kryptum sp. nov. The new species readily produces perithecia with short necks and reniform ascospores, and has Hyalorhinocladiella and Leptographium-like anamorphs. Circumstantial evidence suggests that Tetropium spp. act as vectors of O. kryptum. O. minus and O. kryptum represent additional examples of morphologically similar, yet genetically and ecologically distinct species in the genus Ophiostoma. The new combination, O. crenulatum comb. nov. (syn. CeratocYstiopsis crenulata), is also made.

A new Leptographium species associated with Tomicus piniperda in south-western China.

Tomicus species (Coleoptera: Scolytidae) are serious pests of pines with a wide distribution in Europe, Asia and America. In Yunnan, south-western China,T. piniperda has destroyed more than 0.5 million ha ofPinus yunnanensis in the past 15 years. A blue stain fungus belonging to the genusLeptographium is associated with both the shoot-feeding and trunk-attacking stages of the beetles life cycle. The fungus is morphologically similar to the anamorph ofOphiostoma crassivaginatum and toL. pyrinum, which are both characterised by short robust conidiophores and hyphae covered by a granular layer. Both these species have been isolated from conifers and are associated with insects. After comparing the fungus fromT. piniperda with similarLeptographium species, using light and scanning electron microscopy, we concluded that it represents a new taxon, which is described here asL. yunnanense sp. nov.

A taxonomic and phylogenetic revision of Penicillium section Aspergilloides.

Species belonging to Penicillium section Aspergilloides have a world-wide distribution with P. glabrum, P. spinulosum and P. thomii the most well-known species of this section. These species occur commonly and can be isolated from many substrates including soil, food, bark and indoor environments. The taxonomy of these species has been investigated several times using various techniques, but species delimitation remains difficult. In the present study, 349 strains belonging to section Aspergilloides were subjected to multilocus molecular phylogenetic analyses using partial β-tubulin (BenA), calmodulin (CaM) and RNA polymerase II second largest subunit (RPB2) sequences. Section Aspergilloides is subdivided into 12 clades and 51 species. Twenty-five species are described here as new and P. yezoense, a species originally described without a Latin diagnosis, is validated. Species belonging to section Aspergilloides are phenotypically similar and most have monoverticillate conidiophores and grow moderately or quickly on agar media. The most important characters to distinguish these species were colony sizes on agar media, growth at 30 °C, ornamentation and shape of conidia, sclerotium production and stipe roughness.

Taxonomic re-evaluation of Leptographium lundbergii based on DNA sequence comparisons and morphology

The genus Leptographium was described in 1927 and currently includes 48 species, with L. lundbergii as the type species. In recent years, the taxonomic status of L. lundbergii has not been uniformly agreed upon and it has been the topic of considerable debate. The problem was compounded by the absence of a type specimen, and the species was epitypified at a later stage. Unfortunately, the whereabouts of the epitype is now unknown. In 1983, Wingfield & Marasas described L. truncatum, which is morphologically similar to L. lundbergii. Based on DNA comparisons and similarities in their morphology, this fungus was reduced to synonymy with L. lundbergii. The loss of the type specimen as well as variation in the morphology of strains identified as L. lundbergii prompted us to re-examine the taxonomic status of this species. A number of strains from various geographic areas were studied. These include a strain of L. lundbergii deposited at CBS by Melin in 1929 (CBS 352.29) as well as the ex-type strain of L. truncatum. The strains were compared based on morphology and comparison of multiple gene sequences. Three genes or genic regions, ITS2 and part of the 28S gene, partial beta-tubulin and partial elongation factor 1-alpha were compared. Strains currently identified as L. lundbergii, represented a complex of species. Strains initially described as L. truncatum clustered separately from other L. lundbergii strains, could be distinguished morphologically and should be treated as a distinct taxon. L. lundbergii is provided with a new and expanded description based on a neotype designated for it. A third group was also identified as separate from the main L. lundbergii clade and had a distinct Hyalorhinocladiella-type anamorph, described here as H. pinicola sp. nov.

Phylogenetic relationships among Phialocephala species and other ascomycetes.

Phialocephala was established for species in the Leptographium complex that produce conidia from phialides at the apices of dark mononematous conidiophores. Some species previously included in Phialocephala were re-allocated to Sporendocladia because they resembled Thielaviopsis in having ring-wall-building conidial development and conidia with two attachment points that emerge in false chains. Despite this significant realignment of the genus, a great deal of morphological heterogeneity remains in Phialocephala. The objective of this study was to consider the heterogeneity among Phialocephala spp. based on comparisons of sequence data derived from the large and small subunits (LSU and SSU) of the rRNA operon of species in Phialocephala. Phialocephala dimorphospora, the type species of the genus, and P. fortinii grouped with genera of the Helotiales in phylogenetic trees generated based on the LSU and SSU datasets. Phialocephala xalapensis and P. fusca clearly are unrelated to Phialocephala sensu stricto and should represent a new genus in the Ophiostomatales. Phialocephala compacta resides with representatives of the Hypocreales, and we believe that it represents a distinct genus. Phialocephala scopiformis and P. repens are not closely related to the other Phialocephala species and group within the Dothideales. The morphological heterogeneity among species of Phialocephala clearly is reflected by phylogenetic analysis of sequence data from two conserved rRNA gene regions. Appropriate genera now need to be found to accommodate these fungi.

Pesotum australi sp. nov. and Ophiostoma quercus associated with Acacia mearnsii trees in Australia and Uganda, respectively

Pesotum accommodates synnematal anamorphs of Ophiostoma spp. with sympodially proliferating conidiogenous cells. These fungi are usually closely associated with wounds on trees and the insects that visit them. During tree disease surveys in Uganda, as well as studies of fungi infecting wounds on Acacia mearnsii trees in Uganda and Australia, many isolates resembling species of Pesotum were collected. The aim of this study was to identify these fungi using both morphological and DNA sequence comparisons. The Pesotum, anamorph of O. quercus was the only species collected from multiple collections in Uganda. Collections from Australia represent a new species of Pesotum described here as P. australi sp. nov.

Characterisation of synnematous bark beetle-associated fungi from China, including Graphium carbonarium sp. nov.

Ophiostomatoid fungi on trees are typically bark beetle associates that cause sapstain in timber and some are pathogens. Very little is known regarding the ophiostomatoid fungi associated with bark beetles in China and the aim of this study was to identify a collection of these fungi with synnematous anamorphs. Micromorphology and DNA sequences of the internal transcribed spacer regions (ITS) of the ribosomal DNA and the partial β-tubulin gene were used for identifications. The isolates could be divided in six morphological groups. DNA sequence comparisons with published data confirmed that these groups represented six species, four in the Ophiostomatales (Sordariomycetidae) and two in the Microascales (Hypocreomycetidae). The majority of these were isolated from conifer hosts. Ophiostoma quercus, O. setosum, Pesotum fragrans (Ophiostomatales) and Graphium pseudormiticum (Microascales) were found on Tsuga dumosa infested by a Pissodes sp. In addition, O. quercus and P. fragrans were found associated with Tomicus yunnanensis on Pinus yunnanensis, P. fragrans with a Pissodes sp. on P. armandi, and O. piceae with Ips subelongatus on Larix olgensis. Only two species, O. quercus and a new species in the Graphium penicilliodes complex, described here as Graphium carbonarium sp. nov., were isolated from Pissodes galleries on Salix babylonica. These results include several new fungus-host and fungus-insect associations, and G. pseudormiticum is reported here for the first time from China.

A new species of Penicillium, P. ramulosum sp. nov., from the natural environment

During a recent survey of Penicillium spp. from fynbos soils in the Western Cape Province of South Africa, several undescribed species were isolated. Similar isolates of one of these species also were collected in the Western Cape from Protea infructescences. These strains were compared morphologically to known species of Penicillium but could not be identified with previously published keys. Morphologically these strains belong to subgenus Biverticillium. They are distinguished by strongly funiculose colonies covered by glutinous exudates and conidiophores with thin acerose phialides (8.5–10[−12] × 2.0–2.5 μm) that give rise to chains of subspheroidal to ellipsoidal conidia (2.5–3.0 × 1.5–2.5 μm). Characteristically short (100–150[−250] μm) determinate synnemata are produced in culture after prolonged incubation with much longer synnemata produced in nature. Based on differences in morphology and molecular characters, the strains are described here as Penicillium ramulosum sp. nov.