Alessia Lucia Pepori

Widespread horizontal transfer of the cerato-ulmin gene between Ophiostoma novo-ulmi and Geosmithia species

Previous work had shown that a sequence homologous to the gene encoding class II hydrophobin cerato-ulmin from the fungus Ophiostoma novo-ulmi, the causal agent of Dutch Elm Disease (DED), was present in a strain of the unrelated species Geosmithia species 5 (Ascomycota: Hypocreales) isolated from Ulmus minor affected by DED. As both fungi occupy the same habitat, even if different ecological niches, the occurrence of horizontal gene transfer was proposed. In the present work we have analysed for the presence of the cerato-ulmin gene 70 Geosmithia strains representing 29 species, isolated from different host plants and geographic locations. The gene was found in 52.1 % of the strains derived from elm trees, while none of those isolated from nonelms possessed it. The expression of the gene in Geosmithia was also assessed by real time PCR in different growth conditions (liquid culture, solid culture, elm sawdust, dual culture with O. novo-ulmi), and was found to be extremely low in all conditions tested. On the basis of these results we propose that the cerato-ulmin gene is not functional in Geosmithia, but can be considered instead a marker of more extensive transfers of genetic material as shown in other fungi.

Ecology of invasive forest pathogens

Invasive forest pathogens are a major threat to forests worldwide, causing increasing damage. The knowledge of both the specific traits underlying the capacity of a pathogen to become invasive, and the attributes predisposing an environment to invasion are to be thoroughly understood in order to deal with forest invasions. This paper summarizes the historical knowledge on this subject. Many aspects of the ecological processes underlying alien forest pathogens invasions are still unknown, which raises several scientific issues that need further study. The introduction of invasive forest pathogens to areas where naïve hosts are found, is mainly due to global plant trade. Rapid transportation and reduced delivery times increase the chances of survival of pathogen propagules and of their successful establishment in new environments. In forest pathogens, the reproduction mode seems not to be a crucial determinant of invasiveness, as highly destructive pathogens have a variety of reproductive strategies. The most important drivers of forest pathogen invasions appear to be (a) great adaptability to new environmental conditions; (b) efficient dispersal over long and short distances, possibly assisted by the capacity to form novel associations with endemic and/or alien insect vectors; (c) the ability to exchange genetic material or hybridize with resident or alien species. Moreover, these features interact with some key traits of the invaded environment, e.g. environmental variability and biodiversity richness. Host resistance and natural enemies may occur as a result of rapid selection/adaptation after the epidemic phase of invasion.

Morphological and molecular characterisation of Geosmithia species on European elms.

Species of the genus Geosmithia are associated with insect species, mainly bark beetles. On Ulmus spp., the same beetles are also vectors of Ophiostoma ulmi s.l., the agent of Dutch elm disease (DED), a worldwide elm disease. Aim of this paper is to characterise Geosmithia species associated with elms and/or elm beetles in Europe. Seventy-two strains representative of all morphological taxonomic units were used to build a phylogenetic tree based on ITS, β-tubulin and elongation factor 1-α gene regions. On the basis of molecular and morpho-physiological traits, seven taxonomic entities were identified. In addition to the species previously known our results assigned strains previously identified as Geosmithia pallida to two separate taxa: Geosmithia sp. 2 and Geosmithia sp. 5. Two new species, Geosmithia omnicola and Geosmithia ulmacea, are described. Two strains were assigned to the partially described species Geosmithia sp. 20. Geosmithia species living on Ulmus do not discriminate between elm species, but between different environments. The association between Ulmus and Geosmithia is common, stable, and seems to be related to specific vectors. The relationship between Geosmithia and Ophiostoma would deserve further investigation, as these fungi share the same vectors and habitat for a significant part of their life cycles.

Comparative transcriptional and metabolic responses of Pinus pinea to a native and a non-native Heterobasidion species

Heterobasidion irregulare is a causal agent of root and butt-rot disease in conifers, and is native to North America. In 1944 it was introduced in central Italy in a Pinus pinea stand, where it shares the same niche with the native species Heterobasidion annosum. The introduction of a non-native pathogen may have significant negative effects on a naïve host tree and the ecosystem in which it resides, requiring a better understanding of the system. We compared the spatio-temporal phenotypic, transcriptional and metabolic host responses to inoculation with the two Heterobasidion species in a large experiment with P. pinea seedlings. Differences in length of lesions at the inoculation site (IS), expression of host genes involved in lignin pathway and in cell rescue and defence, and analysis of terpenes at both IS and 12 cm above the IS (distal site, DS), were assessed at 3, 14 and 35 days post inoculation (dpi). Results clearly showed that both species elicit similar physiological and biochemical responses in P. pinea seedlings. The analysis of host transcripts and total terpenes showed differences between inoculation sites and between pathogen and mock inoculated plants. Both pathogen and mock inoculations induced antimicrobial peptide and phenylalanine ammonia-lyase overexpression at IS beginning at 3 dpi; while at DS all the analysed genes, except for peroxidase, were overexpressed at 14 dpi. A significantly higher accumulation of terpenoids was observed at 14 dpi at IS, and at 35 dpi at DS. The terpene blend at IS showed significant variation among treatments and sampling times, while no significant differences were ever observed in DS tissues. Based on our results, H. irregulare does not seem to have competitive advantages over the native species H. annosum in terms of pathogenicity towards P. pinea trees; this may explain why the non-native species has not widely spread over the 73 years since its putative year of introduction into central Italy.

Geosmithia-Ophiostoma: a New Fungus-Fungus Association

In Europe as in North America, elms are devastated by Dutch elm disease (DED), caused by the alien ascomycete Ophiostoma novo-ulmi. Pathogen dispersal and transmission are ensured by local species of bark beetles, which established a novel association with the fungus. Elm bark beetles also transport the Geosmithia fungi genus that is found in scolytids’ galleries colonized by O. novo-ulmi. Widespread horizontal gene transfer between O. novo-ulmi and Geosmithia was recently observed. In order to define the relation between these two fungi in the DED pathosystem, O. novo-ulmi and Geosmithia species from elm, including a GFP-tagged strain, were grown in dual culture and mycelial interactions were observed by light and fluorescence microscopy. Growth and sporulation of O. novo-ulmi in the absence or presence of Geosmithia were compared. The impact of Geosmithia on DED severity was tested in vivo by co-inoculating Geosmithia and O. novo-ulmi in elms. A close and stable relation was observed between the two fungi, which may be classified as mycoparasitism by Geosmithia on O. novo-ulmi. These results prove the existence of a new component in the complex of organisms involved in DED, which might be capable of reducing the disease impact.

Correction to: Geosmithia-Ophiostoma: a New Fungus-Fungus Association

The article Geosmithia-Ophiostoma: a New Fungus-Fungus Association, written by Alessia L. Pepori, Priscilla P. Bettini, Cecilia Comparini, Sabrina Sarrocco, Anna Bonini, Arcangela Frascella, Luisa Ghelardini, & Aniello Scala, Giovanni Vannacci, Alberto Santini.