Symbioses between Geosmithia fungi and wood-boring and bark beetles seldom result in disease induction within the plant host. Yet, exceptions exist such as Geosmithia morbida, the causal agent of Thousand Cankers Disease (TCD) of walnuts and wingnuts, and Geosmithia sp. 41, the causal agent of Foamy Bark Canker disease of oaks. Isolates of G. obscura were recovered from black walnut trees in eastern Tennessee and at least one isolate induced cankers following artificial inoculation. Due to the putative pathogenicity and lack of recovery of G. obscura from natural lesions, a molecular diagnostic screening tool was developed using microsatellite markers mined from the G. obscura genome. A total of 3256 candidate microsatellite markers were identified (2236, 789, 137 di-, tri-, and tetranucleotide motifs, respectively), with 2011, 703, 101 di-, tri-, and tetranucleotide motifs, respectively, containing markers with primers. From these, 75 microsatellite markers were randomly selected, screened, and optimized, resulting in 28 polymorphic markers that yielded single, consistently recovered bands, which were used in downstream analyses. Five of these microsatellite markers were found to be specific to G. obscura and did not cross-amplify into other, closely related species. Although the remaining tested markers could be useful, they cross-amplified within different Geosmithia species, making them not reliable for G. obscura detection. Five novel microsatellite markers (GOBS9, GOBS10, GOBS41, GOBS43, and GOBS50) were developed based on the G. obscura genome. These species-specific microsatellite markers are available as a tool for use in molecular diagnostics and can assist future surveillance studies.

• Keywords
• Bionectriaceae, beetle-fungus symbiosis, cross-amplification, detection, microsatellite markers,
• MeSH
• Coleoptera * microbiology   MeSH
• Hypocreales * genetics   MeSH
• Juglans * microbiology   MeSH
• Microsatellite Repeats genetics   MeSH
• Plant Diseases * microbiology   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Geographicals
• Tennessee MeSH

Fungi of the genus Geosmithia are frequently associated with bark beetles that feed on phloem on various woody hosts. Most studies on Geosmithia were carried out in North and South America and Europe, with only two species being reported from Taiwan, China. This study aimed to investigate the diversity of Geosmithia species in China. Field surveys in Fujian, Guangdong, Guangxi, Hunan, Jiangsu, Jiangxi, Shandong, Shanghai, and Yunnan yielded a total of 178 Geosmithia isolates from 12 beetle species. The isolates were grouped based on morphology. The internal transcribed spacer, β-tubulin, and elongation factor 1-α gene regions of the representatives of each group were sequenced. Phylogenetic trees were constructed based on those sequences. In total, 12 species were identified, with three previously described species (Geosmithia xerotolerans, G. putterillii, and G. pallida) and nine new species which are described in this paper as G. luteobrunnea, G. radiata, G. brevistipitata, G. bombycina, G. granulata (Geosmithia sp. 20), G. subfulva, G. pulverea (G. sp. 3 and Geosmithia sp. 23), G. fusca, and G. pumila sp. nov. The dominant species obtained in this study were G. luteobrunnea and G. pulverea. This study systematically studied the Geosmithia species in China and made an important contribution to filling in the gaps in our understanding of global Geosmithia species diversity.

• Keywords
• 9 new taxa, Geosmithia, bark beetles, fungal community, symbiosis,
• Publication type
• Journal Article MeSH

Geosmithia species (Hypocreales, Ascomycota) are associates of bark beetles and other arthropods. One species, Geosmithia morbida, is a virulent tree pathogen of Juglans nigra. To date, 10 Geosmithia spp. from conifer-infesting, and at least 23 species from hardwood associated bark beetles have been reported from Europe. The aim of this study was to survey Geosmithia spp. associated with 18 bark and ambrosia beetle species in hardwood ecosystems in Poland. In addition, we evaluated the pathogenicity of the six Geosmithia species by inoculating Acer, Fagus, Quercus, Tilia and Ulmus seedlings. Our surveys yielded a total of 1060 isolates from 2915 beetles and 1887 galleries. We identified isolates using morphology and ITS, β-tubulin and TEF1-α sequences. Altogether we identified 11 species including nine previously known and two new species described here as Geosmithia fagi sp. nov. and G. pazoutovae sp. nov. In addition, a sister species G. longistipitata sp. nov., associated with Picea trees, is described here. Bark beetles from hardwoods, with exeption of Dryocoetes alni, D. villosus, Scolytus ratzeburgi and ambrosia beetles, appear to be regular vectors of Geosmithia spp. Like in other parts of the world, most Geosmithia taxa exhibited a distinct level of vector/host specificity. None of Geosmithia isolates induced any disease symptoms under the conditions of our experiment. This study highlights the need for more intensive surveys across additional areas of Central and Northern Europe, insect vectors and host tree species in order to elucidate the Geosmithia species diversity in this region.

• Keywords
• 3 New Taxa, Ambrosia beetle, Bark beetle, Geosmithia, Hardwoods, Pathogenicity,
• MeSH
• Ambrosia MeSH
• Coleoptera * MeSH
• Ecosystem MeSH
• Phylogeny MeSH
• Hypocreales * MeSH
• Plant Bark MeSH
• Weevils * MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Poland MeSH