“…yamaokae (MAT1-1) and L. gibbsii (MAT1-2) and it will not be possible to attempt to produce sexual structures until strains of opposite mating type have been found. The results of this study explain why these fungi failed to produce sexual states in the study by Duong et al (2012). Goheen and Cobb (1978) reported the discovery of a sexual state in the important conifer root pathogen G. wageneri, which was found in the galleries of Hylastes macer.…”
Section: Discussionmentioning
confidence: 63%
“…This is similar to the situation for various other fungi, thought to be asexual but later shown to be heterothallic and where sexual states have recently been discovered for some of the species (Horn et al, 2009;O'Gorman et al, 2008;Seidl et al, 2009). Duong et al (2012) were able to show that G. alacris is heterothallic by randomly crossing different isolates in all possible combinations. Thus, of the five species in the G. serpens complex , sexual sates have been found only in the case of G. alacris.…”
Section: Discussionmentioning
confidence: 99%
“…Single hyphal tip or single conidium cultures were grown in YM broth (2 % malt extract and 0.2% yeast extract) for 3 to 5 days. Mycelium was harvested by centrifugation and DNA was extracted using PrepMan™ Ultra reagent (Applied Biosystems, California, USA) following the methods described in Duong et al (2012). …”
Section: Cultures Growth Conditions and Dna Extractionmentioning
Species of Leptographium sensu lato (Ophiostomatales, Ascomycetes) are sap-stain fungi vectored by bark beetles (Coleoptera, Scolytinae) and some species cause or are associated with tree diseases. Sexual states have been reported for more than 30 species in this group and these have been treated in the sexual genus Grosmannia. No sexual state is known for at least 59 additional species and these reside in the genus Leptographium. The discovery of sexual states for species of Leptographium relies mainly on the presence of fruiting bodies on host tissue at the time of isolation and/or intensive laboratory mating studies, which commonly have a low levels of success. In this study, markers were developed to diagnose mating-type and to study sexual compatibility of species in Leptographium sensu lato using these markers. To achieve this objective, available mating type sequences for species of Leptographium sensu lato and Ophiostoma were obtained, aligned and used to design primers to amplify MAT genes in Grosmannia and Leptographium species. Using these primers, it was possible to amplify portions of the mating type genes for 42 species and to determine thallism, in many species for the first time. Surprisingly, the results showed that heterothallic and putatively heterothallic species are abundant (39 out of 42 species) in Leptographium sensu lato, and only three species were confirmed to be homothallic. The mating-type markers developed in this study will be useful for future studies concerning mating type and sexual compatibility of species in this genus.
“…yamaokae (MAT1-1) and L. gibbsii (MAT1-2) and it will not be possible to attempt to produce sexual structures until strains of opposite mating type have been found. The results of this study explain why these fungi failed to produce sexual states in the study by Duong et al (2012). Goheen and Cobb (1978) reported the discovery of a sexual state in the important conifer root pathogen G. wageneri, which was found in the galleries of Hylastes macer.…”
Section: Discussionmentioning
confidence: 63%
“…This is similar to the situation for various other fungi, thought to be asexual but later shown to be heterothallic and where sexual states have recently been discovered for some of the species (Horn et al, 2009;O'Gorman et al, 2008;Seidl et al, 2009). Duong et al (2012) were able to show that G. alacris is heterothallic by randomly crossing different isolates in all possible combinations. Thus, of the five species in the G. serpens complex , sexual sates have been found only in the case of G. alacris.…”
Section: Discussionmentioning
confidence: 99%
“…Single hyphal tip or single conidium cultures were grown in YM broth (2 % malt extract and 0.2% yeast extract) for 3 to 5 days. Mycelium was harvested by centrifugation and DNA was extracted using PrepMan™ Ultra reagent (Applied Biosystems, California, USA) following the methods described in Duong et al (2012). …”
Section: Cultures Growth Conditions and Dna Extractionmentioning
Species of Leptographium sensu lato (Ophiostomatales, Ascomycetes) are sap-stain fungi vectored by bark beetles (Coleoptera, Scolytinae) and some species cause or are associated with tree diseases. Sexual states have been reported for more than 30 species in this group and these have been treated in the sexual genus Grosmannia. No sexual state is known for at least 59 additional species and these reside in the genus Leptographium. The discovery of sexual states for species of Leptographium relies mainly on the presence of fruiting bodies on host tissue at the time of isolation and/or intensive laboratory mating studies, which commonly have a low levels of success. In this study, markers were developed to diagnose mating-type and to study sexual compatibility of species in Leptographium sensu lato using these markers. To achieve this objective, available mating type sequences for species of Leptographium sensu lato and Ophiostoma were obtained, aligned and used to design primers to amplify MAT genes in Grosmannia and Leptographium species. Using these primers, it was possible to amplify portions of the mating type genes for 42 species and to determine thallism, in many species for the first time. Surprisingly, the results showed that heterothallic and putatively heterothallic species are abundant (39 out of 42 species) in Leptographium sensu lato, and only three species were confirmed to be homothallic. The mating-type markers developed in this study will be useful for future studies concerning mating type and sexual compatibility of species in this genus.
“…For amplification of part of the beta-tubulin (βT) gene, the primers Bt2a and Bt2b (Glass and Donaldson 1995) were used, which were replaced in some cases with the primer pair Bt2b and T10 (O'Donnell and Cigelnik 1997). PCR amplification of the calmodulin gene region (CAL) was done using the primers CL2F and CL2R (Duong et al 2012) and in cases where amplification was difficult, CL2R2 (Duong et al 2012) was used instead of CL2R. For each marker, PCR reaction components, volumes and conditions followed those described by Musvuugwa et al (2015).…”
Rapanea melanophloeos, an important canopy tree in Afromontane forests, is commonly utilised for medicinal bark harvesting. Wounds created from these activities provide entrance for many fungi, including arthropod-associated members of the Ophiostomatales and Microascales (ophiostomatoid fungi). In this study we assess the diversity of wound-associated Ophiostomatales on storm-damaged R. itsvoense sp. nov., S. rapaneae sp. nov., S. utae) while O. noisomeae groups basal in the Ophiostomatales alongside the S. lignivora complex and Graphilbum. In addition to other taxa known from this host, the present study shows that there is a rich, yet still poorly explored, diversity of Ophiostomatales associated with R. melanophloeos in Afromontane forests. More taxa are likely to be discovered with increased research effort. These must be assessed in terms of pathogenicity towards this ecologically and economically important tree.
“…However, Paciura et al (2010) and Linnakoski et al (2012) showed that the ITS2-LSU and bT gene regions have limitations in distinguishing closely related species in the L. procerum complex. Duong et al (2012) thus sequenced an additional two gene regions, calmodulin (CAL) and actin (ACT) and with sequences for five gene regions, were able to successfully delineate morphologically similar but cryptic species in the neighboring G. serpens complex in Leptographium s. l.…”
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