Mycobiont isolation Dotaz Zobrazit nápovědu
Protoplasts isolated from the mycobiont of a cultured lichen Usnea ghattensis were fused with protoplasts of the fungus Aspergillus nidulans in order to increase the growth rate of the cultured lichen mycobiont in vitro. The maximum protoplast yield (102 x 10(4)/g fresh cell mass) was reached in citrate buffer with 50 mmol/L 2-sulfanylethanol ('2-mercaptoethanol') containing 0.1 % Novozym after 1.5 h at pH 5 and
Seagrasses, a small group of submerged marine macrophytes, were reported to lack mycorrhizae, i.e., the root-fungus symbioses most terrestrial plants use for nutrient uptake. On the other hand, several authors detected fungal endophytes in seagrass leaves, shoots, rhizomes, and roots, and an anatomically and morphologically unique dark septate endophytic (DSE) association has been recently described in the roots of the Mediterranean seagrass Posidonia oceanica. Nevertheless, the global diversity of seagrass mycobionts is not well understood, and it remains unclear what fungus forms the DSE association in P. oceanica roots. We isolated and determined P. oceanica root mycobionts from 11 localities in the northwest Mediterranean Sea with documented presence of the DSE association and compared our results with recent literature. The mycobiont communities were low in diversity (only three species), were dominated by a single yet unreported marine fungal species (ca. 90 % of the total 177 isolates), and lacked common terrestrial and freshwater root mycobionts. Our phylogenetic analysis suggests that the dominating species represents a new monotypic lineage within the recently described Aigialaceae family (Pleosporales, Ascomycota), probably representing a new genus. Most of its examined colonies developed from intracellular microsclerotia occupying host hypodermis and resembling microsclerotia of terrestrial DSE fungi. Biological significance of this hitherto overlooked seagrass root mycobiont remains obscure, but its presence across the NW Mediterranean Sea and apparent root intracellular lifestyle indicate an intriguing symbiotic relationship with the dominant Mediterranean seagrass. Our microscopic observations suggest that it may form the DSE association recently described in P. oceanica roots.
- MeSH
- Alismatales mikrobiologie MeSH
- Ascomycota klasifikace genetika růst a vývoj izolace a purifikace MeSH
- endofyty klasifikace genetika růst a vývoj izolace a purifikace MeSH
- fylogeneze MeSH
- kořeny rostlin mikrobiologie MeSH
- molekulární sekvence - údaje MeSH
- spory hub klasifikace genetika růst a vývoj izolace a purifikace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Středozemní moře MeSH
Despite decades of intensive research (especially from 1970s to 1990s), the ericoid mycorrhizal (ErM) hair root is still largely terra incognita and this simplified guide is intended to revive and promote the study of its mycobiota. Basic theoretical knowledge on the ErM symbiosis is summarized, followed by practical advices on Ericaceae root sample collection and handling, microscopic observations and photo-documentation of root fungal colonization, mycobiont isolation, maintenance and identification and resynthesis experiments with ericoid plants. The necessity of a proper selection of the root material and its surface sterilization prior to mycobiont isolation is stressed, together with the need of including suitable control treatments in inoculation experiments. The culture-dependent approach employing plating of single short (~ 2 mm) hair root segments on nutrient media is substantiated as a useful tool for characterization of Ericaceae root-associated fungal communities; it targets living mycelium and provides metabolically active cultures that can be used in physiological experiments and taxonomic studies, thus providing essential reference material for culture-independent approaches. On the other hand, it is stressed that not every mycobiont isolated from an ericoid hair root necessarily represent an ErM fungus. Likewise, not every intracellular hyphal coil formed in the Ericaceae rhizodermis necessarily represents the ErM symbiosis. Taxonomy of the most important ericoid mycobionts is updated, mutualism in the ErM symbiosis is briefly discussed from the mycobiont perspective, and some interesting lines of possible future research are highlighted.
- MeSH
- Ericaceae * MeSH
- kořeny rostlin MeSH
- mykorhiza * MeSH
- rostliny MeSH
- symbióza MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
The Sebacinales are a monophyletic group of ubiquitous hymenomycetous mycobionts which form ericoid and orchid mycorrhizae, ecto- and ectendomycorrhizae, and nonspecific root endophytic associations with a wide spectrum of plants. However, due to the complete lack of fungal isolates derived from Ericaceae roots, the Sebacinales ericoid mycorrhizal (ErM) potential has not yet been tested experimentally. Here, we report for the first time isolation of a serendipitoid (formerly Sebacinales Group B) mycobiont from Ericaceae which survived in pure culture for several years. This allowed us to test its ability to form ericoid mycorrhizae with an Ericaceae host in vitro, to describe its development and colonization pattern in host roots over time, and to compare its performance with typical ErM fungi and other serendipitoids derived from non-Ericaceae hosts. Out of ten serendipitoid isolates tested, eight intracellularly colonized Vaccinium hair roots, but only the Ericaceae-derived isolate repeatedly formed typical ericoid mycorrhiza morphologically identical to ericoid mycorrhiza commonly found in naturally colonized Ericaceae, but yet different from ericoid mycorrhiza formed in vitro by the prominent ascomycetous ErM fungus Rhizoscyphus ericae. One Orchidaceae-derived isolate repeatedly formed abundant hyaline intracellular microsclerotia morphologically identical to those occasionally found in naturally colonized Ericaceae, and an isolate of Serendipita (= Piriformospora) indica produced abundant intracellular chlamydospores typical of this species. Our results confirm for the first time experimentally that some Sebacinales can form ericoid mycorrhiza, point to their broad endophytic potential in Ericaceae hosts, and suggest possible ericoid mycorrhizal specificity in Serendipitaceae.
Ericoid mycorrhiza is arguably the least investigated mycorrhizal type, particularly when related to the number of potential hosts and the ecosystems they inhabit. Little is known about the global distribution of ericoid mycorrhizal (ErM) fungi, and this holds true even for the prominent ErM mycobiont Rhizoscyphus ericae. Earlier studies suggested R. ericae might be low in abundance or absent in the roots of Southern Hemisphere's Ericaceae, and our previous investigations in two Argentine Patagonian forests supported this view. Here, we revisited the formerly investigated area, albeit at a higher altitude, and screened fungi inhabiting hair roots of Gaultheria caespitosa and Gaultheria pumila at a treeless alpine site using the same methods as previously. We obtained 234 isolates, most of them belonging to Ascomycota. In contrast to previous findings, however, among 37 detected operational taxonomic units (OTUs), OTU 1 (=R. ericae s. str.) comprised the highest number of isolates (87, ∼37 %). Most of the OTUs and isolates belonged to the Helotiales, and 82.5 % of isolates belonged to OTUs shared between both Gaultheria species. At the alpine site, ericoid mycorrhizal fungi dominated, followed by dark septate endophytes and aquatic hyphomycetes probably acting as root endophytes. Our results suggest that the distribution of R. ericae is influenced, among others, by factors related to altitude such as soil type and presence/absence and type of the neighboring vegetation. Our study is the first report on R. ericae colonizing Ericaceae roots in the Southern Hemisphere and extends the known range of this prominent ErM species to NW Patagonia.
- MeSH
- Ericaceae mikrobiologie MeSH
- fylogeneze MeSH
- Glomeromycota klasifikace genetika izolace a purifikace MeSH
- kořeny rostlin mikrobiologie MeSH
- mykorhiza fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Argentina MeSH
This study aimed to elucidate the relationship between ericaceous understorey shrubs and the diversity and abundance of ectomycorrhizal fungi (EcMF) associated with the invasive Pinus strobus and native Pinus sylvestris. Seedlings of both pines were grown in mesocosms and subjected to three treatments simulating different forest microhabitats: (a) grown in isolation and grown with (b) Vaccinium myrtillus or (c) Vaccinium vitis-idaea. Ericaceous plants did not act as a species pool of pine mycobionts and inhibited the ability of the potentially shared species Meliniomyces bicolor to form ectomycorrhizae. Similarly, Ericaceae significantly reduced the formation of Thelephora terrestris ectomycorrhizae in P. sylvestris. EcMF species composition in the mesocosms was strongly affected by both the host species and the presence of an ericaceous neighbour. When grown in isolation, P. strobus root tips were predominantly colonised by Wilcoxina mikolae, whereas those of P. sylvestris were more commonly colonised by Suillus and Rhizopogon spp. Interestingly, these differences were less evident (Suillus + Rhizopogon spp.) or absent (W. mikolae) when the pines were grown with Ericaceae. P. strobus exclusively associated with Rhizopogon salebrosus s.l., suggesting the presence of host specificity at the intrageneric level. Ericaceous plants had a positive effect on colonisation of P. strobus root tips by R. salebrosus s.l. This study demonstrates that the interaction of selective factors such as host species and presence of ericaceous plants may affect the realised niche of the ectomycorrhizal fungi.
- MeSH
- biodiverzita MeSH
- borovice lesní růst a vývoj mikrobiologie fyziologie MeSH
- borovice růst a vývoj mikrobiologie MeSH
- fylogeneze MeSH
- hostitelská specificita MeSH
- houby klasifikace genetika růst a vývoj izolace a purifikace MeSH
- molekulární sekvence - údaje MeSH
- mykorhiza klasifikace genetika růst a vývoj izolace a purifikace MeSH
- semenáček MeSH
- Vaccinium růst a vývoj mikrobiologie MeSH
- zavlečené druhy MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH