RESUMEN
Liverworts, which are amongst the earliest divergent plant lineages and important ecosystem pioneers, often form nutritional mutualisms with arbuscular mycorrhiza-forming Glomeromycotina and fine-root endophytic Mucoromycotina fungi, both of which coevolved with early land plants. Some liverworts, in common with many later divergent plants, harbour both fungal groups, suggesting these fungi may complementarily improve plant access to different soil nutrients. We tested this hypothesis by growing liverworts in single and dual fungal partnerships under a modern atmosphere and under 1500 ppm [CO2 ], as experienced by early land plants. Access to soil nutrients via fungal partners was investigated with 15 N-labelled algal necromass and 33 P orthophosphate. Photosynthate allocation to fungi was traced using 14 CO2 . Only Mucoromycotina fungal partners provided liverworts with substantial access to algal 15 N, irrespective of atmospheric CO2 concentration. Both symbionts increased 33 P uptake, but Glomeromycotina were often more effective. Dual partnerships showed complementarity of nutrient pool use and greatest photosynthate allocation to symbiotic fungi. We show there are important functional differences between the plant-fungal symbioses tested, providing new insights into the functional biology of Glomeromycotina and Mucoromycotina fungal groups that form symbioses with plants. This may explain the persistence of the two fungal lineages in symbioses across the evolution of land plants.
Asunto(s)
Carbono/metabolismo , Glomeromycota/fisiología , Hepatophyta/microbiología , Mucor/fisiología , Nitrógeno/metabolismo , Fósforo/metabolismo , Plantas/microbiología , Simbiosis , Biomasa , Endófitos/ultraestructura , Glomeromycota/ultraestructura , Modelos Lineales , Mucor/ultraestructura , Micelio/metabolismoRESUMEN
In general, Glomeromycotina was thought to be the earliest fungi forming mycorrhiza-like structure (MLS) in land plant evolution. In contrast, because the earliest divergent lineage of extant land plants, i.e. Haplomitriopsida liverworts, associates only with Mucoromycotina mycobionts, recent studies suggested that those fungi are novel candidates for the earliest mycobionts. Therefore, Mucoromycotina-Haplomitriopsida association currently attracts attention as an ancient mycorrhiza-like association. However, mycobionts were identified in only 7 of 16 Haplomitriopsida species and the mycobionts diversity of this lineage is largely unclarified. To clarify the taxonomic composition of mycobionts in Haplomitriopsida, we observed MLSs in the rhizome of Haplomitrium mnioides (Haplomitriopsida), the Asian representative Haplomitriopsida species, and conducted molecular identification of mycobionts. It was recorded for the first time that Glomeromycotina and Mucoromycotina co-occur in Haplomitriopsida as mycobionts. Significantly, the arbuscule-like branching (ALB) of Glomeromycotina was newly described. As the Mucoromycotina fungi forming MLSs in H. mnioides, Endogonaceae and Densosporaceae were detected, in which size differences of hyphal swelling (HS) were found between the fungal families. This study provides a novel evidence in the MLS of Haplomitriopsida, i.e. the existence of Glomeromycotina association as well as the dominant Mucoromycotina association. In addition, since hyphal characteristics of the HS-type MLS were quite similar to those of fine endophytes (FE) of Endogonales in other bryophytes and vascular plants previously described, this MLS is suggested to be included in FE. These results suggest that Glomeromycotina and Mucoromycotina were acquired concurrently as the mycobionts by the earliest land plants evolved into arbuscular mycorrhizae and FE. Therefore, dual association of Haplomitriopsida, with Endogonales and Glomeromycotina will provide us novel insight on how the earliest land plants adapted to terrestrial habitats with fungi.
Asunto(s)
Endófitos/fisiología , Hongos/fisiología , Hepatophyta/fisiología , Micorrizas/fisiología , Simbiosis , Glomeromycota/fisiología , Hepatophyta/microbiologíaRESUMEN
Arbuscular mycorrhizas are widespread in land plants including liverworts, some of the closest living relatives of the first plants to colonize land 500 million years ago (MYA). Previous investigations reported near-exclusive colonization of liverworts by the most recently evolved arbuscular mycorrhizal fungi, the Glomeraceae, indicating a recent acquisition from flowering plants at odds with the widely held notion that arbuscular mycorrhizal-like associations in liverworts represent the ancestral symbiotic condition in land plants. We performed an analysis of symbiotic fungi in 674 globally collected liverworts using molecular phylogenetics and electron microscopy. Here, we show every order of arbuscular mycorrhizal fungi colonizes early-diverging liverworts, with non-Glomeraceae being at least 10 times more common than in flowering plants. Arbuscular mycorrhizal fungi in liverworts and other ancient plant lineages (hornworts, lycopods, and ferns) were delimited into 58 taxa and 36 singletons, of which at least 43 are novel and specific to liverworts. The discovery that early plant lineages are colonized by early-diverging fungi supports the hypothesis that arbuscular mycorrhizas are an ancestral symbiosis for all land plants.
Asunto(s)
Evolución Biológica , Glomeromycota/fisiología , Hepatophyta/microbiología , Micorrizas/fisiología , Simbiosis , Microscopía por Crioelectrón , Glomeromycota/ultraestructura , Hepatophyta/ultraestructura , Microscopía Electrónica de Rastreo , Micorrizas/ultraestructura , FilogeniaRESUMEN
Background and Aims: The rhizoids of leafy liverworts (Jungermanniales, Marchantiophyta) are commonly colonized by the ascomycete fungus Pezoloma ericae. These associations are hypothesized to be functionally analogous to the ericoid mycorrhizas (ErMs) formed by P. ericae with the roots of Ericaceae plants in terms of bi-directional phosphorus for carbon exchange; however, this remains unproven. Here, we test whether associations between the leafy liverwort Cephalozia bicuspidata and P. ericae are mutualistic. Methods: We measured movement of phosphorus and carbon between C. bicuspidata and P. ericae using [33P]orthophosphate and 14CO2 isotope tracers in monoxenic cultures. We also measured leafy liverwort growth, with and without P. ericae. Key Results: We present the first demonstration of nutritionally mutualistic symbiosis between a non-vascular plant and an ErM-forming fungus, showing transfer of fungal-acquired P to the liverwort and of liverwort-fixed C to the fungus alongside increased growth in fungus-colonized liverworts. Conclusions: Thus, this ascomycete-liverwort symbiosis can now be described as mycorrhiza-like, providing further insights into ericoid mycorrhizal evolution and adding Ascomycota fungi to mycorrhizal fungal groups engaging in mutualisms with plants across the land plant phylogeny. As P. ericae also colonizes the rhizoids of Schistochilaceae liverworts, which originated in the Triassic and are sister to all other jungermannialean liverworts associated with fungi, our findings point toward an early origin of ascomycete-liverwort symbioses, possibly pre-dating their evolution in the Ericales by some 150 million years.
Asunto(s)
Ascomicetos/fisiología , Hepatophyta/microbiología , Simbiosis , Dióxido de Carbono/metabolismo , Hepatophyta/fisiología , Fosfatos/metabolismo , Filogenia , Raíces de Plantas/microbiologíaRESUMEN
The Sebacinales belong to a taxonomically, ecologically, and physiologically diverse group of fungi in the Basidiomycota. While historically recognized as orchid mycorrhizae, recent DNA studies have brought to light both their pandemic distribution and the broad spectrum of mycorrhizal types they form. Indeed, ecological studies using molecular-based methods of detection have found Sebacinales fungi in field specimens of bryophytes (moss), pteridophytes (fern) and all families of herbaceous angiosperms (flowering plants) from temperate, subtropical and tropical regions. These natural host plants include, among others, liverworts, wheat, maize and Arabidopsis thaliana, the model plant traditionally viewed as non-mycorrhizal. The orchid mycorrhizal fungus Sebacina vermifera (MAFF 305830) was first isolated from the Australian orchid Cyrtostylis reniformis. Research performed with this strain clearly indicates its plant growth promoting abilities in a variety of plants, while demonstrating a lack of specificity that rivals or even surpasses that of arbuscular mycorrhizae. Indeed, these traits thus far appear to characterize a majority of strains belonging to the so-called "clade B" within the Sebacinales (recently re-classified as the Serendipitaceae), raising numerous basic research questions regarding plant-microbe signaling and the evolution of mycorrhizal symbioses. Further, given their proven beneficial impact on plant growth and their apparent but cryptic ubiquity, sebacinoid fungi should be considered as a previously hidden, but amenable and effective microbial tool for enhancing plant productivity and stress tolerance.
Asunto(s)
Basidiomycota/fisiología , Productos Agrícolas/microbiología , Raíces de Plantas/microbiología , Simbiosis , Basidiomycota/genética , Basidiomycota/crecimiento & desarrollo , Hepatophyta/microbiología , Micorrizas/fisiología , Orchidaceae/microbiología , Filogenia , Brotes de la Planta/microbiología , Triticum/microbiologíaRESUMEN
Strain R33(T), an endophyte recovered from Herbertus sendtneri, was identified as representing a novel species of the genus Paenibacillus by using a polyphasic taxonomic approach. The novel strain was observed to be a Gram-stain positive, aerobic, rod-shaped, motile and endospore-forming bacterium. The major polar lipids of strain R33(T) were identified as diphosphatidylglycerol, phosphatidylethanolamine, along with lesser amounts of phosphatidylglycerol, three unidentified aminophospholipids, two unidentified phospholipids and two unidentified lipids. The predominant isoprenoid quinone was identified as MK-7. The major fatty acids (>8.0 %) were found to be anteiso-C15:0 (40.0 %), C16:1 ω11c (9.4 %), C16:1 ω7c alcohol (8.5 %) and C16:0 (8.2 %). The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. The G+C content of genomic DNA was determined to be 56.9 mol%. The 16S rRNA gene sequence similarities of strain R33(T) to other Paenibacillus species ranged from 91.6 to 97.2 %, with high similarities to Paenibacillus humicus PC-147(T) and Paenibacillus pasadenensis SAFN-007(T). The phylogenetic analyses based on 16S rRNA gene sequences and the partial rpoB gene confirmed that strain R33(T) belongs to the genus Paenibacillus. However, strain R33(T) shows differential molecular characteristics compared to other related Paenibacillus species based on 16S rDNA-RFLP analyses; the DNA-DNA relatedness values between strain R33(T) and P. humicus PC-147(T), and that between strain R33(T) and P. pasadenensis SAFN-007(T), were 35.0 ± 2.0 and 41.4 ± 0.9 %, respectively. Based on its phenotypic, chemotaxonomic and phylogenetic properties, strain R33(T) is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus herberti is proposed (type strain R33(T) = CGMCC 1.15042(T) = DSM 29849(T)).
Asunto(s)
Endófitos/clasificación , Endófitos/aislamiento & purificación , Hepatophyta/microbiología , Paenibacillus/clasificación , Paenibacillus/aislamiento & purificación , Aerobiosis , Composición de Base , Pared Celular/química , Análisis por Conglomerados , Citosol/química , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , ARN Polimerasas Dirigidas por ADN , Ácido Diaminopimélico/análisis , Endófitos/genética , Endófitos/fisiología , Ácidos Grasos/análisis , Locomoción , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Paenibacillus/genética , Paenibacillus/fisiología , Peptidoglicano/análisis , Fosfolípidos/análisis , Filogenia , Polimorfismo de Longitud del Fragmento de Restricción , Quinonas/análisis , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Esporas Bacterianas/citologíaRESUMEN
Three new metabolites, asperfumigatin (1), isochaetominine (10), and 8'-O-methylasterric acid (21), together with nineteen known compounds, were obtained from the culture of Aspergillus fumigatus, an endophytic fungus from the Chinese liverwort Heteroscyphus tener (Steph.) Schiffn. Their structures were established by extensive analysis of the spectroscopic data. The absolute configurations of 1 and 10 were determined by analysis of their respective CD spectra. Cytotoxicity of these isolates against four human cancer cell lines was also determined.
Asunto(s)
Antineoplásicos/química , Aspergillus fumigatus/química , Aspergillus fumigatus/fisiología , Hepatophyta/microbiología , Antineoplásicos/aislamiento & purificación , Antineoplásicos/farmacología , Línea Celular Tumoral , Humanos , Neoplasias/tratamiento farmacológicoRESUMEN
Arbuscular mycorrhizal fungi (AMF) can host Gram-positive endobacteria (BLOs) in their cytoplasm. These have been identified as Mollicutes-related microbes based on an inventory of AMF spores from fungal collections. Bacteria-like organisms (BLOs) of unknown identity have also been reported in the cytoplasm of AMF associated with liverworts, the earliest-diverged extant lineage of land plants. A combination of morphological, molecular and phylogenetic analyses revealed that three samples of two liverwort species (Conocephalum conicum and Lunularia cruciata) growing spontaneously in a botanical garden harboured AMF belonging to Glomerales, and these, in turn, hosted coccoid BLOs. 16S rDNA sequences from these BLOs clustered with the Mollicutes sequences identified from the spore collections but revealed the presence of novel phylotypes. Electron microscopy and fluorescence in situ hybridization (FISH) confirmed the presence of BLOs inside the cytoplasm of AMF hyphae colonizing the liverwort thalli. The high genetic variability of BLOs in liverwort-AMF associations thriving in the same ecological niche raises questions about the mechanisms underlying such diversity.
Asunto(s)
Glomeromycota/fisiología , Hepatophyta/microbiología , Micorrizas/fisiología , Tenericutes/fisiología , Glomeromycota/clasificación , Glomeromycota/genética , Glomeromycota/ultraestructura , Hepatophyta/ultraestructura , Hifa/ultraestructura , Hibridación Fluorescente in Situ , Microscopía Electrónica de Transmisión , Datos de Secuencia Molecular , Micorrizas/clasificación , Micorrizas/genética , Micorrizas/ultraestructura , Filogenia , ARN Ribosómico 16S/genética , ARN Ribosómico 18S/genética , Tenericutes/clasificación , Tenericutes/genéticaRESUMEN
We evaluated the antibacterial activity and cytotoxicity of endophytes isolated from Scapania verrucosa Heeg., which belongs to the liverwort class. A total of 49 endophytic fungi were isolated from S. verrucosa and classified into seven genera and one family in our previous study. In this study, the cytotoxic activity of the endophytes was assessed using the brine shrimp lethality bioassay, seven of which showed potent toxicity against the brine shrimp with 50% lethal concentration values less than 20 µg/mL. T-30 was the most toxic, with a 50% lethal concentration value of 7.15 µg/mL. Moreover, T-27 exhibited the strongest antibacterial activity against Staphylococcus aureus, with minimal inhibitory concentrations below 0.25 and 4 mg/mL, which can inhibit the growth of two standard strains - ATCC 25923 (methicillin-sensitive S. aureus) and ATCC 43300 (methicillin-resistant S. aureus) - in a time-dependent manner, respectively. These results suggest that endophytes in S. verrucosa are the sources for the production of natural bioactive products and thus warrant further investigation.
Asunto(s)
Antibiosis/fisiología , Endófitos/fisiología , Hongos/fisiología , Hepatophyta/microbiología , Animales , Artemia/microbiología , Staphylococcus aureus/crecimiento & desarrolloRESUMEN
Bioactivity-guided fractionation of the cytotoxic extract of Aspergillus niger, an endophytic fungus from the Chinese liverwort Heteroscyphus tener (Steph.) Schiffn., afforded five new naphtho-γ-pyrones, rubrofusarin-6-O-α-D-ribofuranoside (1), (R)-10-(3-succinimidyl)-TMC-256A1 (2), asperpyrone E (3), isoaurasperone A (4), and isoaurasperone F (5), as well as four known ones, dianhydroaurasperone C (6), aurasperone D (7), asperpyrone D (8), and asperpyrone A (9), together with a cytotoxic cyclic pentapeptide, malformin A1 (10). Their structures were determined by extensive spectroscopic analysis. The absolute configurations of dimeric naphtho-γ-pyrones 3-9 were also determined by analysis of their respective CD spectra.
Asunto(s)
Aspergillus niger/química , Hepatophyta/microbiología , Naftalenos/química , Pironas/química , Aspergillus niger/aislamiento & purificación , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Dicroismo Circular , Humanos , Células K562 , Espectroscopía de Resonancia Magnética , Conformación Molecular , Pironas/aislamiento & purificación , Pironas/toxicidadRESUMEN
Arabinogalactan proteins (AGP) are a diverse family of proteoglycans associated with the cell surfaces of plants. AGP have been implicated in a wide variety of plant cell processes, including signaling in symbioses. This study investigates the existence of putative AGP in free-living cyanobacterial cultures of the nitrogen-fixing, filamentous cyanobacteria Nostoc punctiforme and Nostoc sp. strain LBG1 and at the symbiotic interface in the symbioses between Nostoc spp. and two host plants, the angiosperm Gunnera manicata (in which the cyanobacterium is intracellular) and the liverwort Blasia pusilla (in which the cyanobacterium is extracellular). Enzyme-linked immunosorbent assay, immunoblotting, and immunofluorescence analyses demonstrated that three AGP glycan epitopes (recognized by monoclonal antibodies LM14, MAC207, and LM2) are present in free-living Nostoc cyanobacterial species. The same three AGP glycan epitopes are present at the Gunnera-Nostoc symbiotic interface and the LM2 epitope is detected during the establishment of the Blasia-Nostoc symbiosis. Bioinformatic analysis of the N. punctiforme genome identified five putative AGP core proteins that are representative of AGP classes found in plants. These results suggest a possible involvement of AGP in cyanobacterial-plant symbioses and are also suggestive of a cyanobacterial origin of AGP.
Asunto(s)
Regulación Bacteriana de la Expresión Génica/fisiología , Hepatophyta/microbiología , Magnoliopsida/microbiología , Mucoproteínas/metabolismo , Nostoc/metabolismo , Simbiosis/fisiología , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Simulación por Computador , Epítopos , Técnica del Anticuerpo Fluorescente Indirecta , Regulación de la Expresión Génica de las Plantas/fisiología , Hepatophyta/metabolismo , Magnoliopsida/metabolismo , Modelos Biológicos , Mucoproteínas/genética , Nostoc/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
In the present study, we obtained in vitro dual cultures between the liverwort Plagiochasma rupestre and two arbuscular mycorrhizal (AM) fungi: Glomus intraradices and Glomus clarum. Four agarized culture media were tested for optimal growth of P. rupestre. Also, a description of the symbiotic association is provided. Plagiochasma rupestre gametophytes profusely grew axenically in MM with sucrose, and thalli were successfully subcultured under these growth conditions. Arbuscular mycorrhizal fungal hyphae colonized P. rupestre thalli through rhizoids or by forming appresoria in the ventral thallus cells. Arbuscules, mycelia and structures resembling intrathallic spores or vesicles were developed in the internal parenchymatic cells. The pattern of AM colonization in P. rupestre was very similar to the Paris-type. After 100 days of dual culture, the external mycelia of both AM fungal strains formed thousands of small viable spores, suggesting that P. rupestre in vitro culture could be a valuable tool for studying the biology of both symbiotic partners and conserving AM fungi in in vitro germplasm collections.
Asunto(s)
Glomeromycota/fisiología , Hepatophyta/microbiología , Hepatophyta/fisiología , Micorrizas/fisiología , Simbiosis , Medios de Cultivo/química , Glomeromycota/crecimiento & desarrollo , Hepatophyta/crecimiento & desarrollo , Micelio/citología , Micelio/crecimiento & desarrollo , Micorrizas/crecimiento & desarrollo , Esporas Fúngicas/citología , Esporas Fúngicas/crecimiento & desarrolloRESUMEN
A pink-pigmented, facultatively methylotrophic bacterium, designated strain JT1(T), was isolated from a thallus of the liverwort Marchantia polymorpha L. and was analysed by using a polyphasic approach. Comparative 16S rRNA gene sequence analysis placed the strain in a clade with Methylobacterium adhaesivum AR27(T), Methylobacterium fujisawaense DSM 5686(T), Methylobacterium radiotolerans JCM 2831(T) and Methylobacterium jeotgali S2R03-9(T), with which it showed sequence similarities of 97.8, 97.7, 97.2 and 97.4â%, respectively. However, levels of DNA-DNA relatedness between strain JT1(T) and these and the type strains of other closely related species were lower than 70â%. Cells of JT1(T) stained Gram-negative and were motile, rod-shaped and characterized by numerous fimbriae-like appendages on the outer surface of their wall (density up to 200 µm(-2)). Major fatty acids were C(18â:â1)ω7c and C(16â:â0). Based on the morphological, physiological and biochemical data presented, strain JT1(T) is considered to represent a novel species of the genus Methylobacterium, for which the name Methylobacterium marchantiae sp. nov. is proposed. The type strain is JT1(T) (â=âDSM 21328(T) â=âCCUG 56108(T)).
Asunto(s)
Hepatophyta/microbiología , Methylobacterium/clasificación , Methylobacterium/aislamiento & purificación , Técnicas de Tipificación Bacteriana , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácidos Grasos/análisis , Fimbrias Bacterianas/fisiología , Locomoción , Methylobacterium/genética , Methylobacterium/fisiología , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Filogenia , Pigmentos Biológicos/metabolismo , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
Previous studies have shown the leafy liverwort Cephaloziella varians to associate consistently with fungi, typically the ericoid mycorrhizal symbiont Rhizoscyphus ericae, across a wide latitudinal gradient in the maritime and sub-Antarctic. Hitherto, however, there are no quantitative data on the intensity of colonisation of C. varians by fungal structures in the natural environment and how colonisation might vary with changing environmental conditions. A study is hence reported showing that the frequency of colonisation by fungal structures of C. varians alters along a latitudinal transect from South Georgia (54° S, 38° W) to Moutonnée Valley on Alexander Island (71° S, 68° W). The percentage of stem length colonised by dark septate (DS) hyphae increased significantly along the transect, from 30% at South Georgia to 97% at Moutonnée Valley. In contrast, the percentage of stem length colonised by hyaline hyphae decreased significantly, from 85% at South Georgia to 13% at Moutonnée Valley, and that colonised by hyphal coils similarly decreased from 71% at the former location to 15% at the latter. The frequencies of DS hyphae were negatively associated with mean annual and seasonal air temperatures, whereas those of hyaline septate hyphae and hyphal coils were positively associated with air temperatures. Coils at northerly locations were more convoluted than those at southerly locations. The data indicate that hyphal coils, usually associated with nutrient exchange between partners in ericoid mycorrhizas, do form in C. varians tissues in the maritime and sub-Antarctic, but that the frequency of these structures diminishes in colder habitats.
Asunto(s)
Ascomicetos/crecimiento & desarrollo , Ecosistema , Hepatophyta/microbiología , Micorrizas/crecimiento & desarrollo , Regiones Antárticas , Ascomicetos/aislamiento & purificación , Ascomicetos/fisiología , Micorrizas/aislamiento & purificación , Micorrizas/fisiologíaRESUMEN
We investigated in vitro antioxidant activities of 49 endophytic fungi isolated from the liverwort Scapania verrucosa. Based on morphological and molecular identification, the endophytic fungi isolated were classified into seven genera (Hypocrea, Penicillium, Tolypocladium, Chaetomium, Xylaria, Nemania, and Creosphaeria), all belonging to one family (Xylariaceae). By screening with the 2,2'-azino-di(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) decolorization assay, the ethyl acetate extracts of five endophytic fungi (T7, T21, T24, T32, and T38 strains), which exhibited remarkable Trolox equivalent (TE) antioxidant capacity (ranging from 997.06 to 1248.10 µmol TE/g extract), were selected and their antioxidant capacity was further evaluated by assays for 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, hydroxyl radical scavenging, reducing power, and ferrous ion chelating. The ethyl acetate extracts of two endophytic fungi (T24 and T38) were found to have comparable scavenging abilities on both DPPH-free radicals (93.9 and 88.7%, respectively, at 50 µg/mL) and hydroxyl radicals (97.1 and 89.4%, respectively, at 2 mg/mL) when compared with those of the positive controls (ascorbic acid and butylated hydroxytoluene, respectively). Although their reducing powers were similar to that of butylated hydroxytoluene, as indicated by absorbance (0.35 and 0.30 at 50 µg/mL, respectively), only the T38 strain's ethyl acetate extract showed ferrous ion chelating ability (92.9% at 1 mg/mL) comparable to that of the EDTA-2Na control. These endophytic fungi in S. verrucosa are a potential novel source of natural antioxidants.
Asunto(s)
Antioxidantes/farmacología , Endófitos/química , Depuradores de Radicales Libres/farmacología , Hongos/química , Genes Fúngicos , Hepatophyta/microbiología , Acetatos , Antioxidantes/aislamiento & purificación , Benzotiazoles/química , Benzotiazoles/metabolismo , Compuestos de Bifenilo/antagonistas & inhibidores , China , Cromanos/farmacología , Mezclas Complejas/aislamiento & purificación , Mezclas Complejas/farmacología , Endófitos/clasificación , Endófitos/aislamiento & purificación , Depuradores de Radicales Libres/aislamiento & purificación , Radicales Libres/antagonistas & inhibidores , Hongos/clasificación , Hongos/aislamiento & purificación , Genes de ARNr , Radical Hidroxilo/antagonistas & inhibidores , Quelantes del Hierro/aislamiento & purificación , Quelantes del Hierro/farmacología , Oxidación-Reducción , Picratos/antagonistas & inhibidores , Solventes , Ácidos Sulfónicos/química , Ácidos Sulfónicos/metabolismoRESUMEN
Liverworts, the most ancient group of land plants, form a range of intimate associations with fungi that may be analogous to the mycorrhizas of vascular plants. Most thalloid liverworts contain arbuscular mycorrhizal glomeromycete fungi similar to most vascular plants. In contrast, a range of leafy liverwort genera and one simple thalloid liverwort family (the Aneuraceae) have switched to basidiomycete fungi. These liverwort switches away from glomeromycete fungi may be expected to parallel switches undergone by vascular plants that target diverse lineages of basidiomycete fungi to form ectomycorrhizas. To test this hypothesis, we used a cultivation-independent approach to examine the basidiomycete fungi associated with liverworts in varied worldwide locations by generating fungal DNA sequence data from over 200 field collections of over 30 species. Here we show that eight leafy liverwort genera predominantly and consistently associate with members of the Sebacina vermifera species complex and that Aneuraceae thalloid liverworts associate nearly exclusively with Tulasnella species. Furthermore, within sites where multiple liverwort species co-occur, they almost never share the same fungi. Our analyses reveal a strikingly conservative ecological and evolutionary pattern of liverwort symbioses with basidiomycete fungi that is unlike that of vascular plant mycorrhizas.
Asunto(s)
Basidiomycota , Ecosistema , Evolución Molecular , Hepatophyta , Micorrizas , Simbiosis/genética , Basidiomycota/clasificación , Basidiomycota/genética , Basidiomycota/crecimiento & desarrollo , Basidiomycota/ultraestructura , Europa (Continente) , Hepatophyta/clasificación , Hepatophyta/genética , Hepatophyta/crecimiento & desarrollo , Hepatophyta/microbiología , Microscopía Electrónica de Rastreo , Micorrizas/crecimiento & desarrollo , Filogenia , Raíces de Plantas/microbiología , Especificidad de la Especie , Simbiosis/fisiologíaRESUMEN
Aneura pinguis (Aneuraceae) is a cosmopolitan thalloid liverwort that shows a specific mycorrhiza-like interaction with basidiomycetes. To date, tropical specimens have not been studied in great depth. Samples of A. pinguis were collected from 48 individuals in one plot in South Ecuador and 54 individuals in five European countries. Light and transmission electron microscopy and molecular analyses based on nuclear rDNA coding for the ribosomal large subunit (nucLSU) and from the 5.8s-ITS2 regions were carried out to identify the associated mycobionts and to study their phylogenetic relationships. Microscopic and ultrastructural investigations of the fungal colonisation showed a high congruence between the European and the Ecuadorian sites and confirmed previous results. Tulasnellales are the only mycobionts that could be detected from ultrastructural characters with certainty. Molecular phylogenetic analysis indicated the presence of tulasnelloid fungi from at least 13 distinct clades. The composition of the communities of tulasnelloid fungi in A. pinguis differs between Ecuador and Europe. The diversity of tulasnelloid fungal partners was much higher at the Ecuadorian site.
Asunto(s)
Basidiomycota/clasificación , Basidiomycota/aislamiento & purificación , Biodiversidad , Hepatophyta/microbiología , Micorrizas/crecimiento & desarrollo , Basidiomycota/citología , Basidiomycota/genética , ADN de Hongos/química , ADN de Hongos/genética , ADN Ribosómico/química , ADN Ribosómico/genética , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Ecuador , Europa (Continente) , Microscopía , Microscopía Electrónica de Transmisión , Datos de Secuencia Molecular , ARN Ribosómico 5.8S/genética , Análisis de Secuencia de ADNRESUMEN
The leafy liverwort Lophozia excisa, which is colonised by basidiomycete fungi in other biomes and which evidence suggests may be colonised by mycorrhizal fungi in Antarctica, was sampled from Léonie Island in the southern maritime Antarctic (67 degrees 36' S, 68 degrees 21' W). Microscopic examination of plants indicated that fungal hyphae colonised 78% of the rhizoids of the liverwort, apparently by entering the tips of rhizoids prior to growing into their bases, where they formed hyphal coils. Extensive colonisation of stem medullary cells by hyphae was also observed. DNA was extracted from surface-sterilised liverwort tissues and sequenced following nested PCR, using the primer set ITS1F/TW14, followed by a second round of amplification using the ITSSeb3/TW13 primer set. Neighbour-joining analyses showed that the sequences obtained nested in Sebacinales clade B as a 100% supported sister group to Sebacinales sequences from the leafy liverworts Lophozia sudetica, L. incisa and Calypogeia muelleriana sampled from Europe. Direct PCR using the fungal specific primer set ITS1F/ITS4 similarly identified fungi belonging to Sebacinales clade B as the principal colonists of L. excisa tissues. These observations indicate the presence of a second mycothallus in Antarctica and support the previous suggestion that the Sebacinales has a wide geographical distribution.
Asunto(s)
Basidiomycota/crecimiento & desarrollo , Basidiomycota/aislamiento & purificación , Hepatophyta/crecimiento & desarrollo , Hepatophyta/microbiología , Regiones Antárticas , ADN de Hongos/química , ADN de Hongos/genética , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Hifa/crecimiento & desarrollo , Microscopía , Datos de Secuencia Molecular , Filogenia , Tallos de la Planta/microbiología , Análisis de Secuencia de ADNRESUMEN
The filamentous cyanobacterium Nostoc punctiforme forms symbioses with plants. Disruption of the catalytic domain of the N. punctiforme adenylate cyclase (CyaC) significantly increased symbiotic competence, whereas reduced infectivity was observed in a mutant with a disruption close to the N terminus of CyaC. The total cellular cyclic AMP levels were significantly reduced in both mutants.
Asunto(s)
Adenilil Ciclasas/genética , Proteínas Bacterianas/genética , Hepatophyta/microbiología , Mutación , Nostoc/genética , Adenilil Ciclasas/metabolismo , Proteínas Bacterianas/metabolismo , AMP Cíclico/metabolismo , Modelos Genéticos , Nostoc/enzimología , Nostoc/patogenicidad , Enfermedades de las Plantas/microbiología , Virulencia/genéticaRESUMEN
Two species of thalloid liverworts, Blasia pusilla and Cavicularia densa, form stable symbioses with nitrogen-fixing cyanobacteria. Both bryophytes promote the persistence of their cyanobacterial associations by producing specialized gemmae, which facilitate the simultaneous dispersal of the host and its nitrogen-fixing symbionts. Here the genetic diversity of cyanobacterial symbionts of Blasia and Cavicularia is examined. The results indicate that the primary symbionts of both bryophytes are closely related and belong to a specific group of symbiotic Nostoc strains. Related strains have previously been reported from hornworts and cycads, and from many terricolous cyanolichens. The evolutionary origins of all these symbioses may trace back to pre-Permian times. While the laboratory strain Nostoc punctiforme PCC 73102 has been widely used in experimental studies of bryophyte-Nostoc associations, sequence-identical cyanobionts have not yet been identified from thalloid liverworts in the field.