Primer registro de Psilocybe cubensis en un ambiente natural de Chile / First record of Psilocybe cubensis from a natural habitat in Chile

Psilocybe cubensiso también llamado hongo San Isidro, es un basidiomicete de distribución amplia que se encuentra por lo general en zonas de clima tropical y subtropical. Aquí se describe el primer registro de esta especie psicotrópica en condiciones naturales para Chile, desde una zona rural de clima de transición entre mediterráneo y templado. Se discuten los posibles causantes de esta extensión geográfica de la especie.(AU)

Psilocybe cubensis, also known as San Isidro, is a widely distributed basidiomycete, generally found in tropical and subtropical climate zones. Here, we describe the first record of this psychotropic speciesin natural conditions for Chile, from a rural environment in the transition zone between mediterranean and temperate climate. Possible causes for thisgeographic expansion of the species arediscussed.(AU)

Early Effects of Mycorrhizal Fungal Inoculum and Fertilizer on Morphological and Physiological Variables of Nursery-Grown Nothofagus alessandrii Plants.

Nothofagus alessandrii (ruil) is an endangered relict species, endemic to the Mediterranean area of Chile, and one of the most threatened trees in the country. Its natural distribution area has been greatly reduced by the effect of human activities; the remaining fragments are mostly intervened and highly deteriorated as a habitat and refuge for the associated biodiversity. In order to produce healthy and resistant nursery plants for recovery and restoration of N. alessandrii forests, this study evaluates the early effects of mycorrhizal fungal inoculum (MFI) combined with fertilization on the cultivation of seedlings. The experiment was established under a completely randomized design with a factorial arrangement of the mycorrhizal factors (M0 = without mycorrhizal, M1 = Thelephora sp. and M2 = Hebeloma sp.) and fertilization (F1 = standard fertilization and F2 = intensive fertilization), with three replicates of each combination, for each type of plant (P1 = plants from one season and P2 = plants from two seasons). Each experimental unit corresponded to a group of 20 plants, with 720 plants in the test. The results indicate that application of fertilizer and MFI significantly affects some growth and photosynthesis parameters of ruil plants in one and two seasons. The morphological parameters obtained in the study show shoot height values ranging between 67 and 91 cm for P1 and between 96 and 111 cm for P2; while, for shoot diameter, values ranged between 7.91 and 8.24 mm for P1 and between 10.91 and 11.49 mm for P2. Although formation of fully developed mycorrhizal roots was not observed during the assay period, we conclude that inoculation of mycorrhizal fungi combined with fertilization could be an efficient strategy to produce a quality plant, in addition to maintaining a high photosynthetic capacity and, therefore, a higher percentage of survival in the field.

Antarctic Lichens under Long-Term Passive Warming: Species-Specific Photochemical Responses to Desiccation and Heat Shock Treatments.

Climate warming in the Antarctic tundra will affect locally dominant cryptogams. Being adapted to low temperatures and freezing, little is known about the response of the polar lichens' primary photochemistry to warming and desiccation. Since 2008, we have monitored the ecophysiological responses of lichens to the future warming scenario during a long-term warming experiment through open top chambers (OTCs) on Fildes Peninsula. We studied the primary photochemical response (potential Fv/Fm and effective efficiency of photosystem II YPSII) of different lichen taxa and morphotypes under desiccation kinetics and heat shock experiments. As lichens grow slowly, to observe changes during warming we methodologically focused on carbon and nitrogen content as well as on the stable isotope ratios. Endemic Himantormia lugubris showed the strongest effect of long-term warming on primary photochemistry, where PSII activity occurred at a lower %RWC inside the OTCs, in addition to higher Fv/Fm values at 30 °C in the heat shock kinetic treatment. In contrast, Usnea aurantiaco-atra did not show any effect of long-term warming but was active at a thallus RWC lower than 10%. Both Cladonia species were most affected by water stress, with Cladonia aff. gracilis showing no significant differences in primary photochemical responses between the warming and the control but a high sensibility to water deficiency, where, at 60% thallus RWC, the photochemical parameters began to decrease. We detected species-specific responses not only to long-term warming, but also to desiccation. On the other hand, the carbon content did not vary significantly among the species or because of the passive warming treatment. Similarly, the nitrogen content showed non-significant variation; however, the C/N ratio was affected, with the strongest C/N decrease in Cladonia borealis. Our results suggest that Antarctic lichens can tolerate warming and high temperature better than desiccation and that climate change may affect these species if it is associated with a decrease in water availability.

Diversity and Host Relationships of the Mycoparasite Sepedonium (Hypocreales, Ascomycota) in Temperate Central Chile.

We present the first major survey of regional diversity, distribution and host-association of Sepedonium. Whereas the rather scarce worldwide records of this mycoparasitic fungus suggested no specific distribution pattern of most species before, we provide new evidence of endemic and specific host-parasite guilds of Sepedonium in Southern South America, including the description of a new species. The corresponding inventory was performed in temperate central Chile. The regional landscape, a mosaic of exotic timber plantations and remnants of native Nothofagus forests, facilitates a unique combination of endemic and adventitious Boletales hosts. During a two-year survey, 35 Sepedonium strains were isolated and cultured from infected basidiomata of allochthonous Chalciporus piperatus, Paxillus involutus, Rhizopogon spp. and Suillus spp., as well as from the native Boletus loyita, B. loyo, B. putidus and Gastroboletus valdivianus. Taxonomic diagnosis included morphology of conidia and conidiophores, sequences of ITS, RPB2 and EF1 molecular markers and characteristics of in vitro cultures. Phylogenetic reconstructions were performed using Bayesian methods. Four Sepedonium species could be identified and characterized, viz.: S. ampullosporum, S. chrysospermum, S. laevigatum and the newly described species S. loyorum. The most frequent species on introduced Boletales was S. ampullosporum, followed by S. chrysospermum and S. laevigatum. S. loyorum sp. nov. was found exclusively on native boletacean hosts, separated from its closest relative S. chalcipori by micromorphological and molecular attributes. Species descriptions and identification keys are provided. Ecological and biogeographical aspects of endemic and allochthonous symbiotic units consisting of mycoparasite, ectomycorrhizal fungal host and respective mycorrhizal tree are discussed.

Inhibition of Primary Photosynthesis in Desiccating Antarctic Lichens Differing in Their Photobionts, Thallus Morphology, and Spectral Properties.

Five macrolichens of different thallus morphology from Antarctica (King George Island) were used for this ecophysiological study. The effect of thallus desiccation on primary photosynthetic processes was examined. We investigated the lichens' responses to the relative water content (RWC) in their thalli during the transition from a wet (RWC of 100%) to a dry state (RWC of 0%). The slow Kautsky kinetics of chlorophyll fluorescence (ChlF) that was recorded during controlled dehydration (RWC decreased from 100 to 0%) and supplemented with a quenching analysis revealed a polyphasic species-specific response of variable fluorescence. The changes in ChlF at a steady state (Fs), potential and effective quantum yields of photosystem II (FV/FM, ΦPSII), and nonphotochemical quenching (NPQ) reflected a desiccation-induced inhibition of the photosynthetic processes. The dehydration-dependent fall in FV/FM and ΦPSII was species-specific, starting at an RWC range of 22-32%. The critical RWC for ΦPSII was below 5%. The changes indicated the involvement of protective mechanisms in the chloroplastic apparatus of lichen photobionts at RWCs of below 20%. In both the wet and dry states, the spectral reflectance curves (SRC) (wavelength 400-800 nm) and indices (NDVI, PRI) of the studied lichen species were measured. Black Himantormia lugubris showed no difference in the SRCs between wet and dry state. Other lichens showed a higher reflectance in the dry state compared to the wet state. The lichen morphology and anatomy data, together with the ChlF and spectral reflectance data, are discussed in relation to its potential for ecophysiological studies in Antarctic lichens.

HPTLC-DESI-HRMS-Based Profiling of Anthraquinones in Complex Mixtures-A Proof-of-Concept Study Using Crude Extracts of Chilean Mushrooms.

High-performance thin-layer chromatography (HPTLC) coupled with negative ion desorption electrospray ionization high-resolution mass spectrometry (DESI-HRMS) was used for the analysis of anthraquinones in complex crude extracts of Chilean dermocyboid Cortinarii. For this proof-of-concept study, the known anthraquinones emodin, physcion, endocrocin, dermolutein, hypericin, and skyrin were identified by their elemental composition. HRMS also allowed the differentiation of the investigated anthraquinones from accompanying compounds with the same nominal mass in the crude extracts. An investigation of the characteristic fragmentation pattern of skyrin in comparison with a reference compound showed, exemplarily, the feasibility of the method for the determination of these coloring, bioactive and chemotaxonomically important marker compounds. Accordingly, we demonstrate that the coupling of HPTLC with DESI-HRMS represents an advanced and efficient technique for the detection of anthraquinones in complex matrices. This analytical approach may be applied in the field of anthraquinone-containing food and plants such as Rheum spp. (rhubarb), Aloe spp., Morinda spp., Cassia spp. and others. Furthermore, the described method can be suitable for the analysis of anthraquinone-based colorants and dyes, which are used in the food, cosmetic, and pharmaceutical industry.

Nor-guanacastepene pigments from the Chilean mushroom Cortinarius pyromyxa.

For the first time, the pigment composition of basidiocarps from the Chilean mushroom Cortinarius pyromyxa was studied under various aspects like phylogeny, chemistry and antibiotic activity. A molecular biological study supports the monotypic position of C. pyromyxa in subgenus Myxacium, genus Cortinarius. Four undescribed diterpenoids, named pyromyxones A-D, were isolated from fruiting bodies of C. pyromyxa. Their chemical structures were elucidated based on comprehensive one- and two-dimensional NMR spectroscopic analysis, ESI-HRMS measurements, as well as X-ray crystallography. In addition, the absolute configurations of pyromyxones A-D were established with the aid of JH,H, NOESY spectra and quantum chemical CD calculation. The pyromyxones A-D possess the undescribed nor-guanacastane skeleton. Tested pyromyxones A, B, and D exhibit only weak activity against gram-positive Bacillus subtilis and gram-negative Aliivibrio fischeri as well as the phytopathogenic fungi Botrytis cinerea, Septoria tritici and Phytophthora infestans.

Diversity and growth-effects of ectomycorrhizal fungi of a Nothofagus pumilio forest in the Andes of Southern Chile / (Diversidad y efectos de crecimiento de hongos ectomicorrízicos en un bosque de Nothofagus pumilio en los andes del sur de Chile

Chilean temperate rainforests have unique climatic, edaphic and biotic conditions, constituting pre-industrial blueprint ecosystems. Mycorrhizal associations play a central role in the biogeochemical processes of these ecosystems´ functioning. Baseline forest ecology studies are necessary in order to better understand diversity patterns, specifically regarding mycorrhizal symbiosis. Therefore, here we describe the vegetation characteristics and the mycorrhizal relationships of vascular plants in a Nothofagus pumilio forest. We also describe, via morphological methods, the ectomycorrhizal diversity present in this forest. Additionally, we determined whether ectomycorrhizal inoculation confers positive growth effects on N. pumilio seedlings. We found that from 46 vascular plant species identified in this study, 42 (91%) were mycorrhizal and of these 33 (72%) were associated with arbuscular mycorrhizae (AM), two (the dominant trees N. pumilio and N. dombeyi) were forming ectomycorrhizae (EM), five were associated with ericoid mycorrhizae, two with orchid mycorrhizae, and four were nonmycorrhizal. Additionally, 26 EM species were detected of which 15 belong to Cortinarius. Finally, there were clear differences in the growth of N. pumilio seedlings inoculated with the ectomycorrhizal fungus Laccaria laccata compared to noninoculated plants. We suggest that mycorrhizal fungi play a key role in seedling colonization of harsh environments such as the Andean treeline.

Los bosques templados lluviosos de Chile tienen condiciones climáticas, edáficas y bióticas únicas, constituyendo ecosistemas preindustriales modelo. Las asociaciones micorrízicas juegan un rol central en los procesos biogeoquímicos del funcionamiento de estos ecosistemas. Por lo tanto, en este estudio describimos las características de la vegetación y las relaciones micorrízicas de las plantas vasculares de un bosque de Nothofagus pumilio. También describimos, vía métodos morfológicos, la diversidad ectomicorrízica presente en este bosque. Adicionalmente, determinamos si inoculaciones ectomicorrízicas confieren efectos de crecimiento positivos a semilleros de N. pumilio. Encontramos que, de 46 especies de plantas vasculares identificadas en este estudio, 42 (91%) son micorrízicas, y de estas, 33 (72%) están formando micorrizas arbusculares (AM), dos (los árboles dominantes N. pumilio y N. dombeyi) están asociados con ectomicorrizas (EM), cinco están asociadas con micorrizas ericoides, dos con micorrizas orquioides, y cuatro fueron nomicorrizadas. Adicionalmente, 26 especies de EM fueron detectadas, de las cuales 15 pertenecen a Cortinarius. Finalmente, hubo claras diferencias en el crecimiento de los semilleros de N. pumilio inoculados con el hongo ectomicorrízico Laccaria laccata, comparados a plantas no inoculadas. Sugerimos que los hongos micorrízicos juegan un rol clave en la colonización de ambientes severos por juveniles, como en el límite altitudinal andino.

Diversity of culturable filamentous Ascomycetes in the eastern South Pacific Ocean off Chile.

Our study reports the diversity of culturable mycoplankton in the eastern South Pacific Ocean off Chile to contribute with novel knowledge on taxonomy of filamentous fungi isolated from distinct physicochemical and biological marine environments. We characterized spatial distribution of isolates, evaluated their viability and assessed the influence of organic substrate availability on fungal development. Thirty-nine Operational Taxonomic Units were identified from 99 fungal strains isolated from coastal and oceanic waters by using Automatic Barcode Gap Discovery. All Operational Taxonomic Units belonged to phylum Ascomycota and orders Eurotiales, Dothideales, Sordariales and Hypocreales, mainly Penicillium sp. (82%); 11 sequences did not match existing species in GenBank, suggesting occurrence of novel fungal taxa. Our results suggest that fungal communities in the South Pacific Ocean off Chile appear to thrive in a wide range of environmental conditions in the ocean and that substrate availability may be a factor influencing fungal viability in the ocean.

Chilenopeptins A and B, Peptaibols from the Chilean Sepedonium aff. chalcipori KSH 883.

The Chilean Sepedonium aff. chalcipori strain KSH 883, isolated from the endemic Boletus loyo Philippi, was studied in a polythetic approach based on chemical, molecular, and biological data. A taxonomic study of the strain using molecular data of the ITS, EF1-α, and RPB2 barcoding genes confirmed the position of the isolated strain within the S. chalcipori clade, but also suggested the separation of this clade into three different species. Two new linear 15-residue peptaibols, named chilenopeptins A (1) and B (2), together with the known peptaibols tylopeptins A (3) and B (4) were isolated from the semisolid culture of strain KSH 883. The structures of 1 and 2 were elucidated on the basis of HRESIMS(n) experiments in conjunction with comprehensive 1D and 2D NMR analysis. Thus, the sequence of chilenopeptin A (1) was identified as Ac-Aib(1)-Ser(2)-Trp(3)-Aib(4)-Pro(5)-Leu(6)-Aib(7)-Aib(8)-Gln(9)-Aib(10)-Aib(11)-Gln(12)-Aib(13)-Leu(14)-Pheol(15), while chilenopeptin B (2) differs from 1 by the replacement of Trp(3) by Phe(3). Additionally, the total synthesis of 1 and 2 was accomplished by a solid-phase approach, confirming the absolute configuration of all chiral amino acids as l. Both the chilenopeptins (1 and 2) and tylopeptins (3 and 4) were evaluated for their potential to inhibit the growth of phytopathogenic organisms.

Productividad del hongo nativo cortinarius austroturmalis en bosques de nothofagus siempreverde y caducifolio de las reservas nacionales altos de lircay y los ruiles de la VII región, Chile / Productivity of the native mushroom cortinarius austroturmalis in evergreen and deciduous nothofagus forests of the national reserves altos de lircay and los ruiles of the VII region, Chile

Los bosques de Nothofagus de la zona mediterránea de Chile, que incluye la VII Región, están constituidos por especies caducifolias y siempreverdes, las cuales se encuentran asociadas a diversos hongos ectomicorrícicos dentro de los que destaca el hongo nativo Cortinarius austroturmalis Moser & Horak (Agaricales, Basidiomycota). Este hongo constituye un Producto Forestal No Maderero (PFNM) poco conocido, cuya productividad no ha sido determinada, lo que implica que la población rural cercana a las Reservas Nacionales Altos de Lircay y Los Ruiles de la VII Región, no lo identifique como un hongo apto para el consumo humano y, por consiguiente, no lo coseche. El objetivo de esta investigación fue conocer la productividad de basidiomas del hongo nativo Cortinarius austroturmalis asociado a especies de Nothofagus siempreverdes y caducifolias en las Reservas Nacionales Altos de Lircay y Los Ruiles de la VII Región. Las hipótesis propuestas fueron: 1) La productividad de basidiomas de Cortinarius austroturmalis en la Reserva Nacional Los Ruiles es mayor que en la Reserva Nacional Altos de Lircay, 2) La productividad de basidiomas de Cortinarius austroturmalis asociados a especies de Nothofagus siempreverdes es mayor que la productividad de este hongo asociado a especies de Nothofagus caducifolias en ambas reservas estudiadas.

Nothofagus forests in the Mediterranean region of Chile, including VII Region consist of deciduous and evergreen species, which are associated with different ectomycorrhizal fungi within which highlights the native mushroom Cortinarius austroturmalis Moser & Horak (Agaricales, Basidiomycota). This fungus is a Non-Wood Forest Product (NWFP) obscure, whose productivity has not been determined, implying that close to the National Reserves Altos de Lircay and Los Ruiles, VII Region, rural population does not identify it a fungus suitable for human consumption and therefore not harvested. The objective of this research was to determine the productivity of fruit body of the native mushroom Cortinarius austroturmalis associated with Nothofagus species of evergreen and deciduous in the National Reserves Altos de Lircay and Los Ruiles, VII Region. The proposed hypotheses were: 1) Productivity of Cortinarius austroturmalis in the National Reserve Los Ruiles is higher than in the National Reserve Altos de Lircay, 2) Productivity of Cortinarius austroturmalis associated with Nothofagus species evergreen is greater than the productivity of this fungus associated with deciduous Nothofagus species in both reserves studied.

Using common mycorrhizal networks for controlled inoculation of Quercus spp. with Tuber melanosporum: the nurse plant method.

The high cost and restricted availability of black truffle spore inoculum for controlled mycorrhiza formation of host trees produced for truffle orchards worldwide encourage the search for more efficient and sustainable inoculation methods that can be applied globally. In this study, we evaluated the potential of the nurse plant method for the controlled inoculation of Quercus cerris and Quercus robur with Tuber melanosporum by mycorrhizal networks in pot cultures. Pine bark compost, adjusted to pH 7.8 by liming, was used as substrate for all assays. Initially, Q. robur seedlings were inoculated with truffle spores and cultured for 12 months. After this period, the plants presenting 74 % mycorrhizal fine roots were transferred to larger containers. Nurse plants were used for two treatments of two different nursling species: five sterilized acorns or five 45-day-old, axenically grown Q. robur or Q. cerris seedlings, planted in containers around the nurse plant. After 6 months, colonized nursling plant root tips showed that mycorrhiza formation by T. melanosporum was higher than 45 % in the seedlings tested, with the most successful nursling combination being Q. cerris seedlings, reaching 81 % colonization. Bulk identification of T. melanosporum mycorrhizae was based on morphological and anatomical features and confirmed by sequencing of the internal transcribed spacer region of the ribosomal DNA of selected root tips. Our results show that the nurse plant method yields attractive rates of mycorrhiza formation by the Périgord black truffle and suggest that establishing and maintaining common mycorrhizal networks in pot cultures enables sustained use of the initial spore inoculum.

Synthesis and establishment of Tuber melanosporum Vitt. ectomycorrhizae on two Nothofagus species in Chile.

Axenically germinated seedlings of two species of Southern beech (Nothofagus obliqua, N. glauca) from Chile were inoculated with spores of the Périgord black truffle (Tuber melanosporum). Ectomycorrhizal development was monitored for 6 months in the greenhouse and compared to the performance of the natural host species Quercus ilex and Quercus robur. Seedling survival and mycorrhization showed major differences in both Nothofagus species: T. melanosporum readily formed ectomycorrhizae with seedlings of N. obliqua, although at a lower rate than with Q. ilex but at a proportion very similar to Q. robur; survival and colonization rates were high, and seedling growth was not visibly affected by the high soil pH required by T. melanosporum. In contrast, more than 50% of N. glauca seedlings died after inoculation, and mycorrhiza formation was very sparse. In both species, no colonization by adventive ectomycorrhizal fungi could be observed, whereas both species of Quercus showed minor colonization by another fungus, probably Inocybe or Hebeloma. Our results show that it is possible to infect N. obliqua with the Périgord black truffle under greenhouse conditions, which opens up the possibility of cultivating this truffle as a secondary crop during reforestation with N. obliqua in Chile.

The mycorrhizal community in a forest chronosequence of Sitka spruce [Picea sitchensis (Bong.) Carr.] in Northern England.

Demography and fungal diversity of the belowground ectomycorrhizal community in a chronosequence of Sitka spruce [Picea sitchensis (Bong.) Carr.] in Northumberland, Northern England, were analysed; mycorrhizal root samples were taken from 6-, 12-, 30- and 40-year-old stands, and fungal fruiting bodies were collected in autumn to complement the survey. Naturally germinated seedlings less than 1 year of age (taken from the 30-year-old stand) were also examined. A total of 118,000 mycorrhizal root tips were extracted from 40 soil cores (ten per age class) and from the complete root systems of 25 seedlings and separated into active and senescent root tips according to their morphology and anatomy. Active tips were distinguished according to their mycobionts which were characterised and identified microscopically. Although almost 100% of all fine roots were mycorrhizal, EM fungal diversity throughout the chronosequence was low, consisting of a total of 16 species of which three were only found as fruiting bodies. Of the six mycobionts found most regularly below ground, Tylospora fibrillosa was the most common, colonising about 70% of all root tips and more than 90% of those of seedlings and young trees. Root density and mycorrhizal diversity increased, but percentage of vital root tips decreased with increasing tree age, levelling off in the 30- and 40-year-old stand. Among the five subdominant fungal species, Dermocybe crocea was found to have its peak of distribution in the 12-year-old stand and Russula emetica, Lactarius rufus, Hymenoscyphus ericae agg. and the unidentified Piceirhiza sulfo-incrustata in the 30- and 40-year-old stands. The possible correlations between the mycorrhizal community structure and biotic and abiotic factors are discussed.