Systematic revision of species of Atractilina and Spiropes hyperparasitic on Meliolales (Ascomycota) in the tropics.

Atractilina Dearn. & Barthol. and Spiropes Cif. are genera of asexual fungi that comprise species mainly hyperparasitic on black mildews (Meliolales, Ascomycota). Although a common group of anamorphic fungi, they have been described up to now only by morphology and their systematic position is unknown. The present study provides a morphological treatise of all known species of Atractilina and Spiropes hyperparasitic on Meliolales, including insights into their systematic position, based on DNA sequences generated here for the first time. The study was conducted, based on 33 herbarium specimens and 23 specimens recently collected in Benin and Panama. The obtained DNA sequence data (28S rDNA and ITS rDNA) of A.parasitica and of two species of Spiropes show systematic placements in the Dothideomycetes and Leotiomycetes, respectively. The sequence data of the two Spiropes spp. do not group together. Moreover, the anamorph-teleomorph connection between Atractilinaparasitica and Malacariameliolicola, a pseudothecioid fungus, is confirmed. Three species in the genus Spiropes are proposed as new to science, namely S.angylocalycis, S.carpolobiae and S.croissantiformis. Four species are reported for Benin for the first time, three species for Panama and one species for mainland America. Atractilina and Spiropes are currently two genera with highly heterogeneous species and they might have to be split in the future, once the taxonomic concepts are validated by morphology and molecular sequence data.

Genetic diversity and population differentiation in Earliella scabrosa, a pantropical species of Polyporales.

Earliella scabrosa is a pantropical species of Polyporales (Basidiomycota) and well-studied concerning its morphology and taxonomy. However, its pantropical intraspecific genetic diversity and population differentiation is unknown. We initiated this study to better understand the genetic variation within E. scabrosa and to test if cryptic species are present. Sequences of three DNA regions, the nuclear ribosomal internal transcribed spacer (ITS), the large subunit ribosomal DNA (LSU), and the translation elongation factor (EF1α) were analysed for 66 samples from 15 geographical locations. We found a high level of genetic diversity (haplotype diversity, Hd = 0.88) and low nucleotide diversity (π = 0.006) across the known geographical range of E. scabrosa based on ITS sequences. The analysis of molecular variance (AMOVA) indicates that the genetic variability is mainly found among geographical populations. The results of Mantel tests confirmed that the genetic distance among populations of E. scabrosa is positively correlated with the geographical distance, which indicates that geographical isolation is an important factor for the observed genetic differentiation. Based on phylogenetic analyses of combined dataset ITS-LSU-EF1α, the low intraspecific divergences (0-0.3%), and the Automated Barcode Gap Discovery (ABGD) analysis, E. scabrosa can be considered as a single species with five different geographical populations. Each population might be in the process of allopatric divergence and in the long-term they may evolve and become distinct species.

Molecular-Based Diversity Studies and Field Surveys Are Not Mutually Exclusive: On the Importance of Integrated Methodologies in Mycological Research.

Understanding and describing the diversity of living organisms is a great challenge. Fungi have for a long time been, and unfortunately still are, underestimated when it comes to taxonomic research. The foundations were laid by the first mycologists through field observations. These important fundamental works have been and remain vital reference works. Nevertheless, a non-negligible part of the studied funga escaped their attention. Thanks to modern developments in molecular techniques, the study of fungal diversity has been revolutionized in terms of tools and knowledge. Despite a number of disadvantages inherent to these techniques, traditional field-based inventory work has been increasingly superseded and neglected. This perspective aims to demonstrate the central importance of field-based research in fungal diversity studies, and encourages researchers not to be blinded by the sole use of molecular methods.

Hyperparasitic Fungi on Black Mildews (Meliolales, Ascomycota): Hidden Fungal Diversity in the Tropics.

Hyperparasitism on plant-parasitic fungi is a widespread but rarely studied phenomenon. Here, for the first time, we compile in a checklist information provided by peer-reviewed literature for fungi growing on colonies of black mildews (Meliolales, Ascomycota), a species-rich group of tropical and subtropical plant-parasitic microfungi. The checklist contains information on 189 species of contact-biotrophic microfungi in 82 genera. They belong to seven morphological groups: dematiaceous hyphomycetes, moniliaceous hyphomycetes, pycnidioid, perithecioid, catathecioid, and apothecioid fungi. By the fact that species accumulation curves do not reach saturation for any tropical country, it is evident that the knowledge of the diversity of hyperparasitic fungi on Meliolales is incomplete. A network analysis of records of hyperparasitic fungi, their host fungi and host plants shows that genera of hyperparasitic fungi are generalists concerning genera of Meliolales. However, most species of hyperparasitic fungi are restricted to meliolalean hosts. In addition to hyperparasitic fungi, diverse further microorganisms use meliolalean colonies as ecological niche. Systematic positions of most species are unknown because DNA sequence data are lacking for species of fungi hyperparasitic on Meliolales. We discuss the specific challenges of obtaining DNA sequence data from hyperparasitic fungi. In order to better understand the diversity, evolution and biology of hyperparasitic fungi, it is necessary to increase sampling efforts and to undertake further morphological, molecular, and ecological studies.

Spatial risk assessment of radiocesium contamination of edible mushrooms - Lessons from a highly frequented recreational area.

The radioactive contamination of edible mushrooms increases human health hazards, especially in high mushroom collection intensity areas. Today only coarse-scale data with low spatial resolution are available, which prevents us from predicting human risk. To reduce the risk for human health, we need spatially explicit recommendations at landscape-scale. We used the Bavarian Forest National Park, a famous mushroom hunting location in Europe, as a model system. We aimed to increase the predictability of the contamination of the two most prominent mushroom species, bay boletus (Imleria badia) and ceps (Boletus edulis), and provide an efficient evidence-based risk assessment at landscape-scale. We revealed a high and a low 137Cs-activity impact area based on soil samples, which is also reflected by the mushroom species. 137Cs-activity of Imleria badia is about five times higher than the contamination of Boletus edulis; with one-third of the Imleria badia samples being over the statutory limit. The difference of contamination between species is more pronounced in the high-impact area. Elevation is a strong predictor in contrast to orientation of slopes. In high-impact areas, mushrooms showed higher 137Cs-activities at lower elevations. Soil analysis revealed that the maximum of the 137Cs-activity is still in the organic layers, indicating further mushroom contamination. We recommend using only Boletus edulis in the low-impact area for diet. We suggest that Boletus edulis bear a lower health risk than Imleria badia due to lower 137Cs-activities. Nevertheless, we need more landscape-scale studies to assess the 137Cs contamination risk for humans. Studies are primarily important in high-impact areas, which can be roughly identified by using soil contamination maps. The focus should be on high accumulating mushroom species like Imleria badia. Our study can serve as a blueprint to rapidly assess human health risks caused by radioactive contamination in landscapes intensively used by mushroom collectors.

Occurrence and chemotaxonomical analysis of amatoxins in Lepiota spp. (Agaricales).

About 95% of fatal mushroom poisonings worldwide are caused by amatoxins and phallotoxins mostly produced by species of Amanita, Galerina, and Lepiota. The genus Lepiota is supposed to include a high number of species producing amatoxins. In this study, we investigated 16 species of Lepiota based on 48 recently collected specimens for the presence of amatoxins by liquid chromatography coupled to a diode-array detector and mass spectrometry (UHPLC-QTOF-MS/MS). By comparing the retention times, UV absorptions, and diagnostic MS fragment ions with data obtained from the benchmark species Amanita phalloides, we detected α-amanitin and γ-amanitin in Lepiota subincarnata, α-amanitin and amaninamide in Lepiota brunneoincarnata, and ß-amanitin and α-amanitin in Lepiota elaiophylla. Phallotoxins have not been detected any of these species. Two possibly undescribed amatoxin derivatives were found in Lepiota boudieri and L. elaiophylla, as well as one further non-amatoxin compound in one specimen of L. cf. boudieri. These compounds might be used to differentiate L. elaiophylla from L. xanthophylla and species within the L. boudieri species complex. No amatoxins were detected in L. aspera, L. castanea, L. clypeolaria, L. cristata, L. erminea, L. felina, L. fuscovinacea, L. lilacea, L. magnispora, L. oreadiformis, L. pseudolilacea, L. sp. (SeSa 5), and L. subalba. By combining the occurrence data of amatoxins with a phylogenetic analysis, a monophyletic group of amatoxin containing species of Lepiota is evident. These chemotaxonomic results highlight the relevance of systematic relationships for the occurrence of amatoxins and expand our knowledge about the toxicity of species of Lepiota.

A new study of Nagrajomyces: with two new species proposed and taxonomic status inferred by phylogenetic methods.

Nagrajomyces (incertae sedis, Ascomycota) is a monotypic genus with a previously unknown systematic position. In this report, two new species are proposed, Nagrajomycesfusiformis and Nagrajomyceslaojunshanensis. These new taxa are proposed based on morphological characteristics evident via light microscopy and molecular data. Multi-locus phylogenetic analyses (ITS rDNA, nrLSU rDNA, RPB2, and TEF1-α) show that specimens recently collected in Yunnan Province, China are closely related to Gnomoniaceae. Both new species and known species were discovered repeatedly in their asexual developmental form exclusively on twigs of Rhododendron spp. (Ericaceae). This indicates a host specificity of Nagrajomyces spp. for species of Rhododendron.

Four new species of Russula subsection Roseinae from tropical montane forests in western Panama.

Species of the genus Russula are key components of ectomycorrhizal ecosystems worldwide. Nevertheless, their diversity in the tropics is still poorly known. This study aims to contribute to the knowledge of the diversity of Russula species classified in subsection Roseinae based on specimens recently collected in tropical montane rainforests in western Panama. A five gene multilocus phylogeny based on the nuclear markers ITS nrDNA, MCM7, RPB1, RPB2 and TEF-1α was constructed to identify the systematic position of 22 collections from Panama. Four new species, Russula cornicolor, Russula cynorhodon, Russula oreomunneae and Russula zephyrovelutipes are formally described and illustrated. None of the four species are sister species and they are more closely related to North American or Asian species. Two of the newly described species were associated with the ectomycorrhizal tree species Oreomunnea mexicana, while the other two species were associated with Quercus species. All four species are so far only known from mountains in western Panama.

Unravelling unexplored diversity of cercosporoid fungi (Mycosphaerellaceae, Mycosphaerellales, Ascomycota) in tropical Africa.

Cercosporoid fungi (Mycosphaerellaceae, Mycosphaerellales, Ascomycota) are one of the largest and most diverse groups of hyphomycetes causing a wide range of diseases of economically important plants as well as of plants in the wild. Although more than 6000 species are known for this group, the documentation of this fungal group is far from complete. Especially in the tropics, the diversity of cercosporoid fungi is poorly known. The present study aims to identify and characterise cercosporoid fungi collected on host plants belonging to Fabaceae in Benin, West Africa. Information on their morphology, host species and DNA sequence data (18S rDNA, 28S rDNA, ITS and tef1) is provided. DNA sequence data were obtained by a simple and non-culture-based method for DNA isolation which has been applied for cercosporoid fungi for the first time in the context of the present study. Among the loci used for the phylogenetic analysis, tef1 provided the best resolution together with the multigene dataset. Species delimitation in many cases, however, was only possible by combining molecular sequence data with morphological characteristics. Based on forty specimens recently collected in Benin, 18 species are presented with morphological descriptions, illustrations and sequence data. Among these, six species in the genus Cercospora and two species in Pseudocercospora are proposed as species new to science. The newly described species are Cercospora (C.) beninensis on Crotalaria macrocalyx, C. parakouensis on Desmodium tortuosum, C. rhynchophora on Vigna unguiculata, C. vignae-subterraneae on Vigna subterranea, C. tentaculifera on Vigna unguiculata, C. zorniicola on Zornia glochidiata, Pseudocercospora sennicola on Senna occidentalis and Pseudocercospora tabei on Vigna unguiculata. Eight species of cercosporoid fungi are reported for Benin for the first time, three of them, namely C. cf. canscorina, C. cf. fagopyri and C. phaseoli-lunati are new for West Africa. The presence of two species of cercosporoid fungi on Fabaceae previously reported from Benin, namely Nothopassalora personata and Passalora arachidicola, is confirmed.

Morphological and genetic diversification of Russula floriformis, sp. nov., along the Isthmus of Panama.

Species of Russula are ubiquitous members of ectomycorrhizal fungal communities in tropical ecosystems. However, an important part of the total tropical diversity of this genus and its biogeographic patterns is unknown due to the lack of studies on Russula in tropical ecosystems. We combined molecular, morphological, ecological, and biogeographic data to elaborate concepts for two new subspecies of R. floriformis (subsection Substriatinae). Russula floriformis subsp. floriformis and R. floriformis subsp. symphoniae are described as new from montane forest dominated by Quercus and/or Oreomunnea (Fagales) from Colombia and Panama, respectively. Phylogenies were constructed using nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS), D1-D2 domains of nuc 28S rDNA (28S), and partial regions of the second largest subunit of RNA polymerase II (rpb2) and translation elongation factor 1-alpha (tef1). Similar environmental conditions, similar morphology, and an ITS sequence similarity higher than 99% with only three different positions indicate that these two subspecies are closely related. Detailed observations of microscopic structures and analyses of further DNA loci, however, revealed morphological and molecular characteristics that allow distinguishing the two subspecies of R. floriformis. Spatial distribution and phylogenetic proximity of the two Russula subspecies and their ectomycorrhizal hosts, i.e., species of Quercus, suggest that their diversification is a result of comigration, adaptation, and geographic isolation along the Isthmus of Panama during the Pliocene and Pleistocene.

Brassicaceous roots as an unexpected diversity hot-spot of helotialean endophytes.

A high number of fungal strains were isolated from roots of Brassicaceae species collected across western and southern Europe, resulting in an unexpectedly rich collection of Cadophora species. These isolates enable us to present a new and comprehensive view of the ecological, morphological, and phylogenetic traits of root-inhabiting members of this helotialean genus. We provide phylogenetic placement of all of our isolates based on a four-gene dataset, analyze their phenotypic traits in relation to their phylogenetic relationships, and infer the potential distribution ranges of the species by sequence comparisons with available databases. We consider seven well supported phylogenetic lineages as species new to science. Six further lineages probably also represent new species but remain undescribed due to the lack of diagnostic morphological characters. Our results show that Cadophora, as currently circumscribed, is paraphyletic and encompasses a broad spectrum of morphologies and lifestyles. Among the new species, only two (C. ferruginea and C. constrictospora) form phialides and conidia typical of Cadophora, three species (C. echinata, C. gamsii and C. variabilis) produce chains of swollen hyphal segments that may function as holoblastic conidia, and one species (C. fascicularis) produces chains of holoblastic ramoconidia and conidia. Ancestral state reconstruction analysis suggests that phialidic conidiogenesis evolved several times in Cadophora s. lat. from a putatively holoblastic common ancestor. Most Cadophora lineages are rare as estimated from the availability of sequence data, in spite of having relatively wide distribution ranges, whereas five lineages may represent endemic relationships given their restricted distributions. Our dataset, probably the most comprehensive available for Cadophora, nevertheless shows knowledge gaps concerning the phylogenetic relationships within this genus and highlights a need for further investigation.

Mapping mycological ignorance - checklists and diversity patterns of fungi known for West Africa.

Scientific information about biodiversity distribution is indispensable for nature conservation and sustainable management of natural resources. For several groups of animals and plants, such data are available, but for fungi, especially in tropical regions like West Africa, they are mostly missing. Here, information for West African countries about species diversity of fungi and fungus-like organisms (other organisms traditionally studied by mycologists) is compiled from literature and analysed in its historical context for the first time. More than 16,000 records of fungi representing 4843 species and infraspecific taxa were found in 860 publications relating to West Africa. Records from the Global Biodiversity Information Facility (GBIF) database (2395 species), and that of the former International Mycological Institute fungal reference collection (IMI) (2526 species) were also considered. The compilation based on literature is more comprehensive than the GBIF and IMI data, although they include 914 and 679 species names, respectively, which are not present in the checklist based on literature. According to data available in literature, knowledge on fungal richness ranges from 19 species (Guinea Bissau) to 1595 (Sierra Leone). In estimating existing species diversity, richness estimators and the Hawksworth 6:1 fungus to plant species ratio were used. Based on the Hawksworth ratio, known fungal diversity in West Africa represents 11.4% of the expected diversity. For six West African countries, however, known fungal species diversity is less than 2%. Incomplete knowledge of fungal diversity is also evident by species accumulation curves not reaching saturation, by 45.3% of the fungal species in the checklist being cited only once for West Africa, and by 66.5% of the fungal species in the checklist reported only for a single country. The documentation of different systematic groups of fungi is very heterogeneous because historically investigations have been sporadic. Recent opportunistic sampling activities in Benin showed that it is not difficult to find specimens representing new country records. Investigation of fungi in West Africa started just over two centuries ago and it is still in an early pioneer phase. To promote proper exploration, the present checklist is provided as a tool to facilitate fungal identification in this region and to aid conceptualisation and justification of future research projects. Documentation of fungal diversity is urgently needed because natural habitats are being lost on a large scale through altered land use and climate change.

Diversity of Fungi in Soils with Different Degrees of Degradation in Germany and Panama.

Soil degradation can have an impact on the soil microbiota, but its specific effects on soil fungal communities are poorly understood. In this work, we studied the impact of soil degradation on the richness and diversity of communities of soil fungi, including three different degrees of degradation in Germany and Panama. Soil fungi were isolated monthly using the soil-sprinkling method for 8 months in Germany and 3 months in Panama, and characterized by morphological and molecular data. Soil physico-chemical properties were measured and correlated with the observed values of fungal diversity. We isolated a total of 71 fungal species, 47 from Germany, and 32 from Panama. Soil properties were not associated with fungal richness, diversity, or composition in soils, with the exception of soil compaction in Germany. The geographic location was a strong determinant of the soil fungal species composition although in both countries there was dominance by members of the orders Eurotiales and Hypocreales. In conclusion, the results of this work do not show any evident influence of soil degradation on communities of soil fungi in Germany or Panama.

Diversity of Trametes (Polyporales, Basidiomycota) in tropical Benin and description of new species Trametes parvispora.

Trametes is a globally distributed genus of white-rot polypores and well sampled in temperate and boreal areas. However, the diversity, taxonomy, and phylogenetic positions of Trametes spp. are poorly known in tropical Africa. This study aims at documenting the diversity of Trametes species in Benin (tropical Africa) and their phylogenetic positions with a focus on the T. elegans species complex. Therefore, we collected specimens of Trametes from different forest types across Benin. To infer phylogenetic relationships between Trametes species, we investigated sequences of five gene regions and added available sequences from GenBank. Using Maximum likelihood and Bayesian phylogeny inference methods, we found eight supported species clades. For the T. elegans species complex, we re-establish the name Trametes palisotii for species previously known as T. elegans in tropical Africa. Furthermore, we propose Trametes parvispora as a species new to science and provide the description of this species. Our molecular phylogeny of Trametes with a focus on tropical Benin contributes to taxonomic clarity of an important wood-decay fungal genus, which is the basis for biodiversity assessments of Trametes in the tropics.

Diversity of Fungi in Soils with Different Degrees of Degradation inGermany and Panama

Soil degradation can have an impact on the soil microbiota, but its specific effects on soilfungal communities are poorly understood. In this work, we studied the impact of soil degradationon the richness and diversity of communities of soil fungi, including three differentdegrees of degradation in Germany and Panama. Soil fungi were isolated monthly using thesoil-sprinkling method for 8 months in Germany and 3 months in Panama, and characterizedby morphological and molecular data. Soil physico-chemical properties were measured andcorrelated with the observed values of fungal diversity. We isolated a total of 71 fungal species,47 from Germany, and 32 from Panama. Soil properties were not associated with fungalrichness, diversity, or composition in soils, with the exception of soil compaction inGermany. The geographic location was a strong determinant of the soil fungal species compositionalthough in both countries there was dominance by members of the ordersEurotiales and Hypocreales. In conclusion, the results of this work do not show any evidentinfluence of soil degradation on communities of soil fungi in Germany or Panama.

Diversity of Fungi in Soils with Different Degrees of Degradation inGermany and Panama

Soil degradation can have an impact on the soil microbiota, but its specific effects on soilfungal communities are poorly understood. In this work, we studied the impact of soil degradationon the richness and diversity of communities of soil fungi, including three differentdegrees of degradation in Germany and Panama. Soil fungi were isolated monthly using thesoil-sprinkling method for 8 months in Germany and 3 months in Panama, and characterizedby morphological and molecular data. Soil physico-chemical properties were measured andcorrelated with the observed values of fungal diversity. We isolated a total of 71 fungal species,47 from Germany, and 32 from Panama. Soil properties were not associated with fungalrichness, diversity, or composition in soils, with the exception of soil compaction inGermany. The geographic location was a strong determinant of the soil fungal species compositionalthough in both countries there was dominance by members of the ordersEurotiales and Hypocreales. In conclusion, the results of this work do not show any evidentinfluence of soil degradation on communities of soil fungi in Germany or Panama.

New neotropical species of Phyllachorales based on molecular, morphological, and ecological data.

Species of tropical tar spot fungi (Phyllachorales, Ascomycota) are obligate biotrophic plant parasitic fungi associated with living leaves of a wide range of families of host plants, mainly in tropical and subtropical regions. In this study, samples of tropical tar spot fungi were collected in forests in Costa Rica and Panamá. To identify taxa, we used morphology and information on host plants and combined multigene phylogeny of four genes: the large subunit nuclear ribosomal DNA (28S rDNA), the small subunit nuclear ribosomal DNA (18S rDNA), the complete internal transcribed spacer region of ribosomal DNA (nuc rDNA ITS1-5.8S-ITS2; ITS), and the translation elongation factor 1-α (tef1). Here we propose one new species in the genus Camarotella and eight new species in Telimena with their morphological descriptions, illustrations, and sequence data. The newly described species are Camarotella licaniae on Licania arborea (Chrysobalanaceae) and in the genus Telimena: T. billiae on Billia rosea (Sapindaceae), T. drymoniae on Drymonia multiflora (Gesneriaceae), T. hydrangeae on Hydrangea sp. (Hydrangeaceae), T. miravallensis on Symplocos panamensis (Symplocaceae), T. protii on Protium sp. (Burseraceae), T. rinoreae on Rinorea sp. (Violaceae), T. semialarii on Semialarium mexicanum (Celastraceae), and T. triseptata on Tapirira mexicana (Anacardiaceae). The new name Telimena nitens on Schlegelia brachyanta (Schlegeliaceae) is presented and 10 species of Phyllachora are transferred to Telimena, leading to the new combinations T. canarii, T. galavisii, T. insueta, T. ruelliae, T. scutiformis, T. serjaniicola, T. spicatae, T. subrepens, T. symploci, and T. symplocicola. Additionally, revisions of tar spot fungi on host families Burseraceae, Sapindaceae, and Symplocaceae are provided, and four new synonyms are proposed.

Panama, a hot spot for Hermatomyces (Hermatomycetaceae, Pleosporales) with five new species, and a critical synopsis of the genus.

Five new species belonging to Hermatomyces (Hermatomycetaceae, Pleosporales) are described based on morphological investigations of specimens collected on rotten twigs and stems of various plants in Panama as well as phylogenetic analyses of sequence data of nuclear ribosomal and protein coding genes (EF1-α, RPB2, ß-TUB). The new species are described as: Hermatomyces bifurcatus, H. constrictus, H. megasporus, H. sphaericoides, and H. verrucosus spp. nov. Previously described species such as H. sphaericus and H. tucumanensis were identified among the studied specimens. The new combination, H. reticulatus, is made for Subicularium reticulatum based on examination of the holotype and fresh collections. Hermatomyces subiculosus, originally described from Thailand, is reduced to synonymy with H. reticulatus; H. tectonae is synonymized under H. sphaericus based on morphological and molecular evidence; and H. chiangmaiensis and H. thailandicus are considered later synonyms of H. krabiensis and H. indicus, respectively. The type material of Scyphostroma mirum was found to be conspecific with H. tucumanensis and, therefore, the generic name Hermatomyces should be conserved or protected against the older name Scyphostroma and the binomial H. tucumanensis against S. mirum. Sixteen species of Hermatomyces are recognized, their distinctive characteristics are highlighted in line drawings and a key is provided for their identification. The peculiar morphology and consistent phylogeny of new and previously known Hermatomyces species supports the recognition of the recently introduced monotypic family Hermatomycetaceae as a well delimited monophyletic taxon within the order Pleosporales.

Georatusin, a Specific Antiparasitic Polyketide-Peptide Hybrid from the Fungus Geomyces auratus.

Georatusin (1), featuring a highly reduced, methylated polyketide moiety fused to a tryptophan by an amide and ester bond forming a 13-membered ring, was produced by the soil fungus Geomyces auratus. An HMQC-COSY spectrum was measured to build up the connectivities despite the overlapping proton signals. DQF-COSY, HETLOC, J-HMBC, and ROESY were implemented to determine the relative configuration of the flexible moiety. Georatusin (1) shows specific antiparasitic activities against Leishmania donovani and Plasmodium falciparum without obvious cytotoxicity. The biosynthesis of 1 was also proposed.