Black Meristematic Fungi Research Articles

Recolonization dynamics of marble monuments after cleaning treatments: A nine-year follow-up study

The prevention and control of biological patinas on outdoor stone monuments represent a demanding challenge for the conservation of cultural heritage also due to some microorganisms, particularly resistant to treatments, such as black meristematic fungi, an eco-physiological group well known for its tolerance to extreme conditions. Even if several methods and eco-friendly products have been proposed as new alternatives, traditional biocides are still far from being completely replaced. Recolonization is a natural process that occurs sooner or later after cleaning. The time that elapses until its occurrence can vary considerably depending on environmental conditions and the used products; unfortunately, the papers describing the effect of treatments over time are rare. This work aims to shed light on the recolonization process of marble surfaces in the ancient monumental cemetery of Bonaria (Cagliari) after nine years from treatments, evaluating the long-term efficiency of two different cleaning methods, namely dimethyl sulfoxide-based gel (DMSO-based gel) and Biotin T (a didecyldimethylammonium chloride-based product-). In this context, the microflora present before treatments and in the following years was assessed by culture-based methods and identified by molecular techniques, with attention on black meristematic fungi, which were used as reference for the most resistant lithobiontic organisms. Different environmental parameters, such as temperature, exposition, dominant winds, and rainfall, were considered, and infrared thermography, portable light microscopy, and image analysis were used. This research evidenced the influence of water availability and lightning in recolonization processes, the transition from the pioneer fungal community versus more resistant black fungal species after Biotin T treatment, and the long-lasting efficiency of the DMSO-based gel. These findings prove that this low-impact method deserves more attention in the conservation of outdoor marble monuments, emphasizing the importance of long-term studies.

Characterization of the Community of Black Meristematic Fungi Inhabiting the External White Marble of the Florence Cathedral

Meristematic black fungi are a highly damaging group of microorganisms responsible for the deterioration of outdoor exposed monuments. Their resilience to various stresses poses significant challenges for removal efforts. This study focuses on the community of meristematic fungi inhabiting the external white marble of the Cathedral of Santa Maria del Fiore, where they contribute to its darkening. Twenty-four strains were isolated from two differently exposed sites of the Cathedral, and their characterization was conducted. Phylogenetic analysis using ITS and LSU rDNA regions revealed a wide diversity of rock-inhabiting fungal strains within the sampled areas. Eight strains, belonging to different genera, were also tested for thermal preferences, salt tolerance, and acid production to investigate their tolerance to environmental stressors and their interaction with stone. All tested strains were able to grow in the range of 5–30 °C, in the presence 5% NaCl, and seven out of eight strains were positive for the production of acid. Their sensitivities to essential oils of thyme and oregano and to the commercial biocide Biotin T were also tested. The essential oils were found to be the most effective against black fungi growth, indicating the possibility of developing a treatment with a low environmental impact.

Knufia obscura sp. nov. and Knufia victoriae sp. nov., two new species from extreme environments.

Six strains of black meristematic fungi were isolated from Antarctic soils, gasoline car tanks and from the marine alga Flabellia petiolata. These fungi were characterized by morphological, physiological and phylogenetic analyses. According to the maximum-likelihood analysis reconstructed with ITS and LSU sequences, these strains belonged to the genus Knufia. Knufia obscura sp. nov. (holotype CBS 148926) and Knufia victoriae sp. nov. (holotype CBS 149015) are proposed as two novel species and descriptions of their morphological, physiological and phylogenetic features are presented. Based on the maximum-likelihood analyses, K. obscura was closely related to Knufia hypolithi (99 % bootstrap support), while K. victoriae clustered in the clade of Knufia cryptophialidica and Knufia perfecta (93 % bootstrap support). Knufia victoriae, recorded in Antarctic soil samples, had a psychrophilic behaviour, with optimal growth between 10 and 15 °C and no growth recorded at 20 °C. Knufia obscura, from a gasoline car tank and algae, displayed optimal growth between 20 and 25 °C and was more tolerant to salinity than K. victoriae.

New records of Oleoguttula mirabilis and Rachicladosporium antarcticum from James Ross Island, Antarctica

A group of seven isolates of black meristematic fungi was collected from rocks within the framework of mycological research focused on the study of diversity of rock-inhabiting fungi on James Ross Island in Antarctica. Their identification was based on ITS rDNA sequence comparisons supported by morphological data. Obtained results assigned four analysed strains as Oleoguttula mirabilis and three isolates as Rachicladosporium antarcticum. To the best of our knowledge this is the second report of isolation of these two psychrophilic species from different parts of James Ross Island which imply that they probably represent a common part of its environmental microbial flora.

Study of the euendolithic activity of black meristematic fungi isolated from a marble statue in the Quirinale Palace’s Gardens in Rome, Italy

The present study analyzed the dark patina present on the surface of a marble statue exposed in the Gardens of the Quirinal Palace in Rome, with a special focus on euendolithic black meristematic fungi. The study of the spatial distribution of microorganisms and their identification were performed by using a multistep approach based on microscopy, cultural analyses, molecular techniques, and embedding resin-casting technique. Black meristematic fungi were observed in the patina. Since morphological features are not sufficient to identify fungi belonging to this group, cultural and molecular analyses were performed. The results highlighted the presence of the species Coniosporium apollinis and of strains related to the genus Knufia (order of Chaetothyriales). The resin-casting embedding technique demonstrated the active penetration of these fungi into the marble grains, thus documenting their true endolithic behavior. The involvement of phototrophic microorganisms present on the surface in the active penetration of the marble was excluded.

Antarctic epilithic lichens as niches for black meristematic fungi.

Sixteen epilithic lichen samples (13 species), collected from seven locations in Northern and Southern Victoria Land in Antarctica, were investigated for the presence of black fungi. Thirteen fungal strains isolated were studied by both morphological and molecular methods. Nuclear ribosomal 18S gene sequences were used together with the most similar published and unpublished sequences of fungi from other sources, to reconstruct an ML tree. Most of the studied fungi could be grouped together with described or still unnamed rock-inhabiting species in lichen dominated Antarctic cryptoendolithic communities. At the edge of life, epilithic lichens withdraw inside the airspaces of rocks to find conditions still compatible with life; this study provides evidence, for the first time, that the same microbes associated to epilithic thalli also have the same fate and chose endolithic life. These results support the concept of lichens being complex symbiotic systems, which offer attractive and sheltered habitats for other microbes.

The Lichen Connections of Black Fungi

Many black meristematic fungi persist on rock surfaces-hostile and exposed habitats where high doses of radiation and periods of desiccation alternate with rain and temperature extremes. To cope with these extremes, rock-inhabiting black fungi show phenotypic plasticity and produce melanin as cell wall pigments. The rather slow growth rate seems to be an additional prerequisite to oligotrophic conditions. At least some of these fungi can undergo facultative, lichen-like associations with photoautotrophs. Certain genera presenting different lifestyles are phylogenetic related among the superclass Dothideomyceta. In this paper, we focus on the genus Lichenothelia, which includes border-line lichens, that is, associations of melanised fungi with algae without forming proper lichen thalli. We provide a first phylogenetic hypothesis to show that Lichenothelia belongs to the superclass Dothideomyceta. Further, culture experiments revealed the presence of co-occurring fungi in Lichenothelia thalli. These fungi are related to plant pathogenic fungi (Mycosphaerellaceae) and to other rock-inhabiting lineages (Teratosphaeriaceae). The Lichenothelia thallus-forming fungi represent therefore consortia of different black fungal strains. Our results suggest a common link between rock-inhabiting meristematic and lichen-forming lifestyles of ascomycetous fungi.

Polyextremotolerant black fungi: oligotrophism, adaptive potential, and a link to lichen symbioses.

Black meristematic fungi can survive high doses of radiation and are resistant to desiccation. These adaptations help them to colonize harsh oligotrophic habitats, e.g., on the surface and subsurface of rocks. One of their most characteristic stress-resistance mechanisms is the accumulation of melanin in the cell walls. This, production of other protective molecules and a plastic morphology further contribute to ecological flexibility of black fungi. Increased growth rates of some species after exposure to ionizing radiation even suggest yet unknown mechanisms of energy production. Other unusual metabolic strategies may include harvesting UV or visible light or gaining energy by forming facultative lichen-like associations with algae or cyanobacteria. The latter is not entirely surprising, since certain black fungal lineages are phylogenetically related to clades of lichen-forming fungi. Similar to black fungi, lichen-forming fungi are adapted to growth on exposed surfaces with low availability of nutrients. They also efficiently use protective molecules to tolerate frequent periods of extreme stress. Traits shared by both groups of fungi may have been important in facilitating the evolution and radiation of lichen-symbioses.

Gynonectria intraspora gen. et sp. nov. (Hypocreales), a perianthicolous phyllosphere fungus with a novel ascospore type

Gynonectria intraspora is described as a new monotypic genus of the Hypocreales (Ascomycetes). Relatively large perithecia form individually within the perianths of the hepatic Odontolejeunea sp. (Lejeuneaceae, Porellales), which grow in the phyllosphere of living vascular plants in the rainforests of Costa Rica and Panama. As in other hyperepiphyllous perianthicolous species, hyphae concentrate in the region of the sporophyte–gametophyte junction. They prevent sporophyte maturation, but do not visibly affect the leafy gametophyte. The most intriguing character of G. intraspora is exhibited by the ascospores. While they are still in the asci, each of them cleaves two new spores within its lumina leaving residual primary spore plasma around them. This novel spore type is comparable to the endoconidia of black meristematic fungi and is called an "intraspore". The formation of intraspores together with other features seem to indicate that the obligate hyperepiphyllous Hypocreales are undergoing an active evolutionary process of diversification, as are their epiphyllous lejeuneaceous hosts.

Two new dothideomycetous endoconidial genera from declining larch

Two endoconidial, black meristematic fungi, Celosporium larixicolum gen. et sp. nov. (Dothideales) and Hispidoconidioma alpina gen. et sp. nov. (Capnodiales) are described from black subicula on twigs of declining larch ( Larix lyallii Parl) trees in Alberta, Canada. Conidioma morphology and phylogenetic analysis of LSU and ITS regions indicate that these taxa are both distinct from each other and from previously described endoconidial genera. Conidiomata of C. larixicolum consist of black cellular clumps (aggregated conidiogenous cells) that are either naked or enveloped by scant to dense mycelium that sometimes organizes into a cupulate peridium. Endoconidia are 1–3 celled, hyaline when released but become pigmented as they age, and very variable in size and shape, e.g., globose, pear-shaped, osteoid, or discoid with an irregular flange. In H. alpina, colonies are three-layered, consisting of a central pseudoparenchymatous layer sandwiched between an upper and a basal hyphal layers, and conidiogenesis occurs in sporadic areas of the central layer. Endoconidia are unicellular, hyaline, and subglobose to ellipsoid. The strong phylogenetic affinities between these newly described taxa and slow-growing, melanized fungi isolated from rocks suggest individual black meristematic fungus lineages may have broad habitat ranges.

Do fungi survive under actual space conditions? Searching for evidence in favour of lithopanspermia

Cryptoendolithic communities and black meristematic fungi from the Antarctic desert have been shown to survive under simulated space and Martian conditions in ground‐based Experiment Verification Tests. An experiment was set to test their resistance under actual space and simulated Martian conditions on the International Space Station in the framework of the EXPOSE‐EuTEF Consortium, LIFE‐ESA project.

Antarctic microfungi as models for exobiology

Microfungi living in different Antarctic environments are generally well adapted to high stress conditions such as low temperatures, wide thermal fluctuations, high UV irradiance, and low water and nutrients availability; for this reason they could be investigated in order to explore limits of microbial life. Microfungi living in simple cryptoendolithic microbial communities inside the porosity of rocks in the Antarctic Dry Valleys, the closest terrestrial analogue of Mars, are particularly suitable for exobiological studies. Until now studies on exobiology focused mainly on prokaryotes; since cryptoendolithic black meristematic fungi are able to tolerate hard desiccation, high UV exposure, extremely low temperatures and wide thermal fluctuations, they are proposed as the best eukaryotic models for the biological exploration of Mars. Indeed, their adaptive strategies could be crucial as predictive tools in investigating the limits of life.

Fungi as Geologic Agents

Although many studies on fungi and geological processes have been published in recent years, books and congress proceedings on geomicrobiology focus mainly on prokaryotes and algae. Therefore, it is the aim of this review to summarize data on the fungal impact on geological processes. These processes include the alteration and weathering of rock and minerals, the accumulation of metals, and the conversion of fossil organic carbon. Fossil records and fungi in subsurface environments are also discussed. This article especially emphasizes the role of epi- and endolithic black meristematic fungi, discussing their deteriorative potential on rock as well as their taxonomy and phylogeny. Moreover, the impact of fungi on weathering of monuments and building materials is described and new methods to study fungi-material interactions are presented. The data summarized here show that "geomycology" is a highly interesting discipline in view of basic geological research, as well as biotechnological application.

Temperature and NaCl-tolerance of rock-inhabiting meristematic fungi.

Black meristematic fungi together with lichens and cyanobacteria dominate the micro-flora of rock surfaces in arid and semi-arid environments of hot and cold deserts. This study shows that rock inhabiting meristematic fungi are extremely tolerant against high temperatures, desiccation and osmotic stress. Their temperature tolerance increases with increasing dehydration of the fungal thallus. Air dried mycelia of black yeasts stand temperatures up to 120 degrees C for at least 0.5 hours. As response to high temperatures multilayered cell walls are developed and trehalose is accumulated whereas the intracellular glycerol regulates the osmotic potential under NaCl stress. Strains from rock in moderate climate (North Germany) show the same tolerance than those isolated from the Mediterranean area. Hortaea werneckii--hitherto only described as agent of human Tinea nigra--is shown to be the most tolerant rock inhabiting species tested. Meristematic fungi cannot be pre-adapted to higher growth temperatures by increased incubation temperatures. Considering the results of this study the justification of the term 'stress' is discussed with regard to rock inhabiting fungi and their natural environment. Consequences for conservation treatments of monuments decayed by meristematic fungi are discussed on the basis of the ecophysiological properties of the fungi.