Didymium arenosum, a myxomycete new to science from the confluence of deserts in northwestern China.

A new myxomycete species, Didymium arenosum, was described based on morphological evidence and phylogenetic analyses. The species was discovered in the arid region at the confluence of the Badain Jaran desert and Tengger desert on the leaves of Betula platyphylla and was cultivated in a moist chamber culture. Morphologically, the species is distinguished by the greenish-yellow calcium carbonate crystals on the surface and the spores covered with small warts, some of which are connected into a short line. A phylogenetic analysis of D. arenosum strongly supports its classification as a separate clade. The spore to spore agar culture of D. arenosum requires 23 days, and this study provides a detailed description of its life cycle.

Tasmaniomyxa umbilicata, a new genus and new species of myxomycete from Tasmania.

A new genus and species of myxomycete, Tasmaniomyxa umbilicata, is described based on numerous observations in Tasmania and additional records from southeastern Australia and New Zealand. The new taxon is characterized by an unusual combination of characters from two families: Lamprodermataceae and Didymiaceae. With Lamprodermataceae the species shares limeless sporocarps, a shining membranous peridium, an epihypothallic stalk, and a cylindrical columella. Like Didymiaceae, it has a soft, flaccid, sparsely branched capillitium, with rough tubular threads that contain fusiform nodes and are firmly connected to the peridium. Other characters of T. umbilicata that also occur in many Didymiaceae are the peridium dehiscing into petaloid lobes, the yellow, motile plasmodium, and the spores ornamented with larger, grouped and smaller, scattered warts. The transitional position of the new taxon is reflected by a three-gene phylogeny, which places T. umbilicata at the base of the branch of all lime-containing Physarales, thus justifying its description as a monotypic genus.

An exceptionally stable and widespread hydrated amorphous calcium carbonate precipitated by the dog vomit slime mold Fuligo septica (Myxogastria).

Biogenic amorphous calcium carbonate (ACC) is typically metastable and can rapidly transform through aging, dehydration, and/or heating to crystalline calcium carbonate. Gaining insight into its structure and properties is typically hampered by its tendency to crystallize over short time periods once isolated from the host organism, and also by the small quantities that are usually available for study. Here we describe an exceptionally stable hydrated ACC (HACC) precipitated by the cosmopolitan slime mold Fuligo septica (L.) F.H. Wigg. (1780). A single slime mold can precipitate up to a gram of HACC over the course of one night. Powder x-ray diffraction (XRD) patterns, transmission electron microscopy images, infrared absorption spectra, together with the lack of optical birefringence are consistent with an amorphous material. XRD simulations, supported by thermogravimetric and evolved gas analysis data, are consistent with an intimate association of organic matter with ~ 1-nm-sized ACC units that have monohydrocalcite- and calcite-like nano-structural properties. It is postulated that this association imparts the extreme stability of the slime mold HACC by inhibiting loss of H2O and subsequent crystallization. The composition, structure, and thermal behavior of the HACC precipitated by F. septica collected over 8000 km apart and in markedly different environments, suggests a common structure, as well as similar biochemical and biomineralization mechanisms.

Didymium pseudonivicola: A new myxomycete from the austral Andes emerges from broad-scale morphological and molecular analyses of D. nivicola collections.

A new nivicolous myxomycete is described as a result of a comprehensive study of Didymium nivicola collections from the entire range of its occurrence. Statistical analysis of 12 morphological characters, phylogenetic analyses of nuc 18S rDNA and elongation factor 1-alpha gene (EF1A), and a delimitation method (automatic barcode gap diversity) have been applied to corroborate the identity of the new species. A preliminary morphological analysis of D. nivicola revealed high variability of South American populations where four types of spore ornamentation were noted. However, results of molecular study and statistical analysis of morphological characters did not support recognition of these four forms but the distinction of two morphotypes. Consequently, two species have been recognized: D. nivicola and the newly proposed D. pseudonivicola. The new species can be distinguished from D. nivicola by distinctly larger and mostly plasmodiocarpic sporophores, which are scattered to gregarious, paler spores, and by the paler, more delicate and more elastic capillitium. Spore ornamentation of D. pseudonivicola is uniform and can be described as distinctly spiny (pilate under scanning electron microscope [SEM]), whereas those of D. nivicola is more variable, where spines (pilae under SEM) are delicate, distinct, or conspicuous. Additionally, whereas D. nivicola is a species distributed worldwide, D. pseudonivicola occurs only in the austral Andes of Argentina and Chile.

Morphological and molecular characterization of the new aethaloid species Didymium yulii.

Five specimens, initially presumed to be Fuligo septica or Mucilago crustacea, were collected from Jilin Province and the Inner Mongolia Autonomous Region in China, but all five turned out to represent a new aethaloid species, Didymium yulii. This new species is characterized by pseudocapillitia without capillitia and an aethalioid fruiting body, features that are morphologically distinct from those of any other species of Didymium. To assess the phylogenetic relationships between D. yulii and other members of Didymium and in the Didymiaceae, sequences from two nonoverlapping targeted portions of nuc 18S rDNA (~450 bp and ~1050 bp) and translation elongation factor 1-alpha (tef1) were obtained and analyzed. The results indicate that D. yulii forms a single clade separate from other species of Didymium and the clade that contains M. crustacea, which strongly supports the identification of the five specimens as a new species.

Fuligo septica Spores Onboard a Stratospheric NASA Balloon and Its Complete In Vitro Life Cycle.

The aim of this study was to demonstrate for the first time Fuligo septica spore viability in the stratosphere through spore germination and its complete life cycle. These protozoan spores were flown by the National Aeronautics and Space Administration (NASA) Columbia Scientific Balloon Facility (CSBF) flight 667NT, launched from its base in Fort Sumner, New Mexico. F. septica spores were exposed to stratospheric conditions on board the NASA/CSBF 667 balloon flight for 9 h. The spores obtained after the flight and those from the control box that stayed at ground maintained the same size and morphology, as will be shown in this work. The spores retained viability, and all life cycle stages were obtained by in vitro culture. Moreover, some life cycle events were observed for the first time in F. septica.

The life cycles of two species of Myxomycetes in Physarales, Physarum rigidum and Didymium squamulosum.

Myxomycetes are eukaryotic microorganisms containing characteristics akin to both fungi and amoebae. They can complete their whole life cycles while being cultured on agar media, and under-laboratory conditions, which favors taxonomic, phylogenetic, and cytological researches. Here, we describe the life cycles of two such species: Didymium squamulosum collected from the field and Physarum rigidum cultured from moist chamber both belonging to the Order Physarales. Three per cent oat-agar media (OAM) was used to culture the plasmodia until they aggregated and were almost starved. Natural light was then applied to the plasmodia to induce fructification. Their life cycles share the same common stages, namely: spore, myxamoebae, swarm cell, plasmodia, and sporulation. In this study, we describe the morphogenesis from spore to spore of two species by differential interference contrast (DIC) and stereoscopic microscopies, as well as discuss the differences between the development of both species and interspecies. We found that the spore germination method of both species was the same. However, there were differences noted in time taken and fruiting body formation. Unlike P. rigidum, the species D. squamulosum did not require natural light stimulation. Moreover, the maturation process of both species had similar color transitions but exhibited distinct morphology in each developmental stage except during the swarm cell stage.

Didymium azorellae, a new myxomycete from cushion plants of cold arid areas of South America.

A new species of Didymium (Myxomycetes), D. azorellae, isolated from plant debris in a moist chamber, collected during studies of cold arid areas of Argentina and Peru, is described. It can be distinguished by its small size, the tightly packed layer of lime crystals on the peridium, the very scant, or absent, capillitium, and the unique spore ornamentation, especially by scanning electron microscopy. The species developed on dead leaves of cushion plants growing in the extremely harsh environments of the central Andean puna at almost 5000 m elevation and the Andino-Patagonian steppe. Morphology was examined with scanning electron microscopy and light microscopy, and micrographs of relevant details are included here. In order to confirm the identity of the new species described in this paper, a molecular study was conducted based on partial sequences of both the 18S rRNA and the elongation factor 1-alpha gene. Phylogenetic analysis including two specimens from different countries of the newly described species, Didymium azorellae, strongly supports the grouping of these specimens as a separate clade from the rest of the analyzed species.

Mycetozoal bloom in a hydroponic culture of garden cress (Lepidium sativum L.).

The role and importance of the true slime moulds (mycetozoans, Mycetozoa, formerly Myxomycetes) for agriculture and food industry are poorly documented, most probably because of a low popularity of these "macroscopic microorganisms" among researchers in the past. Here we report probably for the first time the massive occurrence of true slime moulds in their vegetative, plasmodial form in a product intended for direct consumption, i.e. in a household hydroponic culture of garden cress (Lepidium sativum L.). The plasmodia gradually produced numerous, stalked or sessile sporangia and plasmodiocarps, which made it possible to identify them as Didymium species complex (the stalked sporangia) and Didymium difforme (Pers.) Gray (the sessile sporangia and plasmodiocarps). The mycetozoans were transferred to in vitro culture where they were maintained for several weeks on oat flour. We briefly discuss the importance of this observation from the point of view of biochemical interactions between the plant and the true slime moulds in a general context of slime mould biology. Our observation indicates that the presence of mycetozoan material in food products may be frequent, while its influence on food safety and quality remains unknown.

Sclerotia of the acellular (true) slime mould Fuligo septica as a model to study melanization and anabiosis.

Acellular (true) slime moulds (Myxomycetes) are capable of a transition to the stage of sclerotium - a dormant form of plasmodium produced under unfavourable environmental conditions. In this study, sclerotia of Fuligo septica were analyzed by means of electron paramagnetic resonance (EPR) spectroscopy. The moulds were cultivated in vitro on filter paper, fed with oat flour, and kept until the plasmodia began to produce sclerotia. The obtained sclerotia differed in colour from yellow through orange to dark-brown. The EPR spectra revealed a free radical, melanin-like signal correlated with the depth of the colour; it was strongest in the dark sclerotia. Sclerotization only took place when the plasmodia were starved and very slowly dried. Only the yellow sclerotia were able to regenerate into viable plasmodia. This suggests that myxomycete cytoplasm dehydration is an active process regulated metabolically. Plasmodial sclerotization may therefore serve as a convenient model system to study the regulation of cytoplasmatic water balance, and sclerotia as a convenient material for EPR measurements, combining the quality of plasmodia with the technical simplicity of the measurements characteristic of dry spores. Darkening of the sclerotia is most probably a pathological phenomenon connected with the impairment of water balance during sclerotization.

Description and culture of a new succulenticolous Didymium (Myxomycetes).

A new succulenticolous Myxomycete species, Didymium wildpretii, found on decaying remains of various species of cacti, is described from two arid zones of the world. This species was collected from central Mexico, at the southern limit of the Chihuahuan Desert, and from the Canary Islands (Spain). The new species has small, pale yellow sporocarps, 0.1-0.7 mm high, that are sessile or have short, orange-yellow, calcareous stalks and small, uniformly warted spores. The stability of the taxonomic characters of the species was confirmed with both moist chamber cultures and spore-to-spore culture on agar. Life cycle events are described from germination to sporulation. Myxomycete specimens were examined with scanning electron microscopy and light microscopy, and micrographs of relevant morphological details are included.

Mitochondrial inheritance patterns in Didymium iridis are not influenced by stage of mating competency.

To test whether the timing of transition to mating competency affected mitochondrial transmission patterns in D. iridis. Reciprocal crosses were made by combining mating compatible strains that differed in their competency to mate. The results were compared to crosses where both mating strains were competent at the time of combining and crosses where somatic fusion of plasmodia was allowed. The results show that the mating competency of the parental strains at the time of confronting a compatible mate does not affect mitochondrial transmission patterns, mating efficiency or the likelihood of biparental inheritance. However the timing of plasmodial formation is delayed when precompetent and competent strains are mated compared to when both strains are competent at the time of mixing. We also observed that somatic fusion of plasmodia did not appreciably increase the incidence of biparental inheritance compared to crosses where individual plasmodia were isolated. These results provide additional evidence of the variable nature of mitochondrial inheritance in D. iridis within crosses and between mating trials.

In vivo expression of a group I intron HEG from the antisense strand of Didymium ribosomal DNA.

Two different isolates of the myxomycete Didymium iridis harbour homing endonuclease genes that are expressed from group I introns inserted into identical sites within the small subunit ribosomal DNA. The homing endonuclease proteins are related in sequence, and their gene structures share similar features such as the presence of small spliceosomal introns and functional polyadenylation sites. However, they are transcribed from opposite strands of the ribosomal DNA and presumable by different RNA polymerases. We have previously described the in vivo expression of the I-DirI homing endonuclease from within the ribosomal RNA precursor. In this paper, we demonstrate the in vivo expression of the I-DirII homing endonuclease from the opposite strand of the Didymium rRNA gene. A comparison of the expression strategies of the two genes demonstrates the feasibility of antisense expression and provides insight into nucleolar gene expression.