Fifteen new species from the myxomycete genus Lycogala.

Based on a study of 255 collections from four continents and four floristic kingdoms, we describe 15 new species of the genus Lycogala. The new species, all morphologically close to L. epidendrum, L. exiguum, and L. confusum, differ from each other by the structure of the peridium and, in some cases, also by the color of the fresh spore mass and the ornamentation of the capillitium and spores. Species delimitation is confirmed by two independently inherited molecular markers, as well as previously performed tests of reproductive isolation and genetic distances. We studied authentic material of L. exiguum and L. confusum and found fresh specimens of these species, which allowed us to obtain molecular barcodes and substantiate the separation of new species from these taxa. We propose to retain the name L. epidendrum for the globally most abundant species, for which we provide a more precise description and a neotypification. Two formerly described species, L. leiosporum and L. fuscoviolaceum, we consider to be dubious. We do not recognize the species L. terrestre.

Speciation in progress? A phylogeographic study among populations of Hemitrichia serpula (Myxomycetes).

Myxomycetes (plasmodial slime molds, Amoebozoa) are often perceived as widely distributed, confounding to the "everything is everywhere" hypothesis. To test if gene flow within these spore-dispersed protists is restricted by geographical barriers, we chose the widespread but morphologically unmistakable species Hemitrichia serpula for a phylogeographic study. Partial sequences from nuclear ribosomal RNA genes (SSU) revealed 40 ribotypes among 135 specimens, belonging to three major clades. Each clade is dominated by specimens from a certain region and by one of two morphological varieties which can be differentiated by SEM micrographs. Partial sequences of the protein elongation factor 1 alpha (EF1A) showed each clade to possess a unique combination of SSU and EF1A genotypes. This pattern is best explained assuming the existence of several putative biospecies dominating in a particular geographical region. However, occasional mismatches between molecular data and morphological characters, but as well heterogeneous SSU and heterozygous EF1A sequences, point to ongoing speciation. Environmental niche models suggest that the putative biospecies are rather restricted by geographical barriers than by macroecological conditions. Like other protists, myxomycetes seem to follow the moderate endemicity hypothesis and are in active speciation, which is most likely shaped by limited gene flow and reproductive isolation.

A critical revision of the Tubifera ferruginosa complex.

Based on a combination of morphological and molecular investigations, a critical revision of the widely distributed myxomycete Tubifera ferruginosa is presented. A phylogeny of the morphospecies, based on partial 18S nuc rDNA sequences, displays several clearly distinct clades, all differing by a genetic distance (p distance) of at least 0.15, with the distance within the clades below 0.11. These molecular differences correlate with morphological characters, such as the shape of sporothecal tips, the color of immature fructifications and the ultrastructure of the inner surface of the peridium. The combination of morphological and molecular data provides evidence that T. ferruginosa is actually a species complex, representing at least seven species. These are T. ferruginosa sensu stricto, T. applanata, T. corymbosa, T. dudkae, T. magna, T. montana and T. pseudomicrosperma. Among these T. applanata and T. dudkae (as Reticularia dudkae) were described recently based on morphological characters and the 18S nuc rDNA phylogeny confirmed their separation. Another four species, T. corymbosa, T. magna, T. montana and T. pseudomicrosperma, are described here. We propose an epitype for T. ferruginosa sensu stricto and recognize subsp. ferruginosa and subsp. acutissima within this species. All studied taxa of the T. ferruginosa complex are shown to lack a capillitium. Structures formerly described as capillitium represent the hyphae of fungi occurring within the fructifications.

Perichaena longipes, a new myxomycete from the Neotropics.

A new species of myxomycete, Perichaena longipes, is described from 56 sporocarp specimens that appeared in moist chamber cultures prepared with samples of decaying plant materials collected in Panama, Costa Rica and Brazil. This new species is distinguished from the morphologically similar species P. pedata on the basis of the much longer stipe, lighter peridium and the unique ornamentation of the capillitium. The nuc 18S ribosomal DNA sequences obtained from four specimens of P. longipes support the distinction of this new taxon and its separation from P. pedata. Furthermore, maximum likelihood phylogeny supports earlier evidence that species currently within the genus Perichaena do not form a monophyletic clade. Instead they appear to form three separate branches within the bright-spored clade. The first clade includes P. longipes together with several species of Trichia and Metatrichia, the second includes P. pedata and P. chrysosperma, and the third clade is composed of P. corticalis, P. depressa and P. luteola.

Nivicolous Stemonitales from the Austral Andes: analysis of morphological variability, distribution and phenology as a first step toward testing the large-scale coherence of species and biogeographical properties.

Nivicolous myxomycetes occur at the edge of spring-melting snow in mountainous areas. They are mostly considered cosmopolitan species morphologically and ecologically uniform across their entire distribution ranges. Thus, long-distance dispersal has been suggested to be the main mechanism shaping their ranges and geographical variability patterns. To test this hypothesis we conducted the first detailed analysis of morphological variability, occurrence frequency and phenology of nivicolous myxomycetes collected in the hitherto unexplored Austral Andes of South America (southern hemisphere = SH) in the comparative context of data from the northern hemisphere (NH). We used Stemonitales, the most representative and numerous taxonomic order in nivicolous myxomycetes, as a model. A total of 131 South American collections represented 13 species or morphotypes. One of them, Lamproderma andinum, is new to science and described here. Several others, L. aeneum, L. album, L. pulveratum, "Meriderma aff. aggregatum ad. int.", M. carestiae and "M. spinulosporum ad. int.", were previously unknown from the SH. Lamproderma ovoideum is reported for the first time from South America and Collaria nigricapillitia is new for Argentina. The fine-scale morphological analysis of all species from the study area and reference NH material demonstrated a high intraspecific variability in most of them. This suggests isolation and independent evolutionary processes among remote populations. On the other hand, the uniform morphology of a few species indicates that long-distance dispersal is also an effective mechanism, although not as universal as usually assumed, in some nivicolous myxomycetes. Analysis of nivicolous species assemblages also showed significant differences among major geographic regions in that the Stemonitales were significantly less common in the SH than in the NH. Furthermore, the occurrence of nivicolous species in summer and autumn, out of the typical phenological season, is recognized as a possible distinctive phenomenon for the SH populations.

The genus Alwisia (Myxomycetes) revalidated, with two species new to science.

Based on morphological investigations and a phylogeny constructed with partial sequences of the SSU rDNA gene, we revalidate the genus Alwisia and propose the combination Alwisia bombarda Berk. & Broome to be used against Tubifera bombarda (Berk. & Broome) G.W. Martin. Two new species, Alwisia morula and A. repens, are described based on material collected respectively in Costa Rica and Australia. Both new species lack a capillitium and possess individually stalked subspherical sporothecae. Alwisia repens differs from A. morula by its procumbent stalks and iridescent peridium. A comparison of 83 sequences of species in the genera Lycogala, Reticularia and Tubifera with a recent two-gene phylogeny of the bright-spored myxomycetes resulted in a similar topology of both Bayesian and maximum likelihood trees and placed A. bombarda, A. morula and A. repens in one well delimited clade within Reticulariaceae.

Perichaena megaspora, a new nivicolous species of myxomycete from the Andes.

A new nivicolous species of Perichaena is described from the Andes in Argentina. The most conspicuous characteristics of Perichaena megaspora are the large spores and their ornamentation in the form of flattened warts. The 16-21 µm diam spores make the new species unique in the genus in which all other species have spores rarely reaching 15 µm diam. Twenty-two collections were found in the field during two consecutive years at 10 localities in Mendoza province Argentina, including one collection isolated from a moist chamber culture of ground litter. The new species was examined under stereomicroscope, light microscope and scanning electron microscope and micrographs of relevant details are included.

Photonic simulation method applied to the study of structural color in Myxomycetes.

We present a novel simulation method to investigate the multicolored effect of the Diachea leucopoda (Physarales order, Myxomycetes class), which is a microorganism that has a characteristic pointillistic iridescent appearance. It was shown that this appearance is of structural origin, and is produced within the peridium -protective layer that encloses the mass of spores-, which is basically a corrugated sheet of a transparent material. The main characteristics of the observed color were explained in terms of interference effects using a simple model of homogeneous planar slab. In this paper we apply a novel simulation method to investigate the electromagnetic response of such structure in more detail, i.e., taking into account the inhomogeneities of the biological material within the peridium and its curvature. We show that both features, which could not be considered within the simplified model, affect the observed color. The proposed method is of great potential for the study of biological structures, which present a high degree of complexity in the geometrical shapes as well as in the materials involved.

Description and life cycle of a new Physarum (Myxomycetes) from the Atacama Desert in Chile.

A new species of Physarum (Myxomycetes), Physarum atacamense is described in this paper, and details are provided on its life cycle as observed in spore-to-spore culture in agar. The new species was collected during studies of the Atacama Desert in Chile. It has been collected directly in the field and isolated in moist chamber cultures prepared with material from an endemic cactus. The combination of characters that make this species unique in the genus are its large fusiform nodes of the capillitium, its long, bicolored stalk and the very dark brown and densely warted angular spores. The morphology of specimens of this myxomycete was examined with scanning electron microscopy and light microscopy, and micrographs of relevant details and life cycle stages are included in this paper. The importance of resistant stages in the life cycle of this myxomycete is stressed, and the close association of this myxomycete with its plant substrates is discussed.

Licea eremophila, a new myxomycete from arid areas of South America.

A new stipitate species of myxomycete of the genus Licea is described based on material from arid areas in Argentina and Chile. It was isolated from moist chamber cultures and found fruiting on field collections, usually on the same substrate, Puya sp. (Bromeliaceae). It differs from all described species in the genus in that it has stipitate sporocarps with dehiscence by defined preformed platelets and a smooth inner peridial surface. The new species has polyhedral, yellow spores with a uniform thick spore wall and dense warts except on irregularly dispersed raised bands with fewer warts, visible by SEM, an ornamentation not previously observed in the genus. Life-cycle events are described and illustrated, from germination to sporulation, based on moist chamber and agar cultures. The morphology of the myxomycete specimens was examined with scanning electron microscopy and light microscopy, and both light and SEM micrographs of relevant details are included.

Structural color in Myxomycetes.

In this paper we report evidence of structural color in Myxomycetes, a group of eukaryotic microorganisms with an uncertain taxonomic position. We investigated the Diachea leucopoda, which belongs to the Physarales order, Myxomycetes class, and found that its peridium -protective layer that encloses the mass of spores- is basically a corrugated layer of a transparent material, which produces a multicolored pointillistic effect, characteristic of this species. Scanning (SEM) and transmission (TEM) electron microscopy techniques have been employed to characterize the samples. A simple optical model of a planar slab is proposed to calculate the reflectance. The chromaticity coordinates are obtained, and the results confirm that the color observed is a result of an interference effect.

Two new species of nivicolous Lamproderma (myxomycetes) from the mountains of Europe and America.

As a result of the revision of European and American collections of genus Lamproderma two new nivicolous myxomycete species, Lamproderma argenteobrunneum and L. kowalskii, are described. The new species are characterized by the silvery-brown sporothecae, the areolate peridium and the ferruginous-brown spores in mass. They differ from one another mainly in spore size and ornamentation, the form of the capillitium and the stalk length and ratio of the stalk length to the total height of the sporocarp. The morphology of the new species was examined with light microscopy and scanning electron microscopy (SEM), and micrographs of relevant details are included. Lamproderma argenteobrunneum also was obtained in moist chamber culture, and the mature sporocarps displayed all features typical of field-collected samples. The known geographical distribution of L. argenteobrunneum includes the main ranges of the European alpine system (Alps, Carpathians, Pyrenees) as well as those of North America, while L. kowalskii has been recorded so far from several sites in California in the United States.

In the shadow of Darwin: Anton de Bary's origin of myxomycetology and a molecular phylogeny of the plasmodial slime molds.

In his Origin of Species (John Murray, London, 1859), Charles Darwin described the theory of descent with modification by means of natural selection and postulated that all life may have evolved from one or a few simple kinds of organisms. However, Darwin's concept of evolutionary change is entirely based on observations of populations of animals and plants. He briefly mentioned 'lower algae', but ignored amoebae, bacteria and other micro-organisms. In 1859, Anton de Bary, the founder of mycology and plant pathology, published a seminal paper on the biology and taxonomy of the plasmodial slime molds (myxomycetes). These heterotrophic protists are known primarily as a large composite mass, the plasmodium, in which single nuclei are suspended in a common 'naked' cytoplasm that is surrounded by a plasma membrane. Here we summarize the contents of de Bary's 1859 publication and highlight the significance of this scientific classic with respect to the establishment of the kingdom Protoctista (protists such as amoebae), the development of the protoplasmic theory of the cell, the introduction of the concept of symbiosis and the rejection of the dogma of spontaneous generation. We describe the life cycle of the myxomycetes, present new observations on the myxamoebae and propose a higher-order phylogeny based on elongation factor-1 alpha gene sequences. Our results document the congruence between the morphology-based taxonomy of the myxomycetes and molecular data. In addition, we show that free-living amoebae, common protists in the soil, are among the closest living relatives of the myxomycetes and conclude that de Bary's 'Amoeba-hypothesis' on the evolutionary origin of the plasmodial slime molds may have been correct.

Description and life cycle of a new Didymium (Myxomycetes) from arid areas of Argentina and Chile.

A new species of Didymium (Myxomycetes), D. infundibuliforme, is described herein, and details are provided on its life cycle as observed in spore to spore culture on agar. The new species was recorded during intensive studies of areas of the Monte Desert in Argentina and the Atacama Desert in Chile. It has been collected directly in the field in both countries on several occasions over 4 y and isolated in moist chamber cultures prepared with material from native plant species. The characters that make this species unique in the genus are its funnel-shape sporocarps with white stalks, the apical circumscissile dehiscence of the sporotheca that causes the base to resemble a calyculus and the ornamentation on the spores. The morphology of specimens of this new myxomycete was examined with scanning electron microscopy and light microscopy, and micrographs of relevant details are included in this paper.

Two new species of Perichaena (Myxomycetes) from arid areas of Russia and Kazakhstan.

Two new myxomycete species from dry steppe and desert communities of the Caspian Lowland (Russia) and central Kazakhstan are described and illustrated. They are placed tentatively within genus Perichaena, which does include species with a reduced capillitium and single-layered peridium. Both species were found repeatedly in moist chamber cultures; P. heterospinispora appeared on leaf litter and twigs, whereas P. polygonospora occurred on leaf litter and weathered dung of rodents. Both species have spore ornamentation that is unique for members of genera Licea and Perichaena. The spore ornamentation of the first species includes scattered large, pyramid-like spines 0.9-1.2 microm high that sometimes have enlarged ends. Among these spines the spore surface is covered by evenly and densely distributed warts that are visible only by SEM. The second species is characterized by angular spores with a coarse network of rounded ridges. The areas among these ridges bear scattered composite warts 0.3-0.5 microm high that sometimes coalesce to form clusters but more often are distributed evenly and densely and are visible only by SEM. The stability of the taxonomic characters of both species was confirmed by several collections from different regions obtained in 2 y. The morphology of the fructifications of the two myxomycetes was examined with both scanning electron and light microscopy, and micrographs of all relevant features are presented.

Morphology and life cycle of a new species of Didymium (Myxomycetes) from arid areas of Mexico.

A new species of myxomycete, Didymium umbilicatum, isolated from the bark of Agavaceae, is described from arid zones of Mexico. This species was obtained from moist chamber cultures of Yucca spp. bark, collected in four different years from two states (Puebla and Querétaro) in central Mexico and found in the field from Hidalgo, Oaxaca and Puebla on the dead remains of Agave sp. The new species has small, flat, white sporocarps or short plasmodiocarps, 0.2-1.3 mm diam, and 0.15-0.4 mm tall. They are sessile on a reduced base or have a short, calcareous pale stalk and warted spores, warts fused in an irregular subreticulum by SEM. It is the sixth species of Didymium recently described from arid areas. The stability of the taxonomic characters of the species was confirmed by spore-to-spore culture on agar. Life cycle events are described from germination to sporulation. The morphology of the myxomycete specimens was examined with scanning electron microscopy and light microscopy, and micrographs of relevant details are included.

Labyrinthula terrestris sp. nov., a new pathogen of turf grass.

A species of Labyrinthula that causes 'rapid blight' and death of turfgrass has been isolated and studied. We name this new species Labyrinthula terrestris and briefly summarize morphological characteristics and growth patterns of this pathogen of turfgrass.

Microtubule centers and the interphase microtubule cytoskeleton in amoebae of the cellular slime molds (Mycetozoans) Acytostelium leptosomum and Protostelium mycophaga.

We investigated the microtubule (MT) cytoskeleton and microtubule centers (MTC) in undifferentiated amoebae by indirect immunofluorescence with six monoclonal antitubulin antibodies, and by transmission electron microscopy and immunogold ultracytochemistry. Interphase amoebae of both species contain a distinct cytoplasmic complex of MTs, which is more elaborate in Protostelium mycophaga. In Acytostelium leptosomum amoebae a single MTC is attached to each interphase nucleus at its pointed end, as in the other dictyostelid cellular slime molds Dictyostelium discoideum and Polysphondylium violaceum. Ultrastructurally, MTCs of A. leptosomum also resemble those of these two species: They consist of an electron-opaque core shaped like a stout rod, which is embedded, together with nodules, in a fuzzy matrix. The nodules are the points of origin of the MTs. In most amoebae of P. mycophaga there are two MTCs on opposite sides of and close to the nucleus, but many amoebae also contain a variable number of MTCs that are remote from the nucleus. Nucleus-associated and "remote" MTCs are structurally identical. They consist of a ring-shaped core with inner and outer diameters of ca. 130 nm and 340 nm. A plug sits in the ring, and satellites are connected to the core by fine fibrils. The satellites are the points of origin of MTs. New MTCs are apparently formed during mitosis, the parent MTC probably serving as a template for the genesis of a new ring. The results support the notion that phylogenetically related organisms have similarly constructed MTCs and that these are dissimilar in less closely related organisms.

Spatial patterns in the fruiting bodies of the cellular slime mold Polysphondylium pallidum.

During morphogenesis in the slime mold Polysphondylium pallidum cell masses are periodically pinched off from the base of the developing sorogen. These masses round up and differentiate into secondary sorogens, which become radially ordered arrays of secondary fruiting bodies called whorls. Here we describe the morphogenesis of P. pallidum and characterize the spacing of whorls along the central stalk of the fruiting body and the spacing of sorocarps within whorls. We find both are highly regular. We propose that the linear spacing of whorls can be accounted for satisfactorily by a model that views the periodic release of cell masses from the base of the developing sorogen as the consequence of an imbalance between forces that orient amoebae toward the tip of the culminating sorogen, and cohesive forces between randomly moving cells in the basal region of the sorogen, which act as a retarding force. The orderly arrangement of fruiting bodies within whorls can be explained most easily by models that employ short-range activation and lateral inhibition.

Obligate amoebae of the protostelids: significance for the concept of Eumycetozoa.

Most flagellate Eumycetozoa have a non-flagellate, or obligately amoeboid, trophic state which differentiates from the amoebo-flagellate state and gives rise to the fruiting body. This study examines the morphology, general ultrastructure, and microtubular systems of the obligate amoebae of three flagellate protostelids with typical amoebo-flagellate states. The obligate amoebae of all three species are morphologically distinct indicating that the obligately amoeboid state has evolved independently in the history of each species. Therefore, obligate amoebae may be useful for defining separate evolutionary groups within the Eumycetozoa.