Evidence of fungal decay in petrified legume wood from the Neogene of the Bengal Basin,India

Silicified fossil legume woods of Cynometroxylon Chowdhury & Ghosh collected from the Neogene (late Miocene) sediments of the Bengal Basin, eastern India, exhibit fungal decay seldom found in the fossil record. The wood possesses numerous perforate areas on the surface that seem to be the result of extensive fungal activity. In transverse section, the decayed areas (pockets) appear irregular to ellipsoidal in outline; in longitudinal section these areas of disrupted tissue are somewhat spindle-shaped. Individual pockets are randomly scattered throughout the secondary xylem or are restricted to a narrow zone. The aforesaid patterns of decay in fossil wood show similarities with that of white rot decay commonly produced by higher fungi, specifically basidiomycetes and ascomycetes. The host fossil wood harbors abundant ramifying and septate fungal hyphae with knob like swellings similar to pseudoclamps in basidiomycetes, and three-celled conidia-like reproductive structures. This record expands our current knowledge of wood decaying fungi-host plant interaction in the Neogene tropical forests of Peninsular India.     

Detection and Identification of Decay Fungi in Spruce Wood by Restriction Fragment Length Polymorphism Analysis of Amplified Genes Encoding rRNA

We have developed a DNA-based assay to reliably detect brown rot and white rot fungi in wood at different stages of decay. DNA, isolated by a series of CTAB (cetyltrimethylammonium bromide) and organic extractions, was amplified by the PCR using published universal primers and basidiomycete-specific primers derived from ribosomal DNA sequences. We surveyed 14 species of wood-decaying basidiomycetes (brown-rot and white-rot fungi), as well as 25 species of wood-inhabiting ascomycetes (pathogens, endophytes, and saprophytes). DNA was isolated from pure cultures of these fungi and also from spruce wood blocks colonized by individual isolates of wood decay basidiomycetes or wood-inhabiting ascomycetes. The primer pair ITS1-F (specific for higher fungi) and ITS4 (universal primer) amplified the internal transcribed spacer region from both ascomycetes and basidiomycetes from both pure culture and wood, as expected. The primer pair ITS1-F (specific for higher fungi) and ITS4-B (specific for basidiomycetes) was shown to reliably detect the presence of wood decay basidiomycetes in both pure culture and wood; ascomycetes were not detected by this primer pair. We detected the presence of decay fungi in wood by PCR before measurable weight loss had occurred to the wood. Basidiomycetes were identified to the species level by restriction fragment length polymorphisms of the internal transcribed spacer region.     

Detection and identification of decay fungi in spruce wood by restriction fragment length polymorphism analysis of amplified genes encoding rRNA

We have developed a DNA-based assay to reliably detect brown rot and white rot fungi in wood at different stages of decay. DNA, isolated by a series of CTAB (cetyltrimethylammonium bromide) and organic extractions, was amplified by the PCR using published universal primers and basidiomycete-specific primers derived from ribosomal DNA sequences. We surveyed 14 species of wood-decaying basidiomycetes (brown-rot and white-rot fungi), as well as 25 species of wood-inhabiting ascomycetes (pathogens, endophytes, and saprophytes). DNA was isolated from pure cultures of these fungi and also from spruce wood blocks colonized by individual isolates of wood decay basidiomycetes or wood-inhabiting ascomycetes. The primer pair ITS1-F (specific for higher fungi) and ITS4 (universal primer) amplified the internal transcribed spacer region from both ascomycetes and basidiomycetes from both pure culture and wood, as expected. The primer pair ITS1-F (specific for higher fungi) and ITS4-B (specific for basidiomycetes) was shown to reliably detect the presence of wood decay basidiomycetes in both pure culture and wood; ascomycetes were not detected by this primer pair. We detected the presence of decay fungi in wood by PCR before measurable weight loss had occurred to the wood. Basidiomycetes were identified to the species level by restriction fragment length polymorphisms of the internal transcribed spacer region.     

FUNGI FROM THE LOWER DEVONIAN RHYNIE CHERT: CHYTRIDIOMYCETES

Several different chytridiomycetes are described from the Lower Devonian (Siegenian) Rhynie chert. Included are both eucarpic and apparently holocarpic forms that occur in Palaeonitella, Aglaophyton, Lyonophyton, Horneophyton, and clusters of algal cells, as well as in the surrounding chert matrix. Holocarpic types consist of endobiotic sporangia, each characterized by one discharge tube. Sporangia can be traced from the thallus stage to the discharge of zoospores. Monocentric and polycentric eucarpic chytrids are associated with the miospores of Aglaophyton and various thick-walled fungal spores. In these forms the sporangia are variable in size and shape ranging up to 30 μm. Most appear to be inoperculate and there is evidence that the sporangium ruptured on the distal surface. Some contain zoospores with flagella. One operculate eucarpic form had parasitized the cellular gametophyte emerging from the proximal surface of an Aglaophyton spore. Several of the Rhynie chert chytrids are comparable with a number of extant forms (e.g., Olpidiaceae and Spizellomycetaceae), while others possess features that encompass several groups. These fossil fungi are discussed in the context of their interactions with other organisms in this Lower Devonian freshwater paleoecosystem.     

STUDIES OF PALEOZOIC FUNGI. III. FUNGAL PARASITISM IN A PENNSYLVANIAN GYMNOSPERM

Evidence of fungal parasitism is found in the Pennsylvanian gymnospermous cone, Lasiostrobus polysacci Taylor. Indication of fungal activity is found in the outer cortical region of the axis of the cone and in the fleshy microsporophylls. Specimens exhibit severe tissue disruption, thick-walled, branched, septate hyphae, and possible reproductive structures. Parenchymatous cortical cells may also contain rounded bodies which are continuous with the cell wall. Similar structures are formed in many extant taxa in response to fungal invasion, and are termed wall appositions or callosities. Although their role in extant plants is disputed, they are clearly the product of a living host cell. Such spherical bodies, however, are not restricted to the cell periphery but in some cases occlude the cell lumen. In appearance they resemble resinous remains similar to those found in other coal ball plants. The blockage of entire cells or groups of cells may have served to retard hyphal growth or isolate infected cells. The occurrence of such structures in a Carboniferous plant provides the best evidence to date of parasitism during the Paleozoic.     

Fungal decomposition of woody debris of Castanopsis sieboldii in a subtropical old-growth forest

Fungi, especially basidiomycetes, are the primary agents of woody debris decomposition in terrestrial forest ecosystems. However, quantitative data regarding the abundance and decay activity of wood-inhabiting fungi are lacking, especially for tropical and subtropical areas. This study demonstrates the dynamics of decay columns of wood-inhabiting fungi within decaying woody debris of Castanopsis sieboldii and the wood decay activities of those fungi in a subtropical natural forest. Among six basidiomycetes and two ascomycetes observed as sporocarps on fallen boles of C. sieboldii, Microporus affinis was most abundantly observed in terms of frequency of sporocarps and as percentage area of decay columns within cross-sections of boles, especially those in the early stages of decomposition. In decay columns of M. affinis, both acid-unhydrolyzable residue (AUR) and holocellulose decayed simultaneously, and wood relative density decreased to 45.8% of that of fresh C. sieboldii wood. A pure culture decay test under laboratory conditions showed that M. affinis was a strong decomposer of AUR and holocellulose. These results suggest that M. affinis has a central role in lignocellulose decomposition of wood of C. sieboldii in the early stages of decomposition.     

Inhibition and Stimulation Effects in Communities of Wood Decay Fungi: Exudates from Colonized Wood Influence Growth by Other Species

The effects of exudates from uncolonized and from partly decayed beech wood on the extension rates of 16 later stage decay fungi were investigated. The partly decayed wood had been colonized by the pyrenomycete Eutypa spinosa, or the basidiomycetes Fomes fomentarius, Stereum hirsutum, and Trametes versicolor, all known as common early decay agents in European beech forests. Sterilized wood pieces were placed onto 0.5% malt agar, opposite to small agar plugs containing the test fungi. The latter showed very variable and species-specific growth responses to the various wood types. The presence of uncolonized wood stimulated extension rates in many species, whereas the four previously decayed wood types had variable stimulatory or inhibitory effects. Wood decayed by S. hirsutum resulted in reduced extension rate, delayed growth, or total inhibition in the majority of species, thus it is suggested that this species uses secondary metabolites in a defensive strategy. A single species was, however, stimulated in the presence of S. hirsutum-decayed wood. In contrast, the presence of wood decayed by F. fomentarius was stimulatory to 45% of the species. The other previously decayed wood types generally resulted in more variable responses, depending upon species. The results are discussed in an ecological context and it is suggested that the exudates from the partly decayed wood that are responsible for the reported effects may function as infochemicals, structuring microbial communities in wood.     

Evidence of fungal activity in silicified gymnosperm wood from the Eocene of southern Patagonia (Argentina)

Evidence of fungal activity expressed as typical decay patterns is described from silicified podocarpaceous wood from the Eocene of Patagonia, Argentina. Decay features consist of tracheids of the secondary xylem that are degraded, resulting in thin-celled, lignin-free, translucent, circular to elliptical areas, some of which have cells devoid of all cell wall components including lignin, hemicellulose, and cellulose, and other areas that show only partial simultaneous decay of all cell wall layers. These patterns conform to the white rot and its variant white pocket rot decay patterns produced by basidiomycetes and ascomycetes in gymnosperm and angiosperm wood in modern terrestrial ecosystems. Coagulated opaque bodies in the lumen of some cells and enlarged secondary walls may represent host reactions to infection or remains of metabolic products of fungal enzymatic activity. Similar decay patterns and reaction features have been described from fossil woods ranging in age from the Devonian to the present. This record expands the fossil record of wood rot fungi and underscores their importance as drivers of biological cycles in ancient terrestrial ecosystems.     

An unusual microfungus in a fungal spore from the Lower Devonian Rhynie chert

Abstract: A new fossil microfungus, Kryphiomyces catenulatus gen. et sp. nov., occurs as an endobiotic mycelial thallus in a large spore of a glomeromycotan fungus from the Lower Devonian Rhynie chert. The thallus consists of branched (?pseudo‐)septate hyphae with numerous catenulate swellings. Some hyphal tips produce spherical reproductive structures or propagules. Hyphal morphology in K. catenulatus is reminiscent of that in certain extant Hyphochytridiomycota, Chytridiomycota, and even Ascomycota, but specific diagnostic features that allow assignment of the fossil to modern groups are absent. The discovery of this interfungal association broadens our knowledge about the diversity of microfungi and their intricate associations in early continental ecosystems.     

A blood-brain barrier without tight junctions in the fly central nervous system in the early postembryonic stage

Summary The anatomical basis of the vertebrate blood-brain barrier is a series of tight junctions between endothelial cells of capillaries in the central nervous system. Over two decades ago, tight junctions were also proposed as the basis of the blood-brain barrier in insects. Currently there is a growing understanding that septate junctions might possess barrier properties in various invertebrate epithelial cells. We now examine these two views by studying the blood-brain barrier properties of the early postembryonic larva of a dipteran fly (Delia platura) by transmission electron microscopy. Newly hatched larvae possess a functioning blood-brain barrier that excludes the extracellular tracer, ionic lanthanum. This barrier is intact throughout the second instar stage as well. The ultrastructural correlate of this barrier is a series of extensive septate junctions that pervade the intercellular space between adjacent perineurial cells. No tight junctions were located in either nerve, glial or perineurial cell layers. We suggest that the overall barrier might involve septate junctions within extensive, meandering intercellular clefts.     

Studies on perineural junctional complexes and the sites of uptake of microperoxidase and lanthanum in the cockroach central nervous system

The perineurial junctional complexes in the nerve cord of Periplaneta americana have been shown to consist of septate desmosomes, extensive gap junctions and relatively limited regions of tight junctions. Microperoxidase (M.W. 1,900) undergoes limited intercellular penetration into the septate desmosomes. Lanthanum penetrates both the septate desmosomes and gap junctions. It is concluded that the restricted access of these substances to the underlying extracellular spaces results from the presence of the perineurial tight junctions. These results contrast with those for small peripheral nerves, which lack equivalent junctional complexes, and in which the extracellular spaces are found to be accessible to externally applied lanthanum. The results are discussed in relation to current concepts of the insect blood-brain barrier.     

Assessment of the biocontrol potential of a Trichoderma viride isolate: Part II: Protection against soft rot and basidiomycete decay

A previous paper reported on the establishment of a field and fungal cellar trial set up to determine the biocontrol potential of a specific Trichoderma isolate against wood decay fungi. This paper reports on the analyses used to examine the protective effect of the selected isolate, and presents results indicating an initial protective effect against both basidiomycetes and soft rot fungi. The parameters assessed in the field and fungal cellar trials were soft rot decay, basidiomycete decay, Trichoderma colonisation, moisture content and nitrogen content. The results of these analyses show that the introduction of a biological control agent has had a significant effect on moisture content, decay and nitrogen content. A protective effect has been observed against soft rot and basidiomycete decay fungi in field samples.     

Wood-inhabiting ligninolytic basidiomycetes in soils: Ecology and constraints for applicability in bioremediation

Ligninolytic basidiomycetes associated with wood decay are a physiologically distinct group of saprotrophic fungi capable of decomposition of all major components of wood. Although wood is their natural substrate, several species can survive in soil if suitable substrates are available, the soil-colonizing ability of species in the genera Phanerochaete, Pleurotus and Trametes being comparable to that of the soil-inhabiting basidiomycetes utilizing plant litter. Wood-inhabiting ligninolytic basidiomycetes (WLB) in soil interact with soil microflora, and some species are very efficient competitors affecting both the soil microbial biomass and the composition of the indigenous microbial community. The extracellular enzymes utilized by saprotrophic basidiomycetes for nutrient acquisition participate in the interspecific interactions with other soil biota but are also involved in the transformation of soil organic matter – lignocellulose and humic compounds. Bioremediation research has significantly enriched our knowledge of the ecology of basidiomycetes, but it has also identified several limitations for practical applicability of WLB at a field scale, and has led to the conclusion that natural attenuation or bioaugmentation are todays methods of choice for on site treatment.     

Indoor wood-decay basidiomycetes: damage,causal fungi,physiology, identification and characterization,prevention and control

Indoor wood-decay fungi cause considerable economical damage. Most of the structural damage to the indoors of buildings in Europe and North America is caused by brown-rot fungi that degrade conifer wood; white-rot fungi, which preferentially attack hardwoods, are less common. This review covers the approximately 80 basidiomycetes that commonly occur in buildings. Emphasis was placed on Serpula lacrymans, which is the most common indoor basidiomycete in central Europe. Meruliporia incrassata, the North American pendant to S. lacrymans, has also received considerable attention. In terms of indoor wood decay, moisture and temperature are the most important influences. Wood samples with a low moisture content can be degraded. High temperatures as an alternative control measure do not kill mycelia, with some species surviving in wood samples in the form of heat-resistant arthrospores at temperatures as high 95°C. For refurbishment and scientific purposes, the identity of the causal species should be known. More recently, several molecular techniques have been used to identify fungi; these results are often conflicting with those obtained by other, earlier applied methods. Sequencing of the internal transcribed spacers (ITS) of the rDNA is currently the best molecular tool. Among the other methods available, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) has also been shown to be able to distinguish closely related sister taxa. For further characterization of indoor basidiomycetes, the complete sequences of the 18S, 28S rDNA and the intergenic spacers with the included 5S rDNA have been acquired for some species. If current projects involving whole funal genome sequencing are not taken into account, Antrodia vaillantii is the first basidiomycete for which the complete rDNA sequence has been deposited. The review closes with fundamentals on the prevention and control of indoor wood decay. An erratum to this article can be found at     

The unique ultrastructure of the uterus of the Gyrocotylidea Poche, 1926 (Cestoda) and its phylogenetic implications

The members of the order Gyrocotylidea are monozoic tapeworms and generally considered to be the most primitive group of the Cestoda in terms of the evolution of this platyhelminth class. As part of a series of ultrastructural studies on Gyrocotyle urna (Wagener, 1852), three regions of the uterus were distinguished. The proximal region of the uterus is characterised by underlying perikarya, the presence of septate junctions within the epithelial wall and two types of specialised outer coverings, lamellae and cilia. The middle, syncytial region of the uterus is covered by short lamellae and marked by a concentration of sunken glandular perikarya (uterine glands). Glandular spheroidal granules (c.0.45 μm in diameter) of moderately dense content and a fine fibrillar texture are liberated by migration through the luminal membrane. The epithelium of the sac-shaped, distal region of the uterine duct is interrupted by cytoplasmic processes of sunken epithelial bodies, covered with lamellae and contains septate junctions. A muscular sphincter surrounds the narrow, terminal region of the distal uterine duct. The ultrastructural pattern of the uterus of the Gyrocotylidea has important discriminating traits (i.e. the presence of sunken perikarya along its entire length, septate junctions within the uterine epithelial cytoplasm of the proximal and distal regions, and cilia on the surface of its proximal region) unique among the Neodermata and which may represent autapomorphic character states for the group.     

Mycorrhizal and Endophytic Fungal Colonization in Arrhenatherum elatius L. Roots According to the Soil Contamination in Heavy Metals

The aim of this work was to jointly study non-mycorrhizal (dark septate fungi) and mycorrhizal (arbuscular mycorrhizae) colonization along a large range of heavy metal pollution in soil in order to determine the effective contribution of each type of endophytes in relation to heavy metal uptake and tolerance. Hence, eight sites were chosen in the mining area of northern France with respect both to a large range of heavy metal contamination (Cd, Pb, Zn) and monospecific colonization by Arrhenatherum elatius. Root colonization with both arbuscular mycorrhizae (AM) and dark septate fungi (DSF) as well as spore density in rhizospheric soil were estimated in relation to soil characteristics. Mycorrhizal infestation (hyphae, arbuscules and vesicles) was adversely affected by soil pollution almost to exclusion. The intensity of colonization with DSF was very low in presence of AM in non-contaminated soils but higher in polluted soils. The effect of the fungal colonization on the heavy metal tolerance of Arrhenatherum elatius is discussed.     

HOST SWITCHING PROMOTES DIVERSITY IN HOST‐SPECIALIZED MYCOPARASITIC FUNGI: UNCOUPLED EVOLUTION IN THE BIATOROPSIS‐USNEA SYSTEM

Fungal mycoparasitism—fungi parasitizing other fungi—is a common lifestyle in some basal lineages of the basidiomycetes, particularly within the Tremellales. Relatively nonaggressive mycoparasitic fungi of this group are in general highly host specific, suggesting cospeciation as a plausible speciation mode in these associations. Species delimitation in the Tremellales is often challenging because morphological characters are scant. Host specificity is therefore a great aid to discriminate between species but appropriate species delimitation methods that account for actual diversity are needed to identify both specialist and generalist taxa and avoid inflating or underestimating diversity. We use the Biatoropsis‐Usnea system to study factors inducing parasite diversification. We employ morphological, ecological, and molecular data—methods including genealogical concordance phylogenetic species recognition (GCPSR) and the general mixed Yule‐coalescent (GMYC) model—to assess the diversity of fungi currently assigned to Biatoropsis usnearum. The degree of cospeciation in this association is assessed with two cophylogeny analysis tools (ParaFit and Jane 4.0). Biatoropsis constitutes a species complex formed by at least seven different independent lineages and host switching is a prominent force driving speciation, particularly in host specialists. Combining ITS and nLSU is recommended as barcode system in tremellalean fungi.     

Decay and mineralization of soft tissue in Early Devonian boring ctenostome bryozoans

The Early Devonian of Podolia, Ukraine, has yielded phosphatized colonies of the boring ctenostome bryozoan Podoliapora doroshivi with 3‐D preservation of soft tissues. However, the feeding zooids are not anatomically complete, their preserved soft tissues comprising decay‐resistant structures such as the protective cuticular polypide sacs with presumed parietal muscles inside the wall of the sacs, the setigerous collars, the membranous orificial walls and remains of the muscle tissues. Early diagenetic apatite mineralization occured in numerous feeding zooids of Podoliapora at different stages of decay and may be important for the interpretation of decay processes in these colonial soft‐bodied fossil organisms. A setigerous collar, which is a characteristic of extant ctenostomes, occurs in P. doroshivi in several stages of decay showing progressive collapse and eventual complete loss. This study indicates that the morphological changes of collars induced by decay often resulted in connection with the membranous orificial wall, producing false anatomical structures, unrelated to structures observed in the earlier stages of decay or to the anatomical structures of extant ctenostomes. The most decay‐resistant cuticular polypide sacs mineralized as cryptocrystalline apatite in early stage of decay became degraded in later stages of decay. These data provide evidence that the anatomical interpretation of soft‐bodied fossils preserved only in the later stages of decay may have led to imprecise morphological interpretations.     

Contributions to the diversity in cryptogamic covers in the mid‐Palaeozoic: Nematothallus revisited

Compression fossils from the Silurian and Devonian of southern Britain, composed of cuticles and tubes, were described by W. H. Lang as the genus Nematothallus and placed, with Prototaxites, in Nematophytales, related neither to algae nor tracheophytes. Dispersed cuticles of Nematothallus and perforated forms assigned to Cosmochlaina were frequently recovered in macerates, their affinities being unresolved. New collections from a Lochkovian locality in the Welsh Borderland permitted the reconstruction of the stratified thalli of these nematophytes; they comprise a superficial cortex (which produced the cuticles) overlying a palisade zone composed of septate, parallel tubes, presumed to be hyphae, and a basal zone comprising wefts of randomly interwoven hyphae. Excellent three dimensional preservation allows the erection of a new species of Nematothallus, N. williamii. A similar anatomy is seen in a new group of fossils with either circular incisions in the cortex or complete separation of thickened cortical cells, presumably comprising a developmental sequence. By their stratified organization the nematophytes differ from extant and extinct algae and bryophytes and the enigmatic Spongiophyton. A complex anatomy and septate tubes suggest affinity with lichenized fungi. Limited data support a fungal rather than embryophyte chemistry, but a photobiont is missing. Nematophytes, globally widespread in cryptogamic covers from mid‐Ordovician times, added to the biodiversity in early terrestrial ecosystems and enhanced chemical weathering. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 505–534.