An inexpensive moist chamber culture technique for finding microbiota on live tree bark.

Premise: Traditional moist chamber cultures (MCs) prepared in aseptic laboratory environments using sterile Petri dishes are commonly used to quantify the microbiota of rough-bark tree species and woody vines. MCs are typically expensive and may be difficult to make, so a less expensive option made from easily available supplies was developed. These cost-friendly MCs were compared with standard laboratory methods to demonstrate their efficacy. Methods and Results: Modified MCs were made using inexpensive, store-bought supplies; compared to a standard laboratory setting, the modified MCs are shown to be less expensive with a faster setup time and larger size that facilitates a variety of tree and woody vine species. MC use resulted in the discovery of new species of fungi and myxomycetes with associated locality records. We provide detailed instructions for creating modified MCs, as well as a list of myxomycete species and their associated bark characteristics, pH values, and water-holding capacity. Conclusions: This new, low-cost MC technique makes the study of microbiota more inclusive and accessible for those in research laboratories, classrooms, and homes, including both amateurs and professionals. MCs are easy to prepare, versatile, and applicable for many areas of botany and the biological sciences, potentially allowing exploration into unexplored areas in urban ecosystems.

Autofluorescence and ultrastructure in the Myxomycete Diachea leucopodia (Physarales).

Autofluorescence is reported for the first time in Myxomycete fruiting bodies. Ultrastructure of stalked sporangia of Diachea leucopodia (Didymiaceae, Physarales) was studied using scanning and transmission electron microscopy, energy-dispersive X-ray microanalysis, and fluorescence microscopy. External and internal properties of the peridium that surround the spores and capillitium exhibit autofluorescence. The stalk is composed of calcareous granules and energy-dispersive X-ray microanalysis demonstrates that the elemental composition of the peridium, capillitium, and stalk has varying concentrations of calcium.

Macrofungi associated with vegetation and soils at Ha Ha Tonka State Park, Missouri.

Fungi and vascular plant interactions are necessary components of natural community establishment, productivity and degradation. While many fungal species serve as decomposers of organic matter, others have evolved mutualistic or parasitic relationships with vascular plants. This research focused on characterizing associations among macrofungi, vascular plant communities and soils. Ha Ha Tonka State Park is in central Missouri and has a varying landscape with numerous natural community types that provide diverse habitats and microhabitats that are ideally suited to the investigation of fungal, floral and soil associations. Five communities sampled within the park included glades, open woodlands, flatwoods, closed-canopy forests and karst sinks. Permanent 0.01 ha. plots were surveyed in the 2006 and 2007 growing seasons. Surveys of plots and entire communities yielded 249 fungal taxa and approximately 265 floral taxa. Soils were analyzed to help define specific edaphic components of each community and used to associate soil attributes with plant and fungal communities. Forest communities contained the most ectomycorrhizal (ECM) fungi species. Karst sinks and glades had higher soil pH and phosphorus and fewer ectomycorrhizal fungi. Statistical analyses included non-metric multidimensional scaling, multiresponse permutation procedure and indicator species analysis. Indicator species were identified for flatwood, forest and karst communities, but results were inconclusive for glades and open woodlands.

Aerial reproductive structures of vascular plants as a microhabitat for myxomycetes.

This study explored the occurrence and distribution of myxomycete species on the aerial reproductive structures of vascular plants. Eight species of vascular plants representing five families were sampled. The doubled rope climbing method was used to collect bark and cones from the canopy of Pinus echinata. Bark and aerial seed pods were gathered from Cercis canadensis, follicles and stems from Asclepias syriaca, dried composite inflorescences and stems from Echinacea angustifolia, E. pallida, and E. paradoxa var. paradoxa, and capsules and stems from Yucca glauca and Y. smalliana. Reproductive structures and bark/stems for 202 host plants were separated and cultured in 541 moist chambers, resulting in 118 collections yielding 32 myxomycete species representing 11 genera, seven families and five orders. There was no significant difference in pH values of the reproductive structures and bark/stems of the host plants, however legume pods of C. canadensis (6.9 +/- 1.3) had higher pH than the bark (6.0 +/- 1.1) and had a different composition of myxomycete species. Myxomycete orders have optimal pH ranges. Nonmetric multidimensional scaling, multiresponse permutation procedure and indicator species analysis showed a significant difference in species richness of reproductive structures and bark/stems. The bark of trees had greater mean species richness of myxomycetes than the reproductive structures, but the reproductive structures of herbaceous plants had greater mean species richness of myxomycetes than the stems. A new term, herbicolous myxomycetes, is proposed for a group of myxomycetes frequently associated with herbaceous, perennial, grassland plants. An undescribed species of Arcyria occurred only on cones of P. echinata.

Influence of bark pH on the occurrence and distribution of tree canopy myxomycete species.

This study compares the occurrence and distribution of myxomycete species in the canopy of living trees and neighboring grapevines. Corticolous myxomycetes of three temperate forests in southeastern USA were studied on six tree species (30 trees) and grapevines (30 vines) to determine distribution and occurrence of myxomycete species relating to geographic location, host species, and bark pH. The double-rope climbing technique was used to access the canopy and sample bark up to 16.5 m. Bark samples were examined in 580 moist chamber cultures and 44 myxomycete species were identified representing 21 genera, averaging 3.0 +/- 2.1 species per sample site. Jaccard's coefficient determined community similarity between five individuals of six tree species, Acer saccharum, Fraxinus americana, Liquidambar styraciflua, Liriodendron tulipifera, Platanus occidentalis and Tsuga canadensis, and neighboring grapevines, Vitis aestivalis and V. vulpina. Vertical variation in species richness was significantly different only for Platanus occidentalis and might be attributable to flaking of bark with increasing height in the canopy. Tsuga canadensis and neighboring grapevines had greatest community similarity. Cribraria violacea was observed on all tree and grapevine species except T. canadensis and neighboring grapevines. Occurrence and species assemblages of myxomycetes were associated with bark pH, not geographic location. Bark of V. aestivalis (pH 4.5) was more acidic than neighboring T. canadensis (pH 4.1), compared to grapevines of the same species neighboring other tree species. Results indicated that most species are not regionally restricted, and although some myxomycetes are associated with a certain pH range, others develop on any substratum. Future research protocols for corticolous myxomycetes should emphasize sampling adequate amounts of substrata in a local region from different host species that have a wide range of bark pH, ensuring a representative sample of species for an entire region.

The myxomycete genus Schenella: morphological and DNA sequence evidence for synonymy with the gasteromycete genus Pyrenogaster.

The genus Schenella has proven difficult to classify since its description as a new genus in 1911. Macbride placed it with the Myxomycetes but it was unclear with which myxomycete, if any, it should be grouped. Recent identification of abundant samples of Schenella has aided a re-evaluation of its classification as a myxomycete. Morphological evidence based on light and scanning electron microscopy of recently collected specimens and on the type specimen of Macbride suggested that it might be synonymous with the gasteromycete Pyrenogaster Analysis of DNA sequences from freshly isolated samples indicates that the genus Schenella is related closely to an anciently diverged, monophyletic group of fungi that includes several gasteromycete genera, among them Geastrum, Sphaerobolus and Pseudocolus. Comparisons of the morphology and DNA sequences of authentically identified specimens of Pyrenogaster atrogleba indicate that it is synonymous with Schenella simplex. The nomenclatural implications of this discovery are discussed.

Tree canopy biodiversity in the Great Smoky Mountains National Park: ecological and developmental observations of a new myxomycete species of Diachea.

A survey and inventory of tree canopy bio-diversity for cryptogams (myxomycetes, macrofungi, mosses, liverworts, lichens and ferns) in the Great Smoky Mountains National Park resulted in the discovery of an undescribed myxomycete species. This taxon is classified in the order Physarales, family Di-dymiaceae and genus Diachea. A combination of morphological characteristics distinguishes Diachea arboricola H.W. Keller & M. Skrabal sp. nov. from all other species in the genus: peridium iridescent gold to silvery gray; stalk reddish orange above and whitish below, filled with crystals; capillitial threads stiff, dichotomously branched and arising from the tip of the columella; spore ornamentation uniformly covering the entire spore surface, appearing spiny with light microscopy, with scanning electron microscopy as vertical processes with capitate, clustered, spike-like tips. This type of spore ornamentation has not been found in any other Diachea species. Diachea arboricola is known only from the tree canopy, ranging in height from roughly 3 to 21 m, on three tree species, Fraxinus americana, Juniperus virginiana and Quercus alba. Observations of plasmodial growth and fruiting body development are described based on moist chamber cultures. Tree canopy observations in situ suggest that the plasmodium of this species migrates over extensive vertical areas of tree bark. Ecological factors are discussed that include pH of bark substrata. The species description is based on abundant sporangia from 17 different collections. A key to the species of Diachea is provided to aid in the identification of this taxon.

Vertical distribution and assemblages of corticolous myxomycetes on five tree species in the Great Smoky Mountains National Park.

Corticolous myxomycetes of the Great Smoky Mountains National Park were studied in relation to their association with certain tree species and height of occurrence in the forest canopy. Using the double-rope climbing method, bark was collected at 3 m increments to the tops of trees of five different species. Bark samples from 25 trees were used to prepare 418 moist chamber cultures maintained and observed 4 wk. Eighty-four myxomycete species were identified, including 30 species not known to occur in the park. Tree species, pH, height in tree and water-holding capacity of the bark samples were analyzed to determine the relationships of myxomycete assemblages cultured on the bark. Results suggested that myxomycete community composition among selected tree species were similar, but occurrence and abundance of certain species were related to differences in bark pH. Community similarity values among trees of different species show that trees with the most similar myxomycete communities also have the most similar bark pH. Most myxomycete species in this study have a pH optimum. No variation in species richness was detected at different heights in the trees, and most species were obtained at all heights up to at least 24 m. The water-holding capacity of the bark could not be correlated with species richness or abundance of myxomycetes that inhabit the bark of living trees. This is the first study to characterize myxomycete communities of tree canopies.