Freezing of tissue-limits for the autoradiographic localization of diffusible substances

Frozen thin sections and sections from freeze-dried and embedded tissue are used for the autoradiographic localization of diffusible substances at the electron microscope level. The presence of ice crystals in such sections may limit the autoradiographic resolution. Ice crystals are formed during freezing and may grow during subsequent processing of tissue. The contribution of ice crystal growth to the final image was estimated by measuring the distribution of the ice crystal sizes in freeze-etch replicas and in sections from freeze-dried and embedded tissues. A surface layer (10-15 mu) without visible ice crystals was present in both preparations. Beneath this surface layer the diameter of ice crystals increased towards the interior with the same relationship between crystal size and distance from the surface in the freeze-etch preparation as in the freeze-dry preparation. Ice crystal growth occurring during a much longer time during freeze-drying compared to freeze-etching does not significantly contribute to the final image in the electron microscope. The formation of ice crystals during freezing determines to a large extent the image (and therefore the autoradiographic resolution) of freeze-dry preparations and this probably holds also for thin cryosections of which examples are given.     

Canopy cover and tree regeneration in old-growth cove forests of the Appalachian Mountains

Relationships between canopy cover and tree regeneration were determined for various species in cove forests of the Great Smoky Mountains. Old-growth stands were sampled with six plots covering a total area of 4.8 ha. Each plot was subdivided into contiguous 10×10 m quadrats. Canopy cover overlying each of the 480 quadrats was characterized with three different indices based on visual estimates of cover. Influences of: (1) overlying cover, (2) proximate openings, and (3) total area of proximate openings on quadrat regeneration densities were determined. Most species reproducing by seed and some species reproducing by vegetative means had higher densities in quadrats with openings, but only the intolerants were highly dependent on gaps. Tsuga canadensis, a very shade-tolerant species, was one of the few species with abundant regeneration beneath dense canopy cover. In general, understory areas near gaps had somewhat higher regeneration densities than other areas with overlying cover. Several shade-tolerant species showed a positive regeneration density response to canopy openings and an ability to regenerate in gaps 0.01–0.03 ha in area. These openings were too small for intolerant species. Many species exhibited a positive response to total size of the proximate opening(s). A sharp increase in regeneration density with area of the opening(s) was evident at approximately 0.04 ha for the shade-intolerant species.     

Advances in spatial,individual‐based modelling of forest dynamics

Many individual‐based models of forest dynamics lack spatial complexity. Although, in certain cases, spatially simple models may not be substantially inferior to spatially complex models, advances in vegetation science indicate potential weaknesses, particularly the lack of consideration of propagule availability in horizontal space, and varying patch (or canopy gap) dimensions. Models with vertical and horizontal spatial complexity can address these issues, but, thus far, evidence that they outperform patch (or gap) models is limited. Comparison of projections from models that differ only in their spatial complexity is needed to address the effects of propagule availability in space, spatial pattern of canopy tree mortality, and spatial resolution.     

Dynamics of composition and structure in an old Sequoia sempervirens forest

Abstract. Dynamics of a Sequoia sempervirens forest in northern California were studied with long‐term plot data (1.44 ha) and recent transect data. The study was conducted in an old stand (> 1100 yr) on alluvial flats. Over three decades (1972–2001), changes in the composition and structure of the tree stratum were minor. Sequoia maintained a broad distribution of stem diameters throughout the period. Annual rates of Sequoia mortality (0.0029) and ingrowth (0.0029) were low, reflecting the great longevity of Sequoia and the slow canopy turnover of the study forest. Transect data also indicated a low frequency of canopy gap disturbance (≤ 0.4% of total land area per yr), but gap size was potentially large (> 0.1 ha) and the fraction of area in gaps (ca. 20%) was similar to other temperate forests. Regeneration quadrats sampled along transects, in gap centers, and on logs revealed that Sequoia regeneration is elevated at gap edges. The longevity of Sequoia and its response to gap disturbances ensure that it will remain a dominant species in the study forest.     

Tree species diversity in temperate and tropical forest gaps: The role of lottery recruitment

How canopy gaps promote tree species coexistence in temperate and tropical forests is reviewed. Given that evidence for traditional resource-based niche partitioning of canopy gaps is weak, lottery recruitment, whereby colonization of vacant living space is random with respect to species identity, may be key to maintaining diversity. Gap formation events are not highly predictable and species’ propagules tend to be patchily distributed. For these reasons the predictability of gap-phase recruitment is low and lottery principles apply. Recruitment limitation from discontinuities in species’ propagule supplies in space and time may permit lottery recruitment in temperate and tropical forests. However, the relative importance of recruitment limitation in species-rich versus species-poor communities is unclear. Although lottery models with purely random recruitment in vacant sites can be applied, relatively complex models appear to better capture the essential features of forest community dynamics. For example, models with recruitment probabilities weighted by species abundance can produce the non-random trajectories often observed in communities. Models with local competitive displacement by individuals of dominant species can also produce non-random trajectories. Models with spatial structure, localized competition, and dispersal limitations may provide further insight into the effects of biotic interactions and recruitment limitation on forest dynamics.     

Composition,structure and diversity of cove forest stands in the Great Smoky Mountains: a patch dynamics perspective

Abstract. Cove forests of the Great Smoky Mountains are North American examples of old-growth temperate forest. Ecological attributes of seven stands were studied using one 0.6 - 1.0 ha plot per stand. Stand basal area (39 - 55 m²/ha) and biomass (326 - 471 Mg/ha) were high for temperate deciduous forest. Density ranged from 577 to 1075 stems/ha. All stands had a mixture of deciduous canopy species. Only rarely did a single species comprise more than half of the stand by density, basal area or biomass. Shade-intolerant species were present at low levels (1 - 5 % of total stand density). A wide range of stem diameters was characteristic of most species. However, some species lacked small stems, indicating discontinuous regeneration. Stands tended to have 10 - 20 tree species per ha and at least five species had biomass levels > 10 Mg/ha, indicating high evenness. Canopy gaps covered 10 % of the total area (2 - 21 % by stand). Gaps and conspecific patches of canopy trees > 0.05 ha in size were infrequent. Spatial analyses revealed a variety of patterns among species at inter-tree distances of 1 to 25 m. When all species were combined, juveniles showed aggregation, and adults were often hyperdispersed. Analyses for individual species confirmed that the mosaic of canopy species is influenced by non-random spatial processes. Adults of several species were aggregated at distances > 10 m. Juveniles of all major species exhibited aggregation. Several species exhibited regeneration near conspecific adults. This pattern suggested limited mobility for such species within the shifting mosaic. A diverse patchwork resulted despite the fact that many species did not exhibit segregation of adults and juveniles. Further understanding of patch dynamics and the potential for compositional steady state in cove forests requires long-term study with spatial data.     

Vapor fixation for immunocytochemistry and X-ray microanalysis on cryoultramicrotome sections

Several organic and inorganic vapor fixatives have been tested for their ability to stabilize the ultrastructure of freeze-dried thin cryosections. The vapors from osmium tetroxide and dry formaldehyde gave a good preservation of the ultrastructure. Fixation in formaldehyde vapor preserved the immunoreactivity of alpha-amylase in exocrine pancreas, as was demonstrated with an indirect labeling technique using anti-alpha-amylase and protein A-gold. A major advantage of the use of vapor fixation is that cryosections from a specimen of fresh-frozen tissue can be used for immunocytochemistry as well as for X-ray microanalysis, as was demonstrated for the exocrine pancreas. This opens the possibility of localizing atomic species (X-ray microanalysis) and molecular species (immunocytochemistry) at the subcellular level on thin cryosections from the same tissue block.     

Hurricane disturbance in a temperate deciduous forest: patch dynamics, tree mortality, and coarse woody detritus

Patch dynamics, tree injury and mortality, and coarse woody detritus were quantified to examine the ecological impacts of Hurricane Fran on an oak-hickory-pine forest near Chapel Hill, NC. Data from long-term vegetation plots (1990–1997) and aerial photographs (1998) indicated that this 1996 storm caused patchy disturbance of intermediate severity (10–50% tree mortality; Woods, J Ecol 92:464–476, 2004). The area in large disturbance patches (>0.1 ha) increased from <1% to approximately 4% of the forested landscape. Of the forty-two 0.1-ha plots that were studied, 23 were damaged by the storm and lost 1–66% of their original live basal area. Although the remaining 19 plots gained basal area (1–15% increase), across all 42 stands basal area decreased by 17% because of storm impacts. Overall mortality of trees >10 cm dbh was 18%. The basal area of standing dead trees after the storm was 0.9 m²/ha, which was not substantially different from the original value of 0.7 m²/ha. In contrast, the volume and mass of fallen dead trees after the storm (129 m³/ha; 55 Mg/ha) were 6.1 and 7.9 times greater than the original levels (21 m³/ha; 7 Mg/ha), respectively. Uprooting was the most frequent type of damage, and it increased with tree size. However, two other forms of injury, severe canopy breakage and toppling by other trees, decreased with increasing tree size. Two dominant oak species of intermediate shade-tolerance suffered the largest losses in basal area (30–41% lost). Before the storm they comprised almost half of the total basal area in a forest of 13% shade-tolerant, 69% intermediate, and 18% shade-intolerant trees. Recovery is expected to differ with respect to vegetation (e.g., species composition and diversity) and ecosystem properties (e.g., biomass, detritus mass, and carbon balance). Vegetation may not revert to its former composition; however, reversion of biomass, detritus mass, and carbon balance to pre-storm conditions is projected to occur within a few decades. For example, the net change in ecosystem carbon balance may initially be negative from losses to decomposition, but it is expected to be positive within a decade after the storm. Repeated intermediate-disturbance events of this nature would likely have cumulative effects, particularly on vegetation properties.