The functioning of ectomycorrhizal fungi in the field: linkages in space and time

Individual trees, either of the same or different species, can be linked spatially and temporally by the hyphae of ectomycorrhizal (ECM) fungi that allow carbon and nutrients to pass among them and promote forest establishment following disturbance. Spatial and temporal linkages between plants influence the function of ECM fungi in the field. Studies indicate that ECM linkages can reduce plant competition for resources, promote forest recovery, and influence the pattern of plant succession. The degree of influence depends on many factors, including the composition and arrangement of the vegetative community and soil and climatic conditions. Management practices that create intense disturbance and loss of organic matter or promote the introduction of non-ectomycorrhizal host species can decrease the ability of plants to form linkages with ECM fungi. Management practices that retain living trees and shrubs and input of organic matter provide the energy source and substrate necessary for ECM linkages. More research is needed to determine the degree to which ECM fungal linkages occur in the field and their role in ecosystem function and long-term health.     

Time linkages between pollination onsets of different taxa over an 11-year period in Perugia,Central Italy

Times of pollination of different taxa in the atmosphere of Perugia (Central Italy) over an 11-year period (1982–1992) were recorded and analysed by means of a 7-day volumetric Hirst-type pollen trap. For some taxa, the pollination period varied from year to year from a chronological and/or quantitative point of view. Several taxa showed a linkage in their starting dates of pollination. Knowledge of this kind of linkage allows us to build a forecasting model.     

Human β-glucuronidase: Assignment of the structural gene to chromosome 7 using somatic cell hybrids

-Glucuronidase (GUS) has become an important enzyme model for the genetic study of molecular disease, enzyme realization, and therapy, and for the biogenesis and function of the lysosome and lysosomal enzymes. The genetics of human -glucuronidase was investigated utilizing 188 primary man-mouse and man-Chinese hamster somatic cell hybrids segregating human chromosomes. Cell hybrids were derived from 16 different fusion experiments involving cells from ten different and unrelated individuals and six different rodent cell lines. The genetic relationship of GUS to 28 enzyme markers representing 19 linkage groups was determined, and chromosome studies on selected cell hybrids were performed. The evidence indicates that the -glucuronidase gene is assigned to chromosome 7 in man. Comparative linkage data in man and mouse indicate that the structural gene GUS is located in a region on chromosome 7 that has remained conserved during evolution. Involvement of other chromosomes whose genes may be important in the final expression of GUS was not observed. A tetrameric structure of human -glucuronidase was demonstrated by the formation of three heteropolymers migrating between the human and mouse molecular forms in chromosome 7 positive cell hybrids. Linkage of GUS to other lysosomal enzyme genes was investigated. -Hexosaminidase HEX _B) was assigned to chromosome 5; acid phosphatase₂ (ACP ₂) and esterase A₄ (ES-A ₄) were assigned to chromosome 11; HEX _A was not linked to GUS; and -galactosidase (-GAL) was localized on the X chromosome. These assignments are consistent with previous reports. Evidence was not obtained for a cluster of lysosomal enzyme structural genes. In demonstrating that GUS was not assigned to chromosome 9 utilizing an X/9 translocation segregating in cell hybrids, the gene coding for human adenylate kinase₁ was confirmed to be located on chromosome 9.Supported by NIH Grants HD 05196, GM 20454, and GM 06321, by NSF Grant BMS 73-07072, and by HEW Maternal and Child Health Service, Project 417.     

Factors influencing glycosylation of Trichoderma reesei cellulases. II: N-glycosylation of Cel7A core protein isolated from different strains

A systematic analysis of the N-glycosylation of the catalytic domain of cellobiohydrolase I (Cel7A or CBH I) isolated from several Trichoderma reesei strains grown in minimal media was performed. Using a combination of chromatographic, electrophoretic, and mass spectrometric methods, the presence of glucosylated and phosphorylated oligosaccharides on the three N-glycosylation sites of Cel7A core protein (from T. reesei strains Rut-C30 and RL-P37) confirms previous findings. With N-glycans isolated from other strains, no end-capping glucose could be detected. Phosphodiester linkages were however found in proteins from each strain and these probably occur on both the alpha1-3 and the alpha1-6 branch of the high-mannose oligosaccharide tree. Evidence is also presented for the occurrence of mannobiosyl units on the phosphodiester linkage. Therefore the predominant N-glycans on Cel7A can be represented as (ManP)(0-1)GlcMan(7-8)GlcNAc2 for the hyperproducing Rut-C30 and RL-P37 mutants and as (Man(1-2)P)(0-1-2)Man(5-6-7)GlcNAc2 for the wild-type strain and the other mutants. As shown by ESI-MS, random substitution of these structures on the N-glycosylation sites explains the heterogeneous glycoform population of the isolated core domains. PAG-IEF separates up to five isoforms, resulting from posttranslational modification of Cel7A with mannosyl phosphodiester residues at the three distinct sites. This study clearly shows that posttranslational phosphorylation of glycoproteins is not atypical for Trichoderma sp. and that, in the case of the Rut-C30 and RL-P37 strains, the presence of an end-capped glucose residue at the alpha1-3 branch apparently hinders a second mannophoshoryl transfer.     

The use of ubiquitin lysine mutants to characterize E2–E3 linkage specificity: Mass spectrometry offers a cautionary “tail”

Oligomeric ubiquitin structures (i.e. ubiquitin “chains”) may be formed through any of seven different lysine residues in the polypeptide, or via the amine group of Met 1. Different types of ubiquitin chains can confer very different biological outcomes to a protein substrate, yet the structural characteristics of E2s and E3s that determine ubiquitin linkage specificity remain poorly understood. In vitro autoubiquitylation assays combined with ubiquitin protein variants bearing individually mutated lysine residues (“K‐to‐R” mutants) have thus been widely used to characterize E2–E3 linkage specificity. However, how this type of assay compares to direct identification of ubiquitin linkage types using mass spectrometry (MS) has not been rigorously tested. Here, we characterize the linkage specificity of 12 different E2–E3 combinations using both approaches. The simple MS‐based method described here is more robust, requires less material and is less prone to bias introduced by, e.g. the use of mutant proteins with unknown effects on E1, E2 or E3 recognition, antibodies with uncharacterized epitopes, the low dynamic range of X‐ray film, and additional sources of experimental error. Indeed, our results suggest that the K‐to‐R assay be approached with some caution.     

Quantifying relationships between traits and explicitly measured gradients of stress and disturbance in early successional plant communities

Questions: How can one explicitly quantify, and separately measure, stress and disturbance gradients? How do these gradients affect functional composition in early successional plant communities and to what extent? Can we accurately predict trait composition from knowledge of these gradients? Location: Southern Quebec, Canada. Methods: Using eight environmental variables measured in 48 early successional plant communities, we estimated stress and disturbance gradients through structural equation modelling. We then measured 10 functional traits on the most abundant species of these 48 communities and calculated their community‐level mean and variance weighted by the relative abundance of each species. Finally, we related these community‐weighted means and variances to the estimated stress and disturbance gradients using general linear models or generalized additive models. Results: We obtained a well‐fitting measurement model of the stress and disturbance gradients existing in our sites. Of the 10 studied traits, only average plant reproductive height was strongly correlated with the stress (r²=0.464) and disturbance (r²=0.543) gradients. Leaf traits were not significantly related to either the stress or disturbance gradients. Conclusions: The well‐fitting measurement model of the stress and disturbance gradients, combined with the generally weak trait–environment linkages, suggests that community assembly in these early successional plant communities is driven primarily by stochastic processes linked to the history of arrival of propagules and not to trait‐based environmental filtering.     

Converging forest community composition along an edaphic gradient threatens landscape‐level diversity

Aim Plant communities across the temperate zone are changing in response to successional processes and human‐induced disturbances. Here, we assess how upland forest under‐ and overstorey community composition has changed along an edaphic gradient. Location Northern Wisconsin, USA. Methods Forest sites initially sampled in the 1950s were resampled for overstorey composition and diversity, basal area, and understorey composition and diversity. We used clustering methods to identify groups of stands based on overstorey composition, and we used similarity indices, ordination and diversity indices to evaluate changes in species abundance and overall community structure. Results Sites clustered into four overstorey groups along the edaphic gradient: ‘hemlock’ sites dominated by hemlock in 1950, ‘mesic’ sites dominated by northern hardwoods, ‘dry’ sites with a significant pine inclusion in the canopy and diverse ‘dry‐mesic’ sites in the middle. Collectively, forests gained maple, ash and cherry while losing pines, birches and red oaks. The hemlock forest sites gained hardwoods, while the dry‐mesic sites shifted towards a more mesic hardwood composition. Only the driest sites have remained relatively stable in species composition. Main conclusions These trends reflect both ‘mesification’ and homogenization among northern forests. Highly diverse mid‐gradient and mesic hemlock‐dominated stands are transitioning to maple dominance. Fire suppression may be favouring invasions of more mesic plants into historically drier sites, while high deer abundance likely limits hemlock regeneration. If current trends continue, maples will dominate the majority of northern forests, with significant losses of local native species richness and substantial shifts in understorey composition.     

Relationships among biological elements (macrophytes, macroinvertebrates and ichthyofauna) for different core river types across Europe at two different spatial scales

The objective of this study was to evaluate differences in correlations among Biological Elements and environmental parameters for different river types, analysed at two different spatial scales. A total of 82 sites, with at least good ecological status, were sampled across Europe, representing three core river types: Mountain rivers (26 sites); Lowland rivers (29 sites) and Mediterranean rivers (17 sites). At each site samples of macrophytes, macroinvertebrates and fishes were taken during spring, following the methodological procedures established by the European STAR project. Environmental parameters were also recorded, based on a site protocol developed by the European projects AQEM and STAR. Environmental parameters were divided into three categories: aquatic habitats (mesohabitat scale), global features (reach scale) and obligatory typology parameters of Water Framework Directive (WFD) (geographical scale). Data were analysed to evaluate at the two scales, first, relationships among biological elements, and second, relationships between biological elements and environmental parameters. Within each river type, correlation matrices (Bray–Curtis distance) were calculated separately for each biological element and for each category of environmental parameters. All biological elements were correlated (p<0.01) to the larger spatial scale: macrophytes and macroinvertebrates are more correlated in lowland and mountain rivers, while in Mediterranean rivers, fish and macrophytes presented higher correlations. These links tend to be consistent for different spatial scales, except if they are weak on a larger regional scale, obligatory parameters of WFD were, in most cases, significantly correlated with the three biological communities (p<0.05). Results at different spatial scales supported the hierarchical theory of river formation. Reach and mesohabitat environmental parameters tend to explain aquatic communities at a lower spatial scale, while geographical parameters tend to explain the communities at a major spatial scale.     

Tracing carbon flow in an arctic marine food web using fatty acid-stable isotope analysis

Global warming and the loss of sea ice threaten to alter patterns of productivity in arctic marine ecosystems because of a likely decline in primary productivity by sea ice algae. Estimates of the contribution of ice algae to total primary production range widely, from just 3 to >50%, and the importance of ice algae to higher trophic levels remains unknown. To help answer this question, we investigated a novel approach to food web studies by combining the two established methods of stable isotope analysis and fatty acid (FA) analysis--we determined the C isotopic composition of individual diatom FA and traced these biomarkers in consumers. Samples were collected near Barrow, Alaska and included ice algae, pelagic phytoplankton, zooplankton, fish, seabirds, pinnipeds and cetaceans. Ice algae and pelagic phytoplankton had distinctive overall FA signatures and clear differences in delta(13)C for two specific diatom FA biomarkers: 16:4n-1 (-24.0+/-2.4 and -30.7+/-0.8 per thousand, respectively) and 20:5n-3 (-18.3+/-2.0 and -26.9+/-0.7 per thousand, respectively). Nearly all delta(13)C values of these two FA in consumers fell between the two stable isotopic end members. A mass balance equation indicated that FA material derived from ice algae, compared to pelagic diatoms, averaged 71% (44-107%) in consumers based on delta(13)C values of 16:4n-1, but only 24% (0-61%) based on 20:5n-3. Our estimates derived from 16:4n-1, which is produced only by diatoms, probably best represented the contribution of ice algae relative to pelagic diatoms. However, many types of algae produce 20:5n-3, so the lower value derived from it likely represented a more realistic estimate of the proportion of ice algae material relative to all other types of phytoplankton. These preliminary results demonstrate the potential value of compound-specific isotope analysis of marine lipids to trace C flow through marine food webs and provide a foundation for future work.