Above-ground space sequestration determines competitive success in juvenile beech and spruce trees

A 2-yr phytotron study was conducted to investigate the intra- and inter-specific competitive behaviour of juvenile beech (Fagus sylvatica) and spruce (Picea abies). Competitiveness was analysed by quantifying the resource budgets that occur along structures and within occupied space of relevance for competitive interaction. Ambient and elevated CO(2) and ozone (O(3)) regimes were applied throughout two growing seasons as stressors for provoking changes in resource budgets, growth and allocation to facilitate the competition analysis. The hypothesis tested was that the ability to sequester space at low structural cost will determine the competitive success. Spruce was a stronger competitor than beech, as displayed by its higher above-ground biomass increments in mixed culture compared with monoculture. A crucial factor in the competitive success of spruce was its ability to enlarge crown volume at low structural costs, supporting the hypothesis. Interspecific competition with spruce resulted in a size-independent readjustment of above-ground allocation in beech (reduced leaf : shoot biomass ratio). The efficient use of resources for above-ground space sequestration proved to be a parameter that quantitatively reflects competitiveness.     

Comparing artificial and natural selection in rate of adaptation to genetic stress in Aspergillus nidulans

In an experimental study of adaptation to negative pleiotropic effects of a major fungicide resistance mutation in the filamentous fungus Aspergillus nidulans we have investigated the relative effectiveness of artificial selection vs. natural selection on the rate of compensatory evolution. Using mycelial growth rate as a fitness measure, artificial selection involved the weekly transfer of the fastest growing sector onto a fresh plate. Natural selection was approximated by transferring random samples of all the spores produced by the mycelium. Fungicide resistant and fungicide sensitive haploid and diploid strains were used in an evolution experiment over 10 weekly transfers, which is equivalent to 1200 cell cycles. Two different environmental conditions were applied: a constant fungicide-free environment and a weekly alternation between presence and absence of fungicide. Results show that for all strains and conditions used the transfer of a random sample of all spores leads to more rapid adaptation than the transfer of the visually 'fittest' sector. The rates of compensatory evolution in the constant and the alternating environment did not differ. Moreover, haploid strains tend to have a higher rate of adaptation than isogenic diploid strains.     

Selfing and resource allocation in Schiedea salicaria (Caryophyllaceae), a gynodioecious species

Abstract Levels of selfing and resource allocation patterns were investigated in Schiedea salicaria (Caryophyllaceae), a gynodioecious species with high levels of inbreeding depression and nuclear control of male sterility. Selfing levels were higher in hermaphrodites than females, especially when adjusted for early acting inbreeding depression. The sexes of S. salicaria were similar in most allocation patterns including number of flowers and capsules per inflorescence, seeds per flower, and seed mass. Seeds produced by females had higher levels of germination than seeds of hermaphrodites, a likely result of high selfing levels and the expression of inbreeding depression in the progeny of hermaphrodites. Invasion of females in populations of S. salicaria is probably related to the expression of inbreeding depression at germination and in later life history stages. Comparisons with related species of Schiedea that also have nuclear control of male sterility suggest that reallocation of resources in hermaphrodites to male function occurs as females increase in frequency, but that resource reallocation is not important for the success of females when they first invade populations.     

Pancratistatin: A natural anti-cancer compound that targets mitochondria specifically in cancer cells to induce apoptosis

The major hurdle in the fight against cancer is the non-specific nature of current treatments. The search for specific drugs that are non-cytotoxic to normal cells and can effectively target cancer cells has lead some researchers to investigate the potential anti-cancer activity of natural compounds. Some natural compounds, such as Taxol, have been shown to posses some anti-cancer potential. Pancratistatin (PST) is a natural compound that was isolated from the spider lily Pancratium littorale and shown to exhibit antineoplastic activity. The specificity of PST to cancer cells and the mechanism of PSTs action remain unknown. This study provides a detailed look at the effect of PST treatment on cancerous and normal cells. Our results indicate that PST induced apoptosis selectively in cancer cells and that the mitochondria may be the site of action of PST in cancer cells. A biochemical target available specifically in cancer cells may lead to the development of new and more effective cancer fighting agents.     

A natural variant of Bacillus licheniformis alpha-amylase isolated from flour mill wastewaters sheds light on the origin of high thermostability

AIMS: Understanding the origin of high thermostability exhibited by the alpha-amylase produced by a natural strain of Bacillus licheniformis. METHODS AND RESULTS: The MSH320 alpha-amylase gene has been cloned from a native strain of B. licheniformis isolated from flour mill wastewaters in Kashan, central Iran, and its nucleotide sequence was determined (GenBank Accession Number AF438149). Whereas previously cloned B. licheniformisalpha-amylase (BLA) genes are nearly identical, the MSH320 gene coding sequence presents only 93% identity with the reference 'wild-type' BLA gene, most of the nucleotide changes leading to silent mutations. Amino acid substitutions occurred at 19 of the 483 residues of the matured protein, distributed all along the protein sequence. Nevertheless, the natural BLA variant presents thermoinactivation kinetics similar to that of the reference BLA. Protein modelling and structural predictions at the substitution sites suggest that half of the mutations may have a significant stabilizing or destabilizing effect on the protein structure. Compensatory mutations thus occurred in the natural variant in order to maintain thermostability to the level of the reference enzyme. CONCLUSIONS: The exceptional high thermostability of BLA, although produced by a nonthermophilic organism, is not fortuitous but subject to a selective pressure still at work in natural environments. SIGNIFICANCE AND IMPACT OF THE STUDY: BLA thermal performances are not naturally maximized and can be substantially improved by protein engineering.     

Recombinant shark natural antibodies to thyroglobulin

As cartilaginous fish are the vertebrates most distal from man to produce antibodies, fundamental information regarding conservation and variation of the antigen binding site should be gained by comparing the properties of antibodies directed against the same antigen from the two species. Since monoclonal cell lines cannot be generated using shark B cells, we isolated antigen binding recombinant single chain Fv antibodies (scFv) comprising of the complete variable regions from shark light and heavy chains. Thyroglobulin was used as the selecting antigen as both sharks and humans express natural antibodies to mammalian thyroglobulin in the absence of purposeful immunization. We report that recombinant sandbar shark (Carcharhinus plumbeus) scFvs that bind bovine thyroglobulin consist of heavy chain variable regions (VH) homologous to those of the human VHIII subset and light chain variable regions (VL) homologous to those of the human Vlambda6 subgroup. The homology within the frameworks is sufficient to enable the building of three-dimensional models of the shark VH/VL structure using established human structures as templates. In natural antibodies of both species, the major variability lies in the third complementarity determining region (CDR3) of both VH and VL.     

Anti-pig antibody adsorption efficacy of {alpha}-Gal carrying recombinant P-selectin glycoprotein ligand-1/immunoglobulin chimeras increases with core 2 {beta}1, 6-N-acetylglucosaminyltransferase expression

We have previously described the construction of a P-selectin glycoprotein ligand-1-mouse immunoglobulin Fc fusion protein, which when transiently coexpressed with the porcine alpha1,3 galactosyltransferase in COS cells becomes a very efficient adsorber of xenoreactive, anti-pig antibodies. To relate the adsorption capacity with the glycan expression of individual fusion proteins produced in different cell lines, stable CHO-K1, COS, and 293T cells producing this fusion protein have been engineered. On alpha1,3 galactosyltransferase coexpression, high-affinity adsorbers were produced by both COS and 293T cells, whereas an adsorber of lower affinity was derived from CHO-K1 cells. Stable coexpression of a core 2 beta1,6 N-acetylglucosaminyltransferase in CHO-K1 cells led to increased alpha-Gal epitope density and improved anti-pig antibody adsorption efficacy. ESI-MS/MS of O-glycans released from PSGL-1/mIgG(2b) produced in an alpha1,3 galactosyl- and core 2 beta1,6 N-acetylglucosaminyltransferase expressing CHO-K1 cell clone revealed a number of structures with carbohydrate sequences consistent with terminal Gal-Gal. In contrast, no O-glycan structures with terminal Gal-Gal were identified on the fusion protein when expressed alone or in combination with the alpha1,3 galactosyltransferase in CHO-K1 cells. In conclusion, the density of alpha-Gal epitopes on PSGL-1/mIgG(2b) was dependent on the expression of O-linked glycans with core 2 structures and lactosamine extensions. The structural complexity of the terminal Gal-Gal expressing O-glycans with both neutral as well as sialic acid-containing structures is likely to contribute to the high adsorption efficacy.     

Monitoring Matters: Examining the Potential of Locally-based Approaches

Monitoring of biodiversity and resource use by professional scientists is often costly and hard to sustain, especially in developing countries, where financial resources are limited. Moreover, such monitoring can be logistically and technically difficult and is often perceived to be irrelevant by resource managers and the local communities. Alternatives are emerging, carried out at a local scale and by individuals with little formal education. The methods adopted span a spectrum, from participatory monitoring where aims and objectives are defined by the community, to ranger-based monitoring in protected areas. What distinguishes these approaches is that local people or local government staff are directly involved in data collection and (in most instances) analysis. In this issue of Biodiversity and Conservation, 15 case studies examine whether these new approaches can address the limitations of professional monitoring in developing countries. The case studies evaluate ongoing locally-based monitoring schemes involving more than 1500 community members in 13 countries. The papers are based on a symposium held in Denmark in April 2004 (www. monitoringmatters.org). Here, we review how the case studies shed light on the following key issues concerning locally-based methods: cost, sustainability, their ability to detect true local or larger-scale trends, their links to management decisions and action, and the empowerment of local constituencies. Locally-based monitoring appears to be consistently cheap relative to the costs of management and of professional monitoring, even though the start-up costs can be high. Most local monitoring schemes are still young and thus their chances of being sustained over the longer term are not yet certain. However, we believe their chances of surviving are better than many professional schemes, particularly when they are institutionalised within existing management structures, and linked to the delivery of ecosystem goods or services to local communities. When properly designed, local schemes yield locally relevant results that can be as reliable as those derived from professional monitoring. Many management decisions emanate from local schemes. The decisions appear to be taken promptly, in response to immediate threats to the environment, and often lead to community-based actions to protect habitats, species or the local flow of ecosystem benefits; however, few local schemes have so far led to actions beyond the local scale. Locally-based monitoring schemes often reinforce existing community-based resource management systems and lead to change in the attitude of locals towards more environmentally sustainable resource management. Locally-derived data have considerable unexplored potential to elucidate global patterns of change in the status of populations and habitats, the services they provide, and the threats they face, but more effort is needed to develop effective modalities for feeding locally-derived data up to national and international levels.     

Does Monitoring Matter? A Quantitative Assessment of Management Decisions from Locally-based Monitoring of Protected Areas

Biodiversity monitoring is criticized for being insufficiently relevant to the needs of managers and ineffective in integrating information into decision-making. We examined conservation management interventions resulting from 2½ years of monitoring by 97 rangers and 350 community volunteers over 1 million hectares of Philippine protected areas. Before this monitoring scheme was established, there was little collaboration between local people and park authorities, and park monitoring was restricted to assessments of the quantity of extracted timber. As a result of the scheme, 156 interventions were undertaken in terrestrial, marine and freshwater ecosystems. In total 98% of these interventions were meaningful and justified, 47% targeted the three most serious threats to biodiversity at the site, and 90% were implemented without external support, suggesting that the interventions were relevant and could be sustained over time at the local level. The mean time from sampling to decision-making was only 97 days, probably because 82% of the interventions were initiated by the same people and institutions that had compiled the underlying data, bypassing potential government bureaucracy. Many of the interventions were jointly undertaken by community members and the management authorities or consisted of local bylaws in support of park management. As a result of the monitoring, indigenous resource use regulation schemes were re-established with government recognition in several parks. The monitoring led to more diversified and realistic management responses on the part of the authorities, including a more socially acceptable and effective approach to enforcement. Of the four field monitoring techniques used, the most participatory one generated more interventions aimed at ensuring a continued resource supply for local communities (χ²₃ = 69.1, p <0.01). Although this suggests that the interest of community members is associated with their possibilities to influence the flow of ecosystem goods and services, the 156 interventions targeted, directly or indirectly, all known globally threatened species of mammals, birds and butterflies in the parks.     

Variation for neutral markers is correlated with variation for quantitative traits in the plant pathogenic fungus Mycosphaerella graminicola

We compared genetic variation and population differentiation at RFLP marker loci with seven quantitative characters including fungicide resistance, temperature sensitivity, pycnidial size, pycnidial density, colony size, percentage of leaves covered by pycnidia (PLACP) and percentage of leaves covered by lesions (PLACL) in Mycosphaerella graminicola populations sampled from four regions. Wide variation in population differentiation was found across the quantitative traits assayed. Fungicide resistance, temperature sensitivity, and PLACP displayed a significantly higher Q(ST) than G(ST), consistent with selection for local adaptation, while pycnidial size, pycnidial density and colony size displayed a lower or significantly lower Q(ST) than G(ST), consistent with constraining selection. There was not a statistical difference between Q(ST) and G(ST) in PLACL. We also found a positive and significant correlation between genetic variation in molecular marker loci and quantitative traits at the multitrait scale, suggesting that estimates of overall genetic variation for quantitative traits in M. graminicola could be derived from analysis of the molecular genetic markers.