Identification of dehydrin-like proteins responsive to chilling in floral buds of blueberry (Vaccinium, section Cyanococcus).

The level of three major polypeptides of 65, 60, and 14 kD increased in response to chilling unit accumulation in floral buds of a woody perennial, blueberry (Vaccinium, section Cynaococcus). The level of the polypeptides increased most dramatically within 300 h of chilling and decreased to the prechilling level with the initiation of budbreak. Cold-hardiness levels were assessed for dormant buds of Vaccinium corymbosum and Vaccinium ashei after different chilling treatments until the resumption of growth. These levels coincided with the level of the chilling-responsive polypeptides. Like some other previously described cold-induced proteins in annual plants, the level of the chilling-induced polypeptides also increased in leaves in response to cold treatment; the chilling-induced polypeptides were heat stable, resisting aggregation after incubation at 95 degrees C for 15 min. By fractionating bud proteins first by isoelectric point (pI) and then by molecular mass, the pI values of the 65- and 60-kD polypeptides were found to be 7.5 to 8.0 and the pI value of the 14-kD polypeptide was judged to be 8.5. Purification of the 65- and 60-kD polypeptides, followed by digestion with endoproteinase Lys-C and sequencing of selected fragments, revealed similarities in amino acid composition between the 65- and 60-kD polypeptides and dehydrins. Indeed, antiserum to the lysine-rich consensus sequence EKKGIMDKIKEKLPG of dehydrin proteins cross-reacted to all three of the major chilling-responsive polypeptides of blueberry, identifying these as dehydrins or dehydrin-like proteins.     

The evolution and ecology of multiple antipredator defences

Prey seldom rely on a single type of antipredator defence, often using multiple defences to avoid predation. In many cases, selection in different contexts may favour the evolution of multiple defences in a prey. However, a prey may use multiple defences to protect itself during a single predator encounter. Such “defence portfolios” that defend prey against a single instance of predation are distributed across and within successive stages of the predation sequence (encounter, detection, identification, approach (attack), subjugation and consumption). We contend that at present, our understanding of defence portfolio evolution is incomplete, and seen from the fragmentary perspective of specific sensory systems (e.g., visual) or specific types of defences (especially aposematism). In this review, we aim to build a comprehensive framework for conceptualizing the evolution of multiple prey defences, beginning with hypotheses for the evolution of multiple defences in general, and defence portfolios in particular. We then examine idealized models of resource trade-offs and functional interactions between traits, along with evidence supporting them. We find that defence portfolios are constrained by resource allocation to other aspects of life history, as well as functional incompatibilities between different defences. We also find that selection is likely to favour combinations of defences that have synergistic effects on predator behaviour and prey survival. Next, we examine specific aspects of prey ecology, genetics and development, and predator cognition that modify the predictions of current hypotheses or introduce competing hypotheses. We outline schema for gathering data on the distribution of prey defences across species and geography, determining how multiple defences are produced, and testing the proximate mechanisms by which multiple prey defences impact predator behaviour. Adopting these approaches will strengthen our understanding of multiple defensive strategies.     

Inferring cultural reproduction from lithic data: A critical review

The cultural reproduction of lithic technology, long an implicit assumption of archaeological theories, has garnered increasing attention over the past decades. Major debates ranging from the origins of the human culture capacity to the interpretation of spatiotemporal patterning now make explicit reference to social learning mechanisms and cultural evolutionary dynamics. This burgeoning literature has produced important insights and methodological innovations. However, this rapid growth has sometimes led to confusion and controversy due to an under-examination of underlying theoretical and methodological assumptions. The time is thus ripe for a critical assessment of progress in the study of the cultural reproduction of lithic technology. Here we review recent work addressing the evolutionary origins of human culture and the meaning of artifact variation at both intrasite and intersite levels. We propose that further progress will require a more extended and context-specific evolutionary approach to address the complexity of real-world cultural reproduction.     

An assessment of three selective media for bifidobacteria in faeces

Three previously described media enumerating Bifidobacterium spp. in faeces were compared with respect to their selectivity and quantitative recovery. The results of this study indicate that of the three media studied, Beerens'agar is the most suitable medium for isolation and enumeration of Bifidobacterium spp. from the gut microflora.     

Purification and characterization of dihydroorotate dehydrogenase A from Lactococcus lactis, crystallization and preliminary X-ray diffraction studies of the enzyme.

Lactococcus lactis is the only organism known to contain two dihydroorotate dehydrogenases, i.e., the A- and B-forms. In this paper, we report the overproduction, purification, and crystallization of dihydroorotate dehydrogenase A. In solution, the enzyme is bright yellow. It is a dimer of subunits (34 kDa) that contain one molecule of flavin mononucleotide each. The enzyme shows optimal function in the pH range 7.5-9.0. It is specific for L-dihydroorotate as substrate and can use dichlorophenolindophenol, potassium hexacyanoferrate (III), and, to a lower extent, also molecular oxygen as acceptors of the reducing equivalents, whereas the pyridine nucleotide coenzymes (NAD+, NADP+) and the respiratory quinones (i.e., vitamins Q6, Q10 and K2) were inactive. The enzyme has been crystallized from solutions of 30% polyethylene glycol, 0.2 M sodium acetate, and 0.1 M Tris-HCl, pH 8.5. The resulting yellow crystals diffracted well and showed little sign of radiation damage during diffraction experiments. The crystals are monoclinic, space group P21 with unit cell dimensions a = 54.19 A, b = 109.23 A, c = 67.17 A, and beta = 104.5 degrees. A native data set has been collected with a completeness of 99.3% to 2.0 A and an Rsym value of 5.2%. Analysis of the solvent content and the self-rotation function indicates that the two subunits in the asymmetric unit are related by a noncrystallographic twofold axis perpendicular to the crystallographic b and c axes.     

Tn552 transposase purification and in vitro activities.

The Staphylococcus aureus transposon Tn552 encodes a protein (p480) containing the 'D,D(35)E' motif common to retroviral integrases and the transposases of a number of bacterial elements, including phage Mu, the integron-containing element Tn5090, Tn7 and IS3. p480 and a histidine-tagged derivative were overexpressed in Escherichia coli and purified by methods involving denaturation and renaturation. DNase I footprinting and gel binding assays demonstrated that p480 binds to two adjacent, directly repeated 23 bp motifs at each end of Tn552. Although donor strand cleavage by p480 was not detected, in vitro conditions were defined for strand transfer activity with transposon end fragments having pre-cleaved 3' termini. Strand transfer was Mn(2+)-dependent and appeared to join a single left or right end fragment to target DNA. The importance of the terminal dinucleotide CA-3' was demonstrated by mutation. The in vitro activities of p480 are consistent with its proposed function as the Tn552 transposase.     

Stream-dwelling insects and extremely low frequency electromagnetic fields: a ten-year study

Changes in the structure of benthic insect communities at an experimental site and at a reference site in the Ford River, Michigan were monitored over a 10-year period to determine whether extremely low frequency electromagnetic fields (ELF) affected those communities. Five of 10 biotic parameters monitored are presented: taxon evenness (J), richness (S), numerical dominance of chironomids, and total insect mass. Data were separated into three seasons because coefficient of variation values were lower in the summer than in the spring and fall. Two-way ANOVA tests for the biotic variables were often significantly different between sites and among years, but the interaction terms were less frequently significant. Biotic parameters were regressed against stream discharge, water temperatures, years, and ELF cumulative ground field exposures. At the experimental site, discharge accounted for more variation than did water temperature or years for all biotic parameters except chironomid numerical dominance in the fall. Intervention analyses, using the B.A.C.I parametric or the R.I.A non-parametric showed significant differences in three of 15 cases; namely, for the highly varying chironomid numerical dominance values in the spring and fall and for the low varying total insect mass values in the summer. For those tests, the Before Impact period spanned April 1984 through May 1986. The After Impact period (full ELF power) spanned June 1989 through August 1993. Trend analysis for total insect mass at the experimental site in the summer showed discharge to be more important than water temperatures or ELF ground field exposures. Natural physical factors appear to be more important than the anthropogenic ELF fields in accounting for seasonal and yearly changes in the community.