The active site of an algal prolyl 4-hydroxylase has a large structural plasticity

Prolyl 4-hydroxylases (P4Hs) are 2-oxoglutarate dioxygenases that catalyze the hydroxylation of peptidyl prolines. They play an important role in collagen synthesis, oxygen homeostasis, and plant cell wall formation. We describe four structures of a P4H from the green alga Chlamydomonas reinhardtii, two of the apoenzyme at 1.93 and 2.90 A resolution, one complexed with the competitive inhibitor Zn2+, and one with Zn2+ and pyridine 2,4-dicarboxylate (which is an analogue of 2-oxoglutarate) at 1.85 A resolution. The structures reveal the double-stranded beta-helix core fold (jellyroll motif), typical for 2-oxoglutarate dioxygenases. The catalytic site is at the center of an extended shallow groove lined by two flexible loops. Mutagenesis studies together with the crystallographic data indicate that this groove participates in the binding of the proline-rich peptide-substrates. It is discussed that the algal P4H and the catalytic domain of collagen P4Hs have notable structural similarities, suggesting that these enzymes form a separate structural subgroup of P4Hs different from the hypoxia-inducible factor P4Hs. Key structural differences between these two subgroups are described. These studies provide first insight into the structure-function relationships of the collagen P4Hs, which unlike the hypoxia-inducible factor P4Hs use proline-rich peptides as their substrates.     

Predictors of plant phenology in a diverse high‐latitude alpine landscape: growth forms and topography

Question: Different plant growth forms may have distinctly different functioning in ecosystems. Association of phenological patterns with growth form will therefore help elucidate the role of phenology in an ecosystem. We ask whether growth forms of common vascular plants differ in terms of vegetative and flowering phenology, and if such phenological differences are consistent across environmental gradients caused by landscape‐scale topography. Location: A high‐latitude alpine landscape in Finnmark County, Norway (70°N). Methods: We assessed vegetative and flowering phenology repeatedly in five growth forms represented by 11 common vascular plant species across an altitudinal gradient and among differing slope aspects. Results: Species phenology clustered well according to growth form, and growth form strongly explained variation in both flowering and vegetative phenology. Altitude and aspect were poor predictors of phenological variation. Vegetative phenology of the growth forms, ranked from slowest to fastest, was in the order evergreen shrubs <sedges‐deciduous shrubs <grasses <forbs, while a reverse ranking was found for flowering phenology. Conclusion: Growth form‐specific phenological patterns are associated with fundamentally different abilities for resource acquisition and resource conservation. The weak effect of landscape‐scale topographic factors indicates that variation within growth forms is mainly influenced by local environmental factors not accounted for in this study. On the basis of these results, we argue that growth forms should be considered as predictors of phenology together with the customary use of topography and normalized difference vegetation index, especially when assessing the role of phenology in an ecosystem.     

Cadmium in insects after ash fertilization

Ash fertilization of forests returns nutrients to forest ecosystems and has a positive effect on soil pH, but it also may elevate Cd concentrations of forest biota. Cadmium concentrations of some forest insects ( Formica ants, carabids and Coleopteran larvae from decaying wood) were investigated in southern Finland, where two plots were fertilized with wood ash, while two other plots represented unfertilized control plots. In ants, mean Cd concentration was 3.6 ± 1.4 mg/kg, with nest workers having significantly higher concentrations than workers trapped in pitfall traps. Concentrations at fertilized and unfertilized plots were similar. In carabid beetles, the average Cd concentration of Carabus glabratus was 0.44 ± 0.36 mg/kg, with no significant difference between control plots and fertilized plots. In another carabid beetle, Pterostichus niger , mean Cd concentration was higher at fertilized plots compared to control plots. We conclude that the variation of Cd concentrations in the insects studied is more efficiently controlled by species-specific differences than fertilization history of the forest floor.     

Process length variation in cysts of a dinoflagellate,Lingulodinium machaerophorum,in surface sediments: Investigating its potential as salinity proxy

A biometrical analysis of the dinoflagellate cyst Lingulodinium machaerophorum [Deflandre, G., Cookson, I.C., 1955. Fossil microplankton from Australia late Mesozoic and Tertiary sediments. Australian journal of Marine and Freshwater Research 6: 242–313.] Wall, 1967 in 144 globally distributed surface sediment samples revealed that the average process length is related to summer salinity and temperature at a water depth of 30 m by the equation (salinity/temperature) = (0.078?average process length + 0.534) with R² = 0.69. This relationship can be used to reconstruct palaeosalinities, albeit with caution. The particular ecological window can be associated with known distributions of the corresponding motile stage Lingulodinium polyedrum (Stein) Dodge, 1989. Confocal laser microscopy showed that the average process length is positively related to the average distance between process bases (R² = 0.78), and negatively related to the number of processes (R² = 0.65). These results document the existence of two end members in cyst formation: one with many short, densely distributed processes and one with a few, long, widely spaced processes, which can be respectively related to low and high salinity/temperature ratios. Obstruction during formation of the cysts causes anomalous distributions of the processes. From a biological perspective, processes function to facilitate sinking of the cysts through clustering.     

Spatial distribution of Holcocerus hippophaecolus (Lepidoptera: Cossidae) pupae in a seabuckthorn (Hippophae rhamnoides) stand

The seabuckthorn carpenter moth, Holcocerus hippophaecolus, which has a generation time of four years, is recently becoming one of the major pests of the seabuckthorn (Hippophae rhamnoides) in Inner Mongolia, Liaoning, Shanxi, Ningxia and Shaanxi of China (Hua et al., 1990). The larvae of the H. hippophaecolus mainly damage the stems and roots of the seabuckthorn, and the mature larvae pupate in the soil. The spatial distribution of the pupae was analyzed by using biostatistics and geostatistics in order to effectively control the insect and further study the spatial distribution of the population. Results show that most of the pupae (90%) had an eclosion time span from early June to the end of July. The sex ratio of the pupae was nearly 1:1 in the woodland samples. In addition, 24.3% of the 971 trees investigated had pupae and it ranged from 0 to 4 per tree within a distance of 1.3 m from the base of the stem. 90% of the pupae were aggregated within a distance of 1 m from the base of the stem. The pupae show intense spatial aggregation in the sampled woodland which had an 11.1 m spatial dependence and a 90.7% intensity in the local spatial continuity. Moreover, the population presented an intensive spotted distribution and many aggregated spots were found in the woodlands. As for the relationship between grid size and variogram of the pupae, the variations in the range, the intensity of local spatial continuity and the sill were all very low or non-existent when the grid size was 5 m, 6 m or 7 m. Whereas, the value of the decisive coefficient was the biggest when the grid size was 5 m making it the ideal grid size.     

Carbon debt and carbon sequestration parity in forest bioenergy production

The capacity for forests to aid in climate change mitigation efforts is substantial but will ultimately depend on their management. If forests remain unharvested, they can further mitigate the increases in atmospheric CO₂ that result from fossil fuel combustion and deforestation. Alternatively, they can be harvested for bioenergy production and serve as a substitute for fossil fuels, though such a practice could reduce terrestrial C storage and thereby increase atmospheric CO₂ concentrations in the near‐term. Here, we used an ecosystem simulation model to ascertain the effectiveness of using forest bioenergy as a substitute for fossil fuels, drawing from a broad range of land‐use histories, harvesting regimes, ecosystem characteristics, and bioenergy conversion efficiencies. Results demonstrate that the times required for bioenergy substitutions to repay the C Debt incurred from biomass harvest are usually much shorter (< 100 years) than the time required for bioenergy production to substitute the amount of C that would be stored if the forest were left unharvested entirely, a point we refer to as C Sequestration Parity. The effectiveness of substituting woody bioenergy for fossil fuels is highly dependent on the factors that determine bioenergy conversion efficiency, such as the C emissions released during the harvest, transport, and firing of woody biomass. Consideration of the frequency and intensity of biomass harvests should also be given; performing total harvests (clear‐cutting) at high‐frequency may produce more bioenergy than less intensive harvesting regimes but may decrease C storage and thereby prolong the time required to achieve C Sequestration Parity.     

Elements within the beta-lactoglobulin gene inhibit expression of human serum albumin cDNA and minigenes in transfected cells but rescue their expression in the mammary gland of transgenic mice.

Two new beta-lactoglobulin (BLG)/human serum albumin (HSA) hybrid gene vectors were constructed and tested for expression in COS-7 cells and in transgenic mice. The HSA sequences were inserted between the second and sixth BLG exons. Transient transfection experiments with these vectors as well as a series of additional vectors with either the BLG 5'- or 3'- intragenic sequences revealed that sequences within BLG exon 1/intron 1/exon 2 abrogated BLG- directed HSA expression in vitro, regardless of the presence of HSA introns or the origin of the 3' polyadenylation signal. In contrast, the same BLG expression cassette enabled the efficient expression of HSA cDNA or minigene in the mammary gland of transgenic mice with subsequent secretion of the corresponding protein into the milk of 56 and 82%, respectively of the mouse strains at levels up to 0.3 mg/ml. Previous attempts to express HSA cDNA inserted into exon 1 of the BLG gene had failed [Shani,M., Barash,I., Nathan,M., Ricca,G., Searfoss,G.H., Dekel,I., Faerman,A., Givol,D. and Hurwitz,D.R. (1992) Transgenic Res. 1, 195- 208]. The new BLG expression cassette conferred more stringent tissue specific expression than previously described BLG/HSA constructs [Barash,I, Faerman,A., Ratovitsky,T, Puzis,R., Nathan,M., Hurwitz,D.R. and Shani, M. (1994) Transgenic Res. 3, 141-151]. However, it was not able to insulate the transgenes from the surrounding host DNA sequences and did not result in copy number dependent expression in transgenics. Together, the in vitro and in vivo results suggest both positive and negative regulatory elements within the BLG intragenic sequences evaluated. The new BLG construct represents an extremely valuable vector for the efficient expression of cDNAs in the mammary gland of transgenic animals.     

Allocation of resources within mountain birch canopy after simulated winter browsing

As a response to browsing, birches are known to produce fewer but larger, more nutritious leaves, with enhanced palatability for herbivores. We simulated winter browsing in ramets of mountain birch ( Betula pubescens ssp. czerepanovii ) to find out whether it decreases subsequent foliage biomass and alters the number and type of shoots. After removal of a considerable proportion of buds (up to 35%) in late winter, the birches were able to compensate for the lost leaf biomass in the following summer; there were no differences in total leaf biomass between winter-clipped and control ramets. This indicates that foliage growth was limited by the total amount of stored resources, not by the number of buds. Depending on the position of the buds removed, different mechanisms were responsible for the compensation. After removal of apical buds, the number of leaves decreased significantly but leaves were larger than in control ramets. Removal of the same mass of basal buds – containing similar amount of carbohydrates and proteins as in the treatment removing apical buds – activated dormant buds, especially in apical locations, so that leaf number was similar as in the controls; consequently, size of individual leaves increased only slightly. Thus, while the total leaf biomass in a tree seems to be limited by resources from source organs, the distribution of resources among different canopy sections is controlled by their relative sink strengths. In terms of leaf biomass, apical parts are able to compensate for bud loss by increasing shoot number, basal parts only by increasing leaf size.