Stable carbon isotopes in tree rings of beech: climatic versus site-related influences

 Stable carbon isotopes in tree rings are a promising tool in palaeoclimate research, provided attempts are made to disentangle climatic from local effects (e.g. soil properties, competition, light). The ¹³C/¹²C variations in cellulose of tree rings of beech (Fagus sylvatica) were determined at several sites in the Swiss Central Plateau covering the last 50 years. We chose sites which differ in moisture conditions and sampled cores from four to six trees per site. The mean ¹³C/¹²C series from the different dry sites (distant by up to 40 km) are closely interrelated suggesting a common external cause. Correlation analysis with climate data proved the total precipitation in the months May, June and July to have the strongest effect on the carbon isotopes (r =  – 0.73). This result is in agreement with the commonly used model which relates the isotope discrimination to the water use efficiency. On the other hand, the isotope series of the wet sites are not as well correlated to the climate. At two of the sites (a dry and a humid) tree ring width suddenly increased. We used this effect as a test-case to study the influence of local growth conditions on the climate-isotope relationship. Received: 17 April 1996 / Accepted: 2 September 1996     

A mathematical model to describe the dynamic response of a spruce tree to the wind

 A mathematical, computer-based, dynamic sway model of a Sitka spruce (Picea sitchensis) tree was developed and tested against measurements of the movement of a tree within a forest. The model tree was divided into segments each with a stiffness, mass and damping parameter. Equations were formulated to describe the response of every segment which together form a system of coupled differential equations. These were solved with the aid of matrices and from the resulting modes, the transfer function of the tree was found and used to calculate the movement of the tree in the wind. Comparison of the modelled movement of a tree in response to the measured wind speed above a forest canopy gave good agreement with the measured movement of the top of the tree but less satisfactory agreement close to the base. The comparison also pointed to the complexity of tree response to the wind and inadequacies in the model. In particular, the branches need to be treated as coupled cantilevers attached to the stem rather than simply as masses lumped together. Received: 18 February 1997 / Accepted: 16 December 1997     

Time constant for water transport in loblolly pine trees estimated from time series of evaporative demand and stem sapflow

 The use of stem sap flow data to estimate diurnal whole-tree transpiration and canopy stomatal conductance depends critically upon knowledge of the time lag between transpiration and water flux through the stem. In this study, the time constant for water movement in stems of 12-year-old Pinus taeda L. individuals was estimated from analysis of time series data of stem water flux and canopy transpiration computed from mean daytime canopy conductance, and diurnal vapor pressure deficit and solar radiation measurements. Water uptake through stems was measured using a constant-heat sapflow probe. Canopy transpiration was correlated to stem uptake using a resistance-capacitance equation that incorporates a time constant parameter. A least-squares auto-regression determined the parameters of the resistance-capacitance equation. The time constants for ten loblolly pine trees averaged 48.0 (SE = 2.0) min and the time lag for the diurnal frequency averaged 47.0 (SE = 2.0) min. A direct-cross correlation analysis between canopy transpiration and sap flow time series showed maximum correlation at an approximately 30 min lag. Residuals (model-predicted minus actual stem flow data) increased with increasing soil moisture depletion. While the time constants did not vary significantly within the range of tree sizes studied, hydraulic resistance and capacitance terms were individually dependent on stem cross-sectional area: capacitance increased and resistance decreased with stem volume. This result may indicate an inverse adjustment of resistance and capacitance to maintain a similar time constant over the range of tree sizes studied.     

Stem-growth response of Pinus sylvestris and Picea abies to nitrogen fertilization as related to needle nitrogen concentration

 Responses of stem-volume growth to N application were evaluated in relation to foliar N concentrations. Data from N-fertilization experiments in 28 Pinus sylvestris stands and 21 Picea abies stands were used. Relative stem-growth responses were negatively related to concentrations of N in current-year needles of unfertilized trees. There appeared to be a threshold value of 15–16 mg (g DM)^–1 N in current-year needles, above which N-application is unlikely to stimulate growth. However, relations were non-significant between N concentrations in current-year needles and the absolute stem-growth response [dm³ ha^–1 (5 years)^–1]. The indicated threshold values are discussed in relation to other variables reflecting the N richness of the environment.--> Received: 20 December 1996 / Accepted: 30 September 1997     

M13 DNA fingerprinting can be used in studies on phenotypic reversions of forest tree mutants

 Solitary revertants which have been observed on single mutant tree individuals have up to now been believed to be grow-through cells belonging to the rootstock on which they are commonly grafted. In this study three different phenotypically visible mutants revealing revertant shoots on the same tree were chosen for genetic analysis. The mutant Quercus robur L. ‘argenteomarginata’ was grafted on a normal rootstock, an individual of Carpinus betulus L. var. quercifolia Desf. as well as an individual of Picea glauca (Moench) Voss. ‘conica’ are supposed to have grown from seeds. By means of a highly specific M13 PCR fingerprinting technique the mutant and revertant tissues were analysed in comparison to different individuals of each of the species. With the grafted mutant, cambium tissue of the rootstock was also investigated. Whereas conspecific individuals could be clearly distinguished from each other, mutant and revertant tissues revealed the same banding patterns for each of the three trees. In case of the grafted mutant, the fingerprint obtained from cambium tissue of the rootstock was clearly different from the pattern of mutant and revertant tissue. Results demonstrate the potential of the tool for genetic differentiation between individuals of three tree species hence in the case of the grafted mutant, the hypothesis that the observed reversion is caused by a grow-through of the rootstock is rejected. Furthermore, identical fingerprints of mutant and revertant tissue support identical genetic background of the tissues excluding the gene(s) responsible of the mutation. Possible causes of mutations and reversions regarding the three mutant trees are discussed. Received: 15 September 1997 / Accepted: 24 November 1997     

Scaling of tree height and trunk diameter as a function of ring number

 Phipps (1967) showed, in suppressed maples and oaks, that on average the cross-sectional area of successive tree rings is independent of ring number. From that empirical result I argued that mean ring width and mean ring diameter scale as the inverse square root and square root, respectively, of ring number (Prothero 1997). Here I give an illustrative argument as to how macroscopic trunk variables may be related quantitatively to microscopic ring variables. First, evidence is presented consistent with the theory of Greenhill (1881) and the empirical evidence of McMahon (1973) that tree height in a diversity of species scales as about the two-thirds power of trunk diameter. Here I report further evidence bearing on the same question. From these combined results I suggest that tree height, and by implication trunk height, and other parameters governing macroscopic tree trunk proportions, can all be expressed as powers of a microscopic variable, namely ring number, where the scaling exponents are all proper fractions. I suggest that at least one of these relationships is, in principle, non-adaptive. Received: 9 June 1998 / Accepted: 4 February 1999     

Immunization of mice with melanoma cells transfected to secrete the superantigen, staphylococcal enterotoxin A

Immunization of mice with a melanoma vaccine coupled with staphylococcal enterotoxin A (SEA) inhibits the growth of primary melanoma tumors in mice. We have now successfully transfected B16 cells with the sea gene and have immunized C57BL/6 mice subcutaneously once per week for 4 weeks prior to tumor challenge with vaccines of irradiated B16 cells or, 4 weeks following tumor challenge of naïve mice with B16 cells, with irradiated B16 cells transfected with the sea gene. Primary tumor growth following both types of treatments was inhibited significantly. To characterize immune responses to these immunogens, we examined the production of antibodies to the B700 melanoma antigen, the stimulation of endogenous IL-2 production, the expression of CD4, CD8, Vβ and CD25 T cell markers, and the induction of NK activity. At 4 weeks following immunization of mice, there was a significant increase (P<0.05) in levels of interleukin-2 production by splenocytes from mice immunized with SEA-secreting B16 cells or with the parental B16 cells, compared to controls. Levels of antibodies to the B700 melanoma antigen were also significantly higher in mice immunized with the SEA-secreting B16 cells, as was expression of CD4, CD8, CD25 and Vβ T cell antigens, particularly CD4. Natural killer cell activity (at various E:T ratios) was tenfold higher in splenocytes of mice immunized with SEA-secreting B16 cells, and fivefold higher in mice immunized with the parental B16 cells, compared to controls.?These data confirm the possibility of using irradiated murine melanoma cells transfected to secrete SEA in vaccines targeted at preventing the development and growth of melanoma.     

A rule-based approach to the modelling of bacterial ecosystems

This paper presents an approach to ecological/evolutionary modelling that is inspired by natural bacterial ecosystems and bacterial evolution. An individual-based artificial ecosystem model is proposed, which is designed to explore the evolvability of adaptive behavioural strategies in artificial bacteria represented by rule-based learning classifier systems. The proposed ecosystem model consists of a n-dimensional environmental grid, which can contain different types of artificial resources in arbitrary arrangements. The resources provide the energy that is necessary for the organisms to sustain life, and can trigger different types of behaviour in the organisms, such as movement towards nutrients and away from toxic substances, growth, and the controlled release of signalling resources. The balance between energy and material is modelled carefully to ensure that the ecosystem is dissipative. Those organisms that are able to efficiently exploit the available resources gradually accumulate enough energy to reproduce (by binary fission) and generate copies of themselves in the environment. Organisms are also able to produce their own resources, which can potentially be used as markers to send signals to other organisms (a behaviour known as quorum sensing). The complex relationships between stimuli and actions in the organisms are stochastically altered by means of mutations, thus enabling the organisms to adapt to their environment and maximise their lifespan and reproductive success. In this paper, the proposed bacterial ecosystem model is defined formally and its structure is discussed in detail. This is followed by results from simulation experiments that illustrate the model's operation and how it can be used in evolutionary modelling/computing scenarios.     

Field comparisons of photosynthesis and leaf conductance in Ziziphus mauritiana and other fruit tree species in Zimbabwe

 Data are presented for gas exchange in exposed leaves in field grown Ziziphus mauritiana (Lamk.) at a highland site and potted seedlings at a hotter lowland site in tropical Zimbabwe, together with indigenous and locally-grown commercial fruit crops. The field trial at the highland site included local Ziziphus mauritiana, introduced Indian Z. mauritiana (cv. Umran), fig and peach. In all species assimilation was highest early in the morning, followed by a gradual decline throughout the remainder of the day. Leaf conductance followed the same trend as assimilation for fig and peach, but in Ziziphus cv. Umran and Ziziphus Musau, conductance tracked irradiance, reaching a maximum in the middle of the day. In all species, sub-stomatal CO₂ concentrations increased with declining assimilation indicating that during high irradiance, assimilation was mainly controlled by mesophyll limitations rather than conductance. At the highland site both Ziziphus cv. Umran and Musau were highly productive, with light saturated assimilation significantly higher than in either fig or peach (P < 0.01). At the warmer lowland site, assimilation and conductance in Ziziphus Musau were also higher than in other indigenous fruit tree species growing under similar conditions. Despite increased assimilation in Ziziphus, when compared to the other species, there was no increase in the assimilation ratio (ratio of assimilation/conductance) which was offset by the high conductance values. The data indicate that under conditions where water was not limiting, young Ziziphus showed no enhanced stomatal control over water loss, but was highly productive (per unit leaf area) relative to the other species. Received: 17 October 1996 / Accepted: 8 January 1997     

Retention and processing of the monoclonal anti-prostate E4 antibody after binding to prostatic adenocarcinoma DU 145 cells

 Cellular retention and processing of a radiolabelled monoclonal anti-prostate antibody were evaluated after binding to prostatic adenocarcinoma DU 145 cells. An endocytosis assay revealed that the rate of release of radioactivity from the cells had an initially rapid phase within the first hours after antibody incubation, which was presumably due to release of monovalently bound antibodies. This was followed by a slower phase, with the possible release of intact bivalently bound antibodies and excretion of degraded internalized antibodies. The relative amount of released radioactivity of high molecular mass was high, indicating that the major part of the antibodies were released without being internalized and degraded. However, when only the radiolabelled antibody that remained cell-associated after 2 h and longer was considered, a substantial part was found to be internalized and radioactive degradation products were excreted. About 30% of the initially cell-associated radioactivity still remained associated with the cells after 48 h, indicating a rather slow antibody processing, which is favourable if the antibody is to be used for targeted radiotherapy. The retention of cell-associated radioiodine was very similar irrespective of whether the antibodies were radiolabelled with the direct chloramine-T method or the indirect ATE (succinimidyl based reagent) method. Since the ATE method can be used to form stable antibody constructs with the therapeutically relevant alpha-emitting radionuclide, astatine-211, this was an interesting finding that will be further evaluated in the future. Received: 20 March 1996 / Accepted: 25 June 1996     

Photosynthetic responses of Eucalyptus nitens (Deane and Maiden) Maiden to green pruning

 Three-year-old Eucalyptus nitens (Deane and Maiden) Maiden trees and 1-year-old ramets of a single clone of E. nitens were pruned to remove 0, 50% or 70% of the green crown length. This was equivalent to removal of 0, 55% or 88% of foliage area of trees, and 0, 77% or 94% of foliage area of ramets. CO₂ assimilation (A) and stomatal conductance (g_s) were measured at constant illumination in five height zones and three foliage-age classes of trees over a 16-month period following pruning. Foliar nitrogen (N) and phosphorus (P) concentrations were determined for each measurement time during the first 12 months of the experiment. In ramets A and g_s were measured in four height zones and two foliage-age classes over a six-week period, and N and P concentrations were measured only once, at the end of the experiment. Rates of A increased by up to 175% following pruning. This response occurred throughout the canopy irrespective of position in the crown or foliage age. The magnitude of the response was generally greater in ramets than in trees, and increased with increasing severity of pruning. The initiation of the response was later, and the duration of the response was longer, in trees than ramets. In the lower crown of trees there was evidence of delayed senescence following pruning. Photosynthetic enhancement was not related to changes in foliar N concentrations. The ratio of A/N increased in many zones following pruning, especially after more severe defoliation. There was no evidence that changes in P concentrations were responsible for the result. The increases in A may have been related to changes in g_s, as maximum values of g_s were greater, and the ratio of A/g_s was generally lower, in pruned than unpruned plants. Received: 31 December 1996 / Accepted: 19 August 1997     

Modification of ricin A chain, by addition of endoplasmic reticulum (KDEL) or Golgi (YQRL) retention sequences, enhances its cytotoxicity and translocation

 A pKK expression system in Escherichia coli was used to produce recombinant ricin A chain (rRTA) and rRTA modified by addition of organelle-specific amino acid retention sequences, including KDEL (an endoplasmic reticulum, ER, lumen retention signal), KKMP (an ER membrane retention signal), YQRL (a trans-Golgi network retention signal) and KFERQ (a lysosome-targeting signal) to the C terminus of rRTA. The toxicities of these RTA mutants were assessed in Jurkat cells following fluid-phase endocytosis. rRTA-KDEL and rRTA-YQRL were significantly more cytotoxic for Jurkat cells than rRTA, rRTA-KKMP or rRTA-KFERQ. This difference did not result from signal(KDEL or YQRL)-mediated binding of these RTA mutants to the cell surface. Reconstituted ER and Golgi vesicles have been employed to assess translocation of rRTA and mutant rRTA. RTA-KDEL and RTA-YQRL respectively exhibited 6.7-fold and 6.1-fold more protection against papain digestion in reconstituted ER vesicles and 2.2-fold and 1.8-fold more protection in reconstituted Golgi vesicles, than unmodified rRTA. These mutants were reassociated with ricin B chain to form holotoxins. The mutant RTA-KDEL and RTA-YQRL holotoxins were 3.8-fold and 1.5-fold more cytotoxic for target cells, respectively, than ricin produced using unmodified rRTA. Our results suggest that both ER and the trans-Golgi network may play important roles in the intracellular trafficking and translocation of ricin A chain. Received: 14 August 1997 / Accepted: 14 October 1997     

Distribution patterns of foliar carbon and nitrogen as affected by tree dimensions and relative light conditions in the canopy of Picea abies

 Variations in the partitioning of foliar carbon and nitrogen in combination with changes in needle and shoot structure were studied in trees of Picea abies along a vertical gradient of relative irradiance (RI). RI was the major determinant of needle morphology, causing all needle linear parameters – width, thickness and length – to increase. Due to the different responsiveness of needle thickness and width in respect of RI, the ratio of total to projected needle area increased with RI. Furthermore, shoot structure was also influenced by RI, and the ratio of shoot silhouette area to total needle area, which characterises the packing of needles and needle area within the shoot, was greater at lower values of irradiance. Needle dry weight per total needle area (LWA_t) was also increased by RI. Similarly, irrespective of the measure for surface area, needle nitrogen content per area, as the product of needle dry weight per area and nitrogen content per needle dry weight (N_m), scaled quasi-linearly with needle weight per area. Thus, the changes in needle and shoot morphology made it possible to invest more photosynthesising weight per unit light-intercepting surface there, where the pay-back due to elevated irradiances was the highest. However, N_m behaved in an entirely different manner, decreasing hyperbolically with LWA_t. Since non-structural (carbon in non-structural carbohydrates), and structural (total minus non-structural) needle carbon per dry weight also increased with LWA_t, N_m was inversely correlated with both non-structural and structural carbon. Total tree height, increasing significantly LWA_t, also influenced needle structure. It appeared that total height did not affect needle thickness or width, but larger trees had greater needle density (dry weight per volume). Because needle density was positively correlated with needle carbon content per dry weight, it was assumed that the greater values of needle carbon content can be attributed to increased lignification and thickening of needle cell walls. Thus, it appeared that the proportion of supporting structures was greater in needles of larger trees. Inasmuch as an increased fraction of supporting structures dilutes other leaf substances, including also leaf compounds responsible for CO₂-assimilation, enhanced requirement for supporting structures may be responsible for lower rates of carbon assimilation per foliage dry weight observed in large trees. Increasing water limitation with increasing tree size is discussed as a possible cause for increased needle supporting costs in large trees. Received: 2 April 1995 / Accepted: 16 February 1996     

A decision tree approach to modelling nitrogen fertiliser use efficiency in New Zealand pastures

Quantifying nitrogen (N) fertiliser use efficiency (NFUE) in pastoral systems has important implications for fertiliser management from both economic and environmental points of view. The potential of a decision tree approach for modelling NFUE in New Zealand pastures was investigated. The decision tree model suggested that the time of applying N fertiliser was the most important factor influencing NFUE, with August or September (early spring in New Zealand) being the best time of application. The interaction of rainfall and temperature, rainfall, phosphorus (P) fertiliser history, soil Olsen P and slope were other important factors influencing NFUE. The model was validated for 11 of the 16 trials tested with a predictive accuracy of 69%. The mechanisms by which these factors influenced NFUE and the uncertainty associated with the model prediction were discussed. It was concluded that this type of modelling approach can be used to predict NFUE and thereby to assist decisions on when and where to apply N fertiliser in pastures for increasing productivity while reducing the environmental impact.     

A new species of gobiid fish, <Emphasis Type="Italic">Cristatogobius rubripectoralis</Emphasis>, from Australia

 A species of the gobiid genus Cristatogobius from northeastern Australia is described as new. This species is distinguishable from other species of the genus in having a higher number of scales in a longitudinal row and in a transverse row and a rounded caudal fin. In addition, there are differences in coloration such as brown reticulation on the upper anterior part of body and a red pectoral fin. A species of Cristatogobius reported from S. Java, Indonesia, is also identified as this species. Received: May 21, 2002 /Revised: November 15, 2002 / Accepted: December 16, 2002     

Limitations of compositional approach to identifying horizontally transferred genes

Genes with atypical G+C content and pattern of codon usage in a certain genome are possibly of exotic origin, and this idea has been applied to identify horizontal events. In this way, it was postulated that a total of 755 genes in the E. coli genome are relics of horizontal events after the divergence of E. coli from the Salmonella lineage 100 million years ago (Lawrence and Ochman, 1998). In this paper we propose a new way to study sequence composition more thoroughly. We found that although the 755 genes differ in composition from other genes in the E. coli genome, the difference is minor. If we accepted that these genes are horizontally transferred, then (1) it would be more likely that they were transferred from genomes evolutionarily closely related to E. coli; but (2) the dating method used by Lawrence and Ochman (1997, 1998) largely underestimated the average age of introduced sequences in the E. coli genome, in particular, most of the 755 genes should be introduced into E. coli before, instead of after, the divergence of E. coli from the Salmonella lineage. Our study reveals that atypical G+C content and pattern of codon usage are not reliable indicators of horizontal gene transfer events. Received: 27 September 2000 / Accepted: 9 April 2001     

Description and familial allocation of the African fluvial genus <Emphasis Type="Italic">Teleogramma</Emphasis> to the Cichlidae

 The external morphology, osteology, and myology of the African fluvial genus Teleogramma are described, and its familial allocation is discussed. Teleogramma is included in the family Cichlidae by loss of a major structural association between adductor mandibulae sections 2 and w, and by having an insertion of a large ventral division of adductor mandibulae section 2 onto the anguloarticular, expanded head of the fourth epibranchial, transversus dorsalis subdivided into four parts, functionally decoupled premaxillae and maxillae, the stomach's extendible blind pouch, the left-hand exit to the anterior intestine, the first intestinal loop at the left side, two epurals, seven branched rays on each upper and lower caudal fin lobe, free first uroneural from a united element of first preural and ural vertebra, and third preural vertebra fused with its haemal spine. Seven synapomorphies supporting the monophyly of Teleogramma are indicated, including the absence of or very low supraoccipital crest, the presence of a nostril tube, nonextended supraoccipital anteriorly, absence of extensive cartilaginous cap on the anterior border of the second epibranchial, presence of a beaklike projection on the cleithrum, caudal branched slip of epaxialis that inserts onto the upper two or three branched rays on the upper lobe of the caudal fin, and flexor dorsalis superior, which inserts onto the lower four unbranched rays on the upper lobe of the caudal fin. Received: September 12, 2001 / Revised: December 17, 2001 / Accepted: December 28, 2001