Constitutive and herbivore-inducible glucosinolate concentrations in oilseed rape (Brassica napus) leaves are not affected by Bt Cry1Ac insertion but change under elevated atmospheric CO2 and O3

Glucosinolates are plant secondary compounds involved in direct chemical defence by cruciferous plants against herbivores. The glucosinolate profile can be affected by abiotic and biotic environmental stimuli. We studied changes in glucosinolate patterns in leaves of non-transgenic oilseed rape (Brassica napus ssp. oleifera) under elevated atmospheric CO₂ or ozone (O₃) concentrations and compared them with those from transgenic for herbivore-resistance (Bacillus thuringiensis Cry1Ac endotoxin), to assess herbivory dynamics. Both elevated CO₂ and O₃ levels decreased indolic glucosinolate concentrations in transgenic and non-transgenic lines, whereas O₃ specifically increased the concentration of an aromatic glucosinolate, 2-phenylethylglucosinolate. The herbivore-inducible indolic glucosinolate response was reduced in elevated O₃ whereas elevated CO₂ altered the induction dynamics of indolic and aliphatic glucosinolates. Herbivore-resistant Bt plants experienced minimal leaf damage after target herbivore Plutella xylostella feeding, but exhibited comparatively similar increase in glucosinolate concentrations after herbivory as non-transgenic plants, indicating that the endogenous glucosinolate defence was not severely compromised by transgenic modifications. The observed differences in constitutive and inducible glucosinolate concentrations of oilseed rape under elevated atmospheric CO₂ and O₃ might have implications for plant–herbivore interactions in Brassica crop-ecosystems in future climate scenarios.     

Evaluation of six process‐based forest growth models using eddy‐covariance measurements of CO2 and H2O fluxes at six forest sites in Europe

Reliable models are required to assess the impacts of climate change on forest ecosystems. Precise and independent data are essential to assess this accuracy. The flux measurements collected by the EUROFLUX project over a wide range of forest types and climatic regions in Europe allow a critical testing of the process‐based models which were developed in the LTEEF project. The ECOCRAFT project complements this with a wealth of independent plant physiological measurements. Thus, it was aimed in this study to test six process‐based forest growth models against the flux measurements of six European forest types, taking advantage of a large database with plant physiological parameters. The reliability of both the flux data and parameter values itself was not under discussion in this study. The data provided by the researchers of the EUROFLUX sites, possibly with local corrections, were used with a minor gap‐filling procedure to avoid the loss of many days with observations. The model performance is discussed based on their accuracy, generality and realism. Accuracy was evaluated based on the goodness‐of‐fit with observed values of daily net ecosystem exchange, gross primary production and ecosystem respiration (gC m^?2 d^?1), and transpiration (kg H₂O m^?2 d^?1). Moreover, accuracy was also evaluated based on systematic and unsystematic errors. Generality was characterized by the applicability of the models to different European forest ecosystems. Reality was evaluated by comparing the modelled and observed responses of gross primary production, ecosystem respiration to radiation and temperature. The results indicated that: Accuracy. All models showed similar high correlation with the measured carbon flux data, and also low systematic and unsystematic prediction errors at one or more sites of flux measurements. The results were similar in the case of several models when the water fluxes were considered. Most models fulfilled the criteria of sufficient accuracy for the ability to predict the carbon and water exchange between forests and the atmosphere. Generality. Three models of six could be applied for both deciduous and coniferous forests. Furthermore, four models were applied both for boreal and temperate conditions. However, no severe water‐limited conditions were encountered, and no year‐to‐year variability could be tested. Realism. Most models fulfil the criterion of realism that the relationships between the modelled phenomena (carbon and water exchange) and environment are described causally. Again several of the models were able to reproduce the responses of measurable variables such as gross primary production (GPP), ecosystem respiration and transpiration to environmental driving factors such as radiation and temperature. Stomatal conductance appears to be the most critical process causing differences in predicted fluxes of carbon and water between those models that accurately describe the annual totals of GPP, ecosystem respiration and transpiration. As a conclusion, several process‐based models are available that produce accurate estimates of carbon and water fluxes at several forest sites of Europe. This considerable accuracy fulfils one requirement of models to be able to predict the impacts of climate change on the carbon balance of European forests. However, the generality of the models should be further evaluated by expanding the range of testing over both time and space. In addition, differences in behaviour between models at the process level indicate requirement of further model testing, with special emphasis on modelling stomatal conductance realistically.     

Determination of cyclic nucleotide phosphodiesterase activity by high-performance liquid chromatography

A new assay for cyclic nucleotide phosphodiesterase activity by high-performance liquid chromatography with on-line radiochemical detection has been developed. The method is based on the measurement of 3H-labeled nucleoside monophosphates formed from cyclic nucleotides by the action of 3',5'-cyclic-nucleotide phosphodiesterase (PDE). The reaction products are determined from the incubation mixture after removal of the protein by injection of an aliquot into the liquid chromatograph. The detection limit with counting efficiency of 30% is 20 fmol of 3H-labeled product, which makes the method suitable for detection of low PDE activities.     

Analysis of radioactive and nonradioactive purine bases,nucleosides, and nucleotides by high-speed chromatography on a single column

The concentrations of radioactive and nonradioactive purine bases, purine nucleosides, purine mono-, di-, and trinucleotides in acid extracts of fibroblasts were determined by anion-exchange column chromatography. The concentrations of nonradioactive components were determined by computerized integration of the signal from a double-beam uv-detector. The radioactive metabolites were quantitated by high-efficiency, continuous liquid scintillation counting, employing a discrete sample transport system.     

Bone Morphogenetic Protein-2 Is a Regulator of Cell Adhesion

Bone morphogenetic proteins (BMPs) are a group of peptide growth factors closely related to transforming growth factors-β. The BMPs are suggested to play an essential role in bone development and they are strong candidate molecules to be used clinically to improve fracture healing. BMPs are also involved in the differentiation of several other tissues during embryogenesis. Here, we show that human recombinant BMP-2 regulates cell–matrix interactions by modifying the expression of integrin-type receptors. The synthesis of α3 integrin was down-regulated by BMP-2 in two osteogenic sarcoma-derived cell lines, Saos-2 and HOS, and also in human fetal chondrocytes. BMP-2 had no effect on the expression of α1, α2, α5, α6, and αV integrins. BMP-2 reduced the expression of α3 integrin subunit at mRNA level. Laminin-5 was shown to be the ligand for α3β1 integrin on Saos cells and BMP-2 decreased the ability of Saos cells to attach to it. These results suggest that BMP-2 has a specific effect on the α3β1 integrin-mediated cell adhesion to laminin-5. Given the fact that BMP-2 is expressed in osteosarcomas, in addition to in bone, this mechanism is putatively important especially in bone development and tumors. We also studied the effect of BMP-2 on a human keratinocyte cell line, HaCaT. In HaCaT cells, the expression of α2 integrin was strongly down-regulated by BMP-2, whereas its effect on the expression of α3 integrin was smaller. We suggest that the effects of BMP-2 may be partially mediated by specifically altered cell adhesion.     

Electrophysiological responses of carrot psyllids (Trioza apicalis), in different phases of their life cycle,to volatile carrot and conifer compounds

Carrot psyllids (Trioza apicalis) pose a constant threat to carrot production in parts of Europe, and treatments with chemical insecticides are common. To be able to develop alternative control tactics, more knowledge about the psyllid's host-finding behaviour and underlying physiology is needed. Using single-sensillum recordings from overwintered and non-overwintered males and females, we found essentially identical responses for the two groups and for both sexes. Using six compounds emanating from carrots or conifers, the psyllids' overwintering plants, and extracts of carrot leaves, five different olfactory sensory neuron classes could be distinguished from our high-quality recordings, viz. #1 responding strongly and exclusively to terpinene-4-ol, #2 responding strongly and most often exclusively to nonanal, #3 responding moderately strongly to terpinolene, #4 responding strongly to (Z)-3-hexenal and most often weaker to terpinolene, #5 responding exclusively to carrot extract.     

Effectiveness of treatment with antihypertensive drugs and trends in mortality from stroke in the community.

A study was made of the changes in the treatment of hypertension, in the effectiveness of that treatment, and in mortality from cerebrovascular stroke in men and women in two counties in Finland. One of these counties was North Karelia, where a comprehensive cardiovascular programme based in the community had been launched in 1972. Cross sectional surveys in large representative samples of the middle aged populations were carried out in 1972, 1977, and 1982. Average annual mortality from stroke adjusted for age was calculated for the two year periods 1971-2, 1977-8, and 1980-1. The proportion of hypertensive men being effectively treated for their hypertension increased in both areas from 1972 to 1977 and further from 1977 to 1982. Mortality from stroke in middle aged men decreased in both areas during both observation periods. The proportion of hypertensive women being effectively treated greatly increased in both areas from 1972 to 1977. This was associated with a large decline in mortality from stroke. In North Karelia the proportion of women being effectively treated was remarkably high but decreased somewhat between 1977 and 1982; this decrease was associated with an increase in mortality from stroke in the women aged 35-64. These results support the idea that effective antihypertensive treatment in the community is a major determinant of mortality from stroke in both men and women.     

Hcp2, a Secreted Protein of the Phytopathogen Pseudomonas syringae pv. Tomato DC3000, Is Required for Fitness for Competition against Bacteria and Yeasts

When analyzing the secretome of the plant pathogen Pseudomonas syringae pv. tomato DC3000, we identified hemolysin-coregulated protein (Hcp) as one of the secreted proteins. Hcp is assumed to be an extracellular component of the type VI secretion system (T6SS). Two copies of hcp genes are present in the P. syringae pv. tomato DC3000 genome, hcp1 (PSPTO_2539) and hcp2 (PSPTO_5435). We studied the expression patterns of the hcp genes and tested the fitness of hcp knockout mutants in host plant colonization and in intermicrobial competition. We found that the hcp2 gene is expressed most actively at the stationary growth phase and that the Hcp2 protein is secreted via the T6SS and appears in the culture medium as covalently linked dimers. Expression of hcp2 is not induced in planta and does not contribute to virulence in or colonization of tomato or Arabidopsis plants. Instead, hcp2 is required for survival in competition with enterobacteria and yeasts, and its function is associated with the suppression of the growth of these competitors. This is the first report on bacterial T6SS-associated genes functioning in competition with yeast. Our results suggest that the T6SS of P. syringae may play an important role in bacterial fitness, allowing this plant pathogen to survive under conditions where it has to compete with other microorganisms for resources.     

Spreading of Trioza apicalis and development of “Candidatus Liberibacter solanacearum” infection on carrot in the field conditions

Carrot cultivation in Europe is suffering from infections with “Candidatus Liberibacter solanacearum” (CLso), a psyllid‐transmitted bacterial pathogen. In this study, field experiments were carried out in Finland to separately measure the effects of psyllid feeding damage and CLso infection on the carrot root growth and to reveal the dynamics of the spreading of CLso within the field. Most of the experiments were carried out during the summers 2016 and 2017, and a follow‐up sampling was performed in 2018. Carrot psyllid (Trioza apicalis) flight activity was monitored and carrots were sampled at 25 points within the field. Early in the season a clear spatial correlation was found between the sampling sites showing the psyllid feeding damage, that is, leaf‐curling, up to the range of 40–60 m, indicating aggregation behaviour of the psyllids. No CLso infections were detected in the first sampling, which was performed before the psyllid flight peak in both years. Later, a positive correlation between the psyllid feeding damage and the CLso titre was observed. An increase in the CLso titre occurred approximately a month after the psyllid flight peak, and this increase correlated with the accumulating effective temperature sum. In 2016, both the psyllid feeding damage and CLso infection had a significant effect on the carrot root weight. The effect of CLso titre on root weight was nonlinear, that is, it intensified rapidly at the highest bacterial titres. During the colder summer of 2017 the CLso titres did not reach high enough levels in the plants to cause substantial visible symptoms and root growth reduction. Thus, it seems that in the Nordic conditions the effect of CLso infection on carrot yield is strongly dependent on the weather conditions during the growing season.     

Analyzing the Environmental Benefits of Industrial Symbiosis

Studies of industrial symbiosis (IS) focus on the physical flows of materials and energy in local industrial systems. In an ideal IS, waste material and energy are shared or exchanged among the actors of the system, thereby reducing the consumption of virgin material and energy inputs, and likewise the generation of waste and emissions. In this study, the environmental impacts of an industrial ecosystem centered around a pulp and paper mill and operating as an IS are analyzed using life cycle assessment (LCA). The system is compared with two hypothetical reference systems in which the actors would operate in isolation. Moreover, the system is analyzed further in order to identify possibilities for additional links between the actors. The results show that of the total life cycle impacts of the system, upstream processes made the greatest overall contribution to the results. Comparison with stand‐alone production shows that in the case studied, the industrial symbiosis results in modest improvements, 5% to 20% in most impact categories, in the overall environmental impacts of the system. Most of the benefits occur upstream through heat and electricity production for the local town. All in all it is recommended that when the environmental impacts of industrial symbiosis are assessed, the impacts occurring upstream should also be studied, not only the impacts within the ecosystem.