Above-ground biomass and productivity in a rain forest of eastern South America

Abstract: The dynamics of tropical forest woody plants was studied at the Nouragues Field Station, central French Guiana. Stem density, basal area, above-ground biomass and above-ground net primary productivity, including the contribution of litterfall, were estimated from two large permanent census plots of 12 and 10 ha, established on contrasting soil types, and censused twice, first in 1992¿1994, then again in 2000¿2002. Mean stem density was 512 stems ha¿1 and basal area, 30m2 ha¿1. Stem mortality rate ranged between 1.51% and 2.06% y¿1. In both plots, stem density decreased over the study period. Using a correlation between wood density and wood hardness directly measured by a Pilodyn wood tester,we found that the mean wood densitywas 0.63 g cm¿3, 12% smaller than the mean of wood density estimated from the literature values for the species occurring in our plot. Above-ground biomass ranged from 356 to 398Mgha¿1 (oven-dry mass), and it increased over the census period. Leaf biomass was 6.47Mg ha¿1. Our total estimate of aboveground net primary productivity was 8.81 MgC ha¿1 y¿1 (in carbon units), not accounting for loss to herbivory, branchfalls, or biogenic volatile organic compounds, whichmay altogether account for an additional 1MgC ha¿1 y¿1. Coarse wood productivity (stem growth plus recruitment) contributed to 4.16 MgC ha¿1 y¿1. Litterfall contributed to 4.65MgC ha¿1 y¿1 with 3.16 MgC ha¿1 y¿1 due to leaves, 1.10 MgC ha¿1 y¿1 to twigs, and 0.39MgC ha¿1 y¿1 to fruits and flowers. The increase in above-ground biomass for both trees and lianas is consistentwith the hypothesis of a shift in the functioning of Amazonian rain forests driven by environmental changes, although alternative hypotheses such as a recovery from past disturbances cannot be ruled out at our site, as suggested by the observed decrease in stem density. Key Words: above-ground biomass, carbon, French Guiana, net primary productivity, tropical fores

Effect of different 3’ flanking regions on the activity of the Vitis vinifera alcohol dehydrogenase 2 promoter

3’untranslated regions (3' UTR) are isogene specific regions which contain sequences likely playing an important role in gene expression. To evaluate the importance of these regions on Vitis vinifera alcohol dehydrogenase 2 (VvAdh2) expression, we designed expression cassettes containing the luciferase reporter gene under the VvAdh2 or CaMV 35S promoters and flanked by different 3’ UTRs. Luciferase activity monitoring was performed through transient expression experiments, using biolistic on Cabernet Sauvignon suspension cells. Results showed that absence of the 3’ region had a strong down-regulating effect on the VvAdh2 promoter activity (but not on the CaMV 35S promoter activity). The nature of the flanking 3' UTR was shown to influence expression cassette activity. Whatever the promoter, VvAdh1 and VvAdh2 terminators had similar effect on expression of luciferase in air leading to an activity level very close to that of CaMV 35S terminator. Under anaerobiosis, luciferase expression was strongly increased with all terminators, VvAdh6 3'-end inducing the highest level of expression. Functional constructs with VvAdh2 promoter and VvAdh terminators designed in this study could be used wherever grapevine-homologous, stress-stimulated cassettes should be of interest

High Resolution Crystal Structures of the Wild Type and Cys-55 right-arrow Ser and Cys-59 right-arrow Ser Variants of the Thioredoxin-like [2Fe-2S] Ferredoxin from Aquifex aeolicus

The [2Fe-2S] ferredoxin (Fd4) from Aquifex aeolicus adopts a thioredoxin-like polypeptide fold that is distinct from other [2Fe-2S] ferredoxins. Crystal structures of the Cys-55 right-arrow Ser (C55S) and Cys-59 right-arrow Ser (C59S) variants of this protein have been determined to 1.25 Å and 1.05 Å resolution, respectively, whereas the resolution of the wild type (WT) has been extended to 1.5 Å. The improved WT structure provides a detailed description of the [2Fe-2S] cluster, including two features that have not been noted previously in any [2Fe-2S] cluster-containing protein, namely, pronounced distortions in the cysteine coordination to the cluster and a Calpha -H-Sgamma hydrogen bond between cluster ligands Cys-55 and Cys-9. These features may contribute to the unusual electronic and magnetic properties of the [2Fe-2S] clusters in WT and variants of this ferredoxin. The structures of the two variants of Fd4, in which single cysteine ligands to the [2Fe-2S] cluster are replaced by serine, establish the metric details of serine-ligated Fe-S active sites with unprecedented accuracy. Both the cluster and its surrounding protein matrix change in subtle ways to accommodate this ligand substitution, particularly in terms of distortions of the Fe2S2 inorganic core from planarity and displacements of the polypeptide chain. These high resolution structures illustrate how the interactions between polypeptide chains and Fe-S active sites reflect combinations of flexibility and rigidity on the part of both partners; these themes are also evident in more complex systems, as exemplified by changes associated with serine ligation of the nitrogenase P cluster

Ethylene and other stimuli affect expression of the UDP glucose-flavonoid 3-O-glucosyltransferase in a non-climacteric fruit

The UDP glucose-flavonoid 3-O-glucoslyltransferase (UFGT) is a key enzyme for biosynthesis and stability of anthocyanin pigments of red grapes. Understanding factors affecting expression of this enzyme is thus important for the control of grape colour. A 1640 bp promoter region of the grapevine ufgt gene was cloned and sequenced. Sequence analysis revealed seven putative ethylene-responsive cis-elements and others related to three major signals known to induce anthocyanin accumulation in plant tissues: light, sugar, and abscisic acid. In order to evaluate the ability of ethylene and other signals to drive expression from the ufgt promoter, we ran transient expression experiments using an anthocyanin-rich grape cell culture, with very low green auto-fluorescence. After biolistic bombardment, the cells were treated with various combinations of the four signals on gfp expression (green fluorescent protein). The comparison of fluorescent light intensity in cells subjected to the various treatments showed that ethylene better stimulates expression of the ufgt promoter in the dark than under light. In addition, results showed that there may be a positive interaction between ethylene and abscisic acid. This system, a promoter of interest driving the gfp expression in cells with low auto-fluorescence, may be a good tool for studies about synergistic or antagonist roles of transcription factors. Moreover, treatment of grape berries with a specific inhibitor of ethylene receptors (1-methylcyclopropene) inhibited ufgt mRNA accumulation. This confirms that the ethylene signal is likely a regulator of grape UFGT expression in a non-climacteric fruit.

Somatic embryogenesis and plant regeneration of Vitis vinifera cultivars 'Macabeo' and 'Tempranillo'

Different experimental conditions have been compared to achieve a high efficiency in embryogenic calli initiation from 'Macabeo' and 'Tempranillo' anthers. Specifically, two stages of anther development were tested (corresponding to tetrad cells or uninucleate pollen), and direct culture of anthers was compared to culture after a cold treatment of inflorescences (4 °C during 48 h). In addition, two induction media (C1 P and B2), mainly differing by microelement and cytokinin levels, were evaluated. Experiment repeatability was also examined with a repetition of anther culture one week later. Callus initiation was similar in all media and treatments for both cultivars, usually starting from the anther filament. A simple protocol for efficient induction of embryogenesis in 'Macabeo' and 'Tempranillo' consisted in:selecting the first inflorescence from hardwood cutting,excising anthers at uninucleate pollen stage without cold treatment of the inflorescences,incubating anthers on C1 P medium.The procedure used for embryo germination and plant regeneration, allowed to obtain a conversion rate up to 75 % in 'Macabeo' and 60 % in 'Tempranillo'. The protocol proposed represents the first regeneration system developed for the Spanish cultivars 'Macabeo' and 'Tempranillo'.

Cryopreservation and cryotherapy of grapevine (Vitis vinifera L.)

This study aimed at testing the efficiency of a droplet-vitrification cryopreservation protocol in eliminating selected grapevine viruses. The cryopreservation protocol led to approximately 50 % recovery with cultivar 'Portan' and five international cultivars tested, but very low recovery was noted with Croatian cultivars. GFLV and GLRaV-3, two (economically important grapevine) viruses were eliminated in a high percentage (up to 100 %) of plants regenerated from cryopreserved shoot tips. Virus sanitation was observed as well in samples before liquid nitrogen exposure. Genetic stability of plants regenerated after cryopreservation was studied using AFLP markers. Polymorphic fragments were observed in non-cryopreserved and cryopreserved samples treated with PVS2 solution, the number of which increased with increasing durations of exposure to PVS2 solution

Clinical impact of anti-inflammatory microglia and macrophage phenotypes at glioblastoma margins

Glioblastoma is a devastating brain cancer for which effective treatments are required. Tumour-associated microglia and macrophages promote glioblastoma growth in an immune-suppressed microenvironment. Most recurrences occur at the invasive margin of the surrounding brain, yet the relationships between microglia/macrophage phenotypes, T cells and programmed death-ligand 1 (an immune checkpoint) across human glioblastoma regions are understudied. In this study, we performed a quantitative immunohistochemical analysis of 15 markers of microglia/macrophage phenotypes (including anti-inflammatory markers triggering receptor expressed on myeloid cells 2 and CD163, and the low-affinity-activating receptor CD32a), T cells, natural killer cells and programmed death-ligand 1, in 59 human IDH1-wild-type glioblastoma multi-regional samples (n = 177; 1 sample at tumour core, 2 samples at the margins: the infiltrating zone and leading edge). Assessment was made for the prognostic value of markers; the results were validated in an independent cohort. Microglia/macrophage motility and activation (Iba1, CD68), programmed death-ligand 1 and CD4+ T cells were reduced, and homeostatic microglia (P2RY12) were increased in the invasive margins compared with the tumour core. There were significant positive correlations between microglia/macrophage markers CD68 (phagocytic)/triggering receptor expressed on myeloid cells 2 (anti-inflammatory) and CD8+ T cells in the invasive margins but not in the tumour core (P < 0.01). Programmed death-ligand 1 expression was associated with microglia/macrophage markers (including anti-inflammatory) CD68, CD163, CD32a and triggering receptor expressed on myeloid cells 2, only in the leading edge of glioblastomas (P < 0.01). Similarly, there was a positive correlation between programmed death-ligand 1 expression and CD8+ T-cell infiltration in the leading edge (P < 0.001). There was no relationship between CD64 (a receptor for autoreactive T-cell responses) and CD8+/CD4+ T cells, or between the microglia/macrophage antigen presentation marker HLA-DR and microglial motility (Iba1) in the tumour margins. Natural killer cell infiltration (CD335+) correlated with CD8+ T cells and with CD68/CD163/triggering receptor expressed on myeloid cells 2 anti-inflammatory microglia/macrophages at the leading edge. In an independent large glioblastoma cohort with transcriptomic data, positive correlations between anti-inflammatory microglia/macrophage markers (triggering receptor expressed on myeloid cells 2, CD163 and CD32a) and CD4+/CD8+/programmed death-ligand 1 RNA expression were validated (P < 0.001). Finally, multivariate analysis showed that high triggering receptor expressed on myeloid cells 2, programmed death-ligand 1 and CD32a expression at the leading edge were significantly associated with poorer overall patient survival (hazard ratio = 2.05, 3.42 and 2.11, respectively), independent of clinical variables. In conclusion, anti-inflammatory microglia/macrophages, CD8+ T cells and programmed death-ligand 1 are correlated in the invasive margins of glioblastoma, consistent with immune-suppressive interactions. High triggering receptor expressed on myeloid cells 2, programmed death-ligand 1 and CD32a expression at the human glioblastoma leading edge are predictors of poorer overall survival. Given substantial interest in targeting microglia/macrophages, together with immune checkpoint inhibitors in cancer, these data have major clinical implications

Regional and seasonal patterns of litterfall in tropical South America

The production of aboveground soft tissue represents an important share of total net primary production in tropical rain forests. Here we draw from a large number of published and unpublished datasets (n=81 sites) to assess the determinants of litterfall variation across South American tropical forests. We show that across old-growth tropical rainforests, litterfall averages 8.61±1.91 Mg ha−1 yr−1 (mean ± standard deviation, in dry mass units). Secondary forests have a lower annual litterfall than old-growth tropical forests with a mean of 8.01±3.41 Mg ha−1 yr−1. Annual litterfall shows no significant variation with total annual rainfall, either globally or within forest types. It does not vary consistently with soil type, except in the poorest soils (white sand soils), where litterfall is significantly lower than in other soil types (5.42±1.91 Mg ha−1 yr−1). We also study the determinants of litterfall seasonality, and find that it does not depend on annual rainfall or on soil type. However, litterfall seasonality is significantly positively correlated with rainfall seasonality. Finally, we assess how much carbon is stored in reproductive organs relative to photosynthetic organs. Mean leaf fall is 5.74±1.83 Mg ha−1 yr−1 (71% of total litterfall). Mean allocation into reproductive organs is 0.69±0.40 Mg ha−1 yr−1 (9% of total litterfall). The investment into reproductive organs divided by leaf litterfall increases with soil fertility, suggesting that on poor soils, the allocation to photosynthetic organs is prioritized over that to reproduction. Finally, we discuss the ecological and biogeochemical implications of these result

Characteristics of air showers created by extremely high energy gamma-rays

The technique of adjoint cascade equations has been applied to calculate the properties of extremely high energy gamma-rays in the energy range 10^18--10^22 eV with taking into account the LPM effect and interactions of gamma-rays with the geomagnetic field. Such characteristics are analysed as the electron and muon contents at the observation level, the electron cascade curves, the lateral distribution functions of photoproduced muons.Comment: 36 pages, 19 figures, submitted to J.Phys.G: Nucl.Part.Phy

Spiral2 cryomodules B tests results

MOP010International audienceAssembly and tests of the SPIRAL2 superconducting linac's cryomodules at CEA/Saclay and IPN/Orsay have now reached cruising speed after having faced a series of problems, among them contamination. 19 cryomodules are composing the whole Linac and IPN Orsay is in charge of the 7 cryomodules B, housing two 88 MHz, beta 0.12 Quarter-Wave Resonators. Threecryomodules have been assembled and successfullytested up to the nominal gradient of 6.5 MV/m for all cavities with also cryogenic losses withinspecifications. Two of them are fully qualified and already delivered to GANIL. The thirdone showed misalignment ofone cavity which could lead to partial disassembly if needed. This paper presents the results of those cryomodules tests as well as the status of the remaining ones