Responses of leaf traits to climatic gradients: adaptive variation versus compositional shifts

Dynamic global vegetation models (DGVMs) typically rely on plant functional types (PFTs), which are assigned distinct environmental tolerances and replace one another progressively along environmental gradients. Fixed values of traits are assigned to each PFT; modelled trait variation along gradients is thus driven by PFT replacement. But empirical studies have revealed "universal" scaling relationships (quantitative trait variations with climate that are similar within and between species, PFTs and communities); and continuous, adaptive trait variation has been proposed to replace PFTs as the basis for next-generation DGVMs. Here we analyse quantitative leaf-trait variation on long temperature and moisture gradients in China with a view to understanding the relative importance of PFT replacement vs. continuous adaptive variation within PFTs. Leaf area (LA), specific leaf area (SLA), leaf dry matter content (LDMC) and nitrogen content of dry matter were measured on all species at 80 sites ranging from temperate to tropical climates and from dense forests to deserts. Chlorophyll fluorescence traits and carbon, phosphorus and potassium contents were measured at 47 sites. Generalized linear models were used to relate log-transformed trait values to growing-season temperature and moisture indices, with or without PFT identity as a predictor, and to test for differences in trait responses among PFTs. Continuous trait variation was found to be ubiquitous. Responses to moisture availability were generally similar within and between PFTs, but biophysical traits (LA, SLA and LDMC) of forbs and grasses responded differently from woody plants. SLA and LDMC responses to temperature were dominated by the prevalence of evergreen PFTs with thick, dense leaves at the warm end of the gradient. Nutrient (N, P and K) responses to climate gradients were generally similar within all PFTs. Area-based nutrients generally declined with moisture; Narea and Karea declined with temperature, but Parea increased with temperature. Although the adaptive nature of many of these trait-climate relationships is understood qualitatively, a key challenge for modelling is to predict them quantitatively. Models must take into account that community-level responses to climatic gradients can be influenced by shifts in PFT composition, such as the replacement of deciduous by evergreen trees, which may run either parallel or counter to trait variation within PFTs. The importance of PFT shifts varies among traits, being important for biophysical traits but less so for physiological and chemical traits. Finally, models should take account of the diversity of trait values that is found in all sites and PFTs, representing the "pool" of variation that is locally available for the natural adaptation of ecosystem function to environmental change.

Latitudinal gradients in butterfly population variability are influenced by landscape heterogeneity

The variability of populations over time is positively associated with their risk of local extinction. Previous work has shown that populations at the high-latitude boundary of species’ ranges show higher inter-annual variability, consistent with increased sensitivity and exposure to adverse climatic conditions. However, patterns of population variability at both high- and low-latitude species range boundaries have not yet been concurrently examined. Here, we assess the inter-annual population variability of 28 butterfly species between 1994 and 2009 at 351 and 18 sites in the United Kingdom and Catalonia, Spain, respectively. Local population variability is examined with respect to the position of the species’ bioclimatic envelopes (i.e. whether the population falls within areas of the ‘core’ climatic suitability or is a climatically ‘marginal’ population), and in relation to local landscape heterogeneity, which may influence these range location – population dynamic relationships. We found that butterfly species consistently show latitudinal gradients in population variability, with increased variability in the more northerly UK. This pattern is even more marked for southerly distributed species with ‘marginal’ climatic suitability in the UK but ‘core’ climatic suitability in Catalonia. In addition, local landscape heterogeneity did influence these range location – population dynamic relationships. Habitat heterogeneity was associated with dampened population dynamics, especially for populations in the UK. Our results suggest that promoting habitat heterogeneity may promote the persistence of populations at high-latitude range boundaries, which may potentially aid northwards expansion under climate warming. We did not find evidence that population variability increases towards southern range boundaries. Sample sizes for this region were low, but there was tentative evidence, in line with previous ecological theory, that local landscape heterogeneity may promote persistence in these retracting low-latitude range boundary populations.

Metallicity gradients at large galactocentric radii using the near-infrared Calcium triplet

We describe a new spectroscopic technique for measuring radial metallicity gradients out to large galactocentric radii. We use the DEep Imaging Multi-Object Spectrograph (DEIMOS) on the Keck telescope and the galaxy spectrum extraction technique of Proctor et al. We also make use of the metallicity sensitive near-infrared Calcium ii triplet (CaT) features together with single stellar population models to obtain metallicities. Our technique is applied as a pilot study to a sample of three relatively nearby (< 30 Mpc) intermediate-mass to massive early-type galaxies. Results are compared with previous literature inner region values and generally show good agreement. We also include a comparison with profiles from dissipational disc-disc major merger simulations. Based on our new extended metallicity gradients combined with other observational evidence and theoretical predictions, we discuss possible formation scenarios for the galaxies in our sample. The limitations of our new technique are also discussed.

The spider genus Chrysometa (Araneae, Tetragnathidae) from the Pico da Neblina and Serra do Tapirapeco mountains (Amazonas, Brazil): new species, new records, diversity and distribution along two altitudinal gradients

Eight new species of the spider genus Chrysometa Simon, 1894 (Araneae, Tetragnathidae) are described and illustrated. Chrysometa nubigena n. sp., C. waikoxi n. sp., C. petrasierwaldae n. sp., C. santosi n. sp., C. yanomami n. sp., C. candianii n. sp., C. lomanhungae n. sp., and C. saci n. sp. Those species were collected in a study on the diversity of spider communities along altitudinal gradients in Brazilian Amazonia. C. saci was captured at the Serra do Tapirapeco (Barcelos), while all the other species are from the Pico da Neblina (Sao Gabriel da Cachoeira), the highest mountain in Brazil. We provide new records for C. boraceia, C. flava, C. guttata, C. minuta and C. opulenta, and we describe the male of C. minuta for the first time. We also present the first results on the diversity and altitudinal distribution of the species of Chrysometa at the Pico da Neblina and Serra do Tapirapeco. We sampled the first locality at six different elevations, and obtained 336 specimens distributed in 12 species. Richness and abundance, as well as relative importance peaked at the highest sites sampled (2,000 and 2,400 m). The three most abundant species showed a segregated distribution, being dominant or exclusively distributed in different altitudes. At the Serra do Tapirapeco, sampling at four different elevations up to 1200 m, we only obtained 40 individuals divided in four species, and there was no clear relation to altitude. Most of the new species were found at mid and high altitude sites, while species from lower altitude sites represented widespread species. The comparison with other neotropical spiders inventories highlights the high diversity recorded at Pico da Neblina, which could be assigned to the large environmental variation covered in this work and to the sampling of high-altitude environments. Inventories in the Andean region and other information in the literature also seem to support the association of Chrysometa with high altitude environments.

Aerosol properties, in-canopy gradients, turbulent fluxes and VOC concentrations at a pristine forest site in Amazonia

Aerosol physical and chemical properties were measured in a forest site in central Amazonia (Cuieiras reservation, 2.61S; 60.21W) during the dry season of 2004 (Aug-Oct). Aerosol light scattering and absorption, mass concentration, elemental composition and size distributions were measured at three tower levels (Ground: 2 m; Canopy: 28 m, and Top: 40 m). For the first time, simultaneous eddy covariance fluxes of fine mode particles and volatile organic compounds (VOC) were measured above the Amazonian forest canopy. Aerosol fluxes were measured by eddy covariance using a Condensation Particle Counter (CPC) and a sonic anemometer. VOC fluxes were measured by disjunct eddy covariance using a Proton Transfer Reaction Mass Spectrometer (PTR-MS). At nighttime, a strong vertical gradient of phosphorus and potassium in the aerosol coarse mode was observed, with higher concentrations at Ground level. This suggests a source of primary biogenic particles below the canopy. Equivalent black carbon measurements indicate the presence of light-absorbing aerosols from biogenic origin. Aerosol number size distributions typically consisted of superimposed Aitken (76 nm) and accumulation modes (144 nm), without clear events of new particle formation. Isoprene and monoterpene fluxes reached respectively 7.4 and 0.82 mg m(-2) s(-1) around noon. An average fine particle flux of 0.05 +/- 0.10 10(6) m(-2) s(-1) was calculated, denoting an equilibrium between emission and deposition fluxes of fine mode particles at daytime. No significant correlations were found between VOC and fine mode aerosol concentrations or fluxes. (C) 2009 Elsevier Ltd. All rights reserved.

Electric field gradients in (111)In-doped (Hf/Zr)(3)Al(2) and (Hf/Zr)(4)Al(3) mixed compounds: ab initio calculations, perturbed angular correlation measurements and site preference

The quadrupolar hyperfine interactions of in-diffused (111)In -> (111)Cd probes in polycrystalline isostructural Zr(4)Al(3) and Hf(4)Al(3) samples containing small admixtures of the phases (Zr/Hf)(3)Al(2) were investigated. A strong preference of (111)In solutes for the contaminant (Zr/Hf)(3)Al(2) minority phases was observed. Detailed calculations of the electric field gradient (EFG) at the Cd nucleus using the full-potential augmented plane wave + local orbital formalism allowed us to assign the observed EFG fractions to the various lattice sites in the (Zr/Hf)(3)Al(2) compounds and to understand the preferential site occupation of the minority phases by the (111)In atoms. The effects of the size of the supercell and relaxation around the oversized In and Cd probe atoms were investigated in detail.

Percoll e plasma seminal na preservação do sêmen eqüino a +4º C

O presente estudo visou verificar o efeito sobre alguns parâmetros da seleção por gradiente de Percoll® e da adição de plasma seminal do sêmen eqüino preservado a +4oC. O primeiro experimento avaliou a taxa de recuperação de espermatozóides após seleção por Percoll® em diferentes protocolos de centrifugação. Foram realizadas 5 coletas de sêmen de um garanhão. Imediatamente após a coleta, o sêmen foi avaliado quanto à motilidade, vigor e concentração. Foram retiradas duas amostras de 4 mL, diluídas em leite desnatado UHT com concentrações de 50 e 100 x 106 espermatozóides por mL cada. Cada uma destas amostras foi dividida em 4 alíquotas de 1 mL, que foram então colocadas sobre Percoll® e submetidas a diferentes tempos e velocidades de centrifugação. V1 - 200 g (5 min) + 800 g (10 min); V2 - 800 g (10 min); V3 - 800 g (15 min); V4 - 800 g (20 min). Após esse processo, o sobrenadante foi desprezado e o pellet de cada alíquota ressuspendido com 0,5 mL de leite UHT. As 8 amostras foram novamente avaliadas para concentração, motilidade e vigor. O segundo experimento estudou o efeito da adição de plasma seminal de diferentes qualidades ao sêmen eqüino selecionado por gradiente de Percoll® e resfriado a +4°C por até 72 horas. Foram utilizados 40 ejaculados de 4 garanhões, sendo dois com boa qualidade de sêmen e dois com baixa qualidade de sêmen. Imediatamente após a coleta, o sêmen foi avaliado quanto à motilidade, vigor e concentração e preparadas cinco frações de 100x106 espermatozóides, diluídas 1:1 (v/v) em EDTA-Glicose. Quatro delas, constituídas por 1mL a 2mL, foram depositadas sobre Percoll®. A fração restante foi centrifugada em tubo de vidro de 10 mL, sob as mesmas condições de tempo e velocidade das demais amostras. Foi realizada centrifugação por 5 minutos em 200 g, seguida de 10 minutos em 800 g. O sobrenadante foi descartado e o pellet ressuspendido com leite UHT desnatado compondo os seguintes tratamentos: Sp: 1,5 mL de leite UHT desnatado sem adição de plasma seminal; Hp: 1,425 mL de leite UHT desnatado acrescido de 75 L de plasma seminal homólogo; Ap: 1,425 mL de leite UHT desnatado acrescido de 75 L de plasma seminal do pool de alta qualidade; Bp: 1,425mL de leite UHT desnatado acrescido de 75 L plasma seminal do pool de baixa qualidade; Cc: foi centrifugada sem seleção por Percoll®, teve seu sobrenadante descartado e ressuspendida com 2 mL de leite UHT; C : uma amostra de sêmen diluído em leite UHT foi mantida como controle no processo de armazenamento. As amostras foram resfriadas a +4°C e examinadas a cada 24h até as 72 horas em relação à motilidade, funcionalidade de membrana (teste hiposmótico) e integridade de membrana (CFDA/PI). A seleção por gradiente descontínuo de Percoll® 90/45% mostrou-se efetiva na recuperação de espermatozóides com motilidades progressiva e total. A adição de 5% de plasma seminal ou a ausência de plasma não influenciaram os valores da motilidade das amostras selecionadas por gradiente de Percoll®. A seleção por Percoll não influenciou na percentagem de células com membrana plasmática funcional. Concentrações superiores a 2% de plasma seminal resultaram em decréscimo do número de células com membrana funcional, enquanto que as amostras sem plasma seminal apresentaram os melhores resultados e o grupo controle, que apresentava a maior percentagem de plasma, os piores. A utilização de Percoll® não separou células com alteração de membrana. A percentagem de células com membrana completamente íntegra foi significativamente maior nas amostras que sofreram processo de centrifugação, independentemente da seleção. O processo de seleção por Percoll® foi efetivo na recuperação de espermatozóides de eqüino com motilidade progressiva, mas não selecionou espermatozóides quanto à funcionalidade nem à integridade de membrana. Concentrações inferiores a 2% de plasma seminal melhoraram a funcionalidade de membrana. A ausência de plasma seminal melhorou os resultados de integridade das membranas plasmática e acrossomal; e a adição de plasma de alta qualidade não melhorou a motilidade de espermatozóides selecionados por Percoll®.

Comparative validation using quantitative real-time PCR (qPCR) and conventional PCR of bovine semen centrifuged in continuous density gradient

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chromatin and morphometry alterations of bovine sperm head after Percoll (TM) density gradient centrifugation

The aim of this work was to assess the morphometry and chromatin integrity of bovine sperm head after a three layers discontinuous Percoll (TM) density gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Three ejaculates per bull were evaluated. The semen doses were thawed and two smears were made from each sample before (control) and after the Percoll (TM) centrifugation (Percoll (TM) group). The smears were stained with toluidine blue and grayscale digital images were captured and processed in Scilab environment software. It was observed that chromatin heterogeneity was reduced (P<0.05) and chromatin decondensation was increased (P<0.05) after the Percollm treatment utilized. In addition, it was observed that sperm head length was higher (P<0.05) and the side symmetry was lower (P<0.05) in centrifuged sperm cells. When analyzed separately by subspecies, it was observed that the decrease (P<0.05) in chromatin heterogeneity and the increase (P<0.05) in chromatin decondensation occurred in Zebu sperm heads. In addition, the length and the width:length ratio of sperm heads was affected by Percoll (TM) centrifugation in Zebu semen. In conclusion, the three layers discontinuous Percoll (TM) centrifugation increased the chromatin decondensation and the morphometric alterations of frozen-thawed bovine semen. However, the real implication of these findings in fertility rates of centrifuged sperm must be investigated. (C) 2011 Elsevier B.V. All rights reserved.

Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association

The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.