Timing of breeding in <Emphasis Type="Italic">Ochrotomys nuttalli</Emphasis> and <Emphasis Type="Italic">Peromyscus leucopus</Emphasis> is related to a latitudinal isotherm

Previous comparative studies on patterns of reproduction in small-mammal species focus primarily on latitudinal differences in average litter size. Few studies compare reproductive patterns among northern and southern populations at the landscape scale. Our study compares differences in seasonal patterns of reproduction in northern and southern populations of the golden mouse, Ochrotomys nuttalli, and the white-footed mouse, Peromyscus leucopus. These are remarkably similar species with regard to bioenergetics, body mass, feeding behavior, home-range size, natural history, nest-site preference, and periods of activity. Both species also exhibit very similar intraspecific seasonal patterns of reproduction across their respective geographic ranges. We found that O. nuttalli and P. leucopus switch from a summer breeding season, extending from late spring through early autumn in the north to a winter breeding season extending from late autumn through early spring in the south, near the isotherm where mean annual temperature is 15.6°C (60°F), or approximately 35° N latitude. This latitudinal isotherm provides a geographic benchmark to address future changes in patterns of reproduction attributed to climate change. Findings also suggest that length of the breeding season and patterns of reproduction between species partially explain why P. leucopus is typically more abundant than O. nuttalli in similar habitat types.     

The effect of fire spatial scale on <Emphasis Type="Italic">Bison</Emphasis> grazing intensity

To determine whether fire spatial and temporal scales affect foraging behavior and grazing intensity by Bison (Bison bison), we burned three different patch sizes (225, 900, and 3600 m²) across an otherwise homogeneous grassland landscape. We then monitored grazing intensity for the succeeding 14 months. During the first 5 months after the burn (August–January), the Bison grazing intensity pattern was affected by whether a plot was burned and only marginally affected by plot size. During the next 5 months (January–June), grazing intensity was unaffected by plot size, but was greatest in the unburned 225 and 3600-m² plots. The final 4 months (June–October), grazing intensity was unaffected by treatments other than being higher in the unburned 3600-m² plots. By the final sampling date, biomass was significantly greater in the burned plots and grazing intensity appeared to be responding to the amount of biomass present and the total amount of N present. The pattern displayed within the first 5 months after the burn is congruent with the expectations of optimal foraging theory with overmatching in the smallest plot size of 225 m² (BioScience 37 (1987) 789–799). The next two sampling periods displayed a matching aggregate response relative to biomass availability (Oecologia 100 (1999) 107–117) and total nitrogen mass (g m⁻²). The temporal shift that we found in Bison response to burn patch size is, to our knowledge, the first such examination of both spatial and temporal responses by Bison to landscape heterogeneity. We now have quantitative evidence of how native herbivores can alter their foraging responses to changes in landscape structure over time.     

Landscape characteristics of <Emphasis Type="Italic">Rhizophora mangle</Emphasis> forests and propagule deposition in coastal environments of Florida (USA)

Field dispersal studies are seldom conducted at regional scales even though reliable information on mid-range dispersal distance is essential for models of colonization. The purpose of this study was to examine the potential distance of dispersal of Rhizophora mangle propagules by comparing deposition density with landscape characteristics of mangrove forests. Propagule density was estimated at various distances to mangrove sources (R. mangle) on beaches in southwestern Florida in both high-and low-energy environments, either facing open gulf waters vs. sheltered, respectively. Remote sensing and Geographic Information Systems were used to identify source forests and to determine their landscape characteristics (forest size and distance to deposition area) for the regression analyses. Our results indicated that increasing density of propagules stranded on beaches was related negatively to the distance of the deposition sites from the nearest stands of R. mangle and that deposition was greatly diminished 2 km or more from the source. Measures of fragmentation such as the area of the R. mangle forests were related to propagule deposition but only in low-energy environments. Our results suggest that geographic models involving the colonization of coastal mangrove systems should include dispersal dynamics at mid-range scales, i.e., for our purposes here, beyond the local scale of the forest and up to 5 km distant. Studies of mangrove propagule deposition at various spatial scales are key to understanding regeneration limitations in natural gaps and restoration areas. Therefore, our study of mid-range propagule dispersal has broad application to plant ecology, restoration, and modeling.     

Transient co-expression with three <Emphasis Type="Italic">O</Emphasis>-glycosylation enzymes allows production of GalNAc-<Emphasis Type="Italic">O</Emphasis>-glycosylated Granulocyte-Colony Stimulating Factor in <Emphasis Type="Italic">N. benthamiana</Emphasis>

Background

Expression of economically relevant proteins in alternative expression platforms, especially plant expression platforms, has gained significant interest in recent years. A special interest in working with plants as bioreactors for the production of pharmaceutical proteins is related to low production costs, product safety and quality. Among the different properties that plants can also offer for the production of recombinant proteins, protein glycosylation is crucial since it may have an impact on pharmaceutical functionality and/or stability.

Results

The pharmaceutical glycoprotein human Granulocyte-Colony Stimulating Factor was transiently expressed in Nicotiana benthamiana plants and subjected to mammalian-specific mucin-type O-glycosylation by co-expressing the pharmaceutical protein together with the glycosylation machinery responsible for such post-translational modification.

Conclusions

The pharmaceutical glycoprotein human Granulocyte-Colony Stimulating Factor can be expressed in N. benthamiana plants via agroinfiltration with its native mammalian-specific mucin-type O-glycosylation.

     

Fire severity and seed source influence lodgepole pine (<Emphasis Type="Italic">Pinus contorta</Emphasis> var. <Emphasis Type="Italic">murrayana</Emphasis>) regeneration in the southern cascades,Lassen volcanic National Park,California

Rocky Mountain lodgepole pine, (Pinus contorta var. latifolia) regenerates quickly after high severity fire because seeds from serotinous cones are released immediately post-fire. Sierra lodgepole pine (P. contorta var. murrayana) forests burn with variable intensity resulting in different levels of severity and because this variety of lodgepole pine does not have serotinous cones, little is known about what factors influence post-fire regeneration. This study quantifies tree regeneration in a low, moderate, and high severity burn patch in a Sierra lodgepole forest 24 years after fire. Regeneration was measured in ten plots in each severity type. In each plot, we quantified pre- and post-fire forest structure (basal area, density), counted and aged tree seedlings and saplings of all species, and measured distance to the nearest seed bearing tree. There was no difference in the density of seedlings and saplings among severity classes. Distance and direction to the nearest seed bearing lodgepole pine were the best predictors of lodgepole seedling and sapling density in high severity plots. In contrast to Rocky Mountain lodgepole pine, regeneration of Sierra lodgepole pine appears to rely on in-seeding from surviving trees in low or moderate severity burn patches or live trees next to high severity burn patches. Our data demonstrate that Sierra lodgepole pine follows stand development pathways hypothesized for non-serotinous stands of Rocky Mountain lodgepole pine.     

Shaken but not stirred: multiscale habitat suitability modeling of sympatric marten species (<Emphasis Type="Italic">Martes martes</Emphasis> and <Emphasis Type="Italic">Martes foina</Emphasis>) in the northern Iberian Peninsula

Context

Multispecies and multiscale habitat suitability models (HSM) are important to identify the environmental variables and scales influencing habitat selection and facilitate the comparison of closely related species with different ecological requirements.

Objectives

This study explores the multiscale relationships of habitat suitability for the pine (Martes martes) and stone marten (M. foina) in northern Spain to evaluate differences in habitat selection and scaling, and to determine if there is habitat niche displacement when both species coexist.

Methods

We combined bivariate scaling and maximum entropy modeling to compare the multiscale habitat selection of the two martens. To optimize the HSM, the performance of three sampling bias correction methods at four spatial scales was explored. HSMs were compared to explore niche differentiation between species through a niche identity test.

Results

The comparison among HSMs resulted in the detection of a significant niche divergence between species. The pine marten was positively associated with cooler mountainous areas, low levels of human disturbance, high proportion of natural forests and well-connected forestry plantations, and medium-extent agroforestry mosaics. The stone marten was positively related to the density of urban areas, the proportion and extensiveness of croplands, the existence of some scrub cover and semi-continuous grasslands.

Conclusions

This study outlines the influence of the spatial scale and the importance of the sampling bias corrections in HSM, and to our knowledge, it is the first comparing multiscale habitat selection and niche divergence of two related marten species. This study provides a useful methodological framework for multispecies and multiscale comparatives.

     

The Genus <Emphasis Type="Italic">Fragaria </Emphasis>in Croatia

Only three native species of genus Fragaria (F. moschata, F. vesca and F. viridis) are recorded in three regions of Croatia. These species as well as many of their hybrids, are, or once were, cultivated for their edible fruits. The majority of cultivated strawberries in Europe belong to garden strawberries F. x ananassa (hybrids of F. chiloensis and F. virginiana). The most expanded wild strawberry species in Croatia is a woodland strawberry (F. vesca) whose berries are gathered seasonally as wild edible fruits. They often contain higher amount of nutrients and bioactive compounds in comparison to cultivated varieties. The research on the genus Fragaria species distribution in Croatia has not been carried out, and so is the case with many others wild growing fruit species in Croatia. By summing up a number of individual citations and observations, it is possible to get a perspective regarding the current state of their distribution.     

Isolation of intact sub-dermal secretory cavities from <Emphasis Type="Italic">Eucalyptus</Emphasis>

Background  

The biosynthesis of plant natural products in sub-dermal secretory cavities is poorly understood at the molecular level, largely due to the difficulty of physically isolating these structures for study. Our aim was to develop a protocol for isolating live and intact sub-dermal secretory cavities, and to do this, we used leaves from three species of Eucalyptus with cavities that are relatively large and rich in essential oils.     

Scales of movement by elk (<Emphasis Type="Italic">Cervus elaphus</Emphasis>) in response to heterogeneity in forage resources and predation risk

Animals may respond to spatial and temporal heterogeneity by altering their movement patterns. The time an animal spends in an area of a given size is termed ȁ8first-passage timeȁ9 and can be used to identify the scales at which different movement processes occur. Using first-passage time and 2-h observations, we identified nested spatial scales representing three movement behaviours for elk (Cervus elaphus) – inactive/resting (moves < 50 m), active/foraging (x̄ = 276.7 m, SD = 56.6), and active/relocating (x̄ = 1628.3 m, SD = 436.6). Our ability to identify inactive behaviour was limited by GPS accuracy. The scale separating relocating and foraging behaviour ranged 550–1650 m across individuals and varied quadratically with the mean patch size of cutover forest in an animal’s home range. We classified path segments into the 3 movement behaviours and related behaviours to local environmental conditions. Elk were likely to be inactive in areas having a low predicted use by wolves (Canis lupus), farther than 50 m from anthropogenic linear clearings, and where microclimatic conditions were cool (high shrub cover and north to east-facing slopes). In contrast, elk were most likely to forage in areas having intermediate levels of herbaceous biomass and low movement costs. Elk were most likely to be relocating when in areas of high wolf use, when close to linear clearings, and in energetically costly situations such as moving upslope. We discuss how elk use of potential foraging habitats may be restricted in this landscape by risks imposed by predators, humans, or both.     

Neighbourhood landscape effect on population kinetics of the fossorial water vole (<Emphasis Type="Italic">Arvicola terrestris scherman</Emphasis>)

This paper addresses the issue of whether landscape structure affects A. terrestris population kinetics on a neighbourhood spatial scale, and if so, at what spatial scale is that effect at its maximum. We investigated how the growth of A. terrestris populations is influenced by the landscape context of parcels used for hay production in the French Jura Mountains. Five landscape metrics (relative area of grassland, mean patch area of grassland, patch density of grassland, woodland patch density in grassland, grassland–woodland edge density) were computed over an increasing radius around each parcel (max. 3 km). Redundancy analysis showed that the extent, rate and early onset of A. terrestris population growth were favoured in open grassland areas. Landscape effects on A. terrestris populations as determined by the five metrics are scale-dependent: mean patch area of grassland, patch density of grassland and woodland patch density in grassland had an impact on a grassland parcel within a neighbourhood radius of about 800 m, while relative area of grassland and grassland–woodland edge density had an impact within a neighbourhood radius of about 400 m. Those findings corroborate earlier hypotheses about a multifactorial regulation of A. terrestris populations and a spatial hierarchy of regulating factors. They have potential implications in terms of landscape management and small mammal pest control.     

Edge effect and matrix influence on the nest survival of an old forest specialist,the Brown Creeper (<Emphasis Type="Italic">Certhia americana</Emphasis>)

Conservation strategies should be based on a solid understanding of processes underlying species response to landscape change. In forests fragmented by agriculture, elevated nest predation rates have been reported in many forest bird species, especially near edges. In intensively-managed forest landscapes, timber harvesting might also be associated with negative edge effects or broader “context” effects on some species when the matrix provides additional resources to their major nest predators. In this study, we hypothesized that proximity to a forest edge and proportion of cone-producing plantations will increase nest predation risk in fragments of relatively undisturbed forest. We focused on the Brown Creeper (Certhia americana), an indicator species of late-seral forests. We compared habitat configuration and composition at four spatial scales (0.14, 0.5, 1 and 2 km) around 54 nests and related daily nest survival rate to the distance to the nearest forest edge, mean patch size of late-seral forest (r = 141 m), proportion of non-forested lands (r = 141 m), density of maintained roads (r = 1 km), proportion of cone-producing spruce plantations (r = 2 km), and year. The best model included distance to the nearest edge and proportion of cone-producing plantations. Distance of nests to the nearest edge was the best individual predictor of daily nest survival. A larger sample of nests showed a significant threshold in distance to the nearest forest edge; nests located at least 100 m away were more likely to fledge young. These results suggest that even in managed forest landscapes, matrix effects can be important and some bird species may exhibit negative edge effects.     

A plastome primer set for comprehensive quantitative real time RT-PCR analysis of <Emphasis Type="Italic">Zea mays</Emphasis>: a starter primer set for other <Emphasis Type="Italic">Poaceae</Emphasis> species

Background  

Quantitative Real Time RT-PCR (q2(RT)PCR) is a maturing technique which gives researchers the ability to quantify and compare very small amounts of nucleic acids. Primer design and optimization is an essential yet time consuming aspect of using q2(RT)PCR. In this paper we describe the design and empirical optimization of primers to amplify and quantify plastid RNAs from Zea mays that are robust enough to use with other closely related species.     

High-level diterpene production by transient expression in <Emphasis Type="Italic">Nicotiana benthamiana</Emphasis>

Background

Characterization of plant terpene synthases is typically done by production of recombinant enzymes in Escherichia coli. This is often difficult due to solubility and codon usage issues. Furthermore, plant terpene synthases which are targeted to the plastids, such as diterpene synthases, have to be shortened in a more or less empirical approach to improve expression. We report here an optimized Agrobacterium-mediated transient expression assay in Nicotiana benthamiana for plant diterpene synthase expression and product analysis.

Results

Agrobacterium-mediated transient expression of plant diterpene synthases in N. benthamiana led to the accumulation of diterpenes within 3 days of infiltration and with a maximum at 5 days. Over 50% of the products were exported onto the leaf surface, thus considerably facilitating the analysis by reducing the complexity of the extracts. The robustness of the method was tested by expressing three different plant enzymes, cembratrien-ol synthase from Nicotiana sylvestris, casbene synthase from Ricinus communis and levopimaradiene synthase from Gingko biloba. Furthermore, co-expression of a 1-deoxy-D-xylulose-5-phosphate synthase from tomato and a geranylgeranyl diphosphate synthase from tobacco led to a 3.5-fold increase in the amount of cembratrien-ol produced, with maximum yields reaching 2500 ng/cm².

Conclusion

With this optimized method for diterpene synthase expression and product analysis, a single infiltrated leaf of N. benthamiana would be sufficient to produce quantities required for the structure elucidation of unknown diterpenes. The method will also be of general use for gene function discovery, pathway reconstitution and metabolic engineering of diterpenoid biosynthesis in plants.

     

Divergent landscape effects on genetic differentiation in two populations of the European pine marten (<Emphasis Type="Italic">Martes martes</Emphasis>)

Context

Quantifying gene flow in natural populations is a key topic in both evolutionary and conservation biology. Understanding the extent to which the landscape matrix facilitates or impedes gene flow is becoming a high priority in a context of worldwide habitat loss and fragmentation.

Objectives

Unexpectedly, a lower genetic diversity and a higher genetic structure have been previously observed in the less fragmented and the most forested habitat across four pine marten (Martes martes) populations in France. Our aim was to quantify the effect of landscape on the spatial distribution of genetic diversity in two populations in contrasting habitats.

Methods

We conducted an individual-based landscape genetics analysis in a highly fragmented rural plain (Bresse, n = 126) and in a highly forested (50 %) mountainous area (Ariège, n = 88) in France. We tested for isolation-by-resistance using least-cost distances and used a causal modeling approach on 16,384 landscape and 104 elevation resistance scenarios.

Results

Landscape structure influenced the genetic differentiation in Bresse, with vegetation providing more genetic connectivity over the study area than open areas, while roads and human buildings showed unexpected low resistance to gene flow. In Ariège, genetic differentiation was mainly associated with changes in elevation, with an optimal elevation for gene flow of around 1700 m, likely associated with changes in vegetation structure.

Conclusions

The pine marten seems to be able to cope with human-dominated landscapes and with fragmented forest landscapes, whereas elevation is the major driver of genetic differentiation in our mountainous landscape. Additionally, we highlight the importance of spatial replication in landscape genetics for deriving reliable conservation and management measures over the species distribution.

     

Rapid metabolic profiling of <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> defence responses against <Emphasis Type="Italic">Phytophthora nicotianae</Emphasis> using direct infrared laser desorption ionization mass spectrometry and principal component analysis

Background  

Successful defence of tobacco plants against attack from the oomycete Phytophthora nicotianae includes a type of local programmed cell death called the hypersensitive response. Complex and not completely understood signaling processes are required to mediate the development of this defence in the infected tissue. Here, we demonstrate that different families of metabolites can be monitored in small pieces of infected, mechanically-stressed, and healthy tobacco leaves using direct infrared laser desorption ionization orthogonal time-of-flight mass spectrometry. The defence response was monitored for 1 - 9 hours post infection.     

Protocol: Optimised methodology for isolation of nuclei from leaves of species in the <Emphasis Type="Italic">Solanaceae</Emphasis> and <Emphasis Type="Italic">Rosaceae</Emphasis> families

In this study, a protocol is described for rapid preparation of an enriched, reasonably pure fraction of nuclear proteins from the leaves of tobacco (Nicotiana tabacum), potato (Solanum tuberosum) and apple (Malus domestica). The protocol gives reproducible results and can be carried out quickly in 2 hours. Tissue extracts clarified with filtration were treated with non-ionic detergent (Triton X-100) to lyse membranes of contaminating organelles. Nuclei were collected from a 60% Percoll layer of density gradient following low-speed centrifugation. Western blot analysis using antibodies to marker proteins of organelles indicated that the nuclear protein fractions were highly enriched and free or nearly free of proteins from the endoplasmic reticulum and chloroplasts.     

Factors influencing female home range sizes in elk (<Emphasis Type="Italic">Cervus elaphus</Emphasis>) in North American landscapes

Home range size is a result of individual movements and the spatial distribution of a population. While body size, sex, and age are known to influence the area over which an animal ranges, it remains uncertain how landscape heterogeneity influences home range size. We examined elk (Cervus elaphus) seasonal home range sizes in relation to the quantity and spatial heterogeneity of forage biomass, forest cover, topography, snow–water equivalents, and landscape structure in three study landscapes: Yellowstone National Park, Wyoming, USA; eastern slopes of the Canadian Rockies, Alberta; and northern Wisconsin, USA. We used a 95% fixed kernel estimator to measure the home range size and location of all elk. To identify the scales at which important factors influenced home range sizes, we quantified environmental variables within the estimated home range polygon and within concentric circles with radii of 1000, 2000, 3000, 4000, and 5000 m from the home range center. Results indicate that there was an inverse relationship between forage biomass and summer and winter home range sizes in Alberta and Wisconsin, however the relationship was positive in Yellowstone. The size of summer and winter home ranges was positively related to percent forest cover; however this relationship was significant only when forest cover was quantified within the home range polygon or radii that were greater than or equal to 3000 m. Winter home ranges also had a positive relationship with snow–water equivalents. The predictive strength of summer home range models was greatest when landscape variables were quantified within the concentric circles with a radius of 3000 m or more, whereas the predictive strength of the winter models was greatest within the estimated home range polygon. Results suggest that elk ranging patterns reflected complex trade-offs that affect foraging, group dynamics, movement energetics, predation avoidance and thermal regulation. The multi-scale analysis indicates that elk based home ranging decisions on an area equal to their home range, but areas outside of the estimated home range were also important.     

Chlorophyll <Emphasis Type="Italic">a</Emphasis> fluorescence,under half of the adaptive growth-irradiance,for high-throughput sensing of leaf-water deficit in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> accessions

Background

Non-invasive and high-throughput monitoring of drought in plants from its initiation to visible symptoms is essential to quest drought tolerant varieties. Among the existing methods, chlorophyll a fluorescence (ChlF) imaging has the potential to probe systematic changes in photosynthetic reactions; however, prerequisite of dark-adaptation limits its use for high-throughput screening.

Results

To improve the throughput monitoring of plants, we have exploited their light-adaptive strategy, and investigated possibilities of measuring ChlF transients under low ambient irradiance. We found that the ChlF transients and associated parameters of two contrasting Arabidopsis thaliana accessions, Rsch and Co, give almost similar information, when measured either after ~20 min dark-adaptation or in the presence of half of the adaptive growth-irradiance. The fluorescence parameters, effective quantum yield of PSII photochemistry (Φ_PSII) and fluorescence decrease ratio (R _FD) resulting from this approach enabled us to differentiate accessions that is often not possible by well-established dark-adapted fluorescence parameter maximum quantum efficiency of PSII photochemistry (F _V/F _M). Further, we screened ChlF transients in rosettes of well-watered and drought-stressed six A. thaliana accessions, under half of the adaptive growth-irradiance, without any prior dark-adaptation. Relative water content (RWC) in leaves was also assayed and compared to the ChlF parameters. As expected, the RWC was significantly different in drought-stressed from that in well-watered plants in all the six investigated accessions on day-10 of induced drought; the maximum reduction in the RWC was obtained for Rsch (16%), whereas the minimum reduction was for Co (~7%). Drought induced changes were reflected in several features of ChlF transients; combinatorial images obtained from pattern recognition algorithms, trained on pixels of image sequence, improved the contrast among drought-stressed accessions, and the derived images were well-correlated with their RWC.

Conclusions

We demonstrate here that ChlF transients and associated parameters measured even in the presence of low ambient irradiance preserved its features comparable to that of measured after dark-adaptation and discriminated the accessions having differential geographical origin; further, in combination with combinatorial image analysis tools, these data may be readily employed for early sensing and mapping effects of drought on plant’s physiology via easy and fully non-invasive means.

     

Optimization of Factors Affecting <Emphasis Type="Italic">In vitro</Emphasis> Establishment, <Emphasis Type="Italic">Ex vitro</Emphasis> Rooting and Hardening for Commercial Scale Multiplication of Silk Banana (<Emphasis Type="Italic">Musa</Emphasis> AAB)

Factors concerning aseptic culture establishment and hardening were studied in detail in choicest Silk Banana. Effect of size of sucker (small, medium and large), carbon concentration (10, 20 and 30 g/l), season of initiation (wet and dry) and pre-treatments such as segmentation and incision to the explants were studied during initiation. Further, hardening related factors such as substrates used for primary and secondary hardening, nursery nutrition (source and frequency of application) and pre-treatments for ex vitro rooting were also studied. Results revealed that small suckers were most suited for initiations with the least contamination, maximum establishment and higher percentage of greening. Lower concentration of sucrose, though delayed greening, resulted in 100?% establishment of explants. Initiations performed during the drier period were completely free from the fungal contamination and showed less bacterial contamination than those performed during the rainy season. Segmentation of explant into four parts during first subculture supported maximum shoot proliferation by overcoming apical dominance. Coir pith was observed to be the most congenial substrate during primary hardening, whereas coir pith alone or sand: red earth: coir pith (1:1:1) supported superior performance of plantlets during secondary hardening. Single application of mono ammonium phosphate improved growth of plantlets during secondary hardening. Pre-treatment with mono ammonium phosphate gave the best response in terms of rooting and hardening of un-rooted micro-shoots. Thus, the discussed methodology could help the industries to take up commercial scale propagation of Silk Banana.