Detection of single quantum dots in model organisms with sheet illumination microscopy

Single-molecule detection and tracking is important for observing biomolecule interactions in the microenvironment. Here we report selective plane illumination microscopy (SPIM) with single-molecule detection in living organisms, which enables fast imaging and single-molecule tracking and optical penetration beyond 300 μm. We detected single nanocrystals in Drosophila larvae and zebrafish embryo. We also report our first tracking of single quantum dots during zebrafish development, which displays a transition from flow to confined motion prior to the blastula stage. The new SPIM setup represents a new technique, which enables fast single-molecule imaging and tracking in living systems.     

Climate sensitivity of soil water regime of different Hungarian Chernozem soil subtypes

In this study the possible effects of two predicted climate change scenarios on soil water regime of Hungarian Calcic Chernozem soils has been investigated. Soil profiles classified as Calcic Chernozem — in total 49 — were selected from the MARTHA soil physical database that incorporates soil data at national scale. These profiles were subdivided into three groups (sandy loam, loam and clayey loam) in accordance with their mechanical composition. Soil water retention curves were scaled separately for each of the three textural groups, using similar media scaling in order to represent the variability of soil hydrophysical data with one parameter, the scaling factor (SF). Reference soil profiles were chosen according to the cumulative distribution function of the scaling factor, six for each textural group. Daily downscaled meteorological data from A2 and B2 climate scenarios of the Hadley Centre (2070–2100) and data from a reference period (RF, 1961–1990) were used in this study to characterize different climatic situations. Nine representative years were selected in case of all the three scenarios, using the cumulative probability function of the annual precipitation sum. Scenario analyses were performed, validating the SWAP soil water balance simulation model for the 18 reference soil profiles and 27 representative years in order to evaluate the expected changes in soil water regime under different from the present (RF) climatic conditions (A2 and B2 scenarios). Our results show that the scaling factor could be used as a climate sensitivity indicator of soil water regime. The large climate sensitivity of the majority of Chernozem soil subtypes water regime has been proven.     

Variation in δ13C among species and sexes in the family Restionaceae along a fine‐scale hydrological gradient

Consistent, repeatable segregation of plant species along hydrological gradients is an established phenomenon that must in some way reflect a trade‐off between plants' abilities to tolerate the opposing constraints of drought and waterlogging. In C₃ species tissue carbon isotope discrimination (δ¹³C) is known to vary sensitively in response to stomatal behaviour, reflecting stomatal limitation of photosynthesis during the period of active growth. However, this has not been studied at fine‐spatial scale in natural communities. We tested how δ¹³C varied between species and sexes of individuals in the family Restionaceae growing along a monitored hydrological gradient. Twenty Restionaceae species were investigated using species‐level phylogeny at two sites in the Cape Floristic Region, a biodiversity hotspot. A spatial overlap analysis showed the Restionaceae species segregated significantly (P < 0.001) at both sites. Moreover, there were significant differences in δ¹³C values among the Restionaceae species (P < 0.001) and between male and female individuals of each species (P < 0.01). However, after accounting for phylogeny, species δ¹³C values did not show any significant correlation with the hydrological gradient. We suggest that some other variable (e.g. plant phenology) could be responsible for masking a simple response to water availability.     

光照下菜豆叶片抗氰呼吸与光合作用关系的分析

为了进一步了解光照下植物呼吸作用的内在机理以及呼吸作用和光合作用的关系, 该文研究了在光照下菜豆(Phaseolus vulgaris)叶片抗氰呼吸与光合作用的关系。研究发现, 将黑暗下生长的菜豆幼苗叶片转到光照下10 h, 总呼吸、抗氰呼吸以及抗氰呼吸在总呼吸中的比例均逐步上升; 光照也导致了叶片叶绿体光合放氧和CO₂固定的出现及其速率的增加, 但光合放氧和CO₂固定速率的增加均滞后于抗氰呼吸的增加。将黑暗下生长的叶片转到光照下之前用抗氰呼吸的抑制剂水杨基氧肟酸(SHAM)处理叶片, 发现用SHAM处理并没有导致叶片在光照下光合放氧和CO₂固定速率的明显变化, 这也提示了黑暗下生长的叶片转至光照的过程中, 抗氰呼吸和光合作用没有产生偶联。进一步研究发现, 在黑暗中对叶片施加短时间的光照能够增加抗氰呼吸在总呼吸中的比例, 但短时间的光照对叶片光合CO₂固定速率没有影响。这些结果表明了光照对抗氰呼吸的诱导可以不依赖于光合作用, 光照可能是作为一种直接的信号去诱导抗氰呼吸。     

Evaluating future success of whitebark pine ecosystem restoration under climate change using simulation modeling

Major declines of whitebark pine forests throughout western North America from the combined effects of mountain pine beetle (Dendroctonus ponderosae) outbreaks, fire exclusion policies, and the exotic disease white pine blister rust (WPBR) have spurred many restoration actions. However, projected future warming and drying may further exacerbate the species' decline and possibly compromise long‐term success of today's restoration activities. We evaluated successes of restoration treatments under future climate using a comprehensive landscape simulation experiment. The spatially explicit, ecological process model FireBGCv2 was used to simulate whitebark pine populations on two U.S. Northern Rocky Mountain landscapes over 95 years under two climate, three restoration, and two fire management scenarios. Major findings were that (1) whitebark pine can remain on some high mountain landscapes in a future climate albeit at lower basal areas (50% decrease), (2) restoration efforts, such as thinning and prescribed burning, are vital to ensure future whitebark pine forests, and (3) climate change impacts on whitebark pine vary by local setting. Whitebark pine restoration efforts will mostly be successful in the future but only if future populations are somewhat resistant to WPBR. Results were used to develop general guidelines that address climate change impacts for planning, designing, implementing, and evaluating fine‐scale restoration activities.     

Survival and life expectancy of the tree Protea roupelliae subsp. roupelliae in a montane grassland savanna: Effects of fire regime and plant structure

Survival and life expectancy are key demographic determinants of population dynamics. Using data collected in a field experiment monitored over 14 years in montane grassland of the Ukhahlamba‐Drakensberg Park, South Africa, we determined the effects of components of fire regime and plant structure on the survival and life expectancy of the tree Protea roupelliae subsp. roupelliae (Proteaceae). The field experiment comprised six plots (0.2–0.5 ha in area) from which the survival and life expectancies of 1567 juveniles (non‐reproductives) and 329 adults (reproductives) were estimated in response to differences in fire frequency, biennial seasonal fire, flame height, juvenile height, adult height, basal area and canopy vigour. Juvenile survival and life expectancies were highest when fires were excluded for 8 years. However, a fire after 12 years of fire exclusion and another fire 2 years later eliminated all juveniles. Over the same 14‐year period of biennial fires, juvenile survival was 5%. Juvenile survival and life expectancy were higher after biennial, winter fires than after annual, winter fires. Flame height had no effect on juvenile survival and life expectancy. Both survival and life expectancy of juveniles increased as plants got older and grew taller. Adult survival was unaffected by fire frequency, flame height or tree size, but the survival of adults in response to fire seasonality was inconclusive. Adults with low canopy vigour (<25%) were negatively affected by fire. Juvenile survival and life expectancy are critical bottlenecks in the demography of P. roupelliae. This species is neither a reseeder nor a resprouter. It avoids lethal fire damage by being restricted to rocky habitats with low fire intensities. Biennial winter fires least threaten the survival and life expectancy of P. roupelliae and impact least on its role in the summer feeding and breeding of Gurney's sugarbird.     

Social facilitation is a better predictor of frog reproductive activity than environmental factors

Anuran breeding activity is frequently linked to environmental factors, mainly temperature and rainfall. However, a key feature of anuran reproductive behavior—gathering in choruses and producing loud advertisement calls to attract females—generates a conspicuous social cue that may also facilitate reproductive behavior. Here, I examine the relative importance of environmental and social factors in explaining the intensity of reproductive activity in the Neotropical treefrog Hypsiboas rosenbergi. I show that social cues generally play an important role, but that there are sex differences: male behavior is associated with a combination of environmental and social factors, while female behavior is associated almost exclusively with social cues. I discuss the potential benefits of using social cues in regulating breeding activity, and suggest that conservation efforts may take advantage of the apparently widespread pattern of social facilitation in anuran reproductive ecology.     

Fluorescence Microscopy Gets Faster and Clearer: Roles of Photochemistry and Selective Illumination

Significant advances in fluorescence microscopy tend be a balance between two competing qualities wherein improvements in resolution and low light detection are typically accompanied by losses in acquisition rate and signal-to-noise, respectively. These trade-offs are becoming less of a barrier to biomedical research as recent advances in optoelectronic microscopy and developments in fluorophore chemistry have enabled scientists to see beyond the diffraction barrier, image deeper into live specimens, and acquire images at unprecedented speed. Selective plane illumination microscopy has provided significant gains in the spatial and temporal acquisition of fluorescence specimens several mm in thickness. With commercial systems now available, this method promises to expand on recent advances in 2-photon deep-tissue imaging with improved speed and reduced photobleaching compared to laser scanning confocal microscopy. Superresolution microscopes are also available in several modalities and can be coupled with selective plane illumination techniques. The combination of methods to increase resolution, acquisition speed, and depth of collection are now being married to common microscope systems, enabling scientists to make significant advances in live cell and in situ imaging in real time. We show that light sheet microscopy provides significant advantages for imaging live zebrafish embryos compared to laser scanning confocal microscopy.