Fast Fabrication of Solid-State Nanopores for DNA Molecule Analysis

Solid-state nanopores have been developed as a prominent tool for single molecule analysis in versatile applications. Although controlled dielectric breakdown (CDB) is the most accessible method for a single nanopore fabrication, it is still necessary to improve the fabrication efficiency and avoid the generation of multiple nanopores. In this work, we treated the SiNx membranes in the air–plasma before the CDB process, which shortened the time-to-pore-formation by orders of magnitude. λ-DNA translocation experiments validated the functionality of the pore and substantiated the presence of only a single pore on the membrane. Our fabricated pore could also be successfully used to detect short single-stranded DNA (ssDNA) fragments. Using to ionic current signals, ssDNA fragments with different lengths could be clearly distinguished. These results will provide a valuable reference for the nanopore fabrication and DNA analysis.

Intraspecific niche partition without speciation: individual level web polymorphism within a single island spider population

Early in the process of adaptive radiation, allopatric disruption of gene flow followed by ecological specialization is key for speciation; but, do adaptive radiations occur on small islands without internal geographical barriers? Island populations sometimes harbour polymorphism in ecological specializations, but its significance remains unclear. On one hand, morphs may correspond to ‘cryptic’ species. Alternatively, they could result from population, developmental or behavioural plasticity. The spider Wendilgarda galapagensis (Araneae, Theridiosomatidae) is endemic to the small Isla del Coco and unique in spinning three different web types, each corresponding to a different microhabitat. We tested whether this variation is associated with ‘cryptic’ species or intraspecific behavioural plasticity. Despite analysing 36 803 loci across 142 individuals, we found no relationship between web type and population structure, which was only weakly geographically differentiated. The same pattern holds when looking within a sampling site or considering only F st outliers. In line with genetic data, translocation experiments showed that web architecture is plastic within an individual. However, not all transitions between web types are equally probable, indicating the existence of individual preferences. Our data supports the idea that diversification on small islands might occur mainly at the behavioural level producing an intraspecific niche partition without speciation.

Microfluidic platform enables tailored translocation and reaction cascades in nanoliter droplet networks

In the field of bottom-up synthetic biology, lipid membranes are the scaffold to create minimal cells and mimic reactions and processes at or across the membrane. In this context, we employ here a versatile microfluidic platform that enables precise positioning of nanoliter droplets with user-specified lipid compositions and in a defined pattern. Adjacent droplets make contact and form a droplet interface bilayer to simulate cellular membranes. Translocation of molecules across membranes are tailored by the addition of alpha-hemolysin to selected droplets. Moreover, we developed a protocol to analyze the translocation of non-fluorescent molecules between droplets with mass spectrometry. Our method is capable of automated formation of one- and two-dimensional droplet networks, which we demonstrated by connecting droplets containing different compound and enzyme solutions to perform translocation experiments and a multistep enzymatic cascade reaction across the droplet network. Our platform opens doors for creating complex artificial systems for bottom-up synthetic biology.

Translocation of an arctic seashore plant reveals signs of maladaptation to altered climatic conditions

Ongoing anthropogenic climate change alters the local climatic conditions to which species may be adapted. Information on species’ climatic requirements and their intraspecific variation is necessary for predicting the effects of climate change on biodiversity. We used a climatic gradient to test whether populations of two allopatric varieties of an arctic seashore herb (Primula nutans ssp. finmarchica) show adaptation to their local climates and how a future warmer climate may affect them. Our experimental set-up combined a reciprocal translocation within the distribution range of the species with an experiment testing the performance of the sampled populations in warmer climatic conditions south of their range. We monitored survival, size, and flowering over four growing seasons as measures of performance and, thus, proxies of fitness. We found that both varieties performed better in experimental gardens towards the north. Interestingly, highest up in the north, the southern variety outperformed the northern one. Supported by weather data, this suggests that the climatic optima of both varieties have moved at least partly outside their current range. Further warming would make the current environments of both varieties even less suitable. We conclude that Primula nutans ssp. finmarchica is already suffering from adaptational lag due to climate change, and that further warming may increase this maladaptation, especially for the northern variety. The study also highlights that it is not sufficient to run only reciprocal translocation experiments. Climate change is already shifting the optimum conditions for many species and adaptation needs also to be tested outside the current range of the focal taxon in order to include both historic conditions and future conditions.

Cognitive map–based navigation in wild bats revealed by a new high-throughput tracking system

Seven decades of research on the “cognitive map,” the allocentric representation of space, have yielded key neurobiological insights, yet field evidence from free-ranging wild animals is still lacking. Using a system capable of tracking dozens of animals simultaneously at high accuracy and resolution, we assembled a large dataset of 172 foraging Egyptian fruit bats comprising >18 million localizations collected over 3449 bat-nights across 4 years. Detailed track analysis, combined with translocation experiments and exhaustive mapping of fruit trees, revealed that wild bats seldom exhibit random search but instead repeatedly forage in goal-directed, long, and straight flights that include frequent shortcuts. Alternative, non–map-based strategies were ruled out by simulations, time-lag embedding, and other trajectory analyses. Our results are consistent with expectations from cognitive map–like navigation and support previous neurobiological evidence from captive bats.

Epipelic biofilms respond early to the impacts of urbanization in lowland streams

ABSTRACTMultiple structural and functional endpoints of stream biofilms are employed by water quality monitoring programs to detect both direct and indirect environmental impacts. Since multiple co-occurring stressors influence biofilm development, active biomonitoring (translocation experiments) could provide a useful monitoring tool that reflects the overall water quality of the urbanized sites. The aim of this research was to study the short-term responses of epipelic biofilms caused by their translocation to more polluted reaches in lowland urban streams. Fluvial sediment was translocated in three streams that run through urban areas following an urbanization gradient. The epipelic biofilms in the sediment were sampled to identify any fast occurring changes in their algal and bacterial biomasses, in their respiration and oxygen consumption. The results show that structural changes in the biofilm, such as an increased bacterial density and chlorophyll-a concentration, were measurable after two days of exposure to sites with impaired water quality. These immediate changes in the structure of the biofilm indicate that they are sensitive endpoints that can be employed in fast and inexpensive biomonitoring programs in urbanized streams.

Translocation of an arctic seashore plant reveals signs of maladaptation to altered climatic conditions

Ongoing anthropogenic climate change alters the local climatic conditions to which species may be adapted. Information on species’ climatic requirements and their intraspecific variation is necessary for predicting the effects of climate change on biodiversity. We used a climatic gradient to test whether populations of two allopatric varieties of an arctic seashore herb (Primula nutans ssp. finmarchica) show adaptation to their local climates and how a future warmer climate may affect them. Our experimental set-up combined i) a reciprocal translocation within the distribution range of the species with ii) an experiment testing performance of the sampled populations in warmer climatic conditions south of their range. We monitored survival, size, and flowering over four growing seasons as measures of performance and, thus, proxies of fitness. We found that both varieties performed better in experimental gardens towards the north. Interestingly, highest up in the north, the southern variety outperformed the northern one. Supported by weather data, this suggests that the climatic optima of both varieties has moved at least partly outside their current range. Further warming would make the current environments of both varieties even less suitable. We conclude that Primula nutans ssp. finmarchica is already suffering from adaptation-lag due to climate change, and that further warming may be increase this maladaptation, especially for the northern variety. The study also highlights that it is not sufficient to run only reciprocal translocation experiments. Climate change is already shifting the optimum conditions for many species and adaptation needs also to be tested outside the current range of the focal taxon in order to include both historic conditions and future conditions.