Impact of Burkholderia phytofirmans strain PsJN on the ex vitro acclimatization and in vitro propagation of tissue cultured Dionaea muscipula (Venus fly trap)

Dionaea muscipula, Venus fly trap, is an endangered plant that is propagated primarily through tissue culture. Tissue cultured plants must be acclimatized to their new environments when removed from their test tubes and the ensuing stress can result in plant mortality. A plant growth promoting rhizobacterium, Burkholderia phytofirmans strain PsJN, has been utilized in biotic hardening of tissue cultured plants and in the amelioration of stresses, particularly drought stress in potted plants. We investigated whether this bacterium could improve either propagation or acclimatization of tissue cultured Venus fly traps. Standardized inoculations were performed by adjusting cell density of inoculant by spectrophotometry. In ex vitro acclimatization, inoculation with the bacteria significantly improved survival but only in nonsterile soil media. Initial in vitro inoculations produced negative interactions with significant reduction in propagules and biomass in the plant and bacterial growth on the media. A modified plant tissue culture media using maltose instead of sucrose as a sugar source was created to address this problem. Inoculated plants grown in the maltose media were not significantly different than the mock inoculated controls in terms of biomass, traps produced and propagules produced. Additionally the bacteria did not visibly grow on media supplemented with maltose rather than sucrose as a sugar source. Plant infection was confirmed through fluorescence microscopy. Colony density estimates for both rhizosphere and endosphere populations of the bacteria in tissue cultured plants were generated using plate counting. These findings indicate that Burkholderia phytofirmans strain PsJN may be useful in reduction of mortality during acclimatization of Dionaea muscipula, but only in nonsterile growth situations or when drought stress is expected. The use of alternative tissue culture media using maltose when propagating biotized fly traps in vitro is an improvement over sucrose containing media. Further work must be done to determine whether mortality reduction by strain PsJN is the result of stress reduction by the endophyte or competitive inhibition of an unknown microbial agonist

Fascicles and the interfascicular matrix show adaptation for fatigue resistance in energy storing tendons

Tendon is composed of rope-like fascicles, bound together by interfascicular matrix (IFM). Our previous work shows that the IFM is critical for tendon function, facilitating sliding between fascicles to allow tendons to stretch. This function is particularly important in energy storing tendons, which experience extremely high strains during exercise, and therefore require the capacity for considerable inter-fascicular sliding and recoil. This capacity is not required in positional tendons. Whilst we have previously described the quasi-static properties of the IFM, the fatigue resistance of the IFM in functionally distinct tendons remains unknown. We therefore tested the hypothesis that fascicles and IFM in the energy storing equine superficial digital flexor tendon (SDFT) are more fatigue resistant than those in the positional common digital extensor tendon (CDET). Fascicles and IFM from both tendon types were subjected to cyclic fatigue testing until failure, and mechanical properties were calculated. The results demonstrated that both fascicles and IFM from the energy storing SDFT were able to resist a greater number of cycles before failure than those from the positional CDET. Further, SDFT fascicles and IFM exhibited less hysteresis over the course of testing than their counterparts in the CDET. This is the first study to assess the fatigue resistance of the IFM, demonstrating that IFM has a functional role within tendon and contributes significantly to tendon mechanical properties. These data provide important advances into fully characterising tendon structure-function relationships

Effect of fatigue loading on structure and functional behaviour of fascicles from energy-storing tendons

Tendons can broadly be categorized according to their function: those that act purely to position the limb and those that have an additional function as energy stores. Energy-storing tendons undergo many cycles of large deformations during locomotion, and so must be able to extend and recoil efficiently, rapidly and repeatedly. Our previous work has shown rotation in response to applied strain in fascicles from energy-storing tendons, indicating the presence of helical substructures which may provide greater elasticity and recovery. In the current study, we assessed how preconditioning and fatigue loading affect the ability of fascicles from the energy-storing equine superficial digital flexor tendon to extend and recoil. We hypothesized that preconditioned samples would exhibit changes in microstructural strain response, but would retain their ability to recover. We further hypothesized that fatigue loading would result in sample damage, causing further alterations in extension mechanisms and a significant reduction in sample recovery. The results broadly support these hypotheses: preconditioned samples showed some alterations in microstructural strain response, but were able to recover following the removal of load. However, fatigue loaded samples showed visual evidence of damage and exhibited further alterations in extension mechanisms, characterized by decreased rotation in response to applied strain. This was accompanied by increased hysteresis and decreased recovery. These results suggest that fatigue loading results in a compromised helix substructure, reducing the ability of energy-storing tendons to recoil. A decreased ability to recoil may lead to an impaired response to further loading, potentially increasing the likelihood of injury

Fluorination Influences the Bioisostery of Myo‐Inositol Pyrophosphate Analogs

Inositol pyrophosphates (PP−IPs) are densely phosphorylated messenger molecules involved in numerous biological processes. PP−IPs contain one or two pyrophosphate group(s) attached to a phosphorylated myo-inositol ring. 5PP−IP5 is the most abundant PP−IP in human cells. To investigate the function and regulation by PP−IPs in biological contexts, metabolically stable analogs have been developed. Here, we report the synthesis of a new fluorinated phosphoramidite reagent and its application for the synthesis of a difluoromethylene bisphosphonate analog of 5PP−IP5. Subsequently, the properties of all currently reported analogs were benchmarked using a number of biophysical and biochemical methods, including co-crystallization, ITC, kinase activity assays and chromatography. Together, the results showcase how small structural alterations of the analogs can have notable effects on their properties in a biochemical setting and will guide in the choice of the most suitable analog(s) for future investigations.Swiss National Science Foundation http://dx.doi.org/10.13039/501100001711German Academic Exchange Service http://dx.doi.org/10.13039/100021828Wellcome Trust http://dx.doi.org/10.13039/100010269Peer Reviewe

A Method for Data-Driven Simulations of Evolving Solar Active Regions

We present a method for performing data-driven simulations of solar active region formation and evolution. The approach is based on magnetofriction, which evolves the induction equation assuming the plasma velocity is proportional to the Lorentz force. The simulations of active region coronal field are driven by temporal sequences of photospheric magnetograms from the Helioseismic Magnetic Imager (HMI) instrument onboard the Solar Dynamics Observatory (SDO). Under certain conditions, the data-driven simulations produce flux ropes that are ejected from the modeled active region due to loss of equilibrium. Following the ejection of flux ropes, we find an enhancement of the photospheric horizontal field near the polarity inversion line. We also present a method for the synthesis of mock coronal images based on a proxy emissivity calculated from the current density distribution in the model. This method yields mock coronal images that are somewhat reminiscent of images of active regions taken by instruments such as SDO's Atmospheric Imaging Assembly (AIA) at extreme ultraviolet wavelengths.Comment: Accepted to ApJ; comments/questions related to this article are welcome via e-mail, even after publicatio

Taxonomic shifts in arbuscular mycorrhizal fungal communities with shade and soil nitrogen across conventionally managed and organic coffee agroecosystems

The composition of arbuscular mycorrhizal fungal (AMF) communities should reflect not only responses to host and soil environments, but also differences in functional roles and costs vs. benefits among arbuscular mycorrhizal fungi. The coffee agroecosystem allows exploration of the effects of both light and soil fertility on AMF communities, because of the variation in shade and soil nutrients farmers generate through field management. We used high-throughput ITS2 sequencing to characterize the AMF communities of coffee roots in 25 fields in Costa Rica that ranged from organic management with high shade and no chemical fertilizers to conventionally managed fields with minimal shade and high N fertilization, and examined relationships between AMF communities and soil and shade parameters with partial correlations, NMDS, PERMANOVA, and partial least squares analysis. Gigasporaceae and Acaulosporaceae dominated coffee AMF communities in terms of relative abundance and richness, respectively. Gigasporaceae richness was greatest in conventionally managed fields, while Glomeraceae richness was greatest in organic fields. While total AMF richness and root colonization did not differ between organic and conventionally managed fields, AMF community composition did; these differences were correlated with soil nitrate and shade. OTUs differing in relative abundance between conventionally managed and organic fields segregated into four groups: Gigasporaceae associated with high light and nitrate availability, Acaulosporaceae with high light and low nitrate availability, Acaulosporaceae and a single relative of Rhizophagus fasciculatus with shade and low nitrate availability, and Claroideoglomus/Glomus with conventionally managed fields but uncorrelated with shade and soil variables. The association of closely related taxa with similar shade and light availabilities is consistent with phylogenetic trait conservatism in AM fungi

Fascicles and the interfascicular matrix show decreased fatigue life with ageing in energy storing tendons

Tendon is composed of rope-like fascicles bound together by interfascicular matrix (IFM). The IFM is critical for the function of energy storing tendons, facilitating sliding between fascicles to allow these tendons to cyclically stretch and recoil. This capacity is required to a lesser degree in positional tendons. We have previously demonstrated that both fascicles and IFM in energy storing tendons have superior fatigue resistance compared with positional tendons, but the effect of ageing on the fatigue properties of these different tendon subunits has not been determined. Energy storing tendons become more injury-prone with ageing, indicating reduced fatigue resistance, hence we tested the hypothesis that the decline in fatigue life with ageing in energy storing tendons would be more pronounced in the IFM than in fascicles. We further hypothesised that tendon subunit fatigue resistance would not alter with ageing in positional tendons. Fascicles and IFM from young and old energy storing and positional tendons were subjected to cyclic fatigue testing until failure, and mechanical properties were calculated. The results show that both IFM and fascicles from the SDFT exhibit a similar magnitude of reduced fatigue life with ageing. By contrast, the fatigue life of positional tendon subunits was unaffected by ageing. The age-related decline in fatigue life of tendon subunits in energy storing tendons is likely to contribute to the increased risk of injury in aged tendons. Full understanding of the mechanisms resulting in this reduced fatigue life will aid in the development of treatments and interventions to prevent age-related tendinopathy