TOR acts as a metabolic gatekeeper for auxin-dependent lateral root initiation in Arabidopsis thaliana.

Plant organogenesis requires matching the available metabolic resources to developmental programs. In Arabidopsis, the root system is determined by primary root-derived lateral roots (LRs), and adventitious roots (ARs) formed from non-root organs. Lateral root formation entails the auxin-dependent activation of transcription factors ARF7, ARF19, and LBD16. Adventitious root formation relies on LBD16 activation by auxin and WOX11. The allocation of shoot-derived sugar to the roots influences branching, but how its availability is sensed for LRs formation remains unknown. We combine metabolic profiling with cell-specific interference to show that LRs switch to glycolysis and consume carbohydrates. The target-of-rapamycin (TOR) kinase is activated in the lateral root domain. Interfering with TOR kinase blocks LR initiation while promoting AR formation. The target-of-rapamycin inhibition marginally affects the auxin-induced transcriptional response of the pericycle but attenuates the translation of ARF19, ARF7, and LBD16. TOR inhibition induces WOX11 transcription in these cells, yet no root branching occurs as TOR controls LBD16 translation. TOR is a central gatekeeper for root branching that integrates local auxin-dependent pathways with systemic metabolic signals, modulating the translation of auxin-induced genes.

SNF1-RELATED KINASE 1 and TARGET OF RAPAMYCIN control light-responsive splicing events and developmental characteristics in etiolated Arabidopsis seedlings.

The kinases SNF1-RELATED KINASE 1 (SnRK1) and TARGET OF RAPAMYCIN (TOR) are central sensors of the energy status, linking this information via diverse regulatory mechanisms to plant development and stress responses. Despite the well-studied functions of SnRK1 and TOR under conditions of limited or ample energy availability, respectively, little is known about the extent to which the 2 sensor systems function and how they are integrated in the same molecular process or physiological context. Here, we demonstrate that both SnRK1 and TOR are required for proper skotomorphogenesis in etiolated Arabidopsis (Arabidopsis thaliana) seedlings, light-induced cotyledon opening, and regular development in light. Furthermore, we identify SnRK1 and TOR as signaling components acting upstream of light- and sugar-regulated alternative splicing events, expanding the known action spectra for these 2 key players in energy signaling. Our findings imply that concurring SnRK1 and TOR activities are required throughout various phases of plant development. Based on the current knowledge and our findings, we hypothesize that turning points in the activities of these sensor kinases, as expected to occur upon illumination of etiolated seedlings, instead of signaling thresholds reflecting the nutritional status may modulate developmental programs in response to altered energy availability.

Straightforward Production Methods for Diverse Porous PEDOT:PSS Structures and Their Characterization.

Porous conductive polymer structures, in particular Poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) structures, are gaining in importance due to their versatile fields of application as sensors, hydrogels, or supercapacitors, to name just a few. Moreover, (porous) conducting polymers have become of interest for wearable and smart textile applications due to their biocompatibility, which enables applications with direct skin contact. Therefore, there is a huge need to investigate distinct, straightforward, and textile-compatible production methods for the fabrication of porous PEDOT:PSS structures. Here, we present novel and uncomplicated approaches to producing diverse porous PEDOT:PSS structures and characterize them thoroughly in terms of porosity, electrical resistance, and their overall appearance. Production methods comprise the incorporation of micro cellulose, the usage of a blowing agent, creating a sponge-like structure, and spraying onto a porous base substrate. This results in the fabrication of various porous structures, ranging from thin and slightly porous to thick and highly porous. Depending on the application, these structures can be modified and integrated into electronic components or wearables to serve as porous electrodes, sensors, or other functional devices.

Central clock components modulate plant shade avoidance by directly repressing transcriptional activation activity of PIF proteins.

Light-environment signals, sensed by plant phytochrome photoreceptors, are transduced to target genes through direct regulation of PHYTOCHROME-INTERACTING FACTOR (PIF) transcription factor abundance and activity. Previous genome-wide DNA-binding and expression analysis has identified a set of genes that are direct targets of PIF transcriptional regulation. However, quantitative analysis of promoter occupancy versus expression level has suggested that unknown "trans factors" modulate the intrinsic transcriptional activation activity of DNA-bound PIF proteins. Here, using computational analysis of published data, we have identified PSEUDO-RESPONSE REGULATORS (PRR5 and PRR7) as displaying a high frequency of colocalization with the PIF proteins at their binding sites in the promoters of PIF Direct Target Genes (DTGs). We show that the PRRs function to suppress PIF-stimulated growth in the light and vegetative shade and that they repress the rapid PIF-induced expression of PIF-DTGs triggered by exposure to shade. The repressive action of the PRRs on both growth and DTG expression requires the PIFs, indicating direct action on PIF activity, rather than a parallel antagonistic pathway. Protein interaction assays indicate that the PRRs exert their repressive activity by binding directly to the PIF proteins in the nucleus. These findings support the conclusion that the PRRs function as direct outputs from the core circadian oscillator to regulate the expression of PIF-DTGs through modulation of PIF transcriptional activation activity, thus expanding the roles of the multifunctional PIF-signaling hub.

Development and Characterization of Hybrid, Temperature Sensing and Heating Yarns with Color Change.

A person's body temperature is an important indicator of their health status. A deviation of that temperature by just 2 °C already has or can lead to serious consequences, such as fever or hypothermia. Hence, the development of a temperature-sensing and heatable yarn is an important step toward enabling and improving the monitoring and regulation of a person's body temperature. This technology offers benefits to several industries, such as health care and sports. This paper focuses on the characterization and development of a hybrid yarn, which can measure and visualize temperature changes through a thermoresistive and thermochromic effect. Moreover, the yarn is able to serve as a flexible heating element by connecting to a power source. The structure of the yarn is designed in three layers. Each layer and component ensures the functionality and flexibility of the yarn and additional compatibility with further processing steps. A flexible stainless steel core was used as the heat-sensitive and heat-conducting material. The layer of polyester wrapped around the stainless steel yarn improves the wearing comfort and serves as substrate material for the thermochromic coating. The resulting hybrid yarn has a reproducible sensory function and changes its resistance by 0.15 Ω between 20 and 60 °C for a length of 30 cm. In addition, the yarn has a uniform and reproducible heating power, so that temperature steps can be achieved at a defined length by selecting certain voltages. The thermochromic color change is clearly visible between 28 and 29 °C. Due to its textile structure, the hybrid sensory and actuating yarn can easily be incorporated into a woven fabric or into a textile by means of joining technology sewing.

Textile Based Electrochromic Cells Prepared with PEDOT: PSS and Gelled Electrolyte.

Electrochromic devices can act as passive displays. They change their color when a low voltage is applied. Flexible and bendable hybrid textile-film electrochromic devices with poly-3,4-ethylenedioxythiophene polystyrene sulfonate (PEDOT:PSS) were prepared on polyethylene polyethylene terephthalate (PEPES) membranes using a spray coating technique. The electrolyte consisted of a gelatin glycerol mixture as host matrix and calcium chloride. Titanium dioxide was used as an ion storage layer and a carbon containing dispersion was used for the counter electrode on a polyester rip-stop fabric. The sheet resistance of PEDOT:PSS on PEPES was 500 Ohm/sq. A 5 × 5 electrochromic matrix with individually addressable pixels was successfully designed and assembled. The switching time of the pixels was 2 s at a voltage of 2.0 V directly after assembling. The use of titanium dioxide as ion storage also increased the contrast of the dark-blue reduced electrochromic layer. Coloration was not self-sustaining. The PEDOT:PSS layer needed a constant low voltage of at least 0.5 V to sustain in the dark-blue reduced state. The switching time increased with time. After 12 months the switching time was ~4 s at a voltage of 2.8 V. The addition of glycerol into the electrolyte extended the lifetime of a non-encapsulated textile electrochromic cell, because moisture is retained in the electrolyte. Charge carriers can be transported into and out of the electrochromic layer.

Job Sharing in the Endomembrane System: Vacuolar Acidification Requires the Combined Activity of V-ATPase and V-PPase.

The presence of a large central vacuole is one of the hallmarks of a prototypical plant cell, and the multiple functions of this compartment require massive fluxes of molecules across its limiting membrane, the tonoplast. Transport is assumed to be energized by the membrane potential and the proton gradient established by the combined activity of two proton pumps, the vacuolar H(+)-pyrophosphatase (V-PPase) and the vacuolar H(+)-ATPase (V-ATPase). Exactly how labor is divided between these two enzymes has remained elusive. Here, we provide evidence using gain- and loss-of-function approaches that lack of the V-ATPase cannot be compensated for by increased V-PPase activity. Moreover, we show that increased V-ATPase activity during cold acclimation requires the presence of the V-PPase. Most importantly, we demonstrate that a mutant lacking both of these proton pumps is conditionally viable and retains significant vacuolar acidification, pointing to a so far undetected contribution of the trans-Golgi network/early endosome-localized V-ATPase to vacuolar pH.

Genome Wide Binding Site Analysis Reveals Transcriptional Coactivation of Cytokinin-Responsive Genes by DELLA Proteins.

The ability of plants to provide a plastic response to environmental cues relies on the connectivity between signaling pathways. DELLA proteins act as hubs that relay environmental information to the multiple transcriptional circuits that control growth and development through physical interaction with transcription factors from different families. We have analyzed the presence of one DELLA protein at the Arabidopsis genome by chromatin immunoprecipitation coupled to large-scale sequencing and we find that it binds at the promoters of multiple genes. Enrichment analysis shows a strong preference for cis elements recognized by specific transcription factor families. In particular, we demonstrate that DELLA proteins are recruited by type-B ARABIDOPSIS RESPONSE REGULATORS (ARR) to the promoters of cytokinin-regulated genes, where they act as transcriptional co-activators. The biological relevance of this mechanism is underpinned by the necessity of simultaneous presence of DELLAs and ARRs to restrict root meristem growth and to promote photomorphogenesis.

A quartet of PIF bHLH factors provides a transcriptionally centered signaling hub that regulates seedling morphogenesis through differential expression-patterning of shared target genes in Arabidopsis.

Dark-grown seedlings exhibit skotomorphogenic development. Genetic and molecular evidence indicates that a quartet of Arabidopsis Phytochrome (phy)-Interacting bHLH Factors (PIF1, 3, 4, and 5) are critically necessary to maintaining this developmental state and that light activation of phy induces a switch to photomorphogenic development by inducing rapid degradation of the PIFs. Here, using integrated ChIP-seq and RNA-seq analyses, we have identified genes that are direct targets of PIF3 transcriptional regulation, exerted by sequence-specific binding to G-box (CACGTG) or PBE-box (CACATG) motifs in the target promoters genome-wide. In addition, expression analysis of selected genes in this set, in all triple pif-mutant combinations, provides evidence that the PIF quartet members collaborate to generate an expression pattern that is the product of a mosaic of differential transcriptional responsiveness of individual genes to the different PIFs and of differential regulatory activity of individual PIFs toward the different genes. Together with prior evidence that all four PIFs can bind to G-boxes, the data suggest that this collective activity may be exerted via shared occupancy of binding sites in target promoters.

Interaction with plant transcription factors can mediate nuclear import of phytochrome B.

Phytochromes (phy) are red/far-red-absorbing photoreceptors that regulate the adaption of plant growth and development to changes in ambient light conditions. The nuclear transport of the phytochromes upon light activation is regarded as a key step in phytochrome signaling. Although nuclear import of phyA is regulated by the transport facilitators far red elongated hypocotyl 1 (FHY1) and fhy1-like, an intrinsic nuclear localization signal was proposed to be involved in the nuclear accumulation of phyB. We recently showed that nuclear import of phytochromes can be analyzed in a cell-free system consisting of isolated nuclei of the unicellular green algae Acetabularia acetabulum. We now show that this system is also versatile to elucidate the mechanism of the nuclear transport of phyB. We tested the nuclear transport characteristics of full-length phyB as well as N- and C-terminal phyB fragments in vitro and showed that the nuclear import of phyB can be facilitated by phytochrome-interacting factor 3 (PIF3). In vivo measurements of phyB nuclear accumulation in the absence of PIF1, -3, -4, and -5 indicate that these PIFs are the major transport facilitators during the first hours of deetiolation. Under prolonged irradiations additional factors might be responsible for phyB nuclear transport in the plant.

Textile One-Component Organic Electrochemical Sensor for Near-Body Applications.

The need for more efficient health services and the trend of a healthy lifestyle pushes the development of smart textiles. Since textiles have always been an object of everyday life, smart textiles promise an extensive user acceptance. Thereby, the manufacture of electrical components based on textile materials is of great interest for applications as biosensors. Organic electrochemical transistors (OECTs) are often used as biosensors for the detection of saline content, adrenaline, glucose, etc., in diverse body fluids. Textile-based OECTs are mostly prepared by combining a liquid electrolyte solution with two separate electro-active yarns that must be precisely arranged in a textile structure. Herein, on the other hand, a biosensor based on a textile single-component organic electrochemical transistor with a hardened electrolyte was developed by common textile technologies such as impregnation and laminating. Its working principle was demonstrated by showing that the herein-produced transistor functions similarly to a switch or an amplifier and that it is able to detect ionic analytes of a saline solution. These findings support the idea of using this new device layout of textile-based OECTs as biosensors in near-body applications, though future work must be carried out to ensure reproducibility and selectivity, and to achieve an increased level of textile integration.

Textile Strain Sensor Enhancement by Coating Metal Yarns with Carbon-Filled Silicone.

Flexible and stretchable strain sensors are an important development for measuring various movements and forces and are increasingly used in a wide range of smart textiles. For example, strain sensors can be used to measure the movements of arms, legs or individual joints. Thereby, most strain sensors are capable of detecting large movements with a high sensitivity. Very few are able to measure small movements, i.e., strains of less than 5%, with a high sensitivity, which is necessary to carry out important health measurements, such as breathing, bending, heartbeat, and vibrations. This research deals with the development of strain sensors capable of detecting strain of 1% with a high sensitivity. For this purpose, a total of six commercially available metallic yarns were coated with a carbon-containing silicone coating. The process is based on a vertical dip-coating technology with a self-printed 3D coating bath. Afterwards, the finished yarns were interlooped and stretched by 1% while electrical resistance measurements were carried out. It was shown that, although the coating reduced the overall conductivity of the yarns, it also improved their sensitivity to stress. Conclusively, highly sensitive strain sensors, designed specially for small loads, were produced by a simple coating set-up and interlooping structure of the sensory yarns, which could easily be embedded in greater textile structures for wearable electronics.

Coating of Silicone Monofilaments with Elastic Carbon Black-Silver-Silicone Layers and Their Characterization Especially with Regard to the Change of the Electrical Resistance in Dependence on Strain.

Smart textiles have properties that outperform the conventional protective and decorative function of textiles. By integrating special sensors into clothing, body functions and movements can be detected. Piezoresistive sensors measure a change in electrical resistance due to the application of force in the form of stretching, pressure or bending. In order to manufacture such sensors, conventional non-conductive textile materials need to be made conductive by finishing processes. Therefore, a non-conductive silicone monofilament was coated with a conductive carbon silicone and additional silver-containing components and investigated for its suitability as a strain sensor. The changes in electrical resistance and the gauge factor as a measure of the sensitivity of a sensor were measured and calculated. In this publication, the electrical properties of such a filament-based sensor in the context of particle composition and concentration are discussed. The electrical resistance was already significantly reduced in a first step by coating with conductive carbon silicone (145 kΩ). The addition of silver-containing components further reduced the electrical resistance in a second step. Thereby, flat flakes of silver proved to be much more effective than silver-containing particles (5 kΩ at 20% addition). The former was easier to integrate into the coating and formed contact surfaces with each other at higher concentrations. Stretching the samples increased the resistance by enlarging the distance between the conductive components. With 30% silver-coated glass flakes in the coating, the highest gauge factor of 0.33 was achieved. Consequently, the changes in electrical resistance during stretching can be exploited to detect motion and the gauge factor indicates that even small changes in strain can be detected, so the herein developed coated monofilaments are suggested for use as strain sensors. Future work includes matching the particle composition and concentration to the exact application and investigating the sensors in the field.

Development of Flexible and Functional Sequins Using Subtractive Technology and 3D Printing for Embroidered Wearable Textile Applications.

Embroidery is often the preferred technology when rigid circuit boards need to be connected to sensors and electrodes by data transmission lines and integrated into textiles. Moreover, conventional circuit boards, like Lilypad Arduino, commonly lack softness and flexibility. One approach to overcome this drawback can be flexible sequins as a substrate carrier for circuit boards. In this paper, such an approach of the development of flexible and functional sequins and circuit boards for wearable textile applications using subtractive and additive technology is demonstrated. Applying these techniques, one-sided sequins and circuit boards are produced using wax printing and etching copper-clad foils, as well as using dual 3D printing of conventional isolating and electrically conductive materials. The resulting flexible and functional sequins are equipped with surface mounted devices, applied to textiles by an automated embroidery process and contacted with a conductive embroidery thread.

A cell-free system for light-dependent nuclear import of phytochrome.

Translocation from the cytosol to the nucleus is an essential step in phytochrome (phy) signal transduction. In the case of phytochrome A (phyA), this step occurs with the help of FHY1 (far-red-elongated hypocotyl 1), a specific transport protein. To investigate the components involved in phyA transport, we used a cell-free system that facilitates the controlled addition of transport factors. For this purpose, we isolated nuclei from the unicellular green algae Acetabularia acetabulum. These nuclei are up to 100 mum in diameter and allow easy detection of imported proteins. Experiments with isolated nuclei of Acetabularia showed that FHY1 is sufficient for phyA transport. The reconstituted system demonstrates all the characteristics of phytochrome transport in Arabidopsis thaliana. In addition, FHY1 was also actively exported from the nucleus, consistent with its role as a shuttle protein in plants. Therefore, we believe that isolated Acetabularia nuclei may be used as a general tool to study nuclear transport of plant proteins.

Characterization of carotenoid profiles in goldenberry (Physalis peruviana L.) fruits at various ripening stages and in different plant tissues by HPLC-DAD-APCI-MSn.

Carotenoid profiles of goldenberry (Physalis peruviana L.) fruits differing in ripening states and in different fruit fractions (peel, pulp, and calyx of ripe fruits) were investigated by HPLC-DAD-APCI-MSn. Out of the 53 carotenoids detected, 42 were tentatively identified. The carotenoid profile of unripe fruits is dominated by (all-E)-lutein (51%), whereas in ripe fruits, (all-E)-ß-carotene (55%) and several carotenoid fatty acid esters, especially lutein esters esterified with myristic and palmitic acid as monoesters or diesters, were found. In overripe fruits, carotenoid conversion products and a higher proportion of carotenoid monoesters to diesters compared to ripe fruits were observed. Overripe fruits showed a significant decrease in total carotenoids of about 31% due to degradation. The observed conversion and degradation processes included epoxidation, isomerization, and deesterification. The peel of ripe goldenberries showed a 2.8 times higher total carotenoid content of 332.00 µg/g dw compared to the pulp.

Comparative Aroma Extract Dilution Analysis (cAEDA) of Fat from Tainted Boars, Castrated Male Pigs, and Female Pigs.

The aroma profile of porcine fat from tainted boars, female pigs, and castrated male pigs was investigated by application of comparative aroma extract dilution analysis (cAEDA) on a SAFE distillate of volatiles prepared from porcine back fat samples. The AEDA resulted in a total of 16 aroma active compounds for boar fat with flavor dilution (FD) factors ranging from 2 to 2048, whereas 12 aroma active compounds were found in fat of female pigs and 14 in fat of castrated male pigs, both with FD factors ranging from 2 to 32. Odor activity values (OAVs) of key components for each fat were identified: In boar fat androstenone, skatole, indole, and 2-aminoacetophenone showed highest OAVs, whereas 2,5-dimethylpyrazine, 2,4-decadienal, and δ-decalactone showed highest OAVs in fat of female pigs. Fat of castrated male pigs showed highest OAVs for skatole, indole, 1-octen-3-ol and methional. Finally, the off-flavor attributes of boar fat were successfully simulated by a recombinant of all odorants at their natural concentration level in deodorized sunflower oil.

Polyethylene particle-induced bone resorption in alpha-calcitonin gene-related peptide-deficient mice.

UNLABELLED: This study investigates the impact of alpha-CGRP on bone metabolism after implantation of polyethylene particles. alpha-CGRP knockout mice showed less osteolysis compared with wildtype mice. The local neurogenic microenvironment might be a crucial factor in particle-induced osteolysis. INTRODUCTION: Periprosthetic osteolysis is the major reason for aseptic loosening in joint arthroplasty. This study aimed to investigate the potential impact of alpha-calcitonin gene-related peptide (alpha-CGRP) deficiency on bone metabolism under conditions of polyethylene particle-induced osteolysis. MATERIALS AND METHODS: We used the murine calvarial osteolysis model based on polyethylene particles in 14 C57BL 6 mice and 14 alpha-CGRP-deficient mice divided into four groups of 7 mice each. Groups 1 (C57BL/J 6) and 3 (alpha-CGRP knockout) received sham surgery, and groups 2 (C57BL/J 6) and 4 (alpha-CGRP knockout) were treated with polyethylene particles. Qualitative and quantitative 3D analyses were performed using microCT. In addition, bone resorption was measured within the midline suture by histological examination. The number of osteoclasts was determined by counting the TRACP(+) cells. Calvarial bone was tested for RANKL expression by RT-PCR and immunocytochemistry. RESULTS: Bone resorption was significantly reduced in alpha-CGRP-deficient mice compared with their corresponding wildtype C57BL 6 mice as confirmed by histomorphometric data (p < 0.001) and microCT (p < 0.01). Osteoclast numbers were significantly reduced in group 3 and the particle subgroup compared with group 1 (p < 0.001). We observed a >3-fold increase of basal RANKL mRNA levels within group 1 compared with group 3. Additional low RANKL immunochemistry staining was noted in groups 3 and 4. CONCLUSIONS: In conclusion, alpha-CGRP knockout mice did not show the expected extended osteolysis compared with wildtype mice expressing alpha-CGRP. One of the most reasonable explanations for the observed decrease in osteolysis could be linked to the osteoprotegerin (OPG)/RANK/RANKL system in alpha-CGRP-deficient animals. As a consequence, the fine tuning of osteoclasts mediating resorption in alpha-CGRP-null mice may be deregulated.

Beyond flexibility: controlling stem cells in an ever changing environment.

Developmental plasticity is a defining feature of plants allowing them to colonize a wide range of different ecosystems by promoting environmental adaptation. Their postembryonic development requires life-long maintenance of stem cells, which are embedded into specialized tissues, called meristems. The shoot apical meristem gives rise to all above ground tissues and is a complex and dynamic three-dimensional structure harboring cells of different clonal origins and fates. Functionally divergent subdomains are stably maintained despite permanent cell division, however their relative sizes are modified in response to developmental and environmental signals. In this review, we briefly describe the core regulatory program of the shoot apical meristem and discuss progress in the fields of mechanical and environmental control of its activity.

Analysis and Sensory Evaluation of the Stereoisomers of a Homologous Series (C5-C10) of 4-Mercapto-2-alkanols.

A homologous series of 4-mercapto-2-alkanols (C5-C10) was used to investigate the impact of the stereochemistry on the sensory properties of a class of naturally occurring polyfunctional thiols having a 1,3-oxygen-sulfur functionality. Stereoisomers were obtained via syntheses of racemic mixtures and subsequent lipase-catalyzed kinetic resolutions. Analytical separations of the stereoisomers were achieved by capillary gas chromatography (GC) using chiral stationary phases. The absolute configurations were assigned via NMR analysis. Sensory evaluations by means of GC/olfactometry revealed odor threshold minima for the medium-chain homologues (C7-C9) of the 4-mercapto-2-alkanol stereoisomers. Except for the C5 homologue, the lowest odor thresholds were determined for the (2R,4R)-configured stereoisomers. The variability in odor qualities was mainly determined by the chain length. None of the 4-mercapto-2-alkanol stereoisomers showed consistent odor qualities for all homologues.