Ferulic acid is a putative surrender signal to stimulate programmed cell death in grapevines after infection with Neofusicoccum parvum.

An apoplectic breakdown from grapevine trunk diseases (GTDs) has become a serious challenge to viticulture as a consequence of drought stress. We hypothesize that fungal aggressiveness is controlled by a chemical communication between the host and colonizing fungus. We introduce the new concept of a 'plant surrender signal' accumulating in host plants under stress and facilitating the aggressive behaviour of the strain Neofusicoccum parvum (Bt-67) causing Botryosphaeriaceae-related dieback in grapevines. Using a cell-based experimental system (Vitis cells) and bioactivity-guided fractionation, we identify trans-ferulic acid, a monolignol precursor, as a 'surrender signal'. We show that this signal specifically activates the secretion of the fungal phytotoxin fusicoccin A aglycone. We show further that this phytotoxin, mediated by 14-3-3 proteins, activates programmed cell death in Vitis cells. We arrive at a model showing a chemical communication facilitating fusicoccin A secretion that drives necrotrophic behaviour during Botryosphaeriaceae-Vitis interaction through trans-ferulic acid. We thus hypothesize that channelling the phenylpropanoid pathway from this lignin precursor to the trans-resveratrol phytoalexin could be a target for future therapy.

Endoreplication-Why Are We Not Using Its Full Application Potential?

Endoreplication-a process that is common in plants and also accompanies changes in the development of animal organisms-has been seen from a new perspective in recent years. In the paper, we not only shed light on this view, but we would also like to promote an understanding of the application potential of this phenomenon in plant cultivation. Endoreplication is a pathway for cell development, slightly different from the classical somatic cell cycle, which ends with mitosis. Since many rounds of DNA synthesis take place within its course, endoreplication is a kind of evolutionary compensation for the relatively small amount of genetic material that plants possess. It allows for its multiplication and active use through transcription and translation. The presence of endoreplication in plants has many positive consequences. In this case, repeatedly produced copies of genes, through the corresponding transcripts, help the plant acquire the favorable properties for which proteins are responsible directly or indirectly. These include features that are desirable in terms of cultivation and marketing: a greater saturation of fruit and flower colors, a stronger aroma, a sweeter fruit taste, an accumulation of nutrients, an increased resistance to biotic and abiotic stress, superior tolerance to adverse environmental conditions, and faster organ growth (and consequently the faster growth of the whole plant and its biomass). The two last features are related to the nuclear-cytoplasmic ratio-the greater the content of DNA in the nucleus, the higher the volume of cytoplasm, and thus the larger the cell size. Endoreplication not only allows cells to reach larger sizes but also to save the materials used to build organelles, which are then passed on to daughter cells after division, thus ending the classic cell cycle. However, the content of genetic material in the cell nucleus determines the number of corresponding organelles. The article also draws attention to the potential practical applications of the phenomenon and the factors currently limiting its use.

Subwavelength Grating Waveguide Structures Proposed on the Low-Cost Silica-Titania Platform for Optical Filtering and Refractive Index Sensing Applications.

The sol−gel dip-coating method is a cost-efficient way for the realization of thin films on a planar substrate. In this work, high-quality, low-loss, and low-surface roughness silica−titania thin films are deposited on a glass substrate with the sol−gel dip-coating method. This platform works in the visible to near-IR wavelength ranges and can be useful for several eye-catching photonic components. The paper is comprised of two parts: the first part deals with the development of a low-cost silica−titania waveguide system, whereas the second part provides detail on the numerical modeling of the SWG waveguide filter and SWG waveguide FP-sensor design. The SWG waveguide NIR-stopband filter can achieve an ER of >40 dB and 3-dB bandwidth of 110 nm designed at optimized parameters. The SWG waveguide-FP structure proposed in this work act as a refractive index sensor where the sensitivity is ~120 nm/RIU by reducing the width of the waveguide. This sensitivity can be further enhanced by reducing the waveguide height. We believe that this work is quite important for the realization of low-cost integrated photonic devices based on the silica−titania platform developed via the sol−gel dip-coating method.

Mode Sensitivity Exploration of Silica-Titania Waveguide for Refractive Index Sensing Applications.

In this paper, a novel and cost-effective photonic platform based on silica-titania material is discussed. The silica-titania thin films were grown utilizing the sol-gel dip-coating method and characterized with the help of the prism-insertion technique. Afterwards, the mode sensitivity analysis of the silica-titania ridge waveguide is investigated via the finite element method. Silica-titania waveguide systems are highly attractive due to their ease of development, low fabrication cost, low propagation losses and operation in both visible and near-infrared wavelength ranges. Finally, a ring resonator (RR) sensor device was modelled for refractive index sensing applications, offering a sensitivity of 230 nm/RIU, a figure of merit (FOM) of 418.2 RIU-1, and Q-factor of 2247.5 at the improved geometric parameters. We believe that the abovementioned integrated photonics platform is highly suitable for high-performance and economically reasonable optical sensing devices.

Photonic Integrated Interrogator for Monitoring the Patient Condition during MRI Diagnosis.

In this work, we discuss the idea and practical implementation of an integrated photonic circuit-based interrogator of fiber Bragg grating (FBG) sensors dedicated to monitoring the condition of the patients exposed to Magnetic Resonance Imaging (MRI) diagnosis. The presented solution is based on an Arrayed Waveguide Grating (AWG) demultiplexer fabricated in generic indium phosphide technology. We demonstrate the consecutive steps of development of the device from design to demonstrator version of the system with confirmed functionality of monitoring the respiratory rate of the patient. The results, compared to those obtained using commercially available bulk interrogator, confirmed both the general concept and proper operation of the device.

Melatonin Application Modifies Antioxidant Defense and Induces Endoreplication in Maize Seeds Exposed to Chilling Stress.

The aim of the study was to demonstrate the biostimulating effect of exogenous melatonin (MEL) applied to seeds via hydroconditioning. It was indicated that only well-chosen application technique and MEL dose guarantees success concerning seed germination and young seedlings growth under stress conditions. For maize seed, 50 µM of MEL appeared to be the optimal dose. It improved seed germination and embryonic axes growth especially during chilling stress (5 °C/14 days) and during regeneration after its subsided. Unfortunately, MEL overdosing lowered IAA level in dry seeds and could disrupt the ROS-dependent signal transduction pathways. Very effective antioxidant MEL action was confirmed by low level of protein oxidative damage and smaller quantity of lipid oxidation products in embryonic axes isolated from seeds pre-treated with MEL and then exposed to cold. The stimulatory effects of MEL on antioxidant enzymes: SOD, APX and GSH-PX and on GST-a detoxifying enzyme, was also demonstrated. It was indicated for the first time, that MEL induced defence strategies against stress at the cytological level, as appearing endoreplication in embryonic axes cells even in the seeds germinating under optimal conditions (preventive action), but very intensively in those germinating under chilling stress conditions (intervention action), and after stress removal, to improve regeneration.

Membrane-related hallmarks of kinetin-induced PCD of root cortex cells.

KEY MESSAGE: Changes in cellular membrane potential and their fluidisation are the hallmarks of cell death induction with kinetin in root cortex. Programmed cell death (PCD), one of the essential processes in plant development, is still poorly understood. In this paper, the scientific plant model, V. faba ssp. minor seedling roots after kinetin application which triggers off programmed death of cortex cells, was used to recognise membrane-related aspects of plant cell death. Spectrophotometric, reflectometric and microscopic studies showed that the PCD induced by kinetin is accompanied by higher potassium ions leakage from roots, loss of plasma and ER membrane potentials (expressed by their lower amounts and higher index of fatty acid unsaturation), malformation of nuclear envelope, lower total lipid amount and formation of their peroxides, lower amount of phospholipids and changes in their composition. The results showed that potassium ions leakage, expressed in percentage of their amounts, and loss of plasma and ER membrane potential, expressed in percentage of their fluorescence intensity, together with the nuclear chromatin double staining with ethidium bromide and acridine orange, might be direct and universal methods for detecting specific plant PCD hallmarks and estimation of PCD intensity (percentage of dying and dead cells).

Photosynthesis-related characteristics of the midrib and the interveinal lamina in leaves of the C3-CAM intermediate plant Mesembryanthemum crystallinum.

BACKGROUND AND AIMS: Leaf veins are usually encircled by specialized bundle sheath cells. In C4 plants, they play an important role in CO2 assimilation, and the photosynthetic activity is compartmentalized between the mesophyll and the bundle sheath. In C3 and CAM (Crassulacean acid metabolism) plants, the photosynthetic activity is generally attributed to the leaf mesophyll cells, and the vascular parenchymal cells are rarely considered for their role in photosynthesis. Recent studies demonstrate that enzymes required for C4 photosynthesis are also active in the veins of C3 plants, and their vascular system contains photosynthetically competent parenchyma cells. However, our understanding of photosynthesis in veins of C3 and CAM plants still remains insufficient. Here spatial analysis of photosynthesis-related properties were applied to the midrib and the interveinal lamina cells in leaves of Mesembryanthemum crystallinum, a C3-CAM intermediate plant. METHODS: The midrib anatomy as well as chloroplast structure and chlorophyll fluorescence, diurnal gas exchange profiles, the immunoblot patterns of PEPC (phosphoenolpyruvate carboxylase) and RubisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase), H2O2 localization and antioxidant enzyme activities were compared in the midrib and in the interveinal mesophyll cells in leaves of C3 and CAM plants. KEY RESULTS: Leaf midribs were structurally competent to perform photosynthesis in C3 and CAM plants. The midrib chloroplasts resembled those in the bundle sheath cells of C4 plants and were characterized by limited photosynthetic activity. CONCLUSIONS: The metabolic roles of midrib chloroplasts differ in C3 and CAM plants. It is suggested that in leaves of C3 plants the midrib chloroplasts could be involved in the supply of CO2 for carboxylation, and in CAM plants they could provide malate to different metabolic processes and mediate H2O2 signalling.

The crucial elements of the 'last step' of programmed cell death induced by kinetin in root cortex of V. faba ssp. minor seedlings.

KEY MESSAGE: Kinetin-induced programmed cell death, manifested by condensation, degradation and methylation of DNA and fluctuation of kinase activities and ATP levels, is an autolytic and root cortex cell-specific process. The last step of programmed cell death (PCD) induced by kinetin in the root cortex of V. faba ssp. minor seedlings was explained using morphologic (nuclear chromatin/aggregation) and metabolic (DNA degradation, DNA methylation and kinases activity) analyses. This step involves: (1) decrease in nuclear DNA content, (2) increase in the number of 4',6-diamidino-2-phenylindole (DAPI)-stained chromocenters, and decrease in chromomycin A3 (CMA3)-stained chromocenters, (3) increase in fluorescence intensity of CMA3-stained chromocenters, (4) condensation of DAPI-stained and loosening of CMA3-stained chromatin, (5) fluctuation of the level of DNA methylation, (6) fluctuation of activities of exo-/endonucleolytic Zn(2+) and Ca(2+)/Mg(2+)-dependent nucleases, (7) changes in H1 and core histone kinase activities and (8) decrease in cellular ATP amount. These results confirmed that kinetin-induced PCD was a specific process. Additionally, based on data presented in this paper (DNA condensation and ATP depletion) and previous studies [increase in vacuole, increase in amount of cytosolic calcium ions, ROS production and cytosol acidification "in Byczkowska et al. (Protoplasma 250:121-128, 2013)"], we propose that the process resembles autolytic type of cell death, the most common type of death during development of plants. Lastly, the observations also suggested that regulation of these processes might be under control of epigenetic (methylation/phosphorylation) mechanisms.

Melatonin - This is important to know.

MEL (N-acetyl-5-methoxytryptamine) is a well-known natural compound that controls cellular processes in both plants and animals and is primarily found in plants as a neurohormone. Its roles have been described very broadly, from its antioxidant function related to the photoperiod and determination of seasonal rhythms to its role as a signalling molecule, imitating the action of plant hormones (or even being classified as a prohormone). MEL positively affects the yield and survival of plants by increasing their tolerance to unfavourable biotic and abiotic conditions, which makes MEL widely applicable in ecological farming as a stimulant of growth and development. Thus, it is called a phytobiostimulator. In this review, we discuss the genesis of MEL functions, the presence of MEL at the cellular level and its effects on gene expression and plant development, which can ensure the survival of plants under the conditions they encounter. Moreover, we consider the future application possibilities of MEL in agriculture.

Growth inhibition, oxidative stress and characterisation of mortality in green algae under the influence of beta-blockers and non-steroidal anti-inflammatory drugs.

Bisoprolol and ketoprofen are widely used pharmaceuticals in medical treatment hence these substances are occurring in wastewaters and in water environment. This research investigated the toxic effects of bisoprolol and ketoprofen on two microalgae taxa, Chlorella vulgaris and Desmodesmus armatus. The results showed that both drugs inhibited the growth of the species tested and induced a decrease in chlorophyll a content compared to controls. Ketoprofen turned out to be harmful to algae as the half maximal effective concentration (EC50) values (14 days) were 37.69 mg L-1 for C. vulgaris and 40.93 mg L-1 for D. armatus. On the other hand, for bisoprolol, the EC50 values were greater than the established NOEC, 100 mg L-1. Bisoprolol and ketoprofen induced oxidative stress in the tested microorganisms, as indicated by changes in the activities of antioxidant enzymes. Exposure to 100 mg L-1 of drugs significantly increased the activity of catalase, peroxidase and superoxide dismutase. Fluorescence microscopy showed that both medicaments changed the cells' morphology. There was atrophy of chlorophyll in the cells, moreover, dying multinuclear cells and cells without nuclei were observed. In addition, there were atrophic cells, namely cells that lacked nuclei and chlorophyll. Profile area analyses showed that bisoprolol and ketoprofen treated C. vulgaris cells were approximately 4 and 2 times greater compared to control ones. Our experimental findings highlight the ecotoxicological threats for aquatic primary producers from bisoprolol and ketoprofen and provide insight into the characteristics of their death.

Hormonal crosstalk controls cell death induced by kinetin in roots of Vicia faba ssp. minor seedlings.

Studies of vitality/mortality of cortex cells, as well as of the concentrations of ethylene (ETH), gibberellins (GAs), indolic compounds/auxins (ICs/AUXs) and cytokinins (CKs), were undertaken to explain the hormonal background of kinetin (Kin)-regulated cell death (RCD), which is induced in the cortex of the apical parts of roots of faba bean (Vicia faba ssp. minor) seedlings. Quantification was carried out with fluorescence microscopy, ETH sensors, spectrophotometry and ultrahigh-performance liquid chromatography tandem mass spectrometry (UHPLC‒MS/MS). The results indicated that Kin was metabolized to the transport form, i.e., kinetin-9-glucoside (Kin9G) and kinetin riboside (KinR). KinR was then converted to cis-zeatin (cZ) in apical parts of roots with meristems, to cis-zeatin riboside (cZR) in apical parts of roots without meristems and finally to cis-zeatin riboside 5'-monophosphate (cZR5'MP), which is indicated to be a ligand of cytokinin-dependent receptors inducing CD. The process may be enhanced by an increase in the amount of dihydrozeatin riboside (DHZR) as a byproduct of the pathway of zeatin metabolism. It seems that crosstalk of ETH, ICs/AUXs, GAs and CKs with the cZR5'MP, the cis-zeatin-dependent pathway, but not the trans-zeatin-dependent pathway, is responsible for Kin-RCD, indicating that the process is very specific and offers a useful model for studies of CD hallmarks in plants.

Kinetin induces microtubular breakdown, cell cycle arrest and programmed cell death in tobacco BY-2 cells.

Plant cells can undergo regulated cell death in response to exogenous factors (often in a stress context), but also as regular element of development (often regulated by phytohormones). The cellular aspects of these death responses differ, which implies that the early signalling must be different. We use cytokinin-induced programmed cell death as paradigm to get insight into the role of the cytoskeleton for the regulation of developmentally induced cell death, using tobacco BY-2 cells as experimental model. We show that this PCD in response to kinetin correlates with an arrest of the cell cycle, a deregulation of DNA replication, a loss of plasma membrane integrity, a subsequent permeabilisation of the nuclear envelope, an increase of cytosolic calcium correlated with calcium depletion in the culture medium, an increase of callose deposition and the loss of microtubule and actin integrity. We discuss these findings in the context of a working model, where kinetin, mediated by calcium, causes the breakdown of the cytoskeleton, which, either by release of executing proteins or by mitotic catastrophe, will result in PCD.

Two ways to die: Species dependent PCD modes in grapevine cells.

Programmed cell death (PCD) is considered as a hallmark of strain-specific immunity. In contrast, generic basal immunity is thought to act without PCD. This classical bifurcation has been questioned during recent years. Likewise, the role of jasmonate signalling for these two modes of innate immunity has remained ambiguous. We have addressed both questions using two closely related grapevine cell lines (V. rupestris, V. vinifera cv. 'Pinot Noir') that contrast in their cell-death response to the bacterial elicitor harpin and the hormonal trigger methyl jasmonate (MeJA). We follow different cellular (loss of membrane integrity, mortality), molecular (induction of transcripts for phytoalexin synthesis and for metacaspases), as well as metabolic (sphingolipid profiles) responses to the two triggers in the two cell lines. The role of NADPH oxidases and induction of transcripts for the class-II metacaspases MC5 differ qualitatively between the two cell lines. We tested a possible role of sphingolipid metabolism but can rule this out. We propose a model, where V. rupestris, originating from co-evolution with several biotrophic pathogens, readily activates a hypersensitive cell death in response to harpin, while the context of MeJA-induced cell death in 'Pinot Noir' might not be related to immunity at all. We propose that the underlying signalling is modular, recruiting metacaspases differently depending on upstream signalling.

Silicon-on-insulator chip-to-chip coupling via out-of-plane or vertical grating couplers.

In this work, we study the possibility of chip-to-chip interconnections via out-of-plane grating couplers or vertical grating couplers (VGCs) fabricated on silicon-on-insulator (SOI) technology. Mathematical and experimental results for the efficiency of coupling light between two such grating couplers have been presented. The VGCs are placed vertically against each other in order to study the effect of spatial misalignments on the light coupling efficiency. Waveguides and VGCs are designed according to standard SOI fabrication processes. A two-dimensional finite-difference time-domain method is used for the simulations. Measurements are performed on different combinations of 4 samples on the chip. Light coupling efficiency of -8 dB is achieved and tolerances to misalignments in 3 spatial dimensions are provided.

Influence of bisphenol A on growth and metabolism of Vicia faba ssp. minor seedlings depending on lighting conditions.

The effect of one of anthropogenic pollutants, i.e., 4,4'-isopropylidenediphenol, called 2,2-bis (4-hydroxyphenyl) propane (BPA), at 30 and 120 mg L-1 concentrations in the darkness (DK) or dark/light (DK/LT) on growth and selected elements of metabolism of seedlings and leaf discs of Vicia faba ssp. minor was studied. Treatment with 120 mg L-1 BPA had greater effects which were reflected by increase in the number of necrotic changes in roots and stems as well as in leaf discs and reduction of the length of roots DK and DK/LT, and volume of roots in the DK group. However, minimal and no influence on the fresh and dry weight of roots and stems in plants growing under both types of lighting conditions were observed. In both DK and DK/LT groups these effects were correlated with reduced amounts of storage and cell wall-bound sugars as well as of proteins while in the DK/LT additionally with reduced soluble sugar levels in the roots and increased amounts of hydrogen peroxide and phenols in roots and stems as well as in treatment solutions, where these compounds were released. We suggest that endogenous phenols and BPA can be metabolised in roots and stems to quinones. It seems that TB-1,4-BQ, is the one of that of the five studied quinones. We expect that the results of this paper will help to answer the following question: does the phytomeliorative and phytosanitative V. faba ssp. minor plant is enough to be resistant on negative effects, and to be useful to reduce increasing amount of BPA in the environment?

Biotechnology of the Tree Fern Cyathea smithii (J.D. Hooker; Soft Tree Fern, Katote) II Cell Suspension Culture: Focusing on Structure and Physiology in the Presence of 2,4-D and BAP.

The aim of our research was to describe the structure and growth potential of a cell suspension of the tree fern Cyathea smithii. Experiments were performed on an established cell suspension with ½ MS medium supplemented with 9.05 µM 2,4-D + 0.88 µM BAP. In the experiments, attention was paid to the microscopic description of cell suspension, evaluation of cell growth dependent on the initial mass of cells and organic carbon source in the medium, the length of the passage, the content of one selected flavonoid in the post-culture medium, nuclear DNA content, ethylene production, and the antimicrobial value of the extract. For a better understanding of the cell changes that occurred during the culture of the suspension, the following structures of the cell were observed: nucleus, lipid bodies, tannin deposits, starch grains, cell walls, primary lamina, and the filaments of metabolites released into the medium. The nuclear DNA content (acriflavine-Feulgen staining) of cell aggregates distinctly indicated a lack of changes in the sporophytic origin of the cultured cell suspension. The physiological activity of the suspension was found to be high because of kinetics, intensive production of ethylene, and quercetin production. The microbiological studies suggested that the cell suspension possessed a bactericidal character against microaerobic Gram-positive bacteria. A sample of the cell suspension showed bacteriostatic activity against aerobic bacteria.

Passive Photonic Integrated Circuits Elements Fabricated on a Silicon Nitride Platform.

The fabrication processes for silicon nitride photonic integrated circuits evolved from microelectronics components technology-basic processes have common roots and can be executed using the same type of equipment. In comparison to that of electronics components, passive photonic structures require fewer manufacturing steps and fabricated elements have larger critical dimensions. In this work, we present and discuss our first results on design and development of fundamental building blocks for silicon nitride integrated photonic platform. The scope of the work covers the full design and manufacturing chain, from numerical simulations of optical elements, design, and fabrication of the test structures to optical characterization and analysis the results. In particular, technological processes were developed and evaluated for fabrication of the waveguides (WGs), multimode interferometers (MMIs), and arrayed waveguide gratings (AWGs), which confirmed the potential of the technology and correctness of the proposed approach.

Optical Thin Films Fabrication Techniques-Towards a Low-Cost Solution for the Integrated Photonic Platform: A Review of the Current Status.

In the past few decades, several methods concerning optical thin films have been established to facilitate the development of integrated optics. This paper provides a brief depiction of different techniques for implementing optical waveguide thin films that involve chemical, physical, and refractive index modification methods. Recent advances in these fabrication methods are also been presented. Most of the methods developed for the realization of the thin-films are quite efficient, but they are expensive and require sophisticated equipment. The major interest of the scientists is to develop simple and cost-effective methods for mass production of optical thin films resulting in the effective commercialization of the waveguide technology. Our research group is focused on developing a silica-titania optical waveguide platform via the sol-gel dip-coating method and implementing active and passive optical elements via the wet etching method. We are also exploring the possibility of using nanoimprint lithography (NIL) for patterning these films so that the fabrication process is efficient and economical. The recent developments of this platform are discussed. We believe that silica-titania waveguide technology developed via the sol-gel dip-coating method is highly attractive and economical, such that it can be commercialized for applications such as sensing and optical interconnects.

Short-term cell death in tissues of Pulsatilla vernalis seeds from natural and ex situ conserved populations.

Pulsatilla vernalis is a IUCN listed species that occurs in mountain and lowland habitats. The seeds collected from different populations are remarkably diverse in their viability depending on locality or year of collection. We aim to analyse seed viability, among others, by investigation of the percentage of alive, dying, and dead cells in embryos and endosperm when comparing the seeds from a wild lowland population and ex situ cultivation of plants of lowland and Alpine origin. The cell death was detected by staining with two fluorescence probes, one penetrating only the changed nuclear membranes, the other penetrating also the unchanged cells. 54.5% of Alpine origin seeds were presumably capable of germination if they were sown after collection, however, four months later only 36.4% had healthy embryos. In the case of lowland wild plants it was 31.8% and 18.2%, and from ex situ, 27.3% and 13.6%, respectively. 27.3% of Alpine origin seeds had embryo in torpedo stage (9.1% in the case of lowland seeds). Mean weight of the former was 2.9 mg (2.0 mg in lowland ones). Our results confirm the significance of seed origin and seed weight on viability, and that Pulsatilla seeds have a short 'germination time window'.