Developmental localization and the role of hydroxyproline rich glycoproteins during somatic embryogenesis of banana (Musa spp. AAA)

<p>Abstract</p> <p>Background</p> <p>Hydroxyproline rich glycoproteins (HRGPs) are implicated to have a role in many aspects of plant growth and development but there is limited knowledge about their localization and function during somatic embryogenesis of higher plants. In this study, the localization and function of hydroxyproline rich glycoproteins in embryogenic cells (ECs) and somatic embryos of banana were investigated by using immunobloting and immunocytochemistry with monoclonal JIM11 and JIM20 antibodies as well as by treatment with 3,4-dehydro-L-proline (3,4-DHP, an inhibitor of extensin biosynthesis), and by immunomodulation with the JIM11 antibody.</p> <p>Results</p> <p>Immunofluorescence labelling of JIM11 and JIM20 hydroxyproline rich glycoprotein epitopes was relatively weak in non-embryogenic cells (NECs), mainly on the edge of small cell aggregates. On the other hand, hydroxyproline rich glycoprotein epitopes were found to be enriched in early embryogenic cells as well as in various developmental stages of somatic embryos. Embryogenic cells (ECs), proembryos and globular embryos showed strong labelling of hydroxyproline rich glycoprotein epitopes, especially in their cell walls and outer surface layer, so-called extracellular matrix (ECM). This hydroxyproline rich glycoprotein signal at embryo surfaces decreased and/or fully disappeared during later developmental stages (e.g. pear-shaped and cotyledonary stages) of embryos. In these later developmental embryogenic stages, however, new prominent hydroxyproline rich glycoprotein labelling appeared in tri-cellular junctions among parenchymatic cells inside these embryos. Overall immunofluorescence labelling of late stage embryos with JIM20 antibody was weaker than that of JIM11. Western blot analysis supported the above immunolocalization data. The treatment with 3,4-DHP inhibited the development of embryogenic cells and decreased the rate of embryo germination. Embryo-like structures, which developed after 3,4-DHP treatment showed aberrant non-compact epidermis with discontinuous ECM at the outer surface as well as much less immunolabelling with the JIM11 antibody. This treatment also decreased the plant regeneration capacity in embryogenic banana cultures. Finally, immunomodulation of surface hydroxyproline rich glycoproteins by co-culture of embryos with the JIM11 antibody resulted in a much lower germination capacity of these embryos.</p> <p>Conclusions</p> <p>These results suggest that hydroxyproline rich glycoproteins play an important developmental role, especially in the process of regeneration and germination of embryos during plant regeneration <it>via </it>somatic embryogenesis. Proper content and localization of hydroxyproline rich glycoproteins seem to be essential for the formation and regeneration of banana somatic embryos.</p

WikiGoldSK: Annotated Dataset, Baselines and Few-Shot Learning Experiments for Slovak Named Entity Recognition

Named Entity Recognition (NER) is a fundamental NLP tasks with a wide range of practical applications. The performance of state-of-the-art NER methods depends on high quality manually anotated datasets which still do not exist for some languages. In this work we aim to remedy this situation in Slovak by introducing WikiGoldSK, the first sizable human labelled Slovak NER dataset. We benchmark it by evaluating state-of-the-art multilingual Pretrained Language Models and comparing it to the existing silver-standard Slovak NER dataset. We also conduct few-shot experiments and show that training on a sliver-standard dataset yields better results. To enable future work that can be based on Slovak NER, we release the dataset, code, as well as the trained models publicly under permissible licensing terms at https://github.com/NaiveNeuron/WikiGoldSK.Comment: BSNLP 2023 Workshop at EACL 202

Recent Advances in the Cellular and Developmental Biology of Phospholipases in Plants

Phospholipases (PLs) are lipid-hydrolyzing enzymes known to have diverse signaling roles during plant abiotic and biotic stress responses. They catalyze lipid remodeling, which is required to generate rapid responses of plants to environmental cues. Moreover, they produce second messenger molecules, such as phosphatidic acid (PA) and thus trigger or modulate signaling cascades that lead to changes in gene expression. The roles of phospholipases in plant abiotic and biotic stress responses have been intensively studied. Nevertheless, emerging evidence suggests that they also make significant contributions to plants’ cellular and developmental processes. In this mini review, we summarized recent advances in the study of the cellular and developmental roles of phospholipases in plants

Proteomic Analysis of Arabidopsis pldα1 Mutants Revealed an Important Role of Phospholipase D Alpha 1 in Chloroplast Biogenesis

Phospholipase D alpha 1 (PLDα1) is a phospholipid hydrolyzing enzyme playing multiple regulatory roles in stress responses of plants. Its signaling activity is mediated by phosphatidic acid (PA) production, capacity to bind, and modulate G-protein complexes or by interaction with other proteins. This work presents a quantitative proteomic analysis of two T-DNA insertion pldα1 mutants of Arabidopsis thaliana. Remarkably, PLDα1 knockouts caused differential regulation of many proteins forming protein complexes, while PLDα1 might be required for their stability. Almost one third of differentially abundant proteins (DAPs) in pldα1 mutants are implicated in metabolism and RNA binding. Latter functional class comprises proteins involved in translation, RNA editing, processing, stability, and decay. Many of these proteins, including those regulating chloroplast protein import and protein folding, share common functions in chloroplast biogenesis and leaf variegation. Consistently, pldα1 mutants showed altered level of TIC40 (a major regulator of protein import into chloroplast), differential accumulation of photosynthetic protein complexes and changed chloroplast sizes as revealed by immunoblotting, blue-native electrophoresis, and microscopic analyses, respectively. Our proteomic analysis also revealed that genetic depletion of PLDα1 also affected proteins involved in cell wall architecture, redox homeostasis, and abscisic acid signaling. Taking together, PLDα1 appears as a protein integrating cytosolic and plastidic protein translations, plastid protein degradation, and protein import into chloroplast in order to regulate chloroplast biogenesis in Arabidopsis

Protein-protein interactions in plant antioxidant defense

The regulation of reactive oxygen species (ROS) levels in plants is ensured by mechanisms preventing their over accumulation, and by diverse antioxidants, including enzymes and nonenzymatic compounds. These are affected by redox conditions, posttranslational modifications, transcriptional and posttranscriptional modifications, Ca2+, nitric oxide (NO) and mitogen-activated protein kinase signaling pathways. Recent knowledge about protein-protein interactions (PPIs) of antioxidant enzymes advanced during last decade. The best-known examples are interactions mediated by redox buffering proteins such as thioredoxins and glutaredoxins. This review summarizes interactions of major antioxidant enzymes with regulatory and signaling proteins and their diverse functions. Such interactions are important for stability, degradation and activation of interacting partners. Moreover, PPIs of antioxidant enzymes may connect diverse metabolic processes with ROS scavenging. Proteins like receptor for activated C kinase 1 may ensure coordination of antioxidant enzymes to ensure efficient ROS regulation. Nevertheless, PPIs in antioxidant defense are understudied, and intensive research is required to define their role in complex regulation of ROS scavenging

Biochemical and Genetic Interactions of Phospholipase D Alpha 1 and Mitogen-Activated Protein Kinase 3 Affect Arabidopsis Stress Response

Phospholipase D alpha 1 (PLDα1, AT3G15730) and mitogen-activated protein kinases (MAPKs) participate on signaling-dependent events in plants. MAPKs are able to phosphorylate a wide range of substrates putatively including PLDs. Here we have focused on functional regulations of PLDα1 by interactions with MAPKs, their co-localization and impact on salt stress and abscisic acid (ABA) tolerance in Arabidopsis. Yeast two-hybrid and bimolecular fluorescent assays showed that PLDα1 interacts with MPK3. Immunoblotting analyses likewise confirmed connection between both these enzymes. Subcellularly we co-localized PLDα1 with MPK3 in the cortical cytoplasm close to the plasma membrane and in cytoplasmic strands. Moreover, genetic interaction studies revealed that pldα1mpk3 double mutant was resistant to a higher salinity and showed a higher tolerance to ABA during germination in comparison to single mutants and wild type. Thus, this study revealed importance of new biochemical and genetic interactions between PLDα1 and MPK3 for Arabidopsis stress (salt and ABA) response

Simulation of winter wheat yield and its variability in different climates of Europe: A comparison of eight crop growth models

We compared the performance of eight widely used, easily accessible and well-documented crop growth simulation models (APES, CROPSYST, DAISY, DSSAT, FASSET, HERMES, STICS and WOFOST) for winter wheat (Triticum aestivum L.) during 49 growing seasons at eight sites in northwestern, Central and southeastern Europe. The aim was to examine how different process-based crop models perform at the field scale when provided with a limited set of information for model calibration and simulation, reflecting the typical use of models for large-scale applications, and to present the uncertainties related to this type of model application. Data used in the simulations consisted of daily weather statistics, information on soil properties, information on crop phenology for each cultivar, and basic crop and soil management information. Our results showed that none of the models perfectly reproduced recorded observations at all sites and in all years, and none could unequivocally be labelled robust and accurate in terms of yield prediction across different environments and crop cultivars with only minimum calibration. The best performance regarding yield estimation was for DAISY and DSSAT, for which the RMSE values were lowest (1428 and 1603 kg ha−1) and the index of agreement (0.71 and 0.74) highest. CROPSYST systematically underestimated yields (MBE – 1186 kg ha−1), whereas HERMES, STICS and WOFOST clearly overestimated them (MBE 1174, 1272 and 1213 kg ha−1, respectively). APES, DAISY, HERMES, STICS and WOFOST furnished high total above-ground biomass estimates, whereas CROPSYST, DSSAT and FASSET provided low total above-ground estimates. Consequently, DSSAT and FASSET produced very high harvest index values, followed by HERMES and WOFOST. APES and DAISY, on the other hand, returned low harvest index values. In spite of phenological observations being provided, the calibration results for wheat phenology, i.e. estimated dates of anthesis and maturity, were surprisingly variable, with the largest RMSE for anthesis being generated by APES (20.2 days) and for maturity by HERMES (12.6). The wide range of grain yield estimates provided by the models for all sites and years reflects substantial uncertainties in model estimates achieved with only minimum calibration. Mean predictions from the eight models, on the other hand, were in good agreement with measured data. This applies to both results across all sites and seasons as well as to prediction of observed yield variability at single sites – a very important finding that supports the use of multi-model estimates rather than reliance on single model

Modelling long term effects of cropping and managements systems on soil organic matter, C/N dynamics and crop growth

While simulation of cropping systems over a few years might reflect well the short term effects of management and cultivation, long term effects on soil properties and their consequences for crop growth and matter fluxes are not captured. Especially the effect on soil carbon sequestration/depletion is addressed by this task. Simulations of an ensemble of crop models are performed as transient runs over a period of 120 year using observed weather from three stations in Czech Republic (1961-2010) and transient long time climate change scenarios (2011-2080) from five GCM of the CMIP5 ensemble to assess the effect of different cropping and management systems on carbon sequestration, matter fluxes and crop production in an integrative way. Two cropping systems are regarded comprising two times winter wheat, silage maize, spring barley and oilseed rape. Crop rotations differ regarding their organic input from crop residues, nitrogen fertilization and implementation of catch crops. Models are applied for two soil types with different water holding capacity. Cultivation and nutrient management is adapted using management rules related to weather and soil conditions. Data of phenology and crop yield from the region of the regarded crops were provided to calibrate the models for crops of the rotations. Twelve models were calibrated in this first step. For the transient long term runs results of four models were submitted so far. Outputs are crop yields, nitrogen uptake, soil water and mineral nitrogen contents, as well as water and nitrogen fluxes to the atmosphere and groundwater. Changes in the carbon stocks and the consequences for nitrogen mineralisation, N fertilization and emissions also considered.

Single Amino Acid Exchange in ACTIN2 Confers Increased Tolerance to Oxidative Stress in Arabidopsis <i>der1–3</i> Mutant

Single-point mutation in the ACTIN2 gene of the der1–3 mutant revealed that ACTIN2 is an essential actin isovariant required for root hair tip growth, and leads to shorter, thinner and more randomly oriented actin filaments in comparison to the wild-type C24 genotype. The actin cytoskeleton has been linked to plant defense against oxidative stress, but it is not clear how altered structural organization and dynamics of actin filaments may help plants to cope with oxidative stress. In this study, we characterized root growth, plant biomass, actin organization and antioxidant activity of the der1–3 mutant under oxidative stress induced by paraquat and H2O2. Under these conditions, plant growth was better in the der1–3 mutant, while the actin cytoskeleton in the der1–3 carrying pro35S::GFP:FABD2 construct showed a lower bundling rate and higher dynamicity. Biochemical analyses documented a lower degree of lipid peroxidation, and an elevated capacity to decompose superoxide and hydrogen peroxide. These results support the view that the der1–3 mutant is more resistant to oxidative stress. We propose that alterations in the actin cytoskeleton, increased sensitivity of ACTIN to reducing agent dithiothreitol (DTT), along with the increased capacity to decompose reactive oxygen species encourage the enhanced tolerance of this mutant against oxidative stress