Key Developmental Stages Research Articles

Transcriptome Analyses at Specific Plastochrons Reveal Timing and Involvement of Phytosulfokine in Maize Vegetative Phase Change

Successive developmental stages of representative early and late juvenile, transition, and adult maize leaves were compared using machine-learning-aided analyses of gene expression patterns to characterize vegetative phase change (VPC), including identification of the timing of this developmental transition in maize. We used t-SNE to organize 32 leaf samples into 9 groups with similar patterns of gene expression. oposSOM yielded clusters of co-expressed genes from key developmental stages. TO-GCN supported a sequence of events in maize in which germination-associated ROS triggers a JA response, both relieving oxidative stress and inducing miR156 production, which in turn spurs juvenility. Patterns of expression of MIR395, which regulates sulfur assimilation, led to the hypothesis that phytosulfokine, a sulfated peptide, is involved in the transition to adult patterns of differentiation.

Divergence and convergence in epiphytic and endophytic phyllosphere bacterial communities of rice landraces.

Phyllosphere-associated microbes can significantly alter host plant fitness, with distinct functions provided by bacteria inhabiting the epiphytic (external surface) vs endophytic niches (internal leaf tissue). Hence, it is important to understand the assembly and stability of these phyllosphere communities, especially in field conditions. Broadly, epiphytic communities should encounter more environmental fluctuations and frequent immigration, whereas endophytic microbiota should face stronger host selection. As a result, we expect greater variability in epiphytic than endophytic communities. We analyzed the structure and stability of leaf phyllosphere microbiota of four traditionally cultivated rice landraces and one commercial variety from northeast India grown in the field for 3 consecutive years, supplemented with opportunistic sampling of eight other landraces. Epiphytic and endophytic bacterial communities shared dominant core genera such as Methylobacterium and Sphingomonas. Consistent with an overall strong environmental effect, both communities varied more across sampling years than across host landraces. Seeds sampled from a focal landrace did not support vertical transmission of phyllosphere bacteria, suggesting that both types of communities are assembled anew each generation. Despite these points of convergence, epiphytic communities had distinct composition and significantly higher microbial load and were more rich, diverse, modular, and unstable than endophytic communities. Finally, focused sampling of one landrace across developmental stages showed that the divergence between the two types of communities arose primarily at the flowering stage. Thus, our results show both convergent and divergent patterns of community assembly and composition in distinct phyllosphere niches in rice, identifying key bacterial genera and host developmental stages that may aid agricultural interventions to increase rice yield.IMPORTANCEPhyllosphere (leaf-associated) microbes significantly impact plant fitness, making it crucial to understand how these communities are assembled and maintained. While many studies have analyzed epiphytic (surface) phyllosphere communities, we have a relatively poor understanding of endophytic communities which colonize the very distinct niche formed inside leaf tissues. We found that across several rice landraces, both communities are largely colonized by the same core genera, indicating divergence at the species level across the two leaf niches and highlighting the need to understand the mechanisms underlying this divergence. Surprisingly, both epiphytic and endophytic communities were only weakly shaped by the host landrace, with a much greater role for environmental factors that likely vary over time. Thus, microbiome-based agricultural interventions for increasing productivity could perhaps be generalized across rice varieties but would need to account for the temporal instability of the microbiota. Our results thus highlight the importance of data sets such as ours-with extensive sampling across landraces and years-for understanding phyllosphere microbiota and their applications in the field.

Splicing is dynamically regulated during limb development

Modifications to highly conserved developmental gene regulatory networks are thought to underlie morphological diversification in evolution and contribute to human congenital malformations. Relationships between gene expression and morphology have been extensively investigated in the limb, where most of the evidence for alterations to gene regulation in development consists of pre-transcriptional mechanisms that affect expression levels, such as epigenetic alterations to regulatory sequences and changes to cis-regulatory elements. Here we report evidence that alternative splicing (AS), a post-transcriptional process that modifies and diversifies mRNA transcripts, is dynamic during limb development in two mammalian species. We evaluated AS patterns in mouse (Mus musculus) and opossum (Monodelphis domestica) across the three key limb developmental stages: the ridge, bud, and paddle. Our data show that splicing patterns are dynamic over developmental time and suggest differences between the two mammalian taxa. Additionally, multiple key limb development genes, including Fgf8, are differentially spliced across the three stages in both species, with expression levels of the conserved splice variants, Fgf8a and Fgf8b, changing across developmental time. Our data demonstrates that AS is a critical mediator of mRNA diversity in limb development and provides an additional mechanism for evolutionary tweaking of gene dosage.

Morphogenic, molecular, and cellular adaptations for unidirectional airflow in the chicken lung.

Unidirectional airflow in the avian lung enables gas exchange during both inhalation and exhalation. The underlying developmental process and how it deviates from that of the bidirectional mammalian lung are poorly understood. Sampling key developmental stages with multiscale 3D imaging and single-cell transcriptomics, we delineate morphogenic, molecular, and cellular features that accommodate the unidirectional airflow in the chicken lung. Primary termini of hyper-elongated branches are eliminated via proximal-short and distal-long fusions, forming parabronchi. Neoform termini extend radially through parabronchial smooth muscle to form gas-exchanging alveoli. Supporting this radial alveologenesis, branch stalks halt their proximalization, defined by SOX9-SOX2 transition, and become SOX9 low parabronchi. Primary and secondary vascular plexi interface with primary and neoform termini, respectively. Single-cell and Stereo-seq spatial transcriptomics reveal a third, chicken-specific alveolar cell type expressing KRT14, hereby named luminal cells. Luminal, alveolar type 2, and alveolar type 1 cells sequentially occupy concentric zones radiating from the parabronchial lumen. Our study explores the evolutionary space of lung diversification and lays the foundation for functional analysis of species-specific genetic determinants.

Transcriptomic landscapes reveal development-related physiological processes in the two-spotted spider mite, Tetranychus urticae.

The two-spotted spider mite (Tetranychus urticae Koch, TSSM) is recognized as one of the most problematic spider mite pests. However, the precise gene expression patterns across its key developmental stages remain elusive. Here, we performed a comprehensive transcriptome analysis of TSSM eggs, nymphs and adult females using publicly available RNA sequencing (RNA-seq) data to elucidate the overarching transcriptomic differences between these developmental stages. Principal component analysis and hierarchical clustering analysis unveiled distinct separations among samples across different developmental stages, regardless of their Wolbachia infection status. Differential expression analysis revealed 4,089,2,762, and 1,282 core genes specifically enriched in eggs, nymphs, and adults, respectively. KEGG and GO enrichment analyses showed upregulation of genes in eggs are associated with proteolysis, Wnt signaling pathway, DNA transcription, RNA biosynthetic and metabolic processes, as well as protein folding, sorting, and degradation pathways. Meanwhile, nymphs exhibited increased abundance of genes related to chitin/amino sugar metabolic processes, G protein-coupled receptor signaling pathways, monoatomic ion transport, and neurotransmitter transport pathways. Pathways involving sphingolipid and carbohydrate metabolic processes, proteolysis, lipid transport, and localization were particularly enriched in older females. Altogether, our findings suggest that the egg stage exhibits higher activity in cell differentiation processes, the nymph stage is more involved in chitin development, and the adult stage shows increased metabolic and reproductive activity. This study enhances our understanding of the molecular mechanisms underlying TSSM development and paves the way for further research into the intricate physiological processes of TSSM.

Estimating evapotranspiration and drought dynamics of winter wheat under climate change: A case study in Huang-Huai-Hai region, China

Drought is one of the vital meteorological disasters that influence crop growth. Timely and accurately estimating the drought dynamics of crops is valuable for decision-maker to formulate scientific management measures of agricultural drought risk. In this study, the evapotranspiration and drought dynamics of winter wheat from 1981 to 2020 in the Huang-Huai-Hai (HHH) region of China were evaluated based on long-term multi-source observation data. Four key developmental stages of winter wheat were given attentions: growth before winter stage, overwintering stage, stage of greening-heading, and stage of filling-maturity. The crop water deficit index (CWDI) on a daily scale was established for quantitatively appraising the impacts of drought on winter wheat. Our results indicated that interannual variation in reference crop evapotranspiration (ET0) during the growth season of winter wheat from 1981 to 2020 in the HHH region showed a slight increase trend, with an average of 602.4 mm and obvious spatial differences of decreasing from the Northeast to the Southwest. Over the past forty years, the winter wheat in the HHH region was most severely affected by severe drought, followed by moderate drought, and finally mild drought. In addition, the impacts of drought on winter wheat at different critical growth stages varied greatly. For the growth before winter stage, the winter wheat was mainly threatened by mild, moderate, and severe droughts. For the overwintering stage, the winter wheat was mainly threatened by moderate, severe, and extreme droughts. For the greening-heading stage, the winter wheat was mainly threatened by mild, moderate, severe, and extreme droughts. For the filling-maturity stage, the winter wheat was mainly threatened by mild and moderate droughts. Finally, the impacts of drought on winter wheat during 1981–2020 in the HHH region were revealed to differ extraordinarily in space. In particular, the areas of winter wheat affected by severe drought significantly decreased. However, the areas of winter wheat affected by moderate drought clearly expanded. Our findings provide new insights for further improving climate change impact studies and agricultural drought defense capabilities adapting to continuous environmental change.

Dynamic alternations of three-dimensional chromatin architecture contribute to phenotypic characteristics of breast muscle in chicken

Breast muscle growth rate and intramuscular fat (IMF) content show apparent differences between fast-growing broilers and slow-growing indigenous chickens. However, the underlying genetic basis of these phenotypic characteristics remains elusive. In this study, we investigate the dynamic alterations of three-dimensional genome architecture and chromatin accessibility in breast muscle across four key developmental stages from embryo to starter chick in Arbor Acres (AA) broilers and Yufen (YF) indigenous chickens. The limited breed-specifically up-regulated genes (Bup-DEGs) are embedded in breed-specific A compartment, while a majority of the Bup-DEGs involving myogenesis and adipogenesis are regulated by the breed-specific TAD reprogramming. Chromatin loops allow distal accessible regions to interact with myogenic genes, and those loops share an extremely low similarity between chicken with different growth rate. Moreover, AA-specific loop interactions promote the expression of 40 Bup-DEGs, such as IGF1, which contributes to myofiber hypertrophy. YF-specific loop interactions or distal accessible regions lead to increased expression of 5 Bup-DEGs, including PIGO, PEMT, DHCR7, TMEM38B, and DHDH, which contribute to IMF deposition. These results help elucidate the regulation of breast muscle growth and IMF deposition in chickens.

Quantitative Trait Loci Mapping and Comparative Transcriptome Analysis of Fruit Weight (FW) in Watermelon (Citrullus lanatus L.).

The watermelon (Citrullus lanatus L.) holds substantial economic value as a globally cultivated horticultural crop. However, the genetic architecture of watermelon fruit weight (FW) remains poorly understood. In this study, we used sh14-11 with small fruit and N14 with big fruit to construct 100 recombinant inbred lines (RILs). Based on whole-genome resequencing (WGR), 218,127 single nucleotide polymorphisms (SNPs) were detected to construct a high-quality genetic map. After quantitative trait loci (QTL) mapping, a candidate interval of 31-38 Mb on chromosome 2 was identified for FW. Simultaneously, the bulked segregant analysis (BSA) in the F2 population corroborated the identification of the same interval, encompassing the homologous gene linked to the known FW-related gene fas. Additionally, RNA-seq was carried out across 11 tissues from sh14-11 and N14, revealing expression profiles that identified 1695 new genes and corrected the annotation of 2941 genes. Subsequent differential expression analysis unveiled 8969 differentially expressed genes (DEGs), with 354 of these genes exhibiting significant differences across four key developmental stages. The integration of QTL mapping and differential expression analysis facilitated the identification of 14 FW-related genes, including annotated TGA and NAC transcription factors implicated in fruit development. This combined approach offers valuable insights into the genetic basis of FW, providing crucial resources for enhancing watermelon cultivation.

Genomic and single-cell analyses reveal genetic signatures of swimming pattern and diapause strategy in jellyfish

Jellyfish exhibit innovative swimming patterns that contribute to exploring the origins of animal locomotion. However, the genetic and cellular basis of these patterns remains unclear. Herein, we generated chromosome-level genome assemblies of two jellyfish species, Turritopsis rubra and Aurelia coerulea, which exhibit straight and free-swimming patterns, respectively. We observe positive selection of numerous genes involved in statolith formation, hair cell ciliogenesis, ciliary motility, and motor neuron function. The lineage-specific absence of otolith morphogenesis- and ciliary movement-related genes in T. rubra may be associated with homeostatic structural statocyst loss and straight swimming pattern. Notably, single-cell transcriptomic analyses covering key developmental stages reveal the enrichment of diapause-related genes in the cyst during reverse development, suggesting that the sustained diapause state favours the development of new polyps under favourable conditions. This study highlights the complex relationship between genetics, locomotion patterns and survival strategies in jellyfish, thereby providing valuable insights into the evolutionary lineages of movement and adaptation in the animal kingdom.

Enhanced Genomic and Transcriptomic Resources for Trichinella pseudospiralis and T. spiralis to Underpin the Discovery of Molecular Differences between Stages and Species.

Nematodes of the genus Trichinella are important pathogens of humans and animals. This study aimed to enhance the genomic and transcriptomic resources for T. pseudospiralis (non-encapsulated phenotype) and T. spiralis (encapsulated phenotype) and to explore transcriptional profiles. First, we improved the assemblies of the genomes of T. pseudospiralis (code ISS13) and T. spiralis (code ISS534), achieving genome sizes of 56.6 Mb (320 scaffolds, and an N50 of 1.02 Mb) and 63.5 Mb (568 scaffolds, and an N50 value of 0.44 Mb), respectively. Then, for each species, we produced RNA sequence data for three key developmental stages (first-stage muscle larvae [L1s], adults, and newborn larvae [NBLs]; three replicates for each stage), analysed differential transcription between stages, and explored enriched pathways and processes between species. Stage-specific upregulation was linked to cellular processes, metabolism, and host-parasite interactions, and pathway enrichment analysis showed distinctive biological processes and cellular localisations between species. Indeed, the secreted molecules calmodulin, calreticulin, and calsyntenin-with possible roles in modulating host immune responses and facilitating parasite survival-were unique to T. pseudospiralis and not detected in T. spiralis. These insights into the molecular mechanisms of Trichinella-host interactions might offer possible avenues for developing new interventions against trichinellosis.

P-272 Cannabis negatively impacts female fertility: an in vitro and clinical investigation

Abstract Study question How does tetrahydrocannabinol (THC) impacts human oocyte maturation and female fertility? Summary answer THC alters the oocyte’s RNA profile and affects meiotic spindle formation resulting in an increased rate of aneuploidy in the oocyte and in the blastocyst. What is known already In the past decades, cannabis consumption and legalization has been increasing globally. It is the third most used substance by reproductive aged people and our group demonstrated that tetrahydrocannabinol (THC) reaches the ovarian follicle. THC is known to impact male fertility, but due to the challenges associated with investigating the female gamete in humans, there is no study about the human oocyte. Animal studies have shown that THC impacts oocyte maturation in a concentration dependent manner, but between models, the results are conflicting. Study design, size, duration For the in vitro exposure assessment, GV oocytes naïve to THC were collected from consenting patients undergoing IVF-ICSI from July 2022 to June 2024 (IRB: Veritas #16518). Per patient, oocytes were split into three groups: Control, THC1 (physiological concentration of THC, based on previous human studies) and THC2 (supra-physiological concentration, based on animal studies), and cultured for 24h in our standard clinical in vitro maturation media. Participants/materials, setting, methods Single-cell RNAseq and pathway analysis were conducted to identify key pathways altered by THC treatments (n = 86 from 24 patients). Spindle morphology was assessed using confocal microscopy. Polar body biopsy was performed and ploidy assessed using our clinical low pass whole genome sequencing pipeline. Also, we conducted a multi-year retrospective study (n = 1059 patients) by measuring the concentration of THC in their follicular fluid by mass spectrometry and collecting and comparing IVF outcomes between cases and controls. Main results and the role of chance In vitro, THC treatment marginally increased the maturation rate (46% for Ctrl (n = 103), 51% for THC1 (n = 103) and 58% for THC2 (n = 102)). Using time-lapse imaging, no difference was observed between key developmental stages of oocyte maturation including GVBD and polar body extrusion. Single-cell RNAseq revealed significant alterations to the transcriptome profile of THC treated oocytes. When treated with THC1 (physiological concentration), 89 upregulated and 227 downregulated genes were identified, this was exaggerated when treated with THC2 (supraphysiologic concentration) 402 upregulated and 62 downregulated genes were identified. Pathway analysis showed that differentially expressed genes in THC1 group were associated with immune system and synaptic transmission, while THC2 group showed genes enriched in meiotic spindle formation and apoptosis. Moreover, preliminary results demonstrated abnormal spindle configuration and misaligned chromosomes in THC-treated groups and a concurrent decrease in the number of euploid oocytes. Furthermore, retrospective analysis of clinical IVF outcomes revealed that patients whose FF was positive for THC (62 patients, 6%) also had a slight increased maturation rate (77% vs 73%, p=ns), but a significant reduction of their embryo euploid rate when compared to control patients (59% vs 70%, p < 0.01). Limitations, reasons for caution The in vitro exposure to THC utilized GV oocytes that did not mature after ovarian stimulation, so they might have a reduced capacity of becoming MII. The THC measurement was assessed only on the day of retrieval and we do not know the route, dose, or duration of THC consumption. Wider implications of the findings This is the first study investigating the impact of cannabis on human oocytes. Our data suggests that cannabis consumption may reduce the number of euploid embryos available for transfer. With cannabis legalization increasing worldwide, further investigation into its consequences is critical for clinical consultation and legalization guidelines. Trial registration number Not applicable

The here and now of climate change: Climatic trends throughout Australian cotton regions and implications for the growing season

ContextMuch research has focused on assessing future climatic conditions on cotton production. However, understanding climatic changes that have already occurred throughout Australian cotton regions may provide insights on how existing and new regions are adapting to change. ObjectivesThis study aims to understand climatic changes that have occurred throughout Australian cotton regions, and the implications that these changes may have had for cotton growth and development. MethodsWe utilised historical weather data to undertake an analysis of regional climate using changes in: heat unit accumulation for seasonal assessment of temperature conditions; frequency of extreme weather conditions affecting cotton growth; seasonal potential evapotranspiration (ETO); and the timing of key cotton developmental stages. Nine Australian cotton regions over three time periods from 1961 to 2020 were assessed. We also used the cotton simulation model OZCOT to predict yield, crop evapotranspiration (ETC), and water use efficiency (WUE, lint yield kg/mm ETC) for a crop grown at each location. ResultsOur data shows some reductions in early season cold shocks (≤11 °C min.), and increases in minimum temperature, seasonal heat accumulation, ETO, and the frequency of extremely high temperatures (>40 °C). Overall, time to first flower (FF) has decreased across several locations which may be having implications for less leaf area prior to flowering, but it may be contributing to an opportunity for longer reproductive periods at Emerald and Moree. ConclusionsClimatic changes including decreased frequency of early season cold shocks, increased minimum temperatures and frequency of high temperatures and evapotranspiration, and an increase in season long heat accumulation have occurred across Australian cotton regions from 1961 to 2020, affecting growth and development of cotton. Overall, warmer seasonal temperatures have reduced the time to first flower, increased the time to last effective flower (LEF), and resulted in some changes to last harvestable boll (LHB). ImplicationsClimatic changes have resulted in an opportunity for longer reproductive periods and the potential for increased yields. Recognising that regions have experienced specific changes may help to guide current and future agronomic practice in the different regions.

Clinical characteristics of adolescents and emerging adults presenting for integrated posttraumatic stress and substance use treatment

PurposeAdolescence and emerging adulthood are key developmental stages with high risk for trauma exposure and the development of mental and substance-use disorders (SUDs). This study aims to compare the clinical profiles of adolescents (aged 12–17 years) and emerging adults (aged 18–25 years) presenting for treatment of posttraumatic stress disorder (PTSD) and SUD.Design/methodology/approachData was collected from the baseline assessment of individuals (n = 55) taking part in a randomized controlled trial examining the efficacy of an integrated psychological therapy for co-occurring PTSD and SUDs (PTSD+SUD) in young people.FindingsBoth age groups demonstrated complex and severe clinical profiles, including high-frequency trauma exposure, and very poor mental health reflected on measures of PTSD, SUD, suicidality and domains of social, emotional, behavioral and family functioning. There were few differences in clinical characteristics between the two groups.Research limitations/implicationsSimilarity between the two groups suggests that the complex problems seen in emerging adults with PTSD + SUD are likely to have had their onset in adolescence or earlier and to have been present for several years by the time individuals present for treatment.Originality/valueTo the best of the authors’ knowledge, this is the first study to compare the demographic and clinical profiles of adolescents and emerging adults with PTSD + SUD. These findings yield important implications for practice and policy for this vulnerable group. Evidence-based prevention and early intervention approaches and access to care are critical. Alongside trauma-focused treatment, there is a critical need for integrated, trauma-informed approaches specifically tailored to young people with PTSD + SUD.

Histology Atlas of the Developing Mouse Respiratory System From Prenatal Day 9.0 Through Postnatal Day 30.

Respiratory diseases are one of the leading causes of death and disability around the world. Mice are commonly used as models of human respiratory disease. Phenotypic analysis of mice with spontaneous, congenital, inherited, or treatment-related respiratory tract abnormalities requires investigators to discriminate normal anatomic features of the respiratory system from those that have been altered by disease. Many publications describe individual aspects of normal respiratory tract development, primarily focusing on morphogenesis of the trachea and lung. However, a single reference providing detailed low- and high-magnification, high-resolution images of routine hematoxylin and eosin (H&E)-stained sections depicting all major structures of the entire developing murine respiratory system does not exist. The purpose of this atlas is to correct this deficiency by establishing one concise reference of high-resolution color photomicrographs from whole-slide scans of H&E-stained tissue sections. The atlas has detailed descriptions and well-annotated images of the developing mouse upper and lower respiratory tracts emphasizing embryonic days (E) 9.0 to 18.5 and major early postnatal events. The selected images illustrate the main structures and events at key developmental stages and thus should help investigators both confirm the chronological age of mouse embryos and distinguish normal morphology as well as structural (cellular and organ) abnormalities.

Single-Nucleus RNA-Seq Reveals Spermatogonial Stem Cell Developmental Pattern in Shaziling Pigs.

Normal testicular development ensures the process of spermatogenesis, which is a complex biological process. The sustained high productivity of spermatogenesis throughout life is predominantly attributable to the constant proliferation and differentiation of spermatogonial stem cells (SSCs). The self-renewal and differentiation processes of SSCs are strictly regulated by the SSC niche. Therefore, understanding the developmental pattern of SSCs is crucial for spermatogenesis. The Shaziling pig is a medium-sized indigenous pig breed originating from central China. It is renowned for its superior meat quality and early male sexual maturity. The spermatogenic ability of the boars is of great economic importance to the pig industry. To investigate testicular development, particularly the pattern of SSC development in Shaziling pigs, we used single-cell transcriptomics to identify gene expression patterns in 82,027 individual cells from nine Shaziling pig testes at three key postnatal developmental stages. We generated an unbiased cell developmental atlas of Shaziling pig testicular tissues. We elucidated the complex processes involved in the development of SSCs within their niche in the Shaziling pig. Specifically, we identified potential marker genes and cellular signaling pathways that regulate SSC self-renewal and maintenance. Additionally, we proposed potential novel marker genes for SSCs that could be used for SSC isolation and sorting in Shaziling pigs. Furthermore, by immunofluorescence staining of testicular tissues of different developmental ages using marker proteins (UCHL1 and KIT), the developmental pattern of the spermatogonia of Shaziling pigs was intensively studied. Our research enhances the comprehension of the development of SSCs and provides a valuable reference for breeding Shaziling pigs.

"We all see things through a different lens based on our life experiences": co-production of a web-based implementation toolkit with stakeholders across the health and social care system.

Implementing new innovations across the health and social care system is complex, involving many factors that in recent years have been compounded by Covid-19. While a plethora of implementation tools and frameworks are available, there are limitations in terms of their design and accessibility. Co-production is a valuable mechanism for developing tools that have utility and accessibility for those tasked with using them in health and social care organisations and there is growing acknowledgement of increasing the role of co-production in implementation science. This paper provides novel insight into co-production practices and relevance to implementation science by reporting findings from a study to co-produce a web-based implementation toolkit (WIT) that is accessible, usable and designed to support adaptive implementation across health and social care systems. Key themes relating to the process of co-production are outlined and the value of using co-production in implementation processes are discussed. A web-based survey (n = 36) was conducted with a range of stakeholders across health and social care. Findings identified a need for WIT. Survey respondents were invited to express interest in becoming part of a co-production group and to take part in three online interactive workshops to co-produce WIT. Workshops took place with the group (n = 12) and focused on key developmental stages of WIT. Online co-production workshops were integral to the development and refinement of WIT. Benefits of using this process identified three interrelated themes: (i) Co-designing key features of the toolkit, (ii) Co-producing a toolkit with utility for users across health and social care settings, (iii) Co-producing a toolkit to support the implementation journey. Our approach of undertaking co-production as a dialogic process enabled generation of these themes. To illuminate discussion of these themes we draw upon iterative co-development of the "active ingredients" of key components (e.g., interactive Implementation Wheel) and functions (e.g., interactive "pop-up" definitions of keyword) and features (e.g., case studies) of WIT. Using a co-production approach with a range of end-users across health and social care systems, highlights the benefits of understanding implementation processes for users in these settings. User-centred design and processes for ensuring accessibility readily support the translation of implementation into rapidly changing health and social care systems to benefit outcomes for patients, their families, carers, service users and practitioners.

Developing a simple and efficient modeling solution for predicting key phenological stages of table grapes in a non-traditional viticulture zone in south Asia.

Phenological shifts are one of the most visible signs of climatic variability and change in the biosphere. However, modeling plant phenological responses has always been a key challenge due to climatic variability and plant adaptation. Grapevine is a phenologically sensitive crop and, thus, its developmental stages are affected by the increase in temperature. The goal of this study was to develop a temperature-based grapevine phenology model (GPM) for predicting key developmental stages for different table grape cultivars for a non-traditional viticulture zone in south Asia. Experiments were conducted in two vineyards at two locations (Chakwal and Islamabad) in the Pothawar region of Pakistan during the 2019 and 2020 growing seasons for four cultivars including Perlette, King's Ruby, Sugraone and NARC Black. Detailed phenological observations were obtained starting in January until harvest of the grapes. The Mitscherlich monomolecular equation was used to develop the phenology model for table grapes. There was a strong non-linear correlation between the Eichhorn and Lorenz phenological (ELP) scale and growing degree days (GDD) for all cultivars with coefficient of determinations (R2) ranging from 0.90 to 0.94. The results for model development indicated that GPM was able to predict phenological stages with high skill scores, i.e., a root mean square (RMSE) of 2.14 to 2.78 and mean absolute error (MAE) of 1.86 to 2.26 days. The prediction variability of the model for the onset timings of phenological stages was up to 3 days. The results also reveal that the phenology model based on GDD approach provides an efficient planning tool for viticulture industry in different grape growing regions. The proposed methodology, being a simpler one, can be easily applied to other regions and cultivars as a predictor for grapevine phenology.

Unsupervised Characterization of Prediction Error Markers in Unisensory and Multisensory Streams Reveal the Spatiotemporal Hierarchy of Cortical Information Processing.

Elicited upon violation of regularity in stimulus presentation, mismatch negativity (MMN) reflects the brain's ability to perform automatic comparisons between consecutive stimuli and provides an electrophysiological index of sensory error detection whereas P300 is associated with cognitive processes such as updating of the working memory. To date, there has been extensive research on the roles of MMN and P300 individually, because of their potential to be used as clinical markers of consciousness and attention, respectively. Here, we intend to explore with an unsupervised and rigorous source estimation approach, the underlying cortical generators of MMN and P300, in the context of prediction error propagation along the hierarchies of brain information processing in healthy human participants. The existing methods of characterizing the two ERPs involve only approximate estimations of their amplitudes and latencies based on specific sensors of interest. Our objective is twofold: first, we introduce a novel data-driven unsupervised approach to compute latencies and amplitude of ERP components accurately on an individual-subject basis and reconfirm earlier findings. Second, we demonstrate that in multisensory environments, MMN generators seem to reflect a significant overlap of "modality-specific" and "modality-independent" information processing while P300 generators mark a shift toward completely "modality-independent" processing. Advancing earlier understanding that multisensory contexts speed up early sensory processing, our study reveals that temporal facilitation extends to even the later components of prediction error processing, using EEG experiments. Such knowledge can be of value to clinical research for characterizing the key developmental stages of lifespan aging, schizophrenia, and depression.

Promoter variations of ClERF1 gene determines flesh firmness in watermelon

BackgroundFlesh firmness is a critical factor that influences fruit storability, shelf-life and consumer’s preference as well. However, less is known about the key genetic factors that are associated with flesh firmness in fresh fruits like watermelon.ResultsIn this study, through bulk segregant analysis (BSA-seq), we identified a quantitative trait locus (QTL) that influenced variations in flesh firmness among recombinant inbred lines (RIL) developed from cross between the Citrullus mucosospermus accession ZJU152 with hard-flesh and Citrullus lanatus accession ZJU163 with soft-flesh. Fine mapping and sequence variations analyses revealed that ethylene-responsive factor 1 (ClERF1) was the most likely candidate gene for watermelon flesh firmness. Furthermore, several variations existed in the promoter region between ClERF1 of two parents, and significantly higher expressions of ClERF1 were found in hard-flesh ZJU152 compared with soft-flesh ZJU163 at key developmental stages. DUAL-LUC and GUS assays suggested much stronger promoter activity in ZJU152 over ZJU163. In addition, the kompetitive allele-specific PCR (KASP) genotyping datasets of RIL populations and germplasm accessions further supported ClERF1 as a possible candidate gene for fruit flesh firmness variability and the hard-flesh genotype might only exist in wild species C. mucosospermus. Through yeast one-hybrid (Y1H) and dual luciferase assay, we found that ClERF1 could directly bind to the promoters of auxin-responsive protein (ClAux/IAA) and exostosin family protein (ClEXT) and positively regulated their expressions influencing fruit ripening and cell wall biosynthesis.ConclusionsOur results indicate that ClERF1 encoding an ethylene-responsive factor 1 is associated with flesh firmness in watermelon and provide mechanistic insight into the regulation of flesh firmness, and the ClERF1 gene is potentially applicable to the molecular improvement of fruit-flesh firmness by design breeding.

Variation in Root System Architecture Response to Arsenic during Establishment and Onset of Storage Root Formation in Two Sweetpotato (Ipomoea batatas L.) Cultivars

The primary objective of this work was to generate species-specific information about root architectural adaptations to simulated natural levels of arsenic (As) during the establishment phase and onset of storage root formation in sweetpotato. Cultivars Bayou Belle and Beauregard were grown on sand substrate and provided with 0.5X Hoagland’s nutrient solution with varying levels of As (0, 5, 10, or 15 mg⋅L−1). During the first experiment, entire root systems were sampled at 5, 10, and 15 days, corresponding to key adventitious root developmental stages. Compared with the untreated controls at 15 days, ‘Bayou Belle’ and ‘Beauregard’ provided with 15 mg⋅L−1 As showed respective increases in the following root architectural attributes: 168% and 130% in main root length; 168% and 98% in lateral root length; and 140% and 50% in lateral root density. A second experiment was performed to produce storage root samples at 50 days. Storage root length, width, and length/width ratio did not vary with As levels. The accumulation of As in storage roots increased with increasing As levels. The results support the hypothesis that natural As levels stimulate adventitious root development in sweetpotato in a cultivar-dependent manner. The observations are consistent with findings of other species that show similar growth stimulation at low As levels. This is the first report of sweetpotato root system architecture responses to experimental levels of As that are known to be present in agricultural soils. Standardization of experimental procedures and understanding of root system adaptations to natural levels of As could lead to a more systematic exploitation of genome-wide techniques and characterization of the molecular basis of reduced As uptake in plants.