SARS-CoV-2 RNA levels in Scotland's wastewater.

Nationwide, wastewater-based monitoring was newly established in Scotland to track the levels of SARS-CoV-2 viral RNA shed into the sewage network, during the COVID-19 pandemic. We present a curated, reference dataset produced by this national programme, from May 2020 to February 2022. Viral levels were analysed by RT-qPCR assays of the N1 gene, on RNA extracted from wastewater sampled at 162 locations. Locations were sampled up to four times per week, typically once or twice per week, and in response to local needs. We report sampling site locations with geographical coordinates, the total population in the catchment for each site, and the information necessary for data normalisation, such as the incoming wastewater flow values and ammonia concentration, when these were available. The methodology for viral quantification and data analysis is briefly described, with links to detailed protocols online. These wastewater data are contributing to estimates of disease prevalence and the viral reproduction number (R) in Scotland and in the UK.

Demonstration of protein capture and separation using three-dimensional printed anion exchange monoliths fabricated in one-step.

Three-dimensional printing applications in separation science are currently limited by the lack of materials compatible with chromatographic operations and three-dimensional printing technologies. In this work, we propose a new material for Digital Light Processing printing to fabricate functional ion exchange monoliths in a single step. Through copolymerization of the bifunctional monomer [2-(acryloyloxy)ethyl] trimethylammonium chloride, monolithic structures with quaternary amine ligands were fabricated. The novel formulation was optimized in terms of protein binding and recovery, microporous structure, and its swelling susceptibility by increasing its cross-link density and employing cyclohexanol and dodecanol as pore forming agents. In static conditions, the material demonstrated a maximum binding capacity of 104.2 ± 10.6 mg/mL for bovine serum albumin, in line with commercially available materials. Its anion exchange behavior was validated by separating bovine serum albumin and myoglobin on a monolithic bed with Schoen gyroid morphology. The same column geometry was tested for the purification of C-phycocyanin from clarified as well as cell-laden Arthrospira platensis feedstocks. This represents the first demonstration of one-step printed stationary phases to capture proteins directly from solid-laden feedstocks. We believe that the material presented here represents a significant improvement towards implementation of three-dimensional printed chromatography media in the field of separation science.

A photometric stereo-based 3D imaging system using computer vision and deep learning for tracking plant growth.

BACKGROUND: Tracking and predicting the growth performance of plants in different environments is critical for predicting the impact of global climate change. Automated approaches for image capture and analysis have allowed for substantial increases in the throughput of quantitative growth trait measurements compared with manual assessments. Recent work has focused on adopting computer vision and machine learning approaches to improve the accuracy of automated plant phenotyping. Here we present PS-Plant, a low-cost and portable 3D plant phenotyping platform based on an imaging technique novel to plant phenotyping called photometric stereo (PS). RESULTS: We calibrated PS-Plant to track the model plant Arabidopsis thaliana throughout the day-night (diel) cycle and investigated growth architecture under a variety of conditions to illustrate the dramatic effect of the environment on plant phenotype. We developed bespoke computer vision algorithms and assessed available deep neural network architectures to automate the segmentation of rosettes and individual leaves, and extract basic and more advanced traits from PS-derived data, including the tracking of 3D plant growth and diel leaf hyponastic movement. Furthermore, we have produced the first PS training data set, which includes 221 manually annotated Arabidopsis rosettes that were used for training and data analysis (1,768 images in total). A full protocol is provided, including all software components and an additional test data set. CONCLUSIONS: PS-Plant is a powerful new phenotyping tool for plant research that provides robust data at high temporal and spatial resolutions. The system is well-suited for small- and large-scale research and will help to accelerate bridging of the phenotype-to-genotype gap.

A "Do-It-Yourself" phenotyping system: measuring growth and morphology throughout the diel cycle in rosette shaped plants.

BACKGROUND: Improvements in high-throughput phenotyping technologies are rapidly expanding the scope and capacity of plant biology studies to measure growth traits. Nevertheless, the costs of commercial phenotyping equipment and infrastructure remain prohibitively expensive for wide-scale uptake, while academic solutions can require significant local expertise. Here we present a low-cost methodology for plant biologists to build their own phenotyping system for quantifying growth rates and phenotypic characteristics of Arabidopsis thaliana rosettes throughout the diel cycle. RESULTS: We constructed an image capture system consisting of a near infra-red (NIR, 940 nm) LED panel with a mounted Raspberry Pi NoIR camera and developed a MatLab-based software module (iDIEL Plant) to characterise rosette expansion. Our software was able to accurately segment and characterise multiple rosettes within an image, regardless of plant arrangement or genotype, and batch process image sets. To further validate our system, wild-type Arabidopsis plants (Col-0) and two mutant lines with reduced Rubisco contents, pale leaves and slow growth phenotypes (1a3b and 1a2b) were grown on a single plant tray. Plants were imaged from 9 to 24 days after germination every 20 min throughout the 24 h light-dark growth cycle (i.e. the diel cycle). The resulting dataset provided a dynamic and uninterrupted characterisation of differences in rosette growth and expansion rates over time for the three lines tested. CONCLUSION: Our methodology offers a straightforward solution for setting up automated, scalable and low-cost phenotyping facilities in a wide range of lab environments that could greatly increase the processing power and scalability of Arabidopsis soil growth experiments.

Expression patterns of Passiflora edulis APETALA1/FRUITFULL homologues shed light onto tendril and corona identities.

BACKGROUND: Passiflora (passionflowers) makes an excellent model for studying plant evolutionary development. They are mostly perennial climbers that display axillary tendrils, which are believed to be modifications of the inflorescence. Passionflowers are also recognized by their unique flower features, such as the extra whorls of floral organs composed of corona filaments and membranes enclosing the nectary. Although some work on Passiflora organ ontogeny has been done, the developmental identity of both Passiflora tendrils and the corona is still controversial. Here, we combined ultrastructural analysis and expression patterns of the flower meristem and floral organ identity genes of the MADS-box AP1/FUL clade to reveal a possible role for these genes in the generation of evolutionary novelties in Passiflora. RESULTS: We followed the development of structures arising from the axillary meristem from juvenile to adult phase in P. edulis. We further assessed the expression pattern of P. edulis AP1/FUL homologues (PeAP1 and PeFUL), by RT-qPCR and in situ hybridization in several tissues, correlating it with the developmental stages of P. edulis. PeAP1 is expressed only in the reproductive stage, and it is highly expressed in tendrils and in flower meristems from the onset of their development. PeAP1 is also expressed in sepals, petals and in corona filaments, suggesting a novel role for PeAP1 in floral organ diversification. PeFUL presented a broad expression pattern in both vegetative and reproductive tissues, and it is also expressed in fruits. CONCLUSIONS: Our results provide new molecular insights into the morphological diversity in the genus Passiflora. Here, we bring new evidence that tendrils are part of the Passiflora inflorescence. This points to the convergence of similar developmental processes involving the recruitment of genes related to flower identity in the origin of tendrils in different plant families. The data obtained also support the hypothesis that the corona filaments are likely sui generis floral organs. Additionally, we provide an indication that PeFUL acts as a coordinator of passionfruit development.

Exploring the role of auxin in the androgynophore movement in Passiflora.

The flowers of the species belonging to the genus Passiflorashow a range of features that are thought to have arisen as adaptations to different pollinators. Some Passiflora species belonging to the subgenus Decaloba sect. Xerogona, show touch-sensitive motile androgynophores. We tested the role of auxin polar transport in the modulation of the androgynophore movement by applying auxin (IAA) or an inhibitor of auxin polar transport (NPA) in the flowers. We recorded the movement of the androgynophore during mechano-stimulation and analyzed the duration, speed, and the angle formed by the androgynophore before and after the movement, and found that both IAA and NPA increase the amplitude of the movement in P. sanguinolenta. We hypothesize that auxin might have a role in modulating the fitness of these Decaloba species to different pollination syndromes and demonstrate that an interspecific hybrid between insect- and hummingbird-pollinated Xerogona species present a heterosis effect on the speed of the androgynophore movement.

Rapid touch-stimulated movement in the androgynophore of Passiflora flowers (subgen. Decaloba; Sect. Xerogona): an adaptation to enhance cross-pollination?

Plant touch-sensitive organs have been described since Darwin's observations and are related to a quick response to environment stimuli. Sensitive flower organs have been associated to an increase in the chances of cross pollination but there are few studies regarding this topic. Here we describe for the first time the kinetic of the androgynophore movement of 4 Passiflora species (P. sanguinolenta, P. citrina, P. capsularis, and P. rubra). For that, we collected flowers and recorded the movement after mechano-stimulating the androgynophore. From the recordings, we described the movement regarding its response and sensibility to mechanical stimulus and calculated the duration, speed, and the angle formed by the androgynophore before and after the movement. From our data we were able to propose a link to the pollination habit of these species. The movement of the androgynophore in these Passiflora is a noteworthy floral feature that might lead us to another astonishing example of a mechanism that evolved among angiosperms to assure sexual reproduction.