Exploring aquaporin functions during changes in leaf water potential.

Maintenance of optimal leaf tissue humidity is important for plant productivity and food security. Leaf humidity is influenced by soil and atmospheric water availability, by transpiration and by the coordination of water flux across cell membranes throughout the plant. Flux of water and solutes across plant cell membranes is influenced by the function of aquaporin proteins. Plants have numerous aquaporin proteins required for a multitude of physiological roles in various plant tissues and the membrane flux contribution of each aquaporin can be regulated by changes in protein abundance, gating, localisation, post-translational modifications, protein:protein interactions and aquaporin stoichiometry. Resolving which aquaporins are candidates for influencing leaf humidity and determining how their regulation impacts changes in leaf cell solute flux and leaf cavity humidity is challenging. This challenge involves resolving the dynamics of the cell membrane aquaporin abundance, aquaporin sub-cellular localisation and location-specific post-translational regulation of aquaporins in membranes of leaf cells during plant responses to changes in water availability and determining the influence of cell signalling on aquaporin permeability to a range of relevant solutes, as well as determining aquaporin influence on cell signalling. Here we review recent developments, current challenges and suggest open opportunities for assessing the role of aquaporins in leaf substomatal cavity humidity regulation.

Arabidopsis plasma membrane intrinsic protein (AtPIP2;1) is implicated in a salinity conditional influence on seed germination.

Dynamic changes in aquaporin gene expression occur during seed germination. One example is the ~30-fold increase in Arabidopsis thaliana PIP2;1 transcripts within 24h of seed imbibition. To investigate whether AtPIP2;1 can influence seed germination wild-type Columbia-0, single (Atpip2;1 ) and double (Atpip2;1-Atpip2;2 ) loss-of-function mutants, along with transgenic 2x35S::AtPIP2;1 over-expressing (OE) lines and null-segregant controls, were examined. The various genotypes were germinated in control and saline (75mM NaCl treatment) conditions and tested for germination efficiency, imbibed seed maximum cross sectional (MCS) area, imbibed seed mass, and seed Na+ and K+ content. Seed lacking functional AtPIP2;1 and/or AtPIP2;2 proteins or constitutively over-expressing AtPIP2;1 , had delayed germination in saline conditions relative to wild-type and null-segregant seed, respectively. Exposure to saline germination conditions resulted in Atpip2;1 mutants having greater imbibed seed mass and less accumulated Na+ than wild-type, whereas lines over-expressing AtPIP2;1 had reduced imbibed seed mass and greater seed K+ content than null-segregant control seed. The results imply a role for AtPIP2;1 in seed germination processes, whether directly through its capacity for water and ion transport or H2 O2 signalling, or indirectly through potentially triggering dynamic differential regulation of other aquaporins expressed during germination. Future research will aid in dissecting the aquaporin functions influencing germination and may lead to novel solutions for optimising germination in sub-optimal conditions, such as saline soils.

A high-throughput yeast approach to characterize aquaporin permeabilities: Profiling the Arabidopsis PIP aquaporin sub-family.

Introduction: Engineering membrane transporters to achieve desired functionality is reliant on availability of experimental data informing structure-function relationships and intelligent design. Plant aquaporin (AQP) isoforms are capable of transporting diverse substrates such as signaling molecules, nutrients, metalloids, and gases, as well as water. AQPs can act as multifunctional channels and their transport function is reliant on many factors, with few studies having assessed transport function of specific isoforms for multiple substrates. Methods: High-throughput yeast assays were developed to screen for transport function of plant AQPs, providing a platform for fast data generation and cataloguing of substrate transport profiles. We applied our high-throughput growth-based yeast assays to screen all 13 Arabidopsis PIPs (AtPIPs) for transport of water and several neutral solutes: hydrogen peroxide (H2O2), boric acid (BA), and urea. Sodium (Na+) transport was assessed using elemental analysis techniques. Results: All AtPIPs facilitated water and H2O2 transport, although their growth phenotypes varied, and none were candidates for urea transport. For BA and Na+ transport, AtPIP2;2 and AtPIP2;7 were the top candidates, with yeast expressing these isoforms having the most pronounced toxicity response to BA exposure and accumulating the highest amounts of Na+. Linking putative AtPIP isoform substrate transport profiles with phylogenetics and gene expression data, enabled us to align possible substrate preferences with known and hypothesized biological roles of AtPIPs. Discussion: This testing framework enables efficient cataloguing of putative transport functionality of diverse AQPs at a scale that can help accelerate our understanding of AQP biology through big data approaches (e.g. association studies). The principles of the individual assays could be further adapted to test additional substrates. Data generated from this framework could inform future testing of AQP physiological roles, and address knowledge gaps in structure-function relationships to improve engineering efforts.

Molecular membrane separation: plants inspire new technologies.

Plants draw up their surrounding soil solution to gain water and nutrients required for growth, development and reproduction. Obtaining adequate water and nutrients involves taking up both desired and undesired elements from the soil solution and separating resources from waste. Desirable and undesirable elements in the soil solution can share similar chemical properties, such as size and charge. Plants use membrane separation mechanisms to distinguish between different molecules that have similar chemical properties. Membrane separation enables distribution or retention of resources and efflux or compartmentation of waste. Plants use specialised membrane separation mechanisms to adapt to challenging soil solution compositions and distinguish between resources and waste. Coordination and regulation of these mechanisms between different tissues, cell types and subcellular membranes supports plant nutrition, environmental stress tolerance and energy management. This review considers membrane separation mechanisms in plants that contribute to specialised separation processes and highlights mechanisms of interest for engineering plants with enhanced performance in challenging conditions and for inspiring the development of novel industrial membrane separation technologies. Knowledge gained from studying plant membrane separation mechanisms can be applied to developing precision separation technologies. Separation technologies are needed for harvesting resources from industrial wastes and transitioning to a circular green economy.

Mesophyll conductance is unaffected by expression of Arabidopsis PIP1 aquaporins in the plasmalemma of Nicotiana.

In plants with C3 photosynthesis, increasing the diffusion conductance for CO2 from the substomatal cavity to chloroplast stroma (mesophyll conductance) can improve the efficiencies of both CO2 assimilation and photosynthetic water use. In the diffusion pathway from substomatal cavity to chloroplast stroma, the plasmalemma and chloroplast envelope membranes impose a considerable barrier to CO2 diffusion, limiting photosynthetic efficiency. In an attempt to improve membrane permeability to CO2, and increase photosynthesis in tobacco, we generated transgenic lines in Nicotiana tabacum L. cv Petite Havana carrying either the Arabidopsis PIP1;2 (AtPIP1;2) or PIP1;4 (AtPIP1;4) gene driven by the constitutive dual 2x35S CMV promoter. From a collection of independent T0 transgenics, two T2 lines from each gene were characterized, with western blots confirming increased total aquaporin protein abundance in the AtPIP1;2 tobacco lines. Transient expression of AtPIP1;2-mGFP6 and AtPIP1;4-mGFP6 fusions in Nicotiana benthamiana identified that both AtPIP1;2 and AtPIP1;4 localize to the plasmalemma. Despite achieving ectopic production and correct localization, gas exchange measurements combined with carbon isotope discrimination measurements detected no increase in mesophyll conductance or CO2 assimilation rate in the tobacco lines expressing AtPIP. We discuss the complexities associated with trying to enhance gm through modified aquaporin activity.

Bridges or walls? A metaphorical dichotomy of Pope Francis versus Donald Trump's views of transnational migration.

Within a wider research line on policy-driven institutional discourses on migration by international/national institutions, NGO and political leaders, this contribution is aimed at illustrating the bipolarized social representations of immigrants inspiring 24 speeches by Pope Francis and US President Donald Trump. Statistical analyses using IRAMUTEQ included "specificity analysis" of discursive forms (words) and "cluster analysis." Results show that the Pope's discourse on migration (articulated into four clusters) is richer than the oversimplified Trump's discourse (originating just one cluster): the words "bridges" and "walls" emerge as representational nuclei of their bipolarized views of transnational migration, as metaphorical dichotomies of inclusive/exclusive policies. Emphasizing the need to build walls to protect the Americans, inspired by the sovereign ideology (AMERICA FIRST!), President Trump does not at all suspect that in the globalized interconnected world the AMERICA FIRST may become just AMERICA ALONE!

Mirrored Social Representations of Canadian Caseworkers with Migratory Paths Intervening with Refugees in the Host Country.

In 2015, the resettlement of 25,000 Syrian refugees in Canada placed a strain on social services. Caseworkers employed in these agencies often come from similar migratory trajectories to those of the refugees. This experiential proximity requires an understanding of the subjective perspectives that caseworkers with migratory paths have of refugees in the context of their professional practice. We analyzed fifteen individual interviews with Canadian caseworkers and conducted field observations of resettlement activities in the Ottawa-Gatineau region using inductive reasoning inspired by grounded theory. Adopting a sociogenetic approach to social representation theory, this qualitative study illustrates how the social representation of refugees among foreign-born caseworkers is highly informed by their migratory past experience, as well as by the social identity and social context from which that representation was socio-generated. Our analysis reveals the mirror effect of the caseworkers as a fruitful concept for understanding the identity-otherness dynamics in the encounter between the distant other (refugee) and the self.

A consensus on the Aquaporin Gene Family in the Allotetraploid Plant, Nicotiana tabacum.

Aquaporins (AQPs) are membrane-spanning channel proteins with exciting applications for plant engineering and industrial applications. Translational outcomes will be improved by better understanding the extensive diversity of plant AQPs. However, AQP gene families are complex, making exhaustive identification difficult, especially in polyploid species. The allotetraploid species of Nicotiana tabacum (Nt; tobacco) plays a significant role in modern biological research and is closely related to several crops of economic interest, making it a valuable platform for AQP research. Recently, De Rosa et al., (2020) and Ahmed et al., (2020), concurrently reported on the AQP gene family in tobacco, establishing family sizes of 76 and 88 members, respectively. The discrepancy highlights the difficulties of characterizing large complex gene families. Here, we identify and resolve the differences between the two studies, clarify gene models, and yield a consolidated collection of 84 members that more accurately represents the complete NtAQP family. Importantly, this consensus NtAQP collection will reduce confusion and ambiguity that would inevitably arise from having two different descriptive studies and sets of NtAQP gene names. This report also serves as a case study, highlighting and discussing variables to be considered and refinements required to ensure comprehensive gene family characterizations, which become valuable resources for examining the evolution and biological functions of genes.

The Anthropological and Ethnographic Approaches to Social Representations Theory - an Empirical Meta-Theoretical Analysis.

Part of a larger project aimed at performing an empirical meta-theoretical analysis of the entire corpus of scientific literature on Social Representations Theory (SRT), this research presents the state of the art of the anthropological and ethnographic approaches to SRT. Applying the Grid for Meta-Theoretical Analysis on 295 publications selected from the So.Re.Com"A.S. de Rosa"@-library, we compiled a rich set of meta-data and data illustrative of how SRT was conceptualized and operationalized within the anthropological and ethnographic approaches, as well as its positioning among other theoretical and disciplinary frameworks. The data was submitted to textual analysis, followed by a Hierarchical Clustering on Principal Components analysis. The empirical results suggest that from a theoretical standpoint, the anthropological and ethnographic approaches - inspired by its main exponents Jodelet (1991, 2016) and Duveen and Lloyd, (1986, 1993) - are consistent with the dynamic conceptualization of social representations set out by Moscovici (1961/1976, 1984/2003, 1988, 2000, 2013), as revolutionary paradigm that has shifted the emphasis of social psychology from looking at isolated variables in individuals in the abstract, towards a supra-disciplinary integrative vision of a social science, that investigates the genesis, transformation and negotiation of social representations in the communicative actual contexts (Billig 1991; de Rosa 2013a, b; Sammut et al. 2015a). From an empirical perspective, the variety of qualitative methods employed were open to investigate socio-cultural dimensions and symbolic universes, reflecting the integrative tradition of SRT that bridges diverse neighbouring disciplines in an effort towards a multifaceted perspective on the object of study.

Genome-wide identification and characterisation of Aquaporins in Nicotiana tabacum and their relationships with other Solanaceae species.

BACKGROUND: Cellular membranes are dynamic structures, continuously adjusting their composition, allowing plants to respond to developmental signals, stresses, and changing environments. To facilitate transmembrane transport of substrates, plant membranes are embedded with both active and passive transporters. Aquaporins (AQPs) constitute a major family of membrane spanning channel proteins that selectively facilitate the passive bidirectional passage of substrates across biological membranes at an astonishing 108 molecules per second. AQPs are the most diversified in the plant kingdom, comprising of five major subfamilies that differ in temporal and spatial gene expression, subcellular protein localisation, substrate specificity, and post-translational regulatory mechanisms; collectively providing a dynamic transportation network spanning the entire plant. Plant AQPs can transport a range of solutes essential for numerous plant processes including, water relations, growth and development, stress responses, root nutrient uptake, and photosynthesis. The ability to manipulate AQPs towards improving plant productivity, is reliant on expanding our insight into the diversity and functional roles of AQPs. RESULTS: We characterised the AQP family from Nicotiana tabacum (NtAQPs; tobacco), a popular model system capable of scaling from the laboratory to the field. Tobacco is closely related to major economic crops (e.g. tomato, potato, eggplant and peppers) and itself has new commercial applications. Tobacco harbours 76 AQPs making it the second largest characterised AQP family. These fall into five distinct subfamilies, for which we characterised phylogenetic relationships, gene structures, protein sequences, selectivity filter compositions, sub-cellular localisation, and tissue-specific expression. We also identified the AQPs from tobacco's parental genomes (N. sylvestris and N. tomentosiformis), allowing us to characterise the evolutionary history of the NtAQP family. Assigning orthology to tomato and potato AQPs allowed for cross-species comparisons of conservation in protein structures, gene expression, and potential physiological roles. CONCLUSIONS: This study provides a comprehensive characterisation of the tobacco AQP family, and strengthens the current knowledge of AQP biology. The refined gene/protein models, tissue-specific expression analysis, and cross-species comparisons, provide valuable insight into the evolutionary history and likely physiological roles of NtAQPs and their Solanaceae orthologs. Collectively, these results will support future functional studies and help transfer basic research to applied agriculture.