The natural timing devices of organisms, commonly known as biological clocks, are composed of specific complex folding molecules that interact to regulate the circadian rhythms. Circadian rhythms, the changes or processes that follow a 24-h light–dark cycle, while endogenously programmed, are also influenced by environmental factors, especially in sessile organisms such as plants, which can impact ecosystems and crop productivity. Current knowledge of plant clocks emanates primarily from research on Arabidopsis, which identified the main components of the circadian gene regulation network. Nonetheless, there remain critical knowledge gaps related to the molecular components of circadian rhythms in important crop groups, including the nitrogen-fixing legumes. Additionally, little is known about the synergies and trade-offs between environmental factors and circadian rhythm regulation, especially how these interactions fine-tune the physiological adaptations of the current and future crops in a rapidly changing world. This review highlights what is known so far about the circadian rhythms in legumes, which include major as well as potential future pulse crops that are packed with nutrients, particularly protein. Based on existing literature, this review also identifies the knowledge gaps that should be addressed to build a sustainable food future with the reputed “poor man’s meat”. Circadian rhythms are broadly defined as endogenous biological processes that occur within an oscillation of approximately 24 h. Also referred to as biological clocks, circadian rhythms are subject to both environmental entrainment and temperature compensation . This phArabidopsis thaliana) through a combination of omics approaches [CCA1 and LHY genes. On the other hand, the evening complex (EC) in the evening loop, which is a protein complex consisting of EARLY FLOWERING 3 (ELF3), EARLY FLOWERING 4 (ELF4), and LUX ARRHYTHMO (LUX) proteins, inhibits the expression of PRR7 and PRR9  and lablab (Lablab purpureus), have been promoted as climate-resilient crops, playing a critical role in achieving a suite of the Sustainable Development Goals (SDGs) that focus on tackling global food insecurity and climate change [Leguminosae (or Fabaceae), highlighting key findings in their clock research as well as the gaps in the existing literature. Based on published work on several well-studied leguminous species such as soybean (Glycine max), and common pea (Pisum sativum), we conclude that a more comprehensive understanding of the biological clock in legumes, especially how tissue-specific clocks are coordinated and how these connect to regulate development and physiology, is essential for genotype selection and improving the physiological adaptations of leguminous crops to climate change in the 21st century.With the declaration of 2016 as the International Year of Pulses, the importance of pulse utilization and development in supporting sustainable food systems has been widely recognized . Pulses,e change . The tare change . This reThe circadian system is a highly intricate network, with extensive crosstalk among output pathways that can be influenced by external conditions. External  signals, such as light and temperature, are integrated by the central oscillator that coordinates various physiological processes ,11. A prMedicago truncatula) and birds-foot trefoil (Lotus japonicus), have laid a foundation for the exploration and characterization of flowering genes in a range of other legumes [CCA1 and LHY [TOC1, GI, and ELF3 are present as duplicate copies in some species within the two legume groups. Interestingly, the CONSTANS protein, which promotes flowering in LD plants such as Arabidopsis, is represented by only one co-ortholog in the LD legumes and two in the SD legumes [Flowering time, the period at which a plant produces the first floral bud, is a key factor associated with adaptation and yield responses of a particular species to various environments, locations, and agricultural practices . Althoug legumes ,19,20. W legumes . In most and LHY ,23, whil legumes .FLOWERING LOCUS T/TERMINAL FLOWER 1 (FT/TFL1) gene family, which integrates environmental signaling for floral induction in Arabidopsis, has expanded in legumes and was reported to control the fate of meristems during flowering. Both galegoid and phaseoloid legumes have multiple TFL1 genes and generally have three distinct subclades of FT gene  [SHORT VEGETATIVE PHASE (SVP) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) genes were found [Both genes and environmental factors affect the vegetative-to-reproductive transition in some leguminous species. In a broad sense, flowering time plays a fundamental role in deciding when and how a plant can allocate resources, participating in a complex network of interactions with other developmental processes . The FLOand FTc) . Additiore found ,23.Lhcb (CAB2 and CAB1) transcripts associated with chlorophyll synthesis and photosynthesis. These genes encode light-harvesting chlorophyll-binding proteins that form up to 50% of the thylakoid membrane protein and have cis-elements associated both with light-induced transcription and circadian control [Plants with circadian rhythms that synchronize with their environment are reported to gain growth or metabolic advantages over those that are not synchronized ,26. The  control .Circadian and dial regulation of the levels of phytohormones is widespread in plants, likely involving a complex network of hormone signaling pathways. Circadian oscillations in the major growth-related phytohormones, including ethylene (ET), auxin/indole-3-acetic acid proteins (Aux/IAAs), cytokinins (CKs), gibberellins (GAs), and brassinosteroids (BRs), have been studied in multiple species that demonstrated species- and/or tissue-specific variations ,33,34,35YUCCA8, leading to increased auxin production during the day. In Arabidopsis, the daily rhythms in ABA, BR, IAA, and GA signaling are complex and modulated at many steps [There are several ways in which the plant circadian system can interact with hormone metabolic pathways. The expression of specific genes that encode hormone biosynthesis enzymes in plants has been reported to be clock regulated. For example, many GA biosynthesis genes demonstrated regular daily rhythms in Arabidopsis and common pea ,38, wherny steps . AlthougAtBBX32 in soybean was found to increase the grain yield of multiple transgenic events in field trials [Apart from light and temperature, biotic and abiotic stresses activate plant response pathways that can alter the plant circadian clock. Little is known about the impacts of these stresses on the circadian clock of crop plant species, although major clock genes that are associated with some agronomic traits have been reported. The limited understanding of how different stresses can influence crop circadian clocks is considered one of the key knowledge gaps that impedes the identification of specific circadian traits for further germplasm improvement in breeding programs. Among legumes, abiotic stresses such as heat, drought, and iron deficiency have been reported to change the timing of the expression of core clock genes in soybean, with different stresses causing different physiological effects . Howeverd trials .While the first scientific knowledge of the plant circadian clock was reported nearly 300 years ago, the role of the molecular clock in growth and development is not fully understood and many clock-related components remain to be discovered ,46. ReceMedicago truncatula) is a tiny annual legume that is used widely in legume genomics research. Native to the Mediterranean region, this forage legume has been used to study many aspects of plant biology, especially since the first release of a genome sequence in 2011 [Agrobacterium tumefaciens-mediated and A. rhizogenes-mediated hairy root transformation, have been developed for functional genomic studies in this model legume [Barrel medic  genes have been previously implicated in flowering time regulation and biomass accumulation [CCT genes with flowering time regulation, abiotic stress response, and regulation of growth and development [LHY (LATE ELONGATED HYPOCOTYLS) gene also plays an essential role in the regulation of the endogenous biological clock in barrel medic [MtLHY can severely impact the transcription of several key circadian genes, and its over-expression may result in delayed flowering and hypocotyl elongation. The study also reported reduced nitrogen fixation with altered biological clocks in Mtlhy mutants, causing abnormal nyctinastic leaf movement and biomass reduction. The amenability of barrel medic to be genetically transformed by various methods has made it well-accepted as a model legume species that likely will continue to attract interest from crop researchers across different biological disciplines.Recently, Ma et al.  reportedmulation . Gene clelopment . A recenel medic . FunctioLotus japonicus) is physically small and has a short generation time [Ljmybr, an MYB-related gene, with a possible role in nitrogen fixation regulation. The study identified Ljmybr as a possible CCA1-related gene sharing a conserved SHAQKY domain. MYB transcription factors are known to be associated with plant development, response to environmental signals, and hormone regulation [Zea mays), and Arabidopsis [Similar to barrel medic, birds-foot trefoil , a globally traded legume, serves as a prime source of protein and vegetable oils for human consumption. Deemed the “king of beans”, this major legume is also widely used for animal feed and biofuel production. Recent years have seen rapid improvement in the quantity and quality of soybean seeds due mainly to the advances in genetic and metabolic engineering [Soybean , which is also the model plant used by the father of genetics, Gregor Mendel, to study the laws of inheritance. Since the first description of circadian rhythm in common pea [Another common crop legume is the common pea  and AtELF4, including LATE BLOOMER1 (LATE1) and DIE NEUTRALIS (DNE). According to Liew et al. [STERILE NODES (SN) locus in common pea was among the first photoperiod response genes to be studied, demonstrating the genetic control of flowering-time regulation through long-distance signaling. The study reported that SN formed a complex network with two other circadian clock genes, DNE and HIGH RESPONSE TO PHOTOPERIOD (HR), where HR regulates the expression of SN while SN influences the role of HR and DNE in controlling flowering. The common pea axillary bud transcriptome analysis demonstrated rapid changes in the temporal expression of diurnally regulated genes within the short 170-min time frame, suggesting that future gene expression studies in this crop should take into account the possibility of fast diurnal changes in gene expression [The first research into the circadian clock and photoperiod response mechanism in common pea was reported by Weller et al. , highligw et al. , the STEpression .ELF3 homolog in lentil, along with common pea (HR locus), as the genetic factor underlying flowering time variation [During the past decade, it has been observed that unlocking the potential of underutilized legumes, such as winged bean and lentil, is equally important as the improvement of major legumes such as soybean and common pea ,8. Theseariation . More reThe past two decades have seen enormous progress in understanding the molecular features and architecture of circadian oscillators in plants. Nonetheless, knowledge of the biological and biochemical functions of some core clock components is still lacking, especially in non-model plants. The increasing evidence of novel molecular mechanisms that play a role in circadian rhythmicity calls for more comprehensive research to understand and validate these mechanisms. Recent studies have revealed the presence of tissue-specific clocks in plants that are regulated in a reciprocal fashion, demonstrating more complex and distinct rhythmic properties in the network that had previously been overlooked. More focus should be given to elucidate the molecular mechanisms of these tissue-specific clocks and determine how they communicate with each other to enhance the growth and fitness of plants. Specifically, crop fitness has long been artificially selected through breeding and domestication, and the circadian clock system has been identified as a key to improved fitness through selections. Despite significant progress in Arabidopsis and a few other model crop species, the present knowledge of the crop circadian clock is generally still limited. It is crucial to gain a more precise understanding of the relevant connections between the physiological pathways and the clock’s overall effects on plant development.Early clock research in plants has focused primarily on Arabidopsis—the model plant that has defined many circadian clock components and functions in plants, which suggests that the available knowledge of the overall circadian system may somehow be distorted . For croCircadian clocks play a critical role in plant development and physiology and thus are key components for inclusion in legume crop selection and improvement strategies. Although several core clock genes have been studied in some leguminous species , functio


Lasiurus cinereus). Because it is not possible to monitor population sizes for these solitary and cryptic species using traditional techniques, we must rely on other methods, such as acoustic and genetic monitoring, to provide information regarding population status and trends. Using these approaches, we can build a weight of evidence to assess whether mortality associated with wind turbines is sustainable and determine if and when mitigation measures should be implemented to reduce wind turbine mortality. To this end, we recommend that acoustic data be collected using the North American Bat Monitoring Program (NABat) protocols, and that genetic diversity be monitored at repeated time intervals to document species trends. There are no short-term measures to resolve population-level questions for migratory tree-roosting bats. Nonetheless, we discuss opportunities for relatively short-term investments that will lead to long-term success in reducing uncertainty for these species.Although wind energy provides valuable environmental and economic benefits, it has unintended negative consequences for wildlife, particularly birds and bats that collide with the rotating blades. In North America, for example, there are increasing concerns that wind turbines threaten the persistence of populations of migratory tree-roosting bats, such as the hoary bat (Lasiurus cinereus) in North America. These species are dispersed within their range, making it impractical to estimate census populations size using traditional survey methods. Nonetheless, understanding population size and trends is essential for evaluating and mitigating risk from wind turbine mortality. Using various sampling techniques, including systematic acoustic sampling and genetic analyses, we argue that building a weight of evidence regarding bat population status and trends is possible to (1) assess the sustainability of mortality associated with wind turbines; (2) determine the level of mitigation required; and (3) evaluate the effectiveness of mitigation measures to ensure population viability for these species. Long-term, systematic data collection remains the most viable option for reducing uncertainty regarding population trends for migratory tree-roosting bats. We recommend collecting acoustic data using the statistically robust North American Bat Monitoring Program (NABat) protocols and that genetic diversity is monitored at repeated time intervals to show species trends. There are no short-term actions to resolve these population-level questions; however, we discuss opportunities for relatively short-term investments that will lead to long-term success in reducing uncertainty.Wind turbine-related mortality may pose a population-level threat for migratory tree-roosting bats, such as the hoary bat ( Understanding the structure and dynamics of a population can assist in evaluating the scope and severity of a stressor, and the efficacy of mitigation measures. For most species of bats, population data are limited or nonexistent, making it difficult to assess natural and human-induced stressors ,2,3. DesLasiurus cinereus), eastern red bats , and silver-haired bats (Lasionycteris noctivagans). Note: we elected to use the traditional taxonomy recognized by the taxonomic nomenclature authority and database (www.batnames.org (accessed on 11 November 2021)); [The global expansion of wind energy development represents a potential threat to certain bat populations. In North America, wind energy development primarily impacts migratory tree-roosting species, including hoary bats ( 2021)); ); see [9 2021));  for sugg 2021)); ,13. Give 2021)); ,15, the Placing wind turbine-related bat mortality into context has been challenging because estimating census population size (Nc) using traditional methods is impractical for migratory tree-roosting bats. These species roost individually, are dispersed across the landscape, and migrate over long distances ,17. In lThe lack of available demographic data for many bat species, including hoary bats, emphasizes the importance of collecting data to determine vital rates; however, these studies are labor intensive and may require ≥5 years of data from capturing, marking, and tracking individuals . AlthougThere are no practical, short-term measures to determine Nc for migratory tree-roosting bats, but additional data can be collected to reduce uncertainty and assist in determining population trends ,28. The In 2012, a workshop on bat population monitoring and modeling was convened to design statistically robust and logistically feasible methods for monitoring changes in bat populations . At the 2 cells and probability-based sampling [Pipistrellus pipistrellus), whereas it would take approximately 15 years to detect a 25% decline in this species.For acoustic sampling, NABat uses a systematic grid-based framework of 100 kmsampling ,34,35,36sampling  generatesampling . Becausesampling . Based osampling  reportedMyotis lucifugus). The inference of the study was limited as it was regionally focused and included only 5% of the continental United States; thus, conclusions about the population status of the two species at larger spatial scales could not be determined. Rodhouse et al. [Repeated annual surveys at priority grid cells permits the extension of the model to unsampled areas. As a result, changes in occupancy dynamics, or the probability of a site being unoccupied in one year given it was occupied the previous year, can be estimated . Rodhouse et al.  indicateFor NABat, acoustic surveys can be either mobile or stationary. For stationary surveys, as were used in Rodhouse et al. , each reMobile transects assume each bat pass represents a unique individual because the vehicle is presumably traveling faster than most bats are flying. Since most bats in North America fly between 9 and 32 km/h, mobile transects are unlikely to encounter the same bat twice ,45,46,47Each survey type, stationary vs. mobile, has advantages and biases. Neece et al.  reportedAlthough genetic data cannot provide direct estimates of census population sizes, they are useful for monitoring population status and trends, informing geographic ranges and seasonal movements, and detecting cryptic speciation . These aforementioned aspects of bat biology all require additional investigation and are necessary to properly plan and implement conservation strategies. By combining recent advances in the tools and analytical approaches to monitor genetic variation in wildlife populations with the abundance of bat carcasses that are recovered at certain times of the year in distinct geographic locations across the United States , these data have the potential to be especially informative. In addition, the ability to monitor for changes in genetic diversity over time could provide unique insights that reflect actual fluctuations in census population size. Although it is not possible to isolate changes in genetic diversity due to a single stressor, the scope and severity of these cumulative changes can guide management and mitigation decisions that are necessary for the conservation of migratory tree-roosting bats.Genetic variation in a population is essential for adaptation to environmental change, and it is well established that loss of genetic diversity impedes the long-term survival of species . When laSalmo salar) from Maine revealed significant decreases in Ne and several measures of genetic diversity from 1963 to 2001 [Population genetic analyses also can detect population bottlenecks, estimate historical and current effective population sizes, and identify evolutionarily unique subpopulations . The eff to 2001 . These r to 2001 .Genetic diversity and population structure of migratory tree-roosting bats likely affect resilience to sustained mortality from wind turbines and the likelihood of population persistence. Species with smaller Ne, for example, have lower genetic diversity which may limit their evolutionary potential to respond to selection and avoid inbreeding in the future . AlthougSeveral tools are available to characterize and monitor genetic variation in populations, and recent review papers illustrate how these data can be applied to the conservation of natural populations  provides some guidance on the number of detectors and duration for these surveys . StandarA primary objective for standard preconstruction acoustic surveys was to determine whether preconstruction activity predicts postconstruction mortality. Because no reliable relationship has been established between these variables ,83, it mThere are instances where following the probabilistic sample is not feasible, such as conducting presence/absence surveys to fulfill obligations under Section 7 of the U.S. Endangered Species Act (16 U.S. Code § 1536). The Range-Wide Indiana Bat Survey Guidelines , for exaWhile stationary acoustic data are valuable in determining occupancy probability, mobile acoustic surveys serve as inputs to both occupancy and abundance models. In support of the objective to determine measures of abundance for migratory tree-roosting bats and to place wind turbine mortality in the context of population size and trends, it is recommended to adopt mobile acoustic surveys in high-priority cells. Although offshore transects are not currently part of the NABat protocol, these could be considered, and protocols developed if they are deemed feasible in the offshore environment. It may be possible to conduct boat surveys by adapting the methods used by Whitby et al. . There aData collected in association with wind energy development can increase the quantity and quality of data available for analyses, offering in return data-driven updates to the probability of occupancy, species range maps, indices of abundance, and improved understanding of relationships with habitat features, available resources, and various stressors. To help achieve these goals, standard preconstruction bat data should be contributed to the NABat database, and where possible, transitions to mobile acoustic surveys following the NABat protocol are recommended.There is value in comparative genetic studies across species, geographic regions, and temporal scales. Continued genetic evaluation of opportunistically collected samples, such as those in Cornman et al. , with a For contemporary samples, it will be important to locate sources of existing genetic material . It also will be necessary to identify partners willing to share tissue samples or carcasses from mortality searches at wind turbines during the next several years, with the goal of filling in data gaps in species or geographic coverage that may exist with the samples that are already available.Future research can expand the geographic scope and temporal scale of hoary bat samples. It may be possible to use historical DNA samples to estimate the genetic parameters 10, 20, or even 100 years ago, which can be matched by geographic location to contemporary samples collected from wind energy facilities. The results of these analyses could be used to help answer the following questions: (1) is there evidence of a decline predating the development of wind energy?; (2) do changes in genetic diversity or Ne coincide with the expansion of wind energy development during the past 20–25 years? and (3) how do estimates of Ne and genetic diversity compare among different geographic regions that may be experiencing different stressors, such as wind energy development and climate change impacts?Estimates obtained from population genetic studies will not provide direct estimates of Nc, but they can be used to monitor populations and population impacts over time and to make predictions about relative impacts to different species. Obtaining these estimates for hoary, eastern red, and silver-haired bats can reduce uncertainty regarding the species-specific impacts of wind energy development. It is crucial to invest resources to store and manage genetic samples from bats collected at wind energy facilities during the next decade. This collection should target two different scales that are reflected herein: (1) sample intensively at select geographic areas that have high levels of bat mortality for hoary, eastern red, and silver-haired bats; and (2) sample broadly across North America to capture the range-wide level of impacts for hoary, eastern red, and silver-haired bats. By dedicating collection efforts at these two scales, we will ensure sufficient sample sizes and that we have sampled the entire population for each species.Strigops habroptilus) resulted from a recent population bottleneck associated with European colonization. In another example, Cammen et al. [Halichoerus grypus atlantica) and harbor seals (Phoca vitulina vitulina). These types of studies may also be informative for migratory tree-roosting bats.Taking multiple samples over time allows for monitoring genetic changes associated with various stressors . This tyn et al.  used RADWe recommend sampling several wind energy facilities in each U.S. Fish and Wildlife Service region for continuous long-term monitoring  for the three species of migratory tree-roosting bats with a focus on collecting tissue samples for subsequent genetic analyses. This could be achieved using a low-level monitoring effort, such as searching only gravel pads and roads at selected sites. The collection, processing, and storage of tissue samples and carcasses will require cooperation and coordination during standard postconstruction mortality monitoring efforts. The anticipated cost of tissue collection and storage are not anticipated to be a barrier to participation, but additional funding sources will be necessary to archive the samples and conduct the genetic analyses.Lasiurus ega) and the northern yellow bat (Lasiurus intermedius) using DNA from carcasses collected at wind energy facilities in south Texas, U.S. Yellow bats are in the same genus as hoary bats and eastern red bats, and also collide with wind turbine blades. Wind energy is relatively new in the region and represents a new source of mortality for these species. Chipps et al. [In addition to the strategic monitoring efforts described, we recommend that researchers continue to collect genetic samples from additional bat species that experience wind turbine mortality, especially as wind energy development expands into new areas. For example, Chipps et al.  examineds et al.  detecteds et al. . In addis et al. . FurtherNycticeius humeralis) [Tadarida brasiliensis) [Lasiurus seminolus) [Finally, we recommend the use of readily available and low-cost molecular methods to determine sex ,89,90 anmeralis) , Brazililiensis) ,98, and minolus) . ClearlyWhile migratory tree-roosting bats are experiencing a relatively new source of mortality associated with wind energy development, they are simultaneously experiencing other persistent and emerging stressors, such as climate change ,100, glo


Drosophila germarium over a 24-h cycle, and BMP gradients of several hundred microns establish dorsal-ventral tissue specification in Drosophila, zebrafish and Xenopus embryos in timescales between 30 min and several hours. The mechanisms that shape BMP signaling gradients are also incredibly diverse. Although ligand diffusion plays a dominant role in forming the gradient, a cast of diffusible and non-diffusible regulators modulate gradient formation and confer robustness, including scale invariance and adaptability to perturbations in gene expression and growth. In this Review, we document the diverse ways that BMP gradients are formed and refined, and we identify the core principles that they share to achieve reliable performance.Pattern formation by bone morphogenetic proteins (BMPs) demonstrates remarkable plasticity and utility in several contexts, such as early embryonic development, tissue patterning and the maintenance of stem cell niches. BMPs pattern tissues over many temporal and spatial scales: BMP gradients as short as 1-2 cell diameters maintain the stem cell niche of the  Summary: This Review looks at BMP morphogen gradient patterning in diverse developmental contexts including in Tribolium, Drosophila, Xenopus and zebrafish early embryos, as well as the Drosophila wing imaginal disc and germarium. The core bone morphogenetic protein (BMP) signaling components are largely conserved across metazoans , and the BMP5/6/7/8 orthologs, Screw (Scw) and Glass bottom boat (Gbb). The receptor portfolio is similarly limited, with only two Type I receptors, Thickveins (Tkv) and Saxophone, and two Type II receptors, Punt (Put) and Wishful thinking (Wit). Downstream Dpp/BMP signaling in Drosophila is transmitted through a single R-Smad, Mothers against Dpp (Mad), and a single co-Smad, Medea  components and other immobile binders that impact diffusion range between being completely free with a long range or being hindered and having a short range. This framework is particularly helpful in understanding the Within this framework, reviewed at length in The effect of immobile regulators on morphogen diffusion can be illustrated through the mathematics of reaction-diffusion as demonstrated below:M] is morphogen concentration, x is distance from source, [R] is concentration of immobile regulator and kbinding and kunbinding., lowers effective diffusion and decreased regulator concentration causes greater effective diffusion. This generalized principle manifests in multiple contexts including in the Drosophila germarium, in which ablation of extracellular regulators expands the BMP gradient. The level of regulator, by this simple example, directly tunes the range of BMP distributions and can be modified for scale-invariance, robustness and gradient range over evolutionary time. A much more detailed look at this in the context of scaling is available in where greater regulator concentration, [The relative importance of these factors and mechanisms in gradient formation remains a topic of intense debate  niche and is the site of oocyte production and differentiation. Dpp/BMP signaling is a crucial regulator of GSC recruitment and maintenance DrosophilInteractions with immobile ECM components help to localize Dpp/BMP ligands . For exabag of marbles (bam), a translational repressor of key stem cell maintenance genes   . Dpp/BMPsection) .Drosophila wing imaginal disc  levels in cells close to the morphogen source, allowing longer-range diffusion of the ligand . These cDrosophila embryo, this has the effect of concentrating the Dpp/BMP ligand away from the ventral expression domain of Sog (the Drosophila Chordin homolog), resulting in a Dpp/BMP ligand gradient narrower than the ligand expression domain.All metazoan embryonic patterning systems feature a recurring cast of secreted, diffusible BMP-binding proteins that regulate BMP ligand gradient properties and pattern. However, the mechanistic roles played by these systems are very diverse, and several distinct models have emerged to explain them . The priDrosophila, BMP gradient formation in the zebrafish embryo occurs through a ‘source-sink’ mechanism  at the dorsal midline DrosophilDrosophila, dpp is expressed across the dorsal blastoderm, whereas its heterodimer partner ligand scw is expressed more broadly. A shuttling mechanism acts to concentrate the Dpp-Scw gradient into a sharp peak at the dorsal midline. The ventro-laterally expressed highly diffusive Sog binds the active Dpp-Scw heterodimer and prevents local receptor activation  development in the ignaling . In thisignaling . Other signaling . TherefoDrosophila, in which dpp expression is limited to the dorsal blastoderm, in the flour beetle Tribolium, dpp is expressed uniformly along the putative DV axis Drosophil content .Tribolium and Drosophila accommodate vastly different body plans and morphogen patterning processes. In Tribolium, Dpp gradient formation is not dependent on the intact Dpp-Sog shuttling mechanism observed in Drosophila , the orientation of the DV axis itself and downstream organ development has been inverted in evolution  embryo also found co-localization of Chordin and BMP expression domains , but also confer robustness to the extracellular Dpp/BMP gradient itself , a homolog of TNF-α, and Crossveinless 2 (Cv-2), a membrane-bound Dpp/BMP regulator. Egr provides positive feedback regulation to Dpp/BMP signaling through the JNK pathway. Cv-2 has been shown to both positively and negatively regulate Dpp/BMP signaling  and Pinhead have been identified as another set of coupled regulators of BMP signaling F Yan et. Admp isThe first quarter century of research on the BMP signaling system identified a recurring cast of regulators that shape BMP gradients in a variety of developmental contexts. However, biochemistry and genetics approaches proved limited in generating mechanistic understanding of BMP gradient formation in different systems, as it became increasingly apparent that multiple gradient formation mechanisms are viable given different biophysical properties or regulatory networks. In recent years, new approaches that integrate genetics, embryo perturbations, imaging and high-throughput computational modeling have begun to clarify the complex interplay between biochemical networks that include feedback mechanisms that regulate BMP-mediated morphogen patterning. Currently, most studies of the BMP system evaluate hypothesized mechanisms, either experimentally or computationally, based on their ability to explain the morphogen gradient signaling distribution. This approach may be overlooking several important properties of BMP patterning that could further constrain models and identify underlying regulatory principles.in toto and thus the data are already at hand.In this Review, we have described scale invariance and noise reduction as additional lenses for investigating the robustness of the BMP signaling system. Understanding dynamic changes in the BMP signaling in three-dimensional space over developmental timescales presents another intriguing avenue for future research. In fact, mammalian systems, which have been understudied, have longer developmental periods and greater roles for redundant BMP antagonists . TherefoAs the field has moved from identifying and characterizing the components that interact to form the gradients towards deciphering how the molecules all work in concert as a reliable and robust system, there will continue to be an increasing need for ways to combine the data into mathematical models of the processes. These models should grow to become more user-friendly as a simulation-based aid for testing alternative hypotheses and predicting how a planned experiment may impact robustness, fragility or other emergent behavior. Similarly, as more is known about the limits to a biological system under diverse perturbations, these new data need to be better utilized for identifying the most consistent mathematical models. Multi-objective optimization, MBDOE and large-scale computational screens will be necessary to integrate data from disparate model systems, to generate holistic understanding of the biological mechanisms at play and to identify experiments to expose underlying fragility once key regulatory steps are removed. Developmental biology in general, and the BMP field in particular, has long been at the forefront in integrating systems and engineering approaches to address biological questions. A renewed focus on generating computational methods accessible for all developmental biologists will pay dividends in increasing the pace of scientific progress in understanding development.


Plant diseases are globally causing substantial losses in staple crop production, undermining the urgent goal of a 60% increase needed to meet the food demand, a task made more challenging by the climate changes. Main consequences concern the reduction of food amount and quality. Crop diseases also compromise food safety due to the presence of pesticides and/or toxins. Nowadays, biotechnology represents our best resource both for protecting crop yield and for a science-based increased sustainability in agriculture. Over the last decades, agricultural biotechnologies have made important progress based on the diffusion of new, fast and efficient technologies, offering a broad spectrum of options for understanding plant molecular mechanisms and breeding. This knowledge is accelerating the identification of key resistance traits to be rapidly and efficiently transferred and applied in crop breeding programs. This review gathers examples of how disease resistance may be implemented in cereals by exploiting a combination of basic research derived knowledge with fast and precise genetic engineering techniques. Priming and/or boosting the immune system in crops represent a sustainable, rapid and effective way to save part of the global harvest currently lost to diseases and to prevent food contamination.  With the new millennium, humankind is facing issues for too long postponed. Among the greatest challenges is to meet the food demand for a rapid increase in global population, estimated to exceed 9 billion by 2050 . AdditioR genes) through specific breeding programs, represents alternative methods to combat yield losses to pests. Notably, crop rotation is not always an economically viable strategy ..21].Agrobacterium tumefaciens Ti plasmid can be used to integrate foreign DNA into the plant genome [Some important achievements in plant sciences characterized the second half of the last century: the development of tissue culture and regeneration techniques allowing the use of embryo rescue and doubled haploid, and the genetic engineering technology including chromosome engineering and transgenesis for gene transfer between species distantly related. A further milestone in plant biotechnology was the demonstration that t genome ,23,24. St genome ,26. GeneDuring 1990s, several genetic modified (GM) crops were developed and released on the market, with regulatory approvals of 44 countries reaching more than 400 GM events involving 32 different crops, among which were maize, rice and wheat. in planta production of molecules and development of edible vaccines; (vii) phytoremediation [Among the most important traits introduced in GM crops are: (i) herbicide tolerance—introduced for the first time in soybean by Monsanto—that gave rise to Roundup Ready crops; (ii) biotic and abiotic stress resistance with the introduction of a gene encoding the crystal Bt toxin, providing protection against pests, firstly in soybean and thereafter in other crops like maize; (iii) improved yield and growth; (iv) product quality (the first GM crop released on the market was the Flavr Savr tomato that slow down the fruit softening) ; (v) bioediation . The last 30 years have been full of new discoveries in the area of plant biotechnology, which are more and more used by plant scientists and breeders . Starting from the 1990s, molecular markers assist in plant selection, since many genetic markers associated with loci controlling traits of agronomic interest provided the opportunity to accelerate gain from selection. The development of next-generation sequencing technologies opened the era of Genomic Selection (GS), which allowed simultaneous selection for numerous markers, estimating the effects of all together loci at phenotypic level that would otherwise have no significant effect if individually taken. GS combined with high-throughput phenotyping became a powerful tool for the selection of the best phenotype within a plant population and to discover genes associated with quantitative traits. Several resistance genes have been isolated in potato , wheat , rice 3,33, and In the 2000s, a new approach, the Targeting Induced Local Lesions in Genomes (TILLING) approach, was introduced to identify mutant genotypes harboring mutations in genes of interest . This teIn the last decades, new breeding techniques (NBTs) are rapidly emerging from advances in genomic research and for application in crop traits improvement. They enable precise, targeted, and reliable changes in the genome and do not create multiple, unknown, unintended mutations, unlike chemical or radiation-induced mutagenesis.Genome-editing methods produce defined mutants, thus becoming a potent tool in functional genomics and crop breeding. Zinc Finger Nucleases (ZFN) and Transcription Activator-Like Effector Nucleases (TALENs) were the dominant genome editing tools until the rise of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and Crispr associated protein (Cas). CRISPR-Cas is an antiviral system developed by bacteria: segments of DNA containing short, repetitive base sequences  conserve the memory of intruding nucleic acids. The system is composed of genes encoding Cas nucleases, such as Cas9, and unique spacers (sequences complementary to a target genomic sequence) located in a genomic locus forming the CRISPR array along with crRNAs and trans-activating crRNA (tracrRNA), which are partially complementary to crRNAs. The RNase III processes the transcribed mRNA, releasing crRNA/tracrRNA complexes that activate and guide Cas proteins to target specific genomic loci introducing double strand breaks . For theGn1a, DEP1 and GS3 genes were edited in rice to enhance grain number and grain size [Grain Weight 2 (GW2) gene increases grain weight and yield [EDR1 in wheat results in plants resistant to powdery mildew [SWEET11, SWEET13, and SWEET14 genes [To date, major and minor crops, dicots and monocots, have been edited to improve traits of agronomical interest and with an increasing attention to nutritional and healthy values of derived foods . Yield rain size ; knock-ond yield ,42. Impry mildew ; rice li14 genes .SBEIIb gene; recently, DuPont Pioneer announced intentions to commercialize waxy maize obtained by knock-out of Wx1 gene [Food nutritional quality and safety are essential prerogatives to feed burgeoning world population and to limit malnourishment. Waltz (2016)  knocked Wx1 gene ; the proWx1 gene  and by eWx1 gene .Genome editing techniques have also been used to accelerate the domestication of crops  or to crIn recent years, significant efforts have been made, and results have been obtained in understanding the interplay between plants and their invaders . During Traditionally, PTI and ETI have been considered to act sequentially but independently. However, recent accumulating evidence shows that the distinction between PAMPs and effectors, PRRs and R proteins, therefore between PTI and ETI, cannot strictly be maintained ,71, suggPlant hormones, or phytohormones, are naturally occurring signaling compounds with diverse chemical properties. They play critical roles in the adaptation to environmental changes by driving proper responses, including activation of immunity, to a wide variety of biotic and abiotic stresses. The activity of a given hormone depends on its biosynthesis, conjugation, transport, and degradation as well as hormone activation and inactivation ,73. AlthArabidopsis thaliana EF-Tu (elongation factor thermo unstable) receptor, abbreviated as EFR, was transferred to monocot rice to confer resistance to two Xanthomonas oryzae pv. Oryzae isolates. Rice plants expressing such receptors were able to sense the bacterial ligand of EFR and to elicit an immune response. Moreover, the EFR receptor was able to use components of the rice immune signaling pathway for its function [AtEFR was also expressed in wheat [P. syringae pv. Oryzae. In another study, a lectin receptor-like kinase gene (LecRK) of Haynaldia villosa, a diploid wheat relative, has been transferred to wheat variety Yangmai158, which is powdery mildew susceptible [Knowledge of the plant immune system offers the opportunity to develop new strategies of intervention at the pathogen perception level . Increasfunction . AtEFR win wheat  driven bceptible . TransgePseudomonas syringae pv. Tomatoe and Agrobacterium tumefaciens. As rice Xa21 triggers HR-like responses, its intracellular domain has been used to generate chimeric PRR with rice OsCEPiP ectodomain [Magnaporthe oryzae [A different original approach is represented by engineering novel recombinant PRRs by producing chimeric receptors incorporating the beneficial properties of various RLKs and RLPs ; importatodomain . The rele oryzae .Beyond pathogen-recognition strategies, a better understanding of effectors and their role has allowed interventions at the point of pathogen modulation of host responses. Identification of effector activity targets in plant, for instance, shows which host components are “manipulated” by the invaders to promote disease. In order to interfere with these components of susceptibility, this knowledge was successfully exploited by removing ,106,107 TBF1 gene. Researchers showed that the utilization of “TBF1-cassette” can enhance broad-spectrum disease resistance with minimal adverse effects on plant growth and development [Ideal Plant Architecture 1 (IPA1), known as OsSPL14, in which a naturally occurred allelic variant increased yield and resistance to rice blast  . Three helopment . The timelopment . Therefoce blast . SpecifiR genes [Lr34 and Lr67  transporter with an unknown substrate. Transgenic wheat lines expressing Lr34 gene displayed enhanced resistance to multiple biotrophic pathogens including the leaf rust pathogen and powdery mildew both at seedling and adult stages [Lr67 resistance gene is a specific dominant allele of a hexose transporter that provides resistance to powdery mildew and multiple rusts. Introduction of the Lr34 allele by transformation into rice [Lr67 into barley [Puccinia triticina (wheat leaf rust), P. striiformis f. sp. Tritici (stripe rust), P. graminis f. sp. Tritici (stem rust), Blumeria graminis f. sp. Tritici (powdery mildew), P. hordei (barley leaf rust) and B. graminis f. sp. Hordei (barley powdery mildew), Magnaporthe oryzae (rice blast), P. sorghi (maize rust), and Exserohilum turcicum (northern corn leaf blight) [Lr34 and Lr67 is poorly understood, although it is likely that it provides the activation of biotic or abiotic stress responses allowing the host to limit pathogen development and growth.Adult plant resistance (APR) or “slow rusting” wheat genes represent a class of potential transferable R genes . Differeand Lr67 , have ben cloned ,116. Lr3t stages ,117. Simnto rice , barley nto rice , sorghumnto rice , maize [nto rice , and durnto rice  and of Lo barley  produced blight) ,95,97. THvUGT13248, involved in mycotoxin detoxification [PvPGIP2) and TAXI-III, a xylanase inhibitor [Fusarium graminearum has been observed in wheat plants simultaneously expressing the PvPGIP2 in lemma, palea, rachis, and anthers, whereas the expression of this inhibitor only in the endosperm did not affect FHB symptom development, hinting that further spread of the pathogen in wheat tissues no longer can be blocked once it reaches the endosperm [Wheat resistance to Fusarium species has been greatly improved by expressing either a barley uridine diphosphate-dependent glucosyltransferases (UGT), fication , or pyranhibitor . Interesndosperm .RNA interference (RNAi) was first discovered in plants as a molecular mechanism involved in the recognition and degradation of non-self-nucleic acids, principally directed against virus-derived sequences. In addition to its defensive role, RNAi is essential for endogenous gene expression regulation . InitiatOver the last two decades, RNAi has emerged as a powerful genetic tool for scientific research. In addition to basic studies on the determination of gene function, RNA-silencing technology has been used to develop plants with increased resistance to biotic stresses , Table  123,124124. Geminiviridae family. This virus translates four viral proteins and causes economical losses in wheat and barley when it is transmitted to plants through leafhoppers. Kis et al. [Indeed, the impact of RNAi technology deployed as a GM solution against viruses is clearly demonstrated in different studies ,126,127.Hordeum vulgare) plants, expressing via HIGS a 791 nucleotide (nt) dsRNA (CYP3RNA) targeting all three CYP51 genes  of Fusarium graminearum (Fg) that led to the inhibition of fungal infection [Recently, RNAi has been explored as a strategy to also control fungi and oomycetes. Fungal target genes are obvious candidates for this approach, as disruption is known to be lethal. A biotechnological method, termed host-induced gene silencing (HIGS), has emerged as a promising alternative in plant protection because it combines high selectivity for the target pathogen with minimal side effects, as compared with chemical treatments. Significant effects have been observed in transgenic Arabidopsis and barley , is an effective method to enhance resistance of wheat plants against fungal pathogens. Three hairpin RNAi constructs corresponding to the different regions of Chs3b were found to silence Chs3b in Fg strains. Co-expression of these three RNAi constructs in two independent elite wheat cultivar transgenic lines conferred high levels of stable and consistent resistance (combined type I and II resistance) to both Fusarium Head Blight (FHB) and Fusarium Seedling Blight (FSB).Cheng et al.  reportedvia synthetic biology approaches. In addition, RNAi strategies may be particularly relevant when no pathogen resistance can be identified in natural populations.A better understanding of this process in diverse plant-pathogen interactions may allow to better optimize HIGS strategies providing field-relevant levels of resistance ,131,132.In plant research, NBTs are attracting a lot of attention. NBTs appear to be suitable for many different fields in plant science, such as developmental processes and adaptation/resistance to (a)biotic stresses . NBTs inThe rapid increase in scientific publications documenting the use of CRISPR/Cas highlights how this technique has a greater success rate in gene modification compared to the other available nucleases. Actually, the application of CRISPR/Cas technologies to edit plant genomes is proving to be a powerful tool for future enhancement of agronomic traits in crops, qualitative and health parameters, tolerance to abiotic stress , and alsMLO loci have been targeted by RNA-guided Cas9 endonuclease in bread wheat [MLO encodes a protein with seven transmembrane domains localized in the plasma membrane and is ubiquitously present in monocots and dicots [MLO were susceptibility genes and that homozygous loss-of-function mutants had significantly increased resistance to powdery mildew in barley, Arabidopsis, and tomato [TaMLO-A1) of the three MLO homeoalleles showed improved resistance to Blumeria graminis f. sp. tritici infection, a finding that once again demonstrated the important role of TaMLO genes in powdery mildew disease [TaEDR1 [TaHRC, a gene that encodes a putative histidine-rich calcium-binding protein, is the key determinant of resistance to FHB. Authors have demonstrated that TaHRC encodes a nuclear protein conferring FHB susceptibility and that a CRISPR–Cas9-mediated deletion spanning the start codon of this gene results in FHB resistance. Plant mutants had significantly lower FHB severity than their wild type, suggesting that TaHRC affects FHB susceptibility and that loss of function of TaHRC confers Fhb1 resistance. Plants resistant to rice blast disease were generated through CRISPR/Cas9-mediated disruption of OsERF922 and OsSEC3A genes in rice [Ossec3a mutant plants in a putative subunit of a complex involved in exocytosis revealed a pleiotropic phenotype including improved resistance against Magnaporthe oryzae, higher levels of SA and its related genes, but also dwarf stature [1 and T2 transgene free plants mutated in the ET responsive factor (ERF) 922, a transcription factor involved in multiple stress responses. Mutant plants had a reduced number of blast lesions at both seedling and tillering stages [In a recent study, ad wheat . MLO encd dicots . It had d tomato ,142,143. disease . Another [TaEDR1  have rep in rice ,139. Oss stature . In contOsSWEET13 has been performed in rice to achieve resistance to bacterial blight disease caused by bacterium Xanthomonas oryzae pv. oryzae [OsSWEET13 is a susceptibility gene encoding a sucrose transporter involved in plant-pathogen interaction. X. oryzae produces an effector protein, PthXo2, which induces OsSWEET13 expression in the host and the consequent condition of susceptibility. Zhou et al. [OsSWEET13 in order to better explore PthXo2-dependent disease susceptibility, and resultant mutants were resistant to bacterial blight. Further genome editing strategies for multiplexed recessive resistance using a combination of the major effectors and other R genes will be the next step toward achieving bacterial blight resistance.Relatively few studies have been published on the application of the CRISPR/Cas systems to counteract crop bacterial diseases. CRISPR/Cas9 editing of . oryzae . OsSWEETu et al.  obtainedR genes can be created by a transgenic approach rather than by a traditional crossing approach. An additional advantage of transgenic strategies is that linkage drag can be prevented. In resistance breeding, linkage drag refers to the undesirable reduction in crop yield and quality sometimes associated with selection of genetic resistance to disease. One of the most efficient and sustainable solutions to control plant resistance to pathogens is to use genetic modification and genome editing techniques to complement and extend modern breeding efforts. Our planet is facing unprecedented challenges because of a rising and more affluent world population, while almost half of global cereal production is lost to diseases, biodiversity is diminishing at an alarming pace, and the average temperature on earth continues to rise. To meet the global challenge of food availability, we will need to shift our mentality and lifestyle, increase investments in knowledge creation, and facilitate the usage of innovative technologies, which can shorten timings to reach these goals. On the other hand, agriculture and food production must become more sustainable. The environmental footprint of agriculture needs to diminish, and farming must adapt to the rapidly changing climate. Given that we are witnessing how these aspects are threatening crop yields worldwide, all possible approaches are required to meet these challenges. Mutagenesis had, has and will certainly have an impact on crop genetics and breeding in the attempt to increase stress resistance and productivity. The ability of the DNA to mutate provides genetic variability, which is basilar for plants, as for all organisms, to evolve and adapt to environmental changes. New genotypes with new traits, likely of agronomic interest, can be created in cultivated plants by artificially inducing mutations in their genomes. Crops with improved characteristics can be obtained by both transgenesis and conventional plant breeding, respectively by adding a new gene to the genome of a crop plant or by crossing plants with desirable characteristics and selecting combinations of genes inherited from the two parent lines. With respect to when transgenesis technology was developed, our knowledge on plant gene function(s) and activity has drastically increased, satisfying one prerequisite for transgenesis to be preferable, especially in case the gene of interest do not exist in a species that can be successfully crossed with the crop. Secondly, modern high yielding crop varieties mostly result from careful selection of lines with specific combinations of value-added genes; such combination might be destroyed in the attempt to add by crossing a useful gene variant that has been newly discovered in a wild relative. Nonetheless, additional genes, closely located in the genome to the gene of interest, are almost always transferred as well. It will take several years and different generations to restore those gene combinations, even when using modern molecular breeding techniques, such as marker assisted breeding. These problems can be avoided by introducing a new added-value gene into the high yield crop by direct transformation, therefore by transgenesis. For instance, plant cultivars with Genome engineering techniques have made important advances over the last decades, allowing the capability not only to control but even to edit gene expression in a precise and secure manner, see The application of such biotechnological techniques in agriculture can potentially improve food availability and security by raising crop resistance to pathogens, adverse weather and soil conditions, by enhancing the adaptability of crops to different climates and by improving yields, particularly of staple food crops such as cereals . BiotechAn ambitious target for the future is to continue combining science-based knowledge with biotechnological methods to develop plants that have higher resilience to (a)biotic stresses. This will enable farmers to produce high yields while decreasing the use of chemicals and water.


Luminescent materials are everywhere in our daily life. However, most are not performing at their best performance because of the thorny aggregation-caused fluorescence quenching (ACQ) effect in molecular aggregates. Opposite to ACQ, aggregation-induced emission (AIE) describes the photophysical phenomenon in which non-luminescent molecules in solutions are induced to show bright emission upon aggregate formation, as a result of restriction of intramolecular motions (RIM) caused by intermolecular steric interaction (e.g. tetraphenylethene (TPE) in Fig. Compared to traditional ACQ fluorophores, the greatest advantage of AIE luminogens (AIEgens) is the highly efficient radiative transition in aggregate state for practical applications. Over the past 20 years, many researchers have focused on developing AIEgens, which have demonstrated great potential in different areas from organic electronics to biomedical research and physical process monitoring Fig. . From trAIE liquid crystals with bright emission could be used to fabricate liquid crystal displays without the need for backlight to reduce energy consumption .Similarly, the potential of AIE has been greatly exploited to develop AIE light-up molecular probes, AIE dot probes and AIE theranostic probes for continuous monitoring of biological processes, disease diagnosis, cell tracking, vascular imaging, image-guided surgery and therapy . Recentl with RIM caused by microenvironment changes , an inteDespite the notable success in fundamental study and applications of AIEgens in the past two decades, it should be noted that there are challenges to use of AIEgens. We start with emission, the most distinctive feature of AIEgens. When AIEgens are randomly packed, broad emission is often observed because the torsional structure and changeable conformation can endow AIEgens with abundant energy levels, affecting the color purity in OLEDs. The broad emission can also cause cross-talking in multiplex sensing and imaging. A recent study revealed that adopting fluorophores or quantum dots with very sharp emission bandwidth to form donor-acceptor pairs with AIEgens can offer narrow emission output through Förster resonance energy transfer (FRET) . In addiin vivo applications. Red-shifting the absorption by precisely adjusting the molecular structure is the most straightforward solution to the problem, and one successful example was to integrate rotor structure with twisted intramolecular charge so that AIEgens with more than 700 nm absorption peak were designed for in vivo near-infrared II bioimaging [The short absorption wavelength of most reported AIEgens is often regarded as a disadvantage. This is largely because of their twisted molecular structures, which may not affect their performance in electronics, but can significantly reduce the penetration depth in biomedical research, especially for oimaging . The devoimaging . The excoimaging . In addioimaging .Recent work also revealed that the molecular motions of AIEgens are still inevitable even in aggregate states. Through introducing spacers, molecular motions in the aggregate state have been used to design effective AIE photothermal agents for cancer therapy . This neExploration of AIE applications in healthcare, energy and environmental fields has led to the flourish of many research frontiers. For biomedical research, aggregation-enhanced reactive oxygen species generation of AIE photosensitizers is a very promising strategy to improve photodynamic efficiency in tumor therapy, and the molecular motions in the aggregate state can be well-controlled through molecular design to build photothermal systems with high conversion efficiency for photoacoustic imaging and photothermal therapy. In these applications, red-shifting the absorption of AIEgens to near-infrared range to match the parameters of commercial lasers will shed light on potential clinical translation. Considering the metabolism issue, exploration of clusteroluminescence from non-conjugated AIEgens or even natural products represents another promising pathway to translation. An important parameter not given the same consideration is their absorption. More effective absorption means better light harvesting, and therefore less material may be needed for the same application, which could help improve biocompatibility. On the other hand, mechanoluminescence and organic room-temperature phosphorescence are two new branches closely associated with the concept of AIE, which show great potential in detection, anti-counterfeiting, information encryption, afterglow imaging, etc. These studies require us to be equipped with strong capability in designing molecules that can easily undergo crystallization and may even form different types of crystals. When proper strategies are used to control or narrow down the emission spectra of mechanoluminescence and organic room-temperature phosphorescence, the anti-counterfeiting and information encryption properties can be further improved as a result of minimized overlap among the output signals. Last but not least, chiral AIEgen-based circular polarization LEDs can significantly reduce brightness loss when passing through polarizers, which has great potential to enhance efficiency in 3D display and optical communication.The enrichment of mechanism studies of AIEgens, including anti-Kasha decay, restriction of flip-flop motion, through-space interaction, etc., will continue to play an important role in shaping the AIE field. This will inspire more design principles to be developed for AIEgens and empower them with further improved functions and properties to solve practical problems. It is also important to note that the emergence of new and complementary theories to explain the AIE mechanism is a natural phenomenon. The in-deep mechanism research, coupled with modelling, big data and artificial intelligence will bring AIE research to the next level of excellence in materials innovation.The past 20 years have witnessed rapid development of the AIE field, from both fundamental science and practical applications. In this perspective, we focused on several challenges of AIEgens and discussed potential solutions to address these issues. It is important to note that the future directions mentioned are evolving with time, and integration between AIE and any new emerging research field can create new multi-disciplinary directions at any time. With artificial intelligence powered design and synthesis of AIEgens, the field is expected to have exponential growth in the next decade. By addressing the discussed challenges, AIEgens will become more powerful in addressing national challenges in energy, environment and social security, making an even broader impact on our everyday life.


Investigation of microbial communities has led to many advances in our understanding of ecosystem function, whether that ecosystem is a subglacial lake or the human gut. Within these communities, much emphasis has been placed on interspecific variation and between-species relationships. However, with current advances in sequencing technology resulting in both the reduction in sequencing costs and the rise of shotgun metagenomic sequencing, the importance of intraspecific variation and within-species relationships is becoming realized. Our group conducts multi-omic analyses to understand how spatial structure and resource availability influence diversification within a species and the potential for long-term coexistence of multiple ecotypes within a microbial community. Here, we present examples of ecotypic variation observed in the lab and in the wild, current challenges faced when investigating intraspecies diversity, and future developments that we expect to define the field over the next 5 years. Archaea as a distinct domain of the prokaryotic tree of life but also provided a way to distinguish divergent prokaryotic taxa and assess evolutionary relatedness  bacteria, a collection of 12 ecotypes , 15, incecotypes . Anotherulations , 18.Pseudomonas aeruginosa was reported to rapidly diversify  and nutrient-rich (LB-Miller broth) media rapidly diversify into ecotypes based on location (biofilm versus planktonic) and then utilization of amino acid resources  approach to experimental evolution studies examining how space and stress shape bacterial adaptation. Using a multi-omic approach that combined time-series metagenomic sequencing with genomic, transcriptomic, and exometabolomic analysis of isolated clones, we demonstrated that daily-transferred esources . These eGoing forward, our group will focus more on how environment influences the evolution of ecotypes and if any genetic backgrounds are particularly permissive/inhibitory to the maintenance of subpopulation structure . Beyond To date, the identification of ecotypes and the characterization of their interactions remain challenging. On the genomic level, short reads produced by next-generation sequencing platforms provide the accuracy to confidently identify polymorphisms within a bacterial population but fall short in the ability to phase these polymorphisms across sequencing reads to determine which mutations are present within an individual. To overcome this issue, computational approaches have been developed to assist in the identification of ecotypes, such as clonal reconstruction (a clone-informed maximum-likelihood method to parsing time-series metagenomic sequencing of populations ) or cladOnce ecotypes are identified, characterizing their role within the population or microbiome has additional complications. Microbes often exhibit emergent behavior, and their phenotypic expression may significantly vary in isolation from how they respond in the presence of their community . On a br27–


There is a growing interest in the potential role of adipose tissues in cardiac and renal pathophysiology, and determining the mechanisms by which fat compartments around the heart and kidneys influence cardiovascular disease is of clinical importance in both general and high-risk populations. Epicardial fat and perirenal fat have been associated with adverse outcomes in chronic kidney disease (CKD) patients. Epicardial fat is a rich source of free fatty acids and is capable of secreting inflammatory and pro-atherogenic cytokines that promote atherosclerosis through a local paracrine effect. Recent evidence has demonstrated that perirenal fat has a closer correlation with kidney diseases than other visceral fat deposits in obesity or metabolic disturbances. Moreover, perirenal fat has been reported as an independent risk factor for CKD progression and even associated with cardiorenal dysfunction. Accordingly, these forms of organ-specific fat deposits may act as a connecter between vascular and cardiorenal disease. This review explores the possible links between epicardial and perirenal fat and its significant role as a modulator of cardiorenal dysfunction in CKD patients. Interest in the potential impact of organ-specific adipose tissue on cardiac and renal pathophysiology is growing, and it is clinically important to determine the mechanisms by which fat compartments around the heart and kidneys influence cardiovascular disease (CVD) in both general and high-risk populations , 2. In tThe close anatomical proximity and intercommunication between EF, coronary arteries and myocardium has been reported as a key factor in atherosclerotic disease development and progression . As a riEF represents adipose tissue located below the visceral pericardium in direct contact with the epicardial coronary arteries . EF origAlthough the functional role of EF is complex and currently remains undefined, this tissue participates in multiple areas, such as mechanical, metabolic, thermogenic and endocrine/paracrine/autocrine functions . EF accovia a protein kinase pathway to reduce lipid storage in the myocardium, and also inhibits production of inflammatory mediators to maintain an anti-inflammatory environment in the cardiovascular system. Conversely, under pathological conditions adiponectin production by EF drops, while production and secretion of pro-inflammatory or pro-atherogenic cytokines  are increased , among others , 29 Tab. Moreovevia computed tomography (CT) and magnetic resonance imaging (MRI)  , 38, altng (MRI) . Other sng (MRI) . EF has ng (MRI) . In contRecent evidence has shown that region-specific adipose tissue is an important cardiovascular risk marker in the general and high-risk population. For example, there appears to be a connection between EF and CVD in conditions as common as high blood pressure, metabolic syndrome, diabetes, and cardiorenal disease , 32. RecAlthough studies evaluating EF in CKD are still ongoing, some research has reported a relationship between EF and vascular disease in patients with renal involvement , 43 Tab. In a stRecent studies have identified excess PF as an emerging risk marker of CVD, independently of common metabolic risk factors , 44. OurAs a CVD-related risk factor, diabetes or dyslipidemia-associated PF may also indirectly contribute to CVD. A recent study showed that excess PF was independently associated with hyperinsulinemia and insulin resistance in obese patients, irrespective of other anthropometric or metabolic parameters, suggesting PF as a strong marker of insulin resistance . MoreoveThe physio-pathological processes and association of these specific cardiac and renal adipose tissues allow the hypothesis that an excess of epicardial and PF contributes to CVD. PF is potentially related to EF as both exhibit mesothelial layers like visceral organs enriched in white adipose tissue progenitors which produce adipocytes , 53. ThiElevated serum levels of fibroblast grow factor 23 (FGF23) are strongly associated with increased CVD in CKD patients. Hence, FGF23 levels rise as kidney function declines, with the result that in patients with end state kidney disease (ESKD), FGF23 contributes to hypertension, vascular calcification, and left ventricular hypertrophy by distinct mechanisms . It is cWe postulated that local peri-vascular expression of adiponectin and leptin may dysregulate local FGF-23 production in calcified endothelial cells, suggesting the possibility of an undercover regulatory pathway in a uremic environment. This mechanism may explain why EF has been reported as an independent risk factor for CAC and myocardial dysfunction in CKD patients  reduced EF , and useIn patients with ESKD, use of non-calcium-based phosphate binding agent sevelamer reduced serum cholesterol and markers of inflammation . AdditioRenal-affected patients express extremely high incidence of CVD usually attributed to classical and uremic-related conditions. This suggests that specific adipose tissues may represent an additional cardiovascular risk factor in CKD patients. Finally, organ-specific adipose tissue, specifically EF and PF, represents an interesting and underexplored field to consider in this population.All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Respondents reported several barriers to integrating GBV risk mitigation into their work during COVID-19, including insufficient funding, capacity, knowledge, and guidance. Efforts to mainstream GBV risk mitigation actions should continue and intensify, leveraging the lessons and experiences generated thus far.Available evidence indicates that the COVID-19 pandemic and response measures may lead to increased risk of gender-based violence (GBV), including in humanitarian contexts. This study examined the knowledge, attitudes, and practices of humanitarian practitioners related to GBV risk mitigation approaches during COVID-19 in order to refine current guidance and inform future materials. A global, online cross-sectional survey of humanitarian practitioners was conducted between November 2020 and April 2021. We calculated descriptive statistics and used Chi-square or Fisher’s exact tests to compare knowledge, attitudes, and practices among GBV specialists and non-specialists. Of 170 respondents, 58% were female and 44% were GBV specialists. Almost all (95%) of the respondents agreed or strongly agreed that they have a role to play in GBV risk mitigation. Compared to GBV specialists, a higher proportion of non-specialists reported little to no knowledge on GBV risk mitigation global guidance (38% vs. 7%,  Available evidence demonstrates that the risk of gender-based violence (GBV) increases during humanitarian crises ,2. AlthoRecent reports suggest that efforts to contain the spread of COVID-19, such as quarantine and movement restrictions, may lead to increased risk of GBV, including intimate partner violence (IPV) ,14. ThesGiven the far-reaching economic effects of the pandemic, there is an increased risk of sexual exploitation and abuse (SEA), as households are more likely to face shortages of resources or become dependent on humanitarian assistance . This isprevention efforts that target the root causes of violence and seek to prevent GBV from occurring in the first place; (2) providing specialized response services for survivors of violence, addressing the consequences of GBV after it has happened; and (3) risk mitigation actions which aim to reduce exposure to GBV and ensure that humanitarian response actions and services themselves do not cause harm or increase risk of violence (See Addressing GBV during emergencies includes three pillars of action: (1) implementing ence See  25,26].,26.preveGuidelines for Integrating GBV Interventions in Humanitarian Action (“GBV Guidelines”), a revised version of the first set of guidelines, originally launched in 2005 [Gender-based violence risk mitigation actions are a collective responsibility across the entire humanitarian system, as articulated by the principles of Centrality of Protection and Do No Harm. As such, they should be systematically integrated in the design, implementation, and evaluation of actions to respond to disasters and emergencies, across all sectors, and by all humanitarian practitioners. The foundational guidance document on GBV risk mitigation for all humanitarian sectors and actors is the InterAgency Standing Committee (IASC)’s 2015  in 2005 . The IASThe GBV Guidelines distinguish between ‘GBV Specialists’ and ‘non-GBV specialists’, describing GBV specialists as practitioners with professional training in GBV and/or considerable experience working on GBV programming. The GBV Guidelines are primarily targeted towards non-GBV specialists who work in other non-GBV sectors and who do not have specific expertise in GBV programming, but who could undertake activities in the context of their work to reduce GBV risks for affected populations. The GBV Guidelines describe one role of GBV specialists as assisting non-GBV specialists to undertake GBV risk mitigation efforts in their own sectors and to serve as a technical resource. UNICEF leads the inter-agency efforts on GBV risk mitigation including revision and development of the GBV Guidelines, inter-agency implementation of GBV risk mitigation across the humanitarian system and funding a dedicated inter-agency coordinator for the GBV Guidelines Reference Group, consisting of 15 humanitarian agencies. The GBV Guidelines Implementation Support team has made progress in increasing knowledge, awareness, and uptake of the GBV Guidelines through trainings and other dissemination efforts, though some anecdotal evidence suggests varying levels of uptake by sector.How to support survivors of gender-based violence when a GBV actor is not available in your area (“GBV Pocket Guide”) was developed jointly by the GBV Guidelines Reference Group and the GBV Area of Responsibility (AoR), to help colleagues working in other (non-GBV) sectors to safely and appropriately respond to disclosures of GBV in humanitarian contexts [In addition to the GBV Guidelines, a step-by-step guide for humanitarian practitioners entitled contexts .Identifying & Mitigating Gender-based Violence Risks within the COVID-19 Response [While the existing humanitarian guidance and good practices are relevant in the current context, COVID-19 brings new challenges . Given tResponse . It idenIn order to fill these gaps, and inform future mainstreaming efforts, this research study was carried out to examine how humanitarian practitioners are incorporating GBV risk mitigation actions across all programmatic sectors during COVID-19, their knowledge, attitudes and practices around mitigation approaches and the barriers to integrating these interventions within their day-to-day work. Given that the important role of GBV specialists in providing technical support and serving as a key resource for non-GBV specialists in carrying out GBV risk mitigation actions, we sought to compare knowledge, attitudes and practices and perceived barriers to implementation across these two groups.This is a cross-sectional study nested within the Humanitarian Gender Study, a larger mixed-methods study whose overall aim is to understand gender bias in the humanitarian sector including during active emergencies such as COVID-19. The quantitative component of the Humanitarian Gender Study included a gender bias survey and a GBV risk mitigation and COVID-19 survey. The GBV risk mitigation and COVID-19 survey aimed to better understand humanitarian practitioners’ knowledge, attitudes, and experience in mitigating GBV risks within the context of COVID-19, as well as how humanitarian programming during the pandemic has been adapted and is affecting GBV risks. This survey was conducted between November 2020 and April 2021 by collaborators at the Beth Israel Deaconess Medical Center (BIDMC), Harvard T.H. Chan School of Public Health, Harvard Humanitarian Initiative, Brigham and Women’s Hospital, UNICEF and CARE. The cross-sectional design allowed for rapid deployment and wide-spread dissemination to reach potential respondents around the world through social media platforms and humanitarian networks. The qualitative methods and findings that were part of the Humanitarian Gender Study will be described elsewhere.We implemented this study online and globally through REDCap—a secure, HIPAA compliant, web-based data collection tool hosted at BIDMC . The surwww.humanitariangenderstudy.org, accessed on 18 October 2021). We also published Facebook and LinkedIn advertisements targeted towards employees of major humanitarian organizations. These advertisements linked to the same REDCap consent and survey page and website. Finally, direct outreach was conducted by study team members to humanitarian practitioners within our own networks to raise awareness about the study and to enable further sharing and dissemination. The inclusion criteria included self-identification as age 18 years and older, ability to complete the survey in one of the six available languages, and current or past paid or unpaid work related to humanitarian assistance. Work could be in any role including administrative, programming, research, and policy. Humanitarian assistance was defined as assistance that is intended to save lives, alleviate suffering, and maintain human dignity after man made crises and disasters caused by natural hazards, as well as to prevent and strengthen preparedness for when such situations occur. Respondents were excluded from the sample if they did not meet all of the inclusion criteria or if they had not completed at least one of the non-demographic survey modules.The survey was disseminated primarily through listservs and online networks, including organizations, research groups, professional networks, and social media groups (Facebook and LinkedIn) focused on humanitarian practitioners. Messages sent through these networks included a link to a consent form and the online survey and to the study’s website  regardless of whether they work in protection or non-protection sectors.We analyzed responses collected between 17 November 2020 and 14 April 2021. We received a total of 541 completed quantitative surveys as part of the Humanitarian Gender Study: 371 responses to the gender bias survey and 170 responses to the GBV risk mitigation and COVID-19 survey. Data analysis focused on generating descriptive statistics which are reported as a frequency (percentage). We used Chi-square or Fisher’s exact tests to compare GBV risk mitigation attitudes, experience, and practices among individuals self-describing as GBV specialists and those who reported as non-specialists, regardless of which sector their work was conducted in. Analyses were performed in SAS 9.4 .All of the participants provided informed consent to engage in this research after reviewing a detailed Information Sheet presented in English, Amharic, Arabic, French, Spanish or Swahili at the beginning of the REDCap survey. The study was conducted in accordance with the Declaration of Helsinki, and the protocol was approved by the Institutional Review Board of BIDMC (Protocol 2020P000618). All research staff associated with this study completed CITI Program’s Research, Ethics and Compliance Training.The funders of the study had no role in study design, data collection, data analysis, data interpretation, or writing of the report. The corresponding author had full access to all of the data in the study and had final responsibility for the decision to submit for publication.The sample included 170 humanitarian practitioners  who completed at least one of the survey modules . The majThe majority of respondents in the sample (95%) and across sectors agreed or strongly agreed that as humanitarian professionals, they have a role to play in GBV risk mitigation . Howeverp < 0.001). Similarly, a significantly higher proportion of non-GBV specialists reported little to no knowledge on how to respond to a disclosure of GBV compared to GBV specialists . GBV risk mitigation experience was also significantly higher among GBV specialists. Over 86% of GBV specialists reported having implemented GBV risk mitigation activities at least once during a humanitarian emergency, compared with 50% of non-GBV specialists (p < 0.001).Approximately 24% of the sample reported little to no knowledge on GBV risk mitigation global guidance and this proportion was significantly higher among non-GBV specialists compared to GBV specialists . The most common reasons provided for the limited effectiveness of the GBV risk mitigation interventions include not enough funding (62.4%), COVID-19 restrictions (52.7%), prioritization of other issues (46.2%), insufficient staff capacity or knowledge (43%), not enough guidance or gaps in the guidance (41.9%), lack of organizational commitment to addressing GBV risk (38.7%), insufficient human resources (38.7%), and barriers to accessing guidance (26.9%). The proportion of humanitarians reporting these barriers was generally similar between GBV and non-GBV specialists. However, 52% of non-GBV specialists cited lack of GBV risk mitigation guidance as a barrier compared to only 25.7% of GBV specialists (p = 0.01).Overall, only 31% of practitioners who reported integrating GBV risk mitigation interventions in their sector-specific work, reported that these interventions were very effective or fairly effective . Almost This study finds that while humanitarian practitioners in the sample almost unanimously agree that all humanitarian workers have a role to play in GBV risk mitigation , there are reported gaps in knowledge and experience, and barriers to integration of these actions within day-to-day work during COVID-19. For example, almost 40% of non-GBV specialists in this study reported little or no knowledge of the GBV Guidelines. This signifies that about 60% of interviewed non-GBV specialists do have some knowledge of this foundational guidance document, which is an encouraging sign about efforts to raise awareness. However, almost 45% of the interviewed non-GBV specialists reported insufficient knowledge or skills to carry out GBV risk mitigation within their usual sector-specific work. Even among those who report knowledge and skills, several types of perceived barriers to putting GBV risk mitigation into practice were described.The limited knowledge or skills on GBV risk mitigation reported by practitioners in our study, especially among non-GBV actors, aligns with research conducted before the pandemic. For instance, a series of case studies focused on Central African Republic, South Sudan, Iraq and Sierra Leone found limited understanding of how to operationalize the IASC GBV Guidelines, and lack of consistency in the way that donors, UN agencies and other implementing partners interpreted, prioritized and implemented the guidelines . Gender-For example, practitioners in our study also reported lack of organizational commitment to GBV risk mitigation and prioritization of other areas as barriers to the implementation of GBV risk mitigation interventions. Several factors may underly low commitment and prioritization. For example, lack of GBV expertise at the field level and within senior management within organizations inhibits the prioritization of GBV interventions within humanitarian response plans . Lack ofBoth GBV and non-GBV specialists in our study reported that lack of funding has been a key barrier to integrating effective GBV risk mitigation within their work both before and during COVID-19. This finding is particularly interesting for several reasons. First, higher level commitments to addressing GBV have been visible during the COVID-19 pandemic, including a joint statement by 146 UN member states and observers supporting prioritization of measures to address GBV in national COVID-19 response plans . Our finThe same VOICE and IRC study that assessed funding gaps also highlighted that localization of humanitarian action, including related to GBV, has remained low . LocalizHumanitarian practitioners in this study also shared their perceptions of the changing GBV risk environment during COVID-19 within the contexts in which they work. Most practitioners in this study reported the possibility of increases in risk of IPV during the pandemic; this aligns with growing reports and published research . AdditioOur study sheds important light on the different types of programmatic adaptations that are being made to address GBV risks in emergencies during the pandemic. Notably, changing the modality of consultations with women and girls and of service delivery were the most common adaptations reported, followed closely by incorporating GBV information within COVID-19 education materials. These are similar to examples of adaptations included in other reports . Singh eThis study has several limitations. First, the use of social media and humanitarian networks to recruit study subjects restricts the generalizability of the findings. Several countries, such as Ethiopia, are over-represented in the sample because of the study teams’ larger networks in this setting. Respondents recruited through social media or humanitarian networks may differ from others humanitarian professionals in several ways including literacy, access to technology, and resources. The survey was available in six languages, but practitioners fluent in other languages may not have been able to participate. Data were collected across countries that were at different stages of the pandemic during the study period. This may have limited the examples of GBV risk mitigation adaptations collected as settings with fewer restrictions may not have required adapted approaches. Despite intense efforts to recruit respondents, the sample size for this component of the study was lower than anticipated, and this reduced power to conduct comparisons across the 13 humanitarian sectors. The low sample size was possibly due to the highly specialized topic of the survey and survey fatigue, particularly given the significant increase in online data collection processes that accompanied the COVID-19 response. Despite these limitations, the study has several strengths. To our knowledge, this is the first study to systematically assess GBV risk mitigation in COVID-19 with a focus on non-GBV specialists among a global sample of humanitarian practitioners. The study was able to capture perspectives across 52 countries and 13 humanitarian clusters, demonstrating a wide reach across numerous humanitarian contexts and types of emergencies. The instrument developed for this research was designed to robustly capture a wide range of indicators on GBV risk mitigation including knowledge, attitudes, and practices, among both GBV and non-GBV specialists and will help to strengthen and advance measurement of GBV risk mitigation, an area that has significant gaps .In summary, the study findings suggest that despite increasing efforts to mainstream GBV risk mitigation across the humanitarian system, there are reported knowledge and skills gaps, especially among non-GBV specialists and across sectors, and barriers to implementing GBV risk mitigation interventions both prior to and during the COVID-19 pandemic. Our study provides unique contributions to the expanding field of GBV risk mitigation and highlights the need for the following actions.Efforts to mainstream GBV risk mitigation actions, in particular strengthening knowledge and skills-building, addressing unsupportive attitudes, and overcoming barriers to implementation and effectiveness should not only continue during the COVID-19 pandemic, but should accelerate while leveraging the lessons and experience generated thus far. Complementary to this work, strategies to increase prioritization of GBV risk mitigation in non-GBV sectors should continue to be explored and tested across humanitarian organizations. This may require addressing gender biases and other factors that influence decision-making, promoting women’s leadership in the humanitarian sector and generating evidence to demonstrate how addressing specific GBV risks could also improve sector-specific outcomes ;Misconceptions on GBV risk mitigation, such as the belief that additional and/or separate funding is always required to carry out this work, should be addressed. Additional awareness raising about the range of GBV risk mitigation intervention options, including those that can be implemented with little to no additional funding, would be helpful. For those risk mitigation interventions which do require additional funds, sectors have a responsibility to include budget for GBV risk mitigation within their own funding requests to ensure their programming is safe and accessible during COVID-19 and beyond. Currently across the humanitarian sector, there is no standard way of calculating or tracking GBV risk mitigation expenditure within sectors’ total budgets. Systematizing this type of tracking would be valuable for the field, as it would facilitate understanding and identification of funding gaps and ensure that available resources match the need;Awareness and knowledge of global guidance on GBV risk mitigation was limited among non-GBV specialists in this study, though the stated desire for guidance was high among this group. Therefore, intensifying efforts to promote uptake and use of the available guidance targeting those practitioner groups with less access could help bridge the gap. Donors could also support these efforts by referencing the IASC GBV Guidelines—and/or requesting specific content be included —within project proposals;The range and scope of adaptations to GBV risk mitigation efforts during the COVID-19 pandemic has demonstrated how organizations and practitioners have creatively pivoted during an uncertain, challenging and highly dynamic situation. Further efforts to document the rich learning and experiences of practitioners engaged in this work would be beneficial;As reduced availability and access to GBV services has been reported in conjunction with perceived increases in GBV-related risks due to the COVID-19 pandemic and associated restrictions, ramping up of GBV risk mitigation interventions is more crucial than ever. These efforts must be prioritized and implemented in tandem with GBV prevention and response efforts. In addition, resources such as the GBV Pocket Guide  should bFurther research and evaluations are needed to more robustly assess the effectiveness of specific GBV risk mitigation interventions, both within and outside of the COVID-19 response, and to better understand what works and what does not in different sectors. This will ensure that practitioners who are already overstretched can concentrate their efforts along with the limited available resources on those interventions that will have the greatest impact.