Physiological Constraints Research Articles

Efficient Fermentative Production of β-Alanine from Glucose through Multidimensional Engineering of Escherichia coli.

β-Alanine, a valuable β-type amino acid, is experiencing increased demand due to its multifaceted applications in food flavoring, nutritional supplements, pharmaceuticals, and the chemical industry. Nevertheless, the sustainable biosynthesis of β-alanine currently faces challenges due to the scarcity of robust strains, attributed to the complexities of modulating multiple genes and the inherent physiological constraints. Here, systems metabolic engineering was implemented in Escherichia coli to overcome these limitations. First, an efficient l-aspartate-α-decarboxylase (ADC) was recruited for β-alanine biosynthesis. To conserve phosphoenolpyruvate flux, we subsequently modified the endogenous glucose assimilation system by inactivating the phosphotransferase system (PTS) and introducing an alternative non-PTS system, which increased β-alanine production to 1.70 g/L. The supply of key precursors, oxaloacetate and l-aspartate, was synergistically improved through comprehensive modulation, including strengthening main flux and blocking bypass metabolism, which significantly increased the β-alanine titer to 3.43 g/L. Next, the expression of ADC was optimized by promoter and untranslated region (UTR) engineering. Further transport engineering, which involved disrupting β-alanine importer CycA and heterologously expressing β-alanine exporter NCgI0580, improved β-alanine production to 8.48 g/L. Additionally, corn steep liquor was used to develop a cost-effective medium. The final strain produced 74.03 g/L β-alanine with a yield of 0.57 mol/mol glucose during fed-batch fermentation.

Spatio-temporal organization of the E. coli chromosome from base to cellular length scales.

Escherichia coli has been a vital model organism for studying chromosomal structure, thanks, in part, to its small and circular genome (4.6 million base pairs) and well-characterized biochemical pathways. Over the last several decades, we have made considerable progress in understanding the intricacies of the structure and subsequent function of the E. coli nucleoid. At the smallest scale, DNA, with no physical constraints, takes on a shape reminiscent of a randomly twisted cable, forming mostly random coils but partly affected by its stiffness. This ball-of-spaghetti-like shape forms a structure several times too large to fit into the cell. Once the physiological constraints of the cell are added, the DNA takes on overtwisted (negatively supercoiled) structures, which are shaped by an intricate interplay of many proteins carrying out essential biological processes. At shorter length scales (up to about 1 kb), nucleoid-associated proteins organize and condense the chromosome by inducing loops, bends, and forming bridges. Zooming out further and including cellular processes, topological domains are formed, which are flanked by supercoiling barriers. At the megabase-scale both large, highly self-interacting regions (macrodomains) and strong contacts between distant but co-regulated genes have been observed. At the largest scale, the nucleoid forms a helical ellipsoid. In this review, we will explore the history and recent advances that pave the way for a better understanding of E. coli chromosome organization and structure, discussing the cellular processes that drive changes in DNA shape, and what contributes to compaction and formation of dynamic structures, and in turn how bacterial chromatin affects key processes such as transcription and replication.

Comparative mitogenomic analysis provides evolutionary insights into Formica (Hymenoptera: Formicidae).

Formica is a large genus in the family Formicidae with high diversity in its distribution, morphology, and physiology. To better understand evolutionary characteristics of Formica, the complete mitochondrial genomes (mitogenomes) of two Formica species were determined and a comparative mitogenomic analysis for this genus was performed. The two newly sequenced Formica mitogenomes each included 37 typical mitochondrial genes and a large non-coding region (putative control region), as observed in other Formica mitogenomes. Base composition, gene order, codon usage, and tRNA secondary structure were well conserved among Formica species, whereas diversity in sequence size and structural characteristics was observed in control regions. We also observed several conserved motifs in the intergenic spacer regions. These conserved genomic features may be related to mitochondrial function and their highly conserved physiological constraints, while the diversity of the control regions may be associated with adaptive evolution among heterogenous habitats. A negative AT-skew value on the majority chain was presented in each of Formica mitogenomes, indicating a reversal of strand asymmetry in base composition. Strong codon usage bias was observed in Formica mitogenomes, which was predominantly determined by nucleotide composition. All 13 mitochondrial protein-coding genes of Formica species exhibited molecular signatures of purifying selection, as indicated by the ratio of non-synonymous substitutions to synonymous substitutions being less than 1 for each protein-coding gene. Phylogenetic analyses based on mitogenomic data obtained fairly consistent phylogenetic relationships, except for two Formica species that had unstable phylogenetic positions, indicating mitogenomic data are useful for constructing phylogenies of ants. Beyond characterizing two additional Formica mitogenomes, this study also provided some key evolutionary insights into Formica.

A Multi-Tissue Genome-Scale Model of Populus trichocarpa Elucidates Overexpression Targets for Improving Drought Tolerance

Abstract Populus trichocarpa (poplar) is a fast-growing model tree whose lignocellulosic biomass is a promising biofuel feedstock. Enhancing its viability and yield in non-arable drought-prone lands can reduce biomass cost and accelerate adoption as a biofuel crop. Data from extensive -omics and phenotypic studies were leveraged herein to reconstruct a multi-tissue (root, stem, and leaf) genome-scale model (GSM) of poplar, iPotri3463, encompassing 14,360 reactions, 12,402 metabolites, and 3,463 genes. Two condition-specific GSMs were extracted from iPotri3463: iPotri3016C (control) and iPotri2999D (drought), supported by condition-specific transcript levels and reaction essentiality for growth. Physiological constraints consistent with experimental measurements of drought-stressed plants were imposed to growth, photorespiration, and carbon assimilation rates. Calculated increased flux capacity through the violaxanthin cycle and GABA biosynthetic pathways agree with established key strategies for improving drought tolerance. Differential gene expression analysis was performed on existing transcriptomes of poplar under different watering regimes. Computational flux knockdown was applied to reactions with increased flux capacity under drought which were associated with at least one downregulated gene. Several such reactions were essential for maintaining observed biomass yield and their associated genes are candidates for overexpression to improve drought tolerance. Glutamine synthetase is one whose overexpression in poplar confirms in silico predictions. However, the two most promising candidates are genes encoding ferulate-5-hydroxylase, Potri.007G016400 and Potri.005G117500, as their overexpression in other plant species led to demonstrably improved drought tolerance while previous overexpression in poplar reduced biomass recalcitrance. iPotri3463 is the first poplar-specific whole-plant GSM and the second one available for a woody plant.

Evaluation of extruded starch foam for glucose-supplying biomaterials

The survival rate of mesenchymal stem cells (MSC), a crucial factor in tissue engineering, is highly dependent on glucose supply. The purpose of this paper is to study the potential of starch foams as glucose suppliers. It is investigated through in vitro hydrolysis by amyloglucosidase in conditions that respect physiological constraints (37 °C and pH 7.4), including a duration of 21 days, and no stirring. Nine extruded starch foams with amylose contents ranging from 0 to 74 %, with various cell wall thicknesses (50 to 300 μm), and different crystallinities (0–30 %) were hydrolysed. These kinetics were fitted by a model which shows that the maximum rate of hydrolysis varies from 7 to 100 %, and which allows the rate of hydrolysis at 21 days to be calculated precisely. The results reveal the major role of amylose in glucose delivery kinetics, and the secondary roles of crystallinity and cell wall thickness of the foams. Additional hydrolysis of starch films revealed that thickness positively influences the amylose chain reorganisation during hydrolysis, which, in slows down and limits glucose delivery. A simple glucose delivery kinetics analysis procedure is proposed to select samples for testing as MSC glucose suppliers.

Proteome partitioning constraints in long-term laboratory evolution

Adaptive laboratory evolution experiments provide a controlled context in which the dynamics of selection and adaptation can be followed in real-time at the single-nucleotide level. And yet this precision introduces hundreds of degrees-of-freedom as genetic changes accrue in parallel lineages over generations. On short timescales, physiological constraints have been leveraged to provide a coarse-grained view of bacterial gene expression characterized by a small set of phenomenological parameters. Here, we ask whether this same framework, operating at a level between genotype and fitness, informs physiological changes that occur on evolutionary timescales. Using a strain adapted to growth in glucose minimal medium, we find that the proteome is substantially remodeled over 40 000 generations. The most striking change is an apparent increase in enzyme efficiency, particularly in the enzymes of lower-glycolysis. We propose that deletion of metabolic flux-sensing regulation early in the adaptation results in increased enzyme saturation and can account for the observed proteome remodeling.

Eukaryotic ribosome display for antibody discovery: A review.

Monoclonal antibody biologics have significantly transformed the therapeutic landscape within the biopharmaceutical industry, partly due to the utilisation of discovery technologies such as the hybridoma method and phage display. While these established platforms have streamlined the development process to date, their reliance on cell transformation for antibody identification faces limitations related to library diversification and the constraints of host cell physiology. Cell-free systems like ribosome display offer a complementary approach, enabling antibody selection in a completely in vitro setting while harnessing enriched cellular molecular machinery. This review aims to provide an overview of the fundamental principles underlying the ribosome display method and its potential for advancing antibody discovery and development.

Evolutionary trade-offs in dormancy phenology.

Seasonal animal dormancy is widely interpreted as a physiological response for surviving energetic challenges during the harshest times of the year (the physiological constraint hypothesis). However, there are other mutually non-exclusive hypotheses to explain the timing of animal dormancy, that is, entry into and emergence from hibernation (i.e. dormancy phenology). Survival advantages of dormancy that have been proposed are reduced risks of predation and competition (the 'life-history' hypothesis), but comparative tests across animal species are few. Using the phylogenetic comparative method applied to more than 20 hibernating mammalian species, we found support for both hypotheses as explanations for the phenology of dormancy. In accordance with the life-history hypotheses, sex differences in hibernation emergence and immergence were favored by the sex difference in reproductive effort. In addition, physiological constraint may influence the trade-off between survival and reproduction such that low temperatures and precipitation, as well as smaller body mass, influence sex differences in phenology. We also compiled initial evidence that ectotherm dormancy may be (1) less temperature dependent than previously thought and (2) associated with trade-offs consistent with the life-history hypothesis. Thus, dormancy during non-life-threatening periods that are unfavorable for reproduction may be more widespread than previously thought.

Evolutionary trade-offs in dormancy phenology

Seasonal animal dormancy is widely interpreted as a physiological response for surviving energetic challenges during the harshest times of the year (the physiological constraint hypothesis). However, there are other mutually non-exclusive hypotheses to explain the timing of animal dormancy, that is, entry into and emergence from hibernation (i.e. dormancy phenology). Survival advantages of dormancy that have been proposed are reduced risks of predation and competition (the ‘life-history’ hypothesis), but comparative tests across animal species are few. Using the phylogenetic comparative method applied to more than 20 hibernating mammalian species, we found support for both hypotheses as explanations for the phenology of dormancy. In accordance with the life-history hypotheses, sex differences in hibernation emergence and immergence were favored by the sex difference in reproductive effort. In addition, physiological constraint may influence the trade-off between survival and reproduction such that low temperatures and precipitation, as well as smaller body mass, influence sex differences in phenology. We also compiled initial evidence that ectotherm dormancy may be (1) less temperature dependent than previously thought and (2) associated with trade-offs consistent with the life-history hypothesis. Thus, dormancy during non-life-threatening periods that are unfavorable for reproduction may be more widespread than previously thought.

Aeroponics vertical indoor farming

Aeroponics vertical indoor farming is a farming that involves growing crops in layers that are piled vertically. Aeroponics is a method of growing plants without soil. Vertical indoor farming is climate controlled so it can be done year-round regardless of environmental challenges. Emerging problems of food security, urbanization, farmland shortage, food miles increased greenhouse gas emissions focus on the need for vertical farming physiological constraints that occur within hydroponic systems. Today, agriculture especially in India to concentrate in some situation such as how to increase the productivity and profit. This paper presents a methodology for developing the plants are submerged in a nutrient-rich mist solution. This study challenges is cultivating the plant by organic farming natural nutrition.

Chinook salmon depth distributions on the continental shelf are shaped by interactions between location, season, and individual condition

BackgroundEcological and physical conditions vary with depth in aquatic ecosystems, resulting in gradients of habitat suitability. Although variation in vertical distributions among individuals provides evidence of habitat selection, it has been challenging to disentangle how processes at multiple spatio-temporal scales shape behaviour.MethodsWe collected thousands of observations of depth from >300\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$>~300$$\\end{document} acoustically tagged adult Chinook salmon Oncorhynchus tshawytscha, spanning multiple seasons and years. We used these data to parameterize a machine-learning model to disentangle the influence of spatial, temporal, and dynamic oceanographic variables while accounting for differences in individual condition and maturation stage.ResultsThe top performing machine learning model used bathymetric depth ratio (i.e., individual depth relative to seafloor depth) as a response. We found that bathymetry, season, maturation stage, and spatial location most strongly influenced Chinook salmon depth. Chinook salmon bathymetric depth ratios were deepest in shallow water, during winter, and for immature individuals. We also identified non-linear interactions among covariates, resulting in spatially-varying effects of zooplankton concentration, lunar cycle, temperature and oxygen concentration.ConclusionsOur results suggest Chinook salmon vertical habitat use is a function of ecological interactions, not physiological constraints. Temporal and spatial variation in depth distributions could be used to guide management decisions intended to reduce fishery impacts on Chinook salmon. More generally, our findings demonstrate how complex interactions among bathymetry, seasonality, location, and life history stage regulate vertical habitat selection.

Precipitation variation: a key factor regulating plant diversity in semi-arid livestock grazing lands.

Livestock presence impacts plant biodiversity (species richness) in grassland ecosystems, yet extent and direction of grazing impacts on biodiversity vary greatly across inter-annual periods. In this study, an 8-year (2014-2021) grazing gradient experiment with sheep was conducted in a semi-arid grassland to investigate the impact of grazing under different precipitation variability on biodiversity. The results suggest no direct impact of grazing on species richness in semi-arid Stipa grassland. However, increased grazing indirectly enhanced species richness by elevating community dominance (increasing the sheltering effect of Stipa grass). Importantly, intensified grazing also regulates excessive community biomass resulting from increased inter-annual wetness (SPEI), amplifying the positive influence of annual humidity index on species richness. Lastly, we emphasize that, in water-constrained grassland ecosystems, intra-annual precipitation variability (PCI) was the most crucial factor driving species richness. Therefore, the water-heat synchrony during the growing season may alleviate physiological constraints on plants, significantly enhancing species richness as a result of multifactorial interactions. Our study provides strong evidence for how to regulate grazing intensity to increase biodiversity under future variable climate patterns. We suggest adapting grazing intensity according to local climate variability to achieve grassland biodiversity conservation.

Pushing the limits of C3 intrinsic water use efficiency in Mediterranean semiarid steppes: Responses of a drought‐avoider perennial grass to climate aridification

Abstract Intrinsic water use efficiency (WUEi) reflects the trade‐off between photosynthetic carbon gain and water loss through stomatal conductance and is key for understanding dryland plant responses to climate change. Stipa tenacissima is a perennial tussock C3 grass with an opportunistic, drought‐avoiding water use strategy that dominates arid and semiarid steppes across the western Mediterranean region. However, its ecophysiological responses to aridification and woody shrub encroachment, a major land‐use change in drylands worldwide, are not well‐understood. We investigated the variations in leaf stable isotopes (δ18O, δ13C, δ15N), nutrient concentrations (N, P, K), and culm water content and isotopic composition (δ18O, δ2H) of paired pure‐grass and shrub‐encroached S. tenacissima steppes along a 350 km aridity gradient in Spain (10 sites, 160 individuals). Culm water isotopes revealed that S. tenacissima is a shallow‐rooted grass that depends heavily on recent rainwater for water uptake, which may render it vulnerable to increasingly irregular rainfall combined with faster topsoil drying under climate warming and aridification. With increasing aridity, S. tenacissima enhanced leaf‐level WUEi through more stringent stomatal regulation of plant water flux and carbon assimilation (higher δ13C and δ18O), reaching exceptionally high δ13C values (−23‰ to −21‰) at the most arid steppes. Foliar N concentration was remarkably low across sites regardless of woody shrub encroachment, evidencing severe water and N co‐limitation of photosynthesis and productivity. Shrub encroachment decreased leaf P and K but did not affect S. tenacissima water status. Perennial grass cover decreased markedly with both declining winter rainfall and shrub encroachment suggesting population‐level rather than individual‐level responses of S. tenacissima to these changes. The fundamental physiological constraints of photosynthetic C3 metabolism combined with low foliar N content may hamper the ability of S. tenacissima and other drought‐avoider species with shallow roots to achieve further adaptive improvements in WUEi under increasing climatic stress. A drought‐avoiding water use strategy based on early stomatal closure and photosynthesis suppression during prolonged rainless periods may thus compromise the capacity of semiarid S. tenacissima steppes to maintain perennial grass cover, sustain productivity and cope with ongoing climate aridification at the drier parts of their current distribution. Read the free Plain Language Summary for this article on the Journal blog.

Human–Robot Collaboration in Modern Agriculture: A Review of the Current Research Landscape

Recent years have witnessed an increased utilization of robotics systems in agricultural settings. While fully autonomous farming holds great potential, most systems fall short of meeting the demands of present‐day agricultural operations. The use of human labor or teleoperated robots is also limited due to the physiological constraints of humans and the shortcomings of interfaces used to control robots. To harness the strengths of autonomous capabilities and endurance of robots, as well as the decision‐making capabilities of humans, human–robot collaboration (HRC) has emerged as a viable approach. By identifying the various applications of HRC in current research and the infrastructure employed to develop them, interested parties seeking to utilize collaborative robotics in agriculture can gain a better understanding of the possibilities and challenges they may encounter. In this review, an overview of existing HRC applications in the agricultural domain is provided. Additionally, general trends and weaknesses are identified within the research corpus. This review serves as a presentation of the state‐of‐the‐art research of HRC in agriculture for professionals considering the adoption of HRC. Robotics engineers can utilize this review as a resource for easily accessing information on the hardware, software, and algorithms employed in building HRC systems for agriculture.

The incredible shrinking puffin: Decreasing size and increasing proportional bill size of Atlantic puffins nesting at Machias Seal Island.

Climate change imposes physiological constraints on organisms particularly through changing thermoregulatory requirements. Bergmann's and Allen's rules suggest that body size and the size of thermoregulatory structures differ between warm and cold locations, where body size decreases with temperature and thermoregulatory structures increase. However, phenotypic plastic responses to malnutrition during development can result in the same patterns while lacking fitness benefits. The Gulf of Maine (GOM), located at the southern end of the Labrador current, is warming faster than most of the world's oceans, and many of the marine species that occupy these waters exist at the southern edge of their distributions including Atlantic puffins (Fratercula arctica; hereafter "puffin"). Monitoring of puffins in the GOM, at Machias Seal Island (MSI), has continued annually since 1995. We asked whether changes in adult puffin body size and the proportional size of bill to body have changed with observed rapid ocean warming. We found that the size of fledgling puffins is negatively related to sea surface temperature anomalies (warm conditions = small fledgers), adult puffin size is related to fledgling size (small fledgers = small adults), and adult puffins have decreased in size in recent years in response to malnutrition during development. We found an increase in the proportional size of bill to wing chord, likely in response to some mix of malnutrition during development and increasing air temperatures. Although studies have assessed clinal variation in seabird morphology with temperature, this is the first study addressing changes in seabird morphology in relation to ocean warming. Our results suggest that puffins nesting in the GOM have morphological plasticity that may help them acclimate to ocean warming.

Land use effects on soil microbiome composition and traits with consequences for soil carbon cycling.

The soil microbiome determines the fate of plant-fixed carbon. The shifts in soil properties caused by land use change leads to modifications in microbiome function, resulting in either loss or gain of soil organic carbon (SOC). Soil pH is the primary factor regulating microbiome characteristics leading to distinct pathways of microbial carbon cycling, but the underlying mechanisms remain understudied. Here, the taxa-trait relationships behind the variable fate of SOC were investigated using metaproteomics, metabarcoding, and a 13C-labeled litter decomposition experiment across two temperate sites with differing soil pH each with a paired land use intensity contrast. 13C incorporation into microbial biomass increased with land use intensification in low-pH soil but decreased in high-pH soil, with potential impact on carbon use efficiency in opposing directions. Reduction in biosynthesis traits was due to increased abundance of proteins linked to resource acquisition and stress tolerance. These trait trade-offs were underpinned by land use intensification-induced changes in dominant taxa with distinct traits. We observed divergent pH-controlled pathways of SOC cycling. In low-pH soil, land use intensification alleviates microbial abiotic stress resulting in increased biomass production but promotes decomposition and SOC loss. In contrast, in high-pH soil, land use intensification increases microbial physiological constraints and decreases biomass production, leading to reduced necromass build-up and SOC stabilization. We demonstrate how microbial biomass production and respiration dynamics and therefore carbon use efficiency can be decoupled from SOC highlighting the need for its careful consideration in managing SOC storage for soil health and climate change mitigation.

Barriers and enablers of women carers from culturally and linguistically diverse communities participating in physical activities.

This research article presents a phenomenological evaluation of an organization working within disability and with CALD communities of a project that was supporting women carers who are newly arrived to Australia to be happier and healthier. The initiative aimed to alleviate social isolation, enhance wellbeing, and boost confidence among women from Culturally and Linguistically Diverse (CALD) backgrounds in Metropolitan Melbourne, Victoria. Through a qualitative exploration of the lived experiences of participants, findings were categorized into enablers and barriers, shedding light on the outcomes of physical activity participation for these women. Enablers included improvements in health, skill development, achievement, and enhanced social interaction and support, while barriers encompassed challenges related to access, socio-cultural factors, resource availability, interpersonal dynamics, and physiological constraints. Based on these findings, recommendations are proposed to empower CALD community members in the co-development and co-delivery of future programs, fostering sustainability and community-driven engagement.

Modelling individual tree maximum basal area growth rates of five tall eucalypt species growing in even-aged forests

Inventory plot data were available providing over 87,000 individual tree basal area growth rates from even-aged native forests of three ash eucalypts (Eucalyptus regnans, E. obliqua, and E. delegatensis), from temperate regions, and two other species from more sub-tropical climes (E. grandis and E. pilularis). Models were developed relating maximum observed growth rates for these species in relation to tree size when, presumably, trees were under ideal environmental conditions and without competition from neighbours for site growth resources. These maximum growth rates increased with increasing tree size to a maximum of their own and then declined as tree size (hence age) increased further. The tree sizes, at which these maximum growth rates reached their maxima, were much greater for the ash eucalypts than for the other two species. It is hypothesised that the ash eucalypts may have evolved physiological characteristics that make them more efficient in compensating for the well-known physiological constraints imposed on growth rates as trees grow to great heights and ages.

Modeling and analysis of a multilayer solid tumour with cell physiological age and resource limitations

We study an avascular spherical solid tumour model with cell physiological age and resource constraints in vivo. We divide the tumour cells into three components: proliferating cells, quiescent cells and dead cells in necrotic core. We assume that the division rate of proliferating cells is nonlinear due to the nutritional and spatial constraints. The proportion of newborn tumour cells entering directly into quiescent state is considered, since this proportion can respond to the therapeutic effect of drug. We establish a nonlinear age-structured tumour cell population model. We investigate the existence and uniqueness of the model solution and explore the local and global stabilities of the tumour-free steady state. The existence and local stability of the tumour steady state are studied. Finally, some numerical simulations are performed to verify the theoretical results and to investigate the effects of different parameters on the model.

No evidence for fractal scaling in canopy surfaces across a diverse range of forest types

Abstract Canopy structural complexity is a key emergent property of forest ecosystems that directly relates to their ability to store carbon, cycle water and nutrients, and provide habitat for biodiversity. However, we lack a general framework for quantifying the structural complexity of forest canopies, and it remains to be seen if there are simple rules that explain the huge variation in canopy structure that we observe across the world's forests. A leading candidate for characterizing differences in structural complexity among forest ecosystems are fractal scaling laws, or measures of statistical self‐similarity. If forest canopies were fractal in nature, their structural attributes could be distilled into a single coefficient—their fractal dimension—which could then be used to directly compare structural differences within and across biomes. To test this idea, we used airborne laser scanning (ALS) data acquired across nine landscapes in Australia that span a huge environmental gradient and include everything from dry shrublands, tropical savannas, dense rainforests, to 90 m tall Mountain Ash forests. Using the ALS data, we built high‐resolution 3D canopy height models of each landscape so that we could quantify how closely they followed fractal scaling laws, based on a comprehensive set of fractal dimension estimators. Across all ecosystem types, fractal scaling assumptions were consistently violated, with clear and systematic differences between real‐world forest canopies and simulated fractal surfaces. However, deviations from fractality did vary predictably among sites, with forests in less arid environments, dominated by tall trees with large crowns, exhibiting a higher degree of self‐similarity across scales. Synthesis: Our study conclusively shows that canopy surfaces are not fractal beyond the scale of individual tree crowns. Nevertheless, we still observed ecologically meaningful and generalisable patterns in forest structure across scales, which closely reflect biophysical and physiological constraints on tree size and architecture. These patterns point the way towards a more general framework for characterizing ecological complexity.