AoB PLANTS

O_LIRationale: Plants in urban environments often experience heat stress and responses to heat stress often include vegetative and reproductive traits like rosette width and fruit morphology. However, our understanding of natural variation in vegetative and reproductive traits in urban environments is severely limited. C_LIO_LIMethods: We grew an urban weed, Capsella bursa-pastoris, in common garden environments that simulate an urban heat gradient to determine how heat affected growth, survival and reproduction. Additionally, we used geometric morphometric techniques alongside deterministic techniques to quantify variation in C. bursa-pastoris fruit shape and investigated the predictive relationship between fruit shape and seed production. C_LIO_LIKey results: We found that temperatures above 30C act as an environmental constraint on both C. bursa-pastoris fruit shape and reproduction, resulting in malformed fruits and no seed production. However, leaf number and plant survival were unaffected by high urban heat. C_LIO_LIMain conclusions: While plants may grow and survive in the high urban heat, heat could still limit population persistence. C_LI

Climatic and biogeographic variables are often used as a proxy for tree genetic diversity, but local factors can also influence it. We propose that woodland age, presence of ancient trees, and population size could impact genetic diversity. Using the RBG Kew UK National Tree Seed Project as a study case, we examined how these factors are accounted for during seed collection. We found 42% of tree seed collections come from ancient woodlands and that 8.4% overlap with ancient trees. Sampled forest patches size ranges from few individuals to several thousand. We then carried out a pilot to examine the role of population size on functional traits variation, testing the relationship between population size and seed germination and seedling thermal stress sensitivity in three populations of the Betula pubescens Ehrh. complex. We found that seeds and seedlings from larger populations showed higher fitness and stress resistance. Our results highlight the importance of local factors to predict variation in functional traits, relevant for tree resilience. Existing seed collections of native species stored in conservation seed banks offer a valuable resource to explore these factors and improve our understanding of genetic diversity in tree populations, with implications for biodiversity conservation and forestry production.

Gene flow to marginal populations at a species range edge can facilitate rapid adaptation by increasing genetic diversity, reducing inbreeding depression, and introducing novel alleles. In highly inbred populations, hybrid vigor is often observed in the first generation (F1), but hybrid breakdown may diminish fitness in subsequent generations. Thus, benefits of gene flow may be overestimated when only F1 performance is assessed. We tested whether gene flow among populations of the annual plant Erythranthe laciniata (A. Gray) G.L. Nesom, from similar and contrasting environments, confers persistent fitness advantages across F1 and F2 generations at the high-elevation edge of its range in the California Sierra Nevada. Gene flow was experimentally introduced through pollen transfer between cold-edge populations, between cold edge and central populations, and within local cold edge populations, and compared to self-fertilized offspring, the predominant mating strategy of E. laciniata. For F1 progeny, we measured morphological, phenological, and fitness traits in a common garden located near the cold-climate range limit during 2008-2009, a relatively average year, and for F2 progeny in 2009-2010, a relatively wet year. Although F1 crosses showed no initial performance advantage measured in the previous year, F2 progeny from center-to-edge and edge-to-edge crosses significantly outperformed selfed and locally outcrossed lines in fruit mass, total pedicels, biomass, and height. Our findings demonstrate that gene flow can confer long-term fitness benefits, especially among populations adapted to similar selective pressures, and highlight the potential of assisted gene flow to bolster or rescue peripheral populations facing climate change. SIGNIFICANCE STATEMENTSpecies living at the edges of their geographic ranges often have small, isolated populations with limited genetic diversity, which can restrict their ability to adapt to environmental change. Gene flow from other populations may increase adaptive potential, but its long-term consequences remain uncertain because most studies evaluate only first-generation hybrids. Using experimental crosses in the mountain wildflower Erythranthe laciniata, we show that gene flow can produce stronger fitness benefits in second-generation hybrids than in the first generation at a high-elevation range edge. These results suggest that recombination among populations can generate advantageous genetic combinations that emerge over multiple generations. Our findings highlight the potential for assisted gene flow to enhance adaptation and persistence of range-edge populations under climate change.

O_LIStored seeds are crucial repositories of plant genetic diversity. However, long-term storage inevitably causes seed deterioration and loss of viability, and chemical processes within the seeds during storage can influence germination and seedling establishment. Emerging evidence suggests that seed ageing can also affect traits of adult plants, yet the extent to which this phenomenon is relevant across species, particularly for wild plant species with high genetic variation, remains unclear. C_LIO_LITo address this, we focused on 14 grassland species and subjected their seeds to simulated long-term storage by exposing them to artificial ageing conditions (60% rH, 45{degrees}C). We then compared plants grown from the aged seeds with plants from fresh seeds in a common garden experiment. C_LIO_LIArtificially aged seeds germinated later, the developing seedlings had lower survival rates and reduced growth. Adult plants grown from aged seeds flowered later, produced fewer flowers, and had less biomass by the end of the first vegetation period than those from fresh seeds. The effect of the ageing treatment varied between species, but the trend was overall significant across species, with minor differences between perennials and annuals. Interestingly, in perennial plants, the effects vanished or were inverted in the second growing season, with plants growing from aged seeds flowering earlier and producing more biomass. C_LIO_LISynthesis. Our results show that seed storage affects seedling performance, plant growth, and flowering phenology. These direct storage effects should be considered when using stored seeds for species conservation, ecosystem restoration, or evolutionary research relying on stored seeds. C_LI

Growth kinematics and soil mechanics are key to explain how roots overcome the mechanical resistance of soil, yet few studies are linking these two factors. Formulas for cone penetration tests are typically used to infer the friction experienced by roots, but these fail to consider how growth affects the external forces applied on the root. This study formalised how expansive growth in the root apical meristem can reduce soil friction, and applied the framework to analyse the growth strategy of 6 plant species. The results of the analysis revealed trade-offs between reducing frictions, maintaining a desired growth trajectory and elongation rate. A shorter elongation zone can reduce the fraction of the mechanical energy lost to friction, but this is done at the expense of the elongation rate. A sharper tip or increased radius can help roots maintain the elongation rate at no energetic cost, but these strategies come with the cost of growth instability (tortuous roots) and decrease in specific root length respectively. During establishment, root strategies may therefore occupy a 2-dimensional trait space in which the mechanical efficiency of growth is balanced against the explorative-exploitative trade-off. HighlightsGrowth and form of root tips explain how plants overcome mechanical resistance from the soil Trade-offs link the energy lost by friction, growth stability and elongation rate of roots Larger roots allow faster growth independently of these trade-offs New framework formalises plants strategies to acquire soil resources

Restoration of coastal salt marshes is often limited by their capability to revegetate, either through natural recruitment or active planting methods. Despite the critical need for efficient revegetation methods, direct seeding remains an underrepresented approach in coastal wetland restoration. Additionally, tidal inundation poses special challenges for coastal Seed-Based Restoration (SBR) relative to terrestrial habitats, with tides displacing seeds from the marsh platform. To determine factors potentially influencing successful establishment in coastal wetlands, we conducted a series of greenhouse and lab experiments with Salicornia pacifica (pickleweed), the dominant plant in California marshes. We determined pickleweed seed viability using standardized germination tests. Additionally, we tested factors that influence pickleweed seed viability and germination rates, such as soil moisture, soil type, and sowing depth. We found that pickleweed seeds had an average viability of 22.5%, which increased with larger-sized seeds. We also determined that the most effective dormancy-breaking pretreatments varied by soil type: a one-day cold stratification in freshwater maximized germination in benign soils, whereas a seven-day cold stratification in saltwater maximized germination in stressful soils. Finally, we determined that the optimal conditions for sowing seeds are surface sowing under moderate moisture. Creating conditions to maximize viability and germination is crucial to ensure the greatest chance of successful revegetation post restoration. Our study, which sequentially tested factors affecting different phases of the early life history of a dominant foundation species, can inform SBR for other coastal plants. This approach can help coastal land managers successfully implement SBR for habitat restoration.

O_LIIn fluctuating environments, the kinetics of stomatal opening and closing influence the balance between carbon gain and water loss. Smaller guard cells may respond faster to fluctuating environmental conditions because of their greater surface area for osmolyte flux relative to cell volume. A related hypothesis is that operational stomatal conductance (gop) is often well below its theoretical maximum (gmax) because at this stomatal aperture, guard cell volume is poised to change rapidly with small changes in turgor pressure. C_LIO_LIWe analyzed 2,124 estimates of stomatal closure kinetics in response to an abrupt increase in vapor pressure deficit (VPD) among 29 diverse wild tomato populations in the genus Solanum. C_LIO_LILeaves with small guard cells and a lower gop to gmax ratio (fgmax) closed faster, but explained variation in kinetic parameters at different levels of biological organization. Guard cell size had high phylogenetic heritability and varied relatively little within populations, whereas fgmax varied mostly among individuals and between light intensity treatments. C_LIO_LISmaller stomata can be speedier, but only if stomata are held at an aperture where they are responsive to changing turgor pressure. Selection on stomatal speed may influence not only anatomical traits like guard cell size, but also physiological controls on gop. C_LI

O_LIAquatic algae are key primary producers in the Arctic and Antarctic, yet how cold-water species respond to environmental change is poorly understood. The Polar Regions are increasingly exposed to frequent heat waves, leading to declining ice cover, increased light availability, and decreasing salinity in polar waters. We compared three phylogenetically related but geographically distant polar Chlamydomonas species to test how habitat history shapes algal responses to light, salinity, and temperature stress. C_LIO_LIWe assessed the growth, morphology, and photochemistry of psychrophilic Chlamydomonas acclimated to native-like (lower light, higher salinity) and climate-shifted conditions (higher light, lower salinity). Next, we exposed acclimated cultures to a lethal heat shock and observed how acclimation affects algal temperature stress resilience. C_LIO_LIAll three species acclimated to climate-shifted conditions grew rapidly but showed the greatest sensitivity to temperature stress, with rapid loss of viability and photosynthetic efficiency. In contrast, slow-growing cultures acclimated to native-like conditions exhibited significantly greater resilience to temperature stress. C_LIO_LIOur work is the first to directly link light and salinity acclimation with temperature resilience in psychrophilic algae, suggesting that fast-growing polar green algae may be particularly vulnerable to increasingly frequent heat waves, with major implications for primary productivity in polar environments. C_LI

Urban environments typically experience higher temperatures than surrounding natural landscapes, making urban vegetation crucial for cooling local areas and improving the health of city residents. Impervious urban surfaces limit the absorption and retention of precipitation, potentially limiting tree water access and threatening long-term survival. Here, we measured tree physiology and growth of Lophostemon confertus (Queensland brush box) trees to investigate how a passive irrigation system that stores stormwater affected the performance of young, establishing trees in a hot and dry suburb of western Sydney, Australia. During the 2024-2025 austral summer, three years after planting, the local climate was periodically hot and dry, with a total of 16 days above 35 {degrees}C. Irrigated L. confertus trees had higher water availability (i.e., higher predawn leaf water potential,{Psi} pre), lower water stress (i.e., higher midday leaf water potential,{Psi} mid, more frequently above turgor loss point), greater stomatal conductance (gs) on hot and dry summer days, and reduced leaf temperatures (Tleaf), compared to control trees. No significant differences in growth rates were observed between irrigated and control trees during the first three establishment years, but irrigated trees had greater crown survival during the hot, dry summer. Our results suggest passive irrigation may mitigate periods of short-term heat and drought stress in urban trees by increasing water access to support transpiration that prevents leaves from overheating, improving tree health. Higher tree transpiration may lead to greater ecosystem services by increasing cooling benefits, contributing to mitigation of urban heat island effects.

O_LISubmerged macrophytes are central to freshwater ecosystems functioning but are declining globally under multiple anthropogenic stressors. We aimed to identify general patterns in physiological responses and interaction types, and to assess whether a mechanistic understanding of stressor interactions can be developed from published evidence. C_LIO_LIWe systematically reviewed 12,858 records, identified 172 relevant papers, and extracted effect sizes from 124 experiments included in the meta-analysis. C_LIO_LIMost studies examined combinations of nutrient enrichment, shading, toxic trace metals, warming, and emerging contaminants such as PFAS and microplastics, typically under simplified 2 x 2 factorial laboratory designs. Additive effects dominated (50%), while synergistic interactions were relatively infrequent (14%). Antagonistic interactions often reflected dominance of a single stressor or compensatory responses, whereas synergisms were most frequent with metals combined with co-stressors enhancing bioavailability. C_LIO_LIOur synthesis suggests that accumulated stressors cause negative, but not necessarily amplified, responses, although the limited number of experiments testing more than two stressors means synergistic effects may be underestimated. We propose Stuckenia pectinata as a model organism because of its cosmopolitan distribution, experimental tractability, and available genomic resources, and argue that expanding stressor complexity, duration, and taxonomic breadth will strengthen predictions of macrophyte responses and inform freshwater conservation under global change. C_LI

Winter warming is altering plant exposure to cold events, yet its effects on seasonal cold hardiness dynamics remain poorly understood. Here we quantified bud cold hardiness across four dormant seasons in a boreal peatland forest whole ecosystem warming experiment. Across a +0.00 to +9.00{degrees}C warming gradient, we semi-regularly measured cold hardiness in two overstory (Larix laricina and Picea mariana) and two understory species (Chamaedaphne calyculata and Rhododendron groenlandicum). Warming reduced cold hardiness in fall and spring by delaying acclimation and advancing deacclimation. However, risk was only increased in late winter and spring for three species. Warming reduced snow cover, increasing temperature variability and cold damage to understory shrubs. Together, our results show that cold damage risk depends on species traits, microclimate, and seasonal timing.

Crop improvement can enhance food security, but side effects, such as trade-offs between valuable agronomic traits, are common. Likewise, fertilisation helps ensure high yields, but can devalue mutualisms with soil microbes that would otherwise be essential for nutrient acquisition. If the need for nutritional mutualisms is reduced in crops, mutualisms could be disrupted by selection relaxation or allocation trade-offs. Thus, crops could achieve high yields in spite of, or because of, disruption of nutritional mutualisms. Alternatively, the highest-yielding plants might flourish because they maximise nutrient acquisition from both symbionts and the soil. Here, enhanced mutualism could evolve over the course of agricultural crop improvement. To investigate whether high yields in cultivars and wild accessions are negatively or positively genetically correlated with outcomes in the legume-rhizobia mutualism, we measured whether yield and symbiosis traits trade-off or are positively genetically correlated among cultivars and wild accessions. We also tested whether this relationship differs between accessions released before or after 1950. We measured genetic correlations between yield and mutualism traits in 87 domesticated pea (Pisum sativum) accessions in a common garden agricultural field across three years. Seed yield and N2 fixation (%Ndfa) were positively genetically correlated. While N fixation was more strongly predictive of yield in the pre-1950 accessions than the post-1950 accessions, the underlying positive genetic correlation between the traits did not differ between the groups. The positive genetic correlation between yield and N2 fixation indicates that selection to increase yields has maintained or increased the benefits of the rhizobial mutualism in pea. Our findings predict that breeding to increase yield may continue to produce pea cultivars that get a greater proportion of their N from rhizobia, enhancing symbiotic mutualism and reducing the proportion of N supplied by fertilisation.

CoverCress is a new winter annual oilseed crop developed from field pennycress within the past 20 years. Field pennycress is commonly considered to be self-pollinated but little basic research has been published and there is some misalignment of conclusions. Our experience working with pennycress plant growth in greenhouse and field conditions over the past 13 years suggests that outcrossing is uncommon. We conducted lab, greenhouse, and field experiments to strengthen the body of work. Pollen viability kinetics analysis showed that longevity of pollen viability is negatively impacted by increasing temperatures and by direct exposure to light. Samples treated at 4C declined to 50% viability in 12 hours while it took just 2.5 hrs at 37C, and 1.6 hrs in full sunlight on a cool early April day. Cross-pollination was absent among greenhouse-grown plants flowering inside an agitated plastic pollen-containment covering. Across greenhouse tests, high rates of cross-pollination occurred only in an emasculation treatment that rendered flowers male sterile and opened the pistil to cross-fertilization. Field trials designed to measure pollen flow distance using a trackable fae1 knockout reporter gene failed to show detectable movement of pollen under field conditions in two locations. This data strongly suggests that domesticated field pennycress may be considered a self-pollinated crop and managed as such.

Lack of sexual reproduction limits the fitness and long-term viability of many plant populations. This may pose a particular problem for populations at the edges of species ranges, which are often small and isolated and therefore may be less likely to attract pollinators. But despite the fact that many range-edge populations are of significant conservation concern and value, there is often little information about which visitors are effective pollinators, and few explicit tests of whether range-edge populations experience reduced pollination. Here, we assess which visitors are effective pollinators of sundial lupine (Lupinus perennis), a legume that is threatened in much of its range, and whether pollination success varies between populations in the range core and those at the species northern range edge. Across six populations in the northern USA and southern Canada (Ontario), sundial lupine was visited almost exclusively by bees, but only large bees (Bombus, Xylocopa) could be confirmed as effective pollinators in single-visit experiments. While seed production varied significantly among populations, visitation rates did not. Neither pollinator visitation, pollen receipt, nor seed production declined at sundial lupines northern range edge. We therefore found no evidence that pollination success constrains either performance of at-risk populations of sundial lupine or the species northern range limit.

Forest regeneration depends not only on how many seeds trees produce, but on the physiological quality of those seeds. Yet while climate-driven shifts in seed quantity and masting have received sustained attention, the parallel question of whether climate change degrades seed quality remains poorly resolved. Using a nationwide dataset of seed mass and viability in European beech (Fagus sylvatica L.) collected between 1996 and 2024 (13,349 seed lots from 381 forest districts across Poland), with climate-quality analyses focused on 5,374 freshly harvested seed lots from 353 districts (2004-2023), we asked whether the two components of seed quality respond to different seasonal climatic windows, and whether harvest-year climate also shapes seed performance during long-term cold storage. Seed mass and seed viability were only weakly correlated (Spearmans {rho} = 0.15), acting as two independent dimensions of seed quality. Both revealed substantial temporal variation over the study period, but along distinct trajectories. Seed mass declined markedly between segmented-regression breakpoints in 2009 and 2019, more steeply at higher latitudes, coinciding spatially and temporally with the masting breakdown reported at the species northeastern range margin. Climatic associations were correspondingly divergent. Viability was positively associated with previous summer temperature, consistent with temperature-cued flower initiation, and negatively with spring temperature in the harvest year, plausibly reflecting thermal disruption of early embryogenesis. Seed mass showed no significant association with any seasonal climatic predictor, indicating control by slower or unmeasured processes. Storage duration progressively reduced viability, and this decline was further modulated by climate during seed development, with seeds developing under climatically suboptimal conditions losing viability faster. These results expose a hidden decoupling between seed quantity and seed quality under contemporary climate change, with direct consequences for forest regeneration and for ex situ conservation strategies that assume mast-year seeds will remain viable for decades.

Arabidopsis thaliana naturally occurs across a wide geographic range and displays extensive natural variation in several traits including adaptive responses to the abiotic environment (e.g. temperature, drought, salt). Quantitative techniques like Genome Wide Association Studies (GWAS) enable mapping the genetic basis of such environmental responses and benefits from extensive genetic variation, but the size of the chosen diversity panel is often limited by phenotyping capacity. Most studies therefore use subpanels, often based on maximization of genetic diversity. However, this type of selection may overrepresent cosmopolitan alleles and underrepresent rare environment-specific alleles. Here, we demonstrate that the genetic variation in a GWAS subpanel of Arabidopsis thaliana accessions depends almost entirely on the number of accessions in the panel and very little on the composition of the panel. We present the EcoCore panel designed by grouping accessions of the 1001 genomes (1001G; 1135 accessions) collection, based on their native collection environment and selecting an equal number of accessions from each environment. We assessed hypocotyl lengths of plants grown at control and ambient high temperatures (20{degrees}C and 28{degrees}C) for 913 accessions of the 1001G and mapped these traits with the full 1001G panel versus the EcoCore panel. The EcoCore panel revealed novel genetic associations with hypocotyl length which is attributed to enrichment of alleles from rare environments. We present the EcoCore panel as a manageable resource for studying phenotypic plasticity and the genetic basis of plant-environment interactions.

Belowground, plants are exposed to a wide range of biotic stresses that vary in severity and nature, including tissue damage, disruption of vascular connectivity, and depletion of assimilates. How plants adapt their root systems to cope with different types of belowground biotic stresses is not well known. In this paper we compare above- and belowground plant adaptations to three nematode species with distinct tissue migration and feeding behaviours to study mechanisms underlying tolerance to different types of biotic stresses. We monitored both green canopy growth and changes in root system architecture of Arabidopsis inoculated with Pratylenchus penetrans, Heterodera schachtii, and Meloidogyne incognita. This revealed three distinct phases in aboveground plant responses: (i) initial growth inhibition associated with host invasion and tissue damage, (ii) persistent growth reduction associated with nematode sedentarism, and (iii) late growth stimulus in more advanced stages of infection. Specific adaptations in the root systems further revealed fundamentally different stress coping strategies. Tissue damage and intermittent feeding by P. penetrans in the root cortex did not induce significant changes in root system architecture. Tissue damage to the root cortex and prolonged feeding on host vascular cells by H. schachtii induced secondary root formation compensating for primary root growth inhibition. Prolonged feeding on host vascular cell by M. incognita alone did not induce secondary root formation, but was accompanied by typical local tissue swelling instead. Our data suggest that local secondary root formation and tissue swelling are two distinct compensatory mechanisms underlying tolerance to sedentarism by root-feeding nematodes. HighlightHow plants utilize root system plasticity to cope with different types of biotic stresses by root feeding nematodes remains largely unknown. Here, we report on specific adaptive growth responses in Arabidopsis roots to three nematode species, Pratylenchus penetrans, Heterodera schachtii, and Meloidogyne incognita, with fundamentally different strategies for host invasion, subsequent migration through host tissue, and feeding on host cells.

Lucerne is gaining interest as a living mulch in agroecological productions. However, its vigorous growth can lead to competition with cash crops for light and nutrients, necessitating new ideotypes. This study investigated the genetic basis of traits relevant to ideotype breeding: dormancy, spring regrowth, height, growth habit, leaflet size, stem diameter, and plant structure. Individuals from a diversity panel of 27 accessions and a synthetic population were phenotyped in a spaced plant nursery. Over 100,000 SNP markers were used for genotyping. Genome-wide association study (GWAS) and genomic prediction were conducted, considering population structure. Heritability estimates ranged from moderate to high in diversity panel (h{superscript 2} = 0.36-0.70) but were lower in synthetic population (h{superscript 2} = 0.17-0.33), reflecting reduced genetic variance. Trait correlations differed markedly between populations, indicating the possibility of recombining traits to create new ideotypes. GWAS identified a few QTL (r{superscript 2} up to 0.27) for leaflet size, height, growth habit, and plant structure, with candidate genes linked to growth, stress response, and signalling pathways. Genomic prediction was highly accurate in diversity panel, where broad genetic variation allowed reliable estimation of marker effects, with prediction accuracies exceeding 0.8 for heritable traits, including growth habit and leaflet size. In contrast, accuracies were low in synthetic population, reflecting its limited diversity and small size, whether training was based on the synthetic population itself or on the diversity panel. These results highlight the potential to recombine traits and develop lucerne ideotypes using molecular tools such as QTL detection and genomic prediction.

Flowering time and monocarpic senescence are tightly environmentally and genetically controlled. Typically, early flowering and staygreen traits are associated with opposing life-history strategies; stress avoidance versus adaptation; with flowering time an overarching regulator of crop cycle length. We developed RIL populations segregating for Ppd-1 and NAM-1 variation, which are otherwise isogenic. Multi-year field experiments enabled exploration and uncoupling of the relationship between heading and staygreen traits. Heading date manipulation enabled introduction of staygreen traits to their target breeding environments, characterised by a hot-finish. Under moderate stress, we report a 2.9% and 1.9% increase in grain width (P<0.0001), and 5.8% and 3.7% increase in TGW (P<0.0001), plus significantly greater yield (P<0.1) for late heading staygreen RILs homozygous for NAM-A1, and NAM-D1 missense variants, respectively. Grain yield increases were proportionate to the delay in senescence, being greater for the NAM-A1 than the NAM-D1 variant. For RIL populations segregating for both traits, senescence variation was observed relative to heading-date. Regarding grain yield, the staygreen trait-associated increase in source size could not compensate for the Ppd-1a associated pleiotropic reduction in sink size, even under hypothesised continental target breeding environments, with trait competition identified. Therefore, to maximise the benefits associated with staygreen traits, especially in early-heading favouring environments required targeted manipulation of source-sink dynamics, and we propose multiple strategies. HighlightStaygreen traits were associated with extending grain fill duration, increasing grain width, TGW and grain yield. There appears an antagonist relationship between earlier heading and staygreen traits.

Micro-dwarf tomato cultivars are increasingly considered for urban and controlled-environment agriculture due to their compact architecture and suitability for high-density planting. However, optimal canopy management strategies for these cultivars remain poorly defined. In this study, we evaluated the effects of different leaf removal intensities on leaf-level physiological performance, fruit yield, and fruit quality in three micro-dwarf tomato cultivars (Mohammed, Hahms Gelbe Topftomate, and Red Robin) grown under contrasting seasonal light conditions. Plants were subjected to low (15%), moderate (30%), or severe (90%) leaf removal, and leaf-level gas exchange was measured across canopy layers, along with yield and fruit quality assessments. Severe leaf removal (90%) increased carbon assimilation, transpiration, and stomatal conductance in middle and lower canopy leaves by up to approximately twofold compared with control plants, indicating improved light availability at the leaf level. However, these physiological enhancements did not consistently translate into higher yield, reflecting reduced whole-plant source capacity under excessive leaf removal. Low to moderate leaf removal (15-30%) generally increased or maintained yield and fruit number, whereas severe leaf removal reduced yield in Hahms Gelbe and Red Robin, particularly under low seasonal radiation. In contrast, Mohammed exhibited yield increases of up to 220% under low leaf removal and maintained increased yield even under severe leaf removal under high-light conditions. Fruit quality was largely unaffected by leaf removal, except for total soluble solids, which declined by approximately 12% under severe leaf removal across cultivars, consistent with sugar dilution under source limitation. Overall, these results demonstrate that optimal leaf removal in micro-dwarf tomatoes requires balancing improved canopy light distribution with maintenance of sufficient leaf area for carbon assimilation. Leaf removal thresholds are strongly cultivar- and light-dependent, emphasizing the need for cultivar-specific canopy management strategies in compact tomato systems and controlled-environment agriculture.