American Journal of Botany

Premise Rising temperatures can reduce plant reproductive success by disrupting pollen development, yet little is known about how reproductive heat sensitivity varies among natural populations. We evaluated pollen viability responses to elevated temperature in two closely related taxa of yellow monkeyflowers across multiple geographic scales to determine whether heat sensitivity varies among species, between populations and across elevations. Methods We conducted three common garden growth chamber experiments. First, we compared pollen viability responses to heat stress between Mimulus guttatus and members of the M. tilingii species complex. Second, we tested whether broad elevational categories explained variation in heat responses. Third, we evaluated heat sensitivity among nine eastern Sierra Nevada populations of M. guttatus spanning an elevational gradient. Pollen viability was assessed following exposure to control and heat treatments. Key results Heat exposure reduced pollen viability in all experiments. In the species comparison experiment, pollen viability decreased more strongly in the high-elevation restricted M. tilingii species complex than the widespread M. guttatus. We found substantial population-level variation in pollen viability within M. guttatus but not members of the M. tilingii complex. Broad elevational categories did not explain this variation in pollen viability responses. However, within eastern Sierra Nevada populations, reductions in pollen viability became stronger with increasing source elevation. Several populations deviated from this trend, indicating reproductive responses to heat cannot be explained by elevation alone. Conclusions Pollen heat sensitivity varies substantially among species and populations of yellow monkeyflowers. Elevation alone is insufficient to predict pollen viability responses to elevated temperatures, suggesting that local adaptation or genetic variation may contribute to reproductive heat tolerance. Some populations of M. guttatus may contain greater capacity to maintain reproductive function under future warming, highlighting the importance of considering intraspecific variation when predicting plant responses to climate change.

Premise of the study. Phenotypic plasticity is advantageous for many plants that experience environmental fluctuations, but strong selection pressures can establish limits to plasticity. In deciduous trees, the ability to time leaf elongation in response to spring temperatures can extend the length of the growing season for a given year's climate. However, emergence timing may have undergone strong selection by factors like cold temperatures, limiting the capacity of locally adapted populations to adjust via plasticity to new conditions. Methods. Using a nine-year dataset of 659 families of Quercus lobata in two common gardens, we tested how climate of origin and annual temperatures at the gardens affected variation in phenology. We then examined whether leaf emergence timing was under selection in the gardens by testing its effects on growth. Finally, we tested whether among-year plasticity in leaf emergence timing conferred a growth benefit. Key results. Year-to-year environmental variation explained more phenological variation than genetic differences associated with climate of origin, yet trees from cooler climates were more constrained in their ability to track temperature than trees from warmer climates. We found evidence of weak stabilizing and directional selection on phenology, suggesting that strong selection in favor of earlier phenology is unlikely. Finally, we found no evidence that selection has favored increased plasticity in Q. lobata. Conclusions. Phenological plasticity remains a key driver of plants' response to changing temperatures, but it is constrained by natural selection. As temperatures rise, these processes may disproportionately limit the ability of trees adapted to cooler climates to shift their phenology to match spring temperatures.

Myricaria Desv. (Tamaricaceae) is a genus of shrubs that play a keystone role in montane riparian ecosystems across Eurasia, shaping riverine habitats and supporting ecological function. The genus has a centre of origin in the Qinghai-Tibetan Plateau and exhibits a broad distribution, but many species boundaries are unresolved. Here, we present the first phylogenomic analysis of Myricaria using the Angiosperms353 target enrichment bait set, with sampling across its full geographic range. Off-target plastid loci were extracted from next-generation sequencing data using the CAPTUS bioinformatics pipeline. We infer species relationships using ASTRAL-IV and network analysis with SplitsTree, in conjunction with morphology. We also provide a taxonomic treatment of the genus, integrating our morphological observations and phylogenomic analyses for species delimitations. Based on our analyses, recognise two infrageneric sections: sect. Parallelantherae (containing only M. elegans) and sect. Myricaria (the remainder of the genus, with type species M. germanica). We also propose synonymising M. davurica, M. bracteata and M. squamosa with M. germanica. Although species boundaries in the Asian highlands remain unresolved, we conclude that the species M. elegans, M. germanica, M. laxiflora, M. longifolia, M. platyphylla, and M. pulcherrima are well-supported both phylogenomically and morphologically.

Self-fertilization results in reduction of heterozygosity over time, accelerated by small population sizes, restricted ranges, and fragmented habitats. Yet some species maintain heterozygosity despite these factors. One is Corallorhiza bentleyi, a threatened, mycoheterotrophic, self-pollinating orchid restricted to Virginia and West Virginia, USA. Reduced representation sequencing was employed across five of the ~12 known localities of C. bentleyi, including representatives of sister species C. involuta from Mexico, to quantify patterns of population structure and genomic diversity. Surveys of C. bentleyi revealed a predominance of the cleistogamous (closed-flowered) growth form. Differentiation was observed between the two species but with weak population structure in C. bentleyi. Surprisingly, excess heterozygosity and negative inbreeding were recovered in both species, counter to expectations. Analyses provided evidence against bioinformatic artifacts (collapsed paralogs), small-sample bias (Robertson effect), and accumulation of mutations along chromosomes due to low recombination associated with selfing (Meselson effect) as explanations for excess heterozygosity. Demographic analyses favored a model of post-glacial divergence among C. bentleyi and C. involuta lacking bottlenecks. Finally, simulations under alternative selection scenarios favored models of overdominance vs. those of negative frequency dependent selection. It was concluded that overdominance, namely selection against selfed homozygotes, is the most likely explanation for elevated heterozygosity in C. bentleyi. Conservation concerns are discussed in light of these findings and the threats of extinction for one of the most vulnerable orchid species in North America.

Bulk phloem flow is driven by pressure gradients generated by water drawn from the xylem, coupling source-sink transport with water status. Huanglongbing (HLB), a phloem-affecting bacterial disease of citrus, disrupts phloem transport, and causes canopy water deficit. Although improved water management can enhance HLB-affected tree health, the effects of HLB on water-driven source-sink relations are unknown. Fruits function as both carbon sink and water reservoir, but the role of fruit in regulating whole-tree water status remain poorly understood. We examined leaf-fruit water relations in HLB-affected citrus and evaluated how HLB exposure duration, fruit developmental stage, and source-sink manipulation shape these dynamics. Source-sink relations were manipulated in branches of symptomatic trees with long-term (four years) and recent (six months) HLB exposure. We girdled branches with a single fruit as the sink and defoliated at (0%, 50%, or 75%), and an ungirdled, non-defoliated branch served as control. We measured stomatal conductance, leaf hydraulic traits, leaf and fruit soluble sugars, diurnal fruit shrinkage (to estimate xylem backflow) and fruit water loss. Source-sink manipulation did not affect shoot total soluble sugar. HLB-exposure resulted in less negative osmotic potential, more negative turgor potential and reduced modulus of elasticity and relative water content at turgor loss point. HLB exposure also increased diurnal fruit shrinkage, with shrinkage responses to source-sink manipulation varying across fruit development stages. Collectively, these findings suggest that long-term HLB exposure promotes leaf acclimation to water deficits, however, high sink demand may diminish the leaf water status regulation at Stage II, when fruit sink carbon demand is highest.

The Asparagaceae is an economically and ecologically important angiosperm (monocot) clade, containing iconic garden plants such as asparagus, hyacinth and agave, and arid-adapted keystone species like Joshua tree and dragon blood tree. The approximately 2,500 species in the family are spread across seven subfamilies: Agavoideae, Aphyllanthoideae, Asparagoideae, Brodiaeoideae, Lomandroideae, Nolinoideae and Scilloideae. Relationships among the seven subfamilies are generally unresolved and the sources of gene tree-species tree discordance are largely unknown. Investigating the biology underlying such discordance requires a robust sampling of both taxa and molecular markers, due to the extremely short branches that form the backbone of the Asparagaceae phylogeny. Using targeted sequencing of 1,726 low-copy nuclear genes, and a nearly complete sampling of genera, we interrogate relationships among all seven Asparagaceae subfamilies and investigate the underlying sources of discordance across the phylogeny. Analyses using different phylogenomic approaches (multispecies coalescence vs. nuclear gene supermatrix vs. chloroplast gene supermatrix) each revealed extensive discordance across the Asparagaceae backbone, which may be associated with rapid lineage diversifications that occurred in Africa shortly following the origin of the family ~49 Ma. Specifically, we found that two branches, representing ancestors of Aphyllanthoideae and Nolinoideae, may be responsible for much of the backbone discordance. The placement of Aphyllanthoideae, represented by one extant taxon (Aphyllanthes monspeliensis), is controversial with discordant topologies across many studies. However, multispecies coalescence-based analyses from this study strongly support a sister relationship between Aphyllanthoideae and a larger Agavoideae + Scilloideae + Brodiaeoideae clade. According to gene tree quartet frequencies and ABBA-BABA tests, we cannot reject incomplete lineage sorting (ILS) as the main source of discordance for the placement of Aphyllanthoideae; our results leave open the possibility of an ancient reticulation involving an ancestor of Aphyllanthoideae and an extinct basal Asparagaceae lineage. Regarding Nolinoideae, we found that conflicting support for two possible bifurcating clades (Asparagoideae + Nolinoideae vs. Asparagoideae + Lomandroideae) was associated with skewed support for alternative gene tree topologies, suggesting that phenomena other than ILS may explain the observed discordance. A quartet-based ABBA-BABA analysis, used to test for signatures of introgression, revealed a putative ancient hybridization event between two divergent, ancestral lineages which may have contributed to the origin of the Nolinoideae clade in Africa ~45 Ma. In sum, this study elucidates the long-troublesome backbone of the Asparagaceae phylogeny and advances our understanding of the evolutionary and biogeographical origins of all extant subfamilies. More broadly, this study expands our understanding of the various sources of gene tree discordance that may arise from natural processes associated with rapid lineage radiations across the tree of life.

Streambank erosion degrades both terrestrial and aquatic ecosystems, and riparian buffers are among the most widely used tools to control it. Yet most of what is known about how riparian vegetation controls erosion comes from one or two-species systems, which overlook the diversity of riparian communities and species interactions (competition and facilitation) that may shape the root systems that bind soil. We used a streambank mesocosm experiment to test whether plant species richness alters root traits, and whether root traits and species interactions reduce erosion. We established 85 mesocosms ranging from unvegetated controls to six-species polycultures, measured root biomass, length, volume, and diameter, quantified species interactions with relative yield totals (RYT), and measured erosion rate with a jet erosion test. Vegetated mesocosms eroded significantly less than unvegetated controls (t = 4.05, p = 0.023). Richness did not affect most root traits, although mean root diameter decreased as richness increased ({beta} = -0.0225, p = 0.047). RYT ranged from competition (RYT < 1) to facilitation (RYT >1), yet neither interactions nor any root trait was directly associated with erosion rate. However, RYT was negatively associated with soil moisture ({beta} = -0.196, p = 0.010). Piecewise structural equation models showed that erosion was best explained by a pathway linking species presence, root length, and soil moisture (AIC = 39.27; Fisher's C = 11.27, df = 12, p = 0.506). Panicum virgatum increased root length, whereas Lolium multiflorum and Echinochloa crus-galli reduced it; longer roots were associated with drier soil (standardized {beta} = -0.31, p = 0.005), and wetter soils eroded more slowly (standardized {beta} = -0.39, p < 0.001). Which species were present mattered more than how many, and plants were linked to erosion through soil moisture rather than root traits alone. Species selection for riparian buffers may need to consider effects on soil moisture as well as rooting extent, particularly as droughts followed by intense storms become more common under a changing climate.

Understanding spatial and temporal variation in community composition is central to protecting and monitoring biodiversity. Similarity in the composition of two communities decreases along spatial and temporal gradients, a relationship known as distance decay. Comparing distance-decay relationships across taxonomic, functional, and bioclimatic facets of diversity provides a means of interrogating niche-based versus neutral influences on community assembly. Furthermore, the context in which these facets yield overlapping or complementary information about global change impacts remains poorly understood. In this study, we examined spatiotemporal distance-decay relationships for taxonomic, functional, and bioclimatic facets of alpine plant diversity leveraging elevation gradients on five peaks in the White Mountains, California, USA, sampled at 3-4 time points between 2007 and 2024. Using multivariate Bayesian beta regression models, we estimated the slopes of elevation distance decay over multiple surveys and across taxonomic, functional and bioclimatic diversity to understand how spatial distance-decay relationships are changing through time and across different facets of diversity. We found strong spatial distance decay of taxonomic similarity across all peaks, with the highest elevation peak showing the strongest decay. Functional and bioclimatic similarities decayed more weakly than taxonomic similarity, suggesting environmental filtering shapes trait and niche distributions. Critically, spatial decay patterns showed no temporal signal across facets of diversity, indicating community stasis rather than biotic homogenization or differentiation. This study adds further evidence that xeric alpine plant communities are resisting climate change impacts through time.

Synthetic nitrogen fertilizers enhance maize (Zea mays ssp. mays) productivity but impose substantial environmental and economic costs. Some Mexican maize landraces support biological nitrogen fixation (BNF) in aerial root mucilage, raising the question of how much variation exists in this trait and how domestication, geographic spread, and improvement have shaped BNF-related traits across Zea in the Americas. We evaluated aerial root traits and BNF capacity across the maize domestication continuum using 60 Zea accessions grown in field experiments over two years under low-N conditions. Some accessions of wild Zea, landraces from Mexico and South America, and improved varieties had elevated nitrogen derived from the atmosphere (%Ndfa), ranging from 20 to 40%; some of these also had greater %Ndfa than typical improved varieties. Spread and improvement appear to have significantly reduced the number of aerial root-bearing nodes, with wild Zea and Mexican landraces consistently producing more. Nevertheless, the proportion of nitrogen derived from the atmosphere (%Ndfa) does not vary systematically across the domestication continuum. Thus, crop evolution shaped aerial root phenotypes more than associative BNF capacity. The persistence of substantial within-group variation indicates that the genetic potential for enhanced BNF remains within the Zea gene pool, providing opportunities to encourage and improve nitrogen use efficiency in maize cropping systems.

Fire-dependent or fire-influenced ecosystems harbor species with diverse response strategies, such as smoke-stimulated germination and dormancy release through heat shocks. Although these cues influence germination in many fire-prone ecosystems, their effects on seeds of woody species from tropical dry forests (TDFs) remain poorly understood. We evaluated the germination response of twelve TDF woody species to three heat-shock temperatures (80, 120, and 160C), two exposure times (3 and 5 min), and two smoke conditions (with and without smoke water), plus dry and wet controls. We also assessed the relationship between germination and functional seed traits, including permeability, seed-coat thickness, mass, and shape index. Of the eleven species that germinated, four showed broad tolerance, with no significant reduction in germination even at 160C for 5 min, which was associated with lower permeability, thicker seed coats, and greater mass. Four other species showed restricted tolerance to heat, with significant declines in germination at higher temperatures or longer exposure times, and were associated with higher permeability, thinner seed coats, and lower seed mass. The remaining three species showed heat-stimulated germination between 80 and 120C and tended to have lower shape-index values. Smoke treatments significantly increased germination in three species, Ceiba pentandra, Guazuma ulmifolia, and Samanea saman. In the remaining species, significant differences were either absent or attributable to moisture or imbibition effects rather than to smoke itself. Overall, germination responses to heat were associated with variation in functional seed traits, whereas responses to smoke were less widespread and showed a less consistent functional pattern.

Aim Regional climate-change refugia - areas where abiotic conditions related to hydrology and topography sustain unique plant communities despite an otherwise unsuitable regional macroclimate - have been essential for maintaining biodiversity through past climatic transitions. At fine spatial scales, regional refugia can also contain microrefugia, where canopy structure and topography strongly buffer understory microclimate. We tested whether such microrefugia structure understory plant communities within a regional climate-change refugium, where sheltered, groundwater-fed canyons have sustained a biogeographically distinctive flora since the Last Glacial Maximum. Location Temperate forest in the North American Great Plains, Nebraska, USA. Methods In a 20.2-ha forest inventory plot spanning a sharp topographic gradient, we quantified diversity, composition, and distributions and of understory woody and herbaceous plants across seven functional groups along fine-scale topographic, light, and soil moisture gradients. Results Woody species density and richness were significantly higher within sheltered microrefugia in canyon bottoms and on forested slopes and strongly decreased with increasing canopy openness, whereas C4 grass richness was highest along the exposed, high-light forest-prairie ecotone, reflecting divergent functional group responses to microclimate buffering. Soil moisture structured richness across functional groups: woody plant and C3 grass richness peaked at moisture extremes, sedge richness at intermediate moisture variability, and C4 grass richness declined with increasing moisture variability. Both woody and herbaceous understory plants showed strong variation in composition with topography; in particular, woody seedlings and graminoids exhibited strong species turnover with increasing elevation. Main Conclusions These results demonstrate that topographically driven microrefugia support disproportionate understory biodiversity and suggest that protecting both regional refugia and finer-scale microrefugia will be critical for conserving plant communities, including species documented exclusively in microrefugia, as macroclimate continues to change.

Premise of the study: Doryopteris J.Sm. is a genus of dry-adapted ferns found widely across the southern hemisphere and the tropics. In the Hawaiian Islands, the genus is represented by four endemic species and one putative hybrid. Two of the endemic species (D. decora and D. decipiens) are widespread across all major islands, while two (D. angelica and D. takeuchii) are federally and state-listed endangered single-island endemics.We aimed to resolve the evolutionary relationships and biogeographic history of Hawaiian Doryopteris. Methods: We used a target-capture approach and the GoFlag 408 flagellate land plant probe set to sequence more than 400 nuclear loci for 40 Hawaiian and 29 non-Hawaiian individuals in Doryopteris and the related genera Lytoneuron and Ormopteris. After processing the sequences into alignments and single nucleotide polymorphism sites, we performed phylogenetic inference and population-level principal component analyses. Key results: We infer that Hawaiian Doryopteris are monophyletic and are nested within the pantropical species D. concolor. Within Hawai'i, clades and clusters of individuals correspond to geographical, geological, and climatological factors, more so than species identifications, with a few exceptions. We posit that Hawaiian Doryopteris represents an incipient radiation at the very earliest stages of divergence, and that budding speciation has led to non-monophyly of several taxa. Conclusions Our inference using genome-wide sequencing data sheds light on the complex evolutionary history of Hawaiian Doryopteris. Overall, the phylogeny of Hawaiian Doryopteris shows strong links with geographical, geological, and climatological processes and little with morphological evolution.

A longstanding question in evolution is if similar morphologies shared by distantly related organisms facing similar environmental pressures belie similar functions. The carnivorous pitcher form has evolved in three distinct plant lineages and encompasses a suite of convergent morphological traits, providing a natural experiment to test hypothesized links between morphology and function. Multiple pitcher plant species possess fenestrations (i.e., small windows) that let light pass through the pitcher leaf. While research on one species, Nepenthes aristolochioides, demonstrates that fenestrations manipulate light to create false exits and increase prey capture, whether this convergent morphology serves this convergent function in other pitcher species remains unresolved. We explored the function of fenestrations in two additional species of pitcher plants, Nepenthes klossii and Cephalotus follicularis, where the morphology and function of fenestration were previously unexplored. We inspected micromorphology of pitcher structure, conducted an insect bioassay with live plants to test for links between morphology and function, and quantified traits associated with light manipulation from herbarium specimens. Both species exhibited fenestration and related pitcher morphology similar to N. aristolochioides. Yet our experiments did not reveal a significant effect of light passing through fenestrations on fly capture for these two species. Key differences between these species and the previously investigated species may be the presence of wax crystals and the absence of viscoelastic fluid. While future studies may uncover weak to moderate light manipulation function not detected in our study, our data do not support convergent function in these distantly related, morphologically similar species.

- C2 photosynthesis reduces photorespiratory carbon loss by shuttling glycine from mesophyll (M) to bundle sheath (BS) tissues, where glycine decarboxylase (GDC) releases CO2 that accumulates around rubisco. C2 photosynthesis has been considered a constitutive character, but as a metabolic trait, it may acclimate to the rate of photorespiratory glycine production. - We grew eleven species from two eudicot and two grass lineages spanning C3 to C2 phenotypes at 32 {degrees}C days and 200 or 800 {micro}mol mol-1 CO2. Acclimation was assessed using leaf gas exchange, GDC P-protein immunolocalization, M and BS ultrastructure, photosynthetic enzyme activity, and photosynthesis modelling. - Low relative to high growth CO2 enhanced C2-related phenotypes in sub-C2 and C2 species but not C3 or proto-Kranz species. Sub-C2 species positioned chloroplasts closer to mitochondria and had lower modelled BS conductance. C2 species increased rubisco capacity and GDC allocation to the BS. Aspartate aminotransferase activity increased in both sub-C2 and C2 species. - Acclimation to greater photorespiratory conditions strengthens C2 photosynthesis, but only in species whose BS has crossed a threshold of organelle and GDC investment. We hypothesize that acclimation could expose C2-related traits to selection and facilitate the origin of C4 photosynthesis.

Asian Passifloraceae occupy a distinctive evolutionary and ecological position within the family despite their relatively low species diversity compared with the Neotropics. Many Asian representatives are highly localized, small and often ephemeral climbers that persist seasonally through rhizomes, tuberous stems or dormant seeds, while considerable morphological plasticity may obscure species limits and evolutionary relationships. Xishuangbanna, southwestern China, lies at the convergence of Southeast Asian, Indian-Himalayan and southern Chinese floristic elements and represents an important regional centre of Passifloraceae diversity, with two species of Adenia and five species of Passiflora, including the recently described P. xishuangbannaensis and P. menghaiensis. We investigate the evolutionary relationships, plastome evolution and population genomics of these taxa using complete plastome assemblies, a concatenated dataset of 68 selected plastid protein-coding genes, 356 reconstructed nuclear Coding Sequences (CDS), and Genotyping-By-Sequencing (GBS) across the known wild distribution of P. xishuangbannaensis. Plastid, concatenated nuclear and nuclear coalescent phylogenies were strongly incongruent and differed from published classifications based on morphology and earlier phylogenies (ITS, ncpGS, ctyGS, ndhF and trnL-F). Gene-tree discordance and phylogenetic-network inference indicated widespread incomplete lineage sorting together with introgression. The two statistically best-supported network models placed P. menghaiensis, which showed high genetic similarity to the Indian P. napalensis, as a sister lineage involved in independent reticulation events associated with P. altebilobata, P. xishuangbannaensis and P. sumatrana. A pronounced biogeographic pattern was evident within Passiflora supersection Disemma, particularly in the P. siamica-P. cochinchinensis lineage across the Hua Line, whereas relationships among other Asian Passiflora blurred this boundary. This pattern suggests that diversification and reticulation within the Asian lineage may have overlapped temporally with the development of the biogeographic divide represented by the Hua Line. The concentration of early-diverging lineages around Xishuangbanna and the eastern Himalayan-Indian region further points to this area as an important centre in the early diversification of Asian Passiflora. Within this context, the highly localized P. xishuangbannaensis may represent either a relictual of an early Asian lineage or the surviving product of an ancient reticulate complex, contrasting with lineages such as P. siamica that subsequently expanded farther across Southeast Asia. Comparative plastome analyses revealed extensive structural and gene-content evolution within subgenus Decaloba, including pseudogenization, gene and intron losses, and repeated Inverted-Repeat (IR) expansions and contractions; substantial plastome modification also characterized supersection Disemma. Passiflora sumatrana exhibited a particularly distinctive IR expansion extending into the accD-pafII region, potentially characterizing this lineage. Adenia plastomes were generally more structurally conservative, although A. penangiana displayed an IR-boundary contraction involving the rps19-rpl2 region. Mitochondrial CDS and non-coding regions provided the best-resolved phylogenetic signal and revealed strong discordance with the chloroplast phylogeny, consistent with maternal mitochondrial inheritance and variable plastid transmission, potentially involving paternal or biparental inheritance and heteroplasmy. GBS analyses of P. xishuangbannaensis using complementary distant-reference, de novo and closely related-reference approaches showed that mapping to P. organensis provided the clearest SNP distribution and population resolution. ADMIXTURE, PCA and FST analyses revealed two strongly geographically structured populations with additional substructure and substantial differentiation (FST = 0.112). Linkage disequilibrium based demographic reconstruction using GONE2 indicated declining effective population size (Ne) toward the present in both populations, with a particularly abrupt recent decline in Pop1 and a more gradual decline in Pop2. Few candidate Self-Incompatibility (SI) genes could be confidently reconstructed, including limited recovery of highly variable docking regions. However, the intracellular region of an S-locus Receptor Kinases (SRK) contained a distinctive non-synonymous substitution predicted to alter protein configuration without major disruption of overall folding, potentially contributing to compatibility differences between populations. Together, these results reveal a complex evolutionary history combining historical dispersal, incomplete lineage sorting, reticulation, morphological plasticity, plastome structural evolution and strong population differentiation. The narrow distributions and recent demographic decline of these newly recognized Xishuangbanna taxa emphasize their conservation importance, while broader Asian taxon sampling and deeper genomic sequencing will be essential to resolve the evolutionary history of this discreet and poorly known Passifloraceae radiation.

Global change has a major impact on rivers and their associated terrestrial compartment, the riparian forest. The black poplar (Populus nigra L.), the dominant species of the European riparian forest, is considered an indicator of the biodiversity dynamics of these ecosystems, but is threatened with extinction in several parts of its range. The species is dioecious and shows wide variability in many traits, enabling it to adapt to a broad range of environmental conditions. A detailed characterization of this natural variability is therefore a key issue both for the conservation of natural populations and for biological engineering aimed at restoring riverbanks. Phenology is both an indicator of climate change and an adaptation lever for populations, particularly in perennial species such as trees. In this study, we characterized the spatial and temporal variability in flowering phenology by studying 13 natural black poplar populations along the Loire River, France. Flowering date and duration showed substantial variability both within and between populations, and males generally flowered earlier than females. Despite this variability, the ranking of individual trees' flowering dates was highly consistent across years. Most male-female pairs shared several days of overlapping flowering each year, and this overlap was moderately stable across years. Flowering dates advanced by about five to six days per degree Celsius during early spring, with no detectable difference in sensitivity between sexes. This work contributes to the characterization of flowering variability in an economically important tree species and may inform both the conservation of genetic resources and breeding.

Stemflow transports water and biological material from forest canopies to the stem base, yet fungal studies of this flux have largely relied on microscopy of conidia. We used ITS2 metabarcoding to characterize fungal environmental DNA (eDNA) in stemflow from 15 trees - five each of American beech (Fagus grandifolia), sugar maple (Acer saccharum), and tulip poplar (Liriodendron tulipifera) in a closed-canopy forest in northeastern Ohio, USA. Stemflow was sampled during eight rain events in the leafless season (November 2024-May 2025), along with simulated bark and gutter washes to assess near-collar and collector signals. Natural stemflow communities were compositionally distinct from both controls and occupied greater ordination space. Ascomycota dominated across all samples, with Parafenestella, Cytospora, Microcera, Nectria, and Aureobasidium among the most abundant resolved genera and 13 of the 25 most abundant taxa differed significantly among host species. Woody-plant-associated endophytes, canker fungi, and saprobes dominated ecological guild composition. Storm event explained 28.6% of taxonomic variation, host tree species 24.1%, and their interaction 10.0%. Guild composition was more strongly structured by host species (35.4%) than storm event (20.4%). Lichenized fungi and lichen-associated fungi were detected. Thus, leafless-season stemflow integrates broad bark-associated fungal eDNA, including recurrent lichen-derived material, beyond the morphologically distinctive conidia captured by traditional methods. Stemflow eDNA therefore offers a minimally invasive means of monitoring canopy fungal diversity and the strong host and temporal filtering of DNA exported from woody surfaces.

Forest canopy height is sensitive to shifts in demography (which affect tree size distributions and abundances of short-vs. tall-statured species) and allometry. Demographic and allometric effects on canopy height have not previously been studied across a large region. We analyzed more than two million individual tree measurements from about 60,000 remeasured inventory plots in non-plantation forests across 19 ecoregions of the conterminous United States to quantify changes in aboveground live tree biomass (AGB), canopy height, and its demographic and allometric components from ~2003-2025. We tested the hydraulic limitation hypothesis, which predicts decreasing tree height with increasing hydraulic stress. AGB was strongly correlated with canopy height across ecoregions, and AGB change was strongly correlated with canopy height change over time. Canopy height change was positive in nearly all eastern ecoregions but varied in sign across western ecoregions. Demographic shifts led to increased canopy height in most eastern ecoregions but decreased canopy height in most western ecoregions. In contrast, allometric shifts, averaged across species, caused either positive or no significant change in ecoregion-mean canopy height. Our results were consistent with several predictions of the hydraulic limitation hypothesis: (i) intraspecific allometric shifts were more positive for understory than for canopy trees, (ii) across ecoregions, canopy height was positively correlated with precipitation and negatively correlated with vapor pressure deficit, and (iii) canopy height change over time was positively correlated with precipitation change. However, our results suggest modifications to the hydraulic limitation hypothesis, because allometric shifts tended to be positive, even in ecoregions with decreasing precipitation and increasing vapor pressure deficits. Such allometric shifts, which seem counter to the hydraulic limitation hypothesis, may be caused by CO2-driven increases in water-use efficiency. Whatever their cause, allometric shifts should be accounted for to avoid biased estimates of canopy height change and biomass change.

In many flowering plants, self-incompatibility (SI) is a genetically controlled recognition system that prevents self-fertilization, thereby promoting outcrossing and maintaining genetic diversity within populations. The gametophytic SI system in grasses (Poaceae) is governed by two multiallelic loci, S and Z, each locus comprising of three genes governing the initial recognition between pollen and stigma. Identifying these genes establishes the molecular basis of SI in grasses, but how its unique two-locus architecture has shaped the evolution of the underlying loci and recognition genes remains largely unexplored. Here, we report targeted sequence capture of SI genes in 362 genotypes across nine forage grass species, revealing extensive allelic richness and sequence diversity. The SI genes exhibited elevated nucleotide diversity ({pi}) (0.134 < {pi} < 0.403) and great amino acid sequence divergence, ranging between 48.2-58.0% for the female determinants (sS and sZ) and 66.5-83.4% for the male determinants (SI-DUF247). The SI-DUF247 proteins also displayed trans-generic polymorphism, indicating that alleles predate speciation. Phylogenetic analysis of SI determinants showed congruence at both S- and Z-locus, consistent with co-evolution of recognition partners. Our results demonstrate that the unique two-locus SI system in grasses shares evolutionary characteristics with other SI systems, suggesting that their reproductive function is shaping their evolution. These findings also indicate that the grass SI loci are under long-term balancing selection to maintain recognition specificity while creating sufficient allelic diversity. Our study provides comprehensive sequence resources for SI genes and new insights into the evolution of the unique two-locus SI system of grasses.

Whether phenotypes and environments predict performance under stress is central to ecology and evolution and can reveal species' vulnerability to global change. We sampled Mimulus guttatus range-wide to test whether reproductive, morphological, and physiological traits predict drought responses. Surprisingly, we find that stomatal response, a key mediator of water balance, did not change as drought intensified. Climate, however, predicted survival after turgor loss: plants from regions with large daily relative to seasonal temperature ranges survived wilting longer. Stress responses also depended on life history: where summers are dry, perennials withstood lower water potentials, whereas annuals increased leaf cooling as they wilted. Known trait associations were not fixed: although many baseline traits tracked climate, none predicted drought responses. Drought performance therefore cannot be inferred from traits measured under well-watered conditions. Lacking stomatal regulation, riparian drought escapers like M. guttatus may be vulnerable if shorter growing seasons bring drought before reproduction.