Annals of Botany

Telomeres have the crucial role of protecting chromosome ends and its length varies significantly between organisms. What evolutionary factor shapes this natural variation has been a long standing question, with answers providing insights into chromosome function and evolution. In animals life-history variation relating to senescence or tumorigenesis explains telomere length variation. For plants, however, we lack knowledge of what factor is driving their length variation in nature. Is life-history differences also driving telomere length variation among plant species? We answer this question by analyzing the telomeres of 257 plant species spanning the major orders of the green plant kingdom. Telomere lengths were estimated for species that vary significantly in their life-history, geographical distribution, and trait characters. Results showed telomere length increased with the evolution of terrestrialization, where Chlorophytes had the shortest telomeres and Spermatophytes had the longest telomeres. Surprisingly, plants with different life-histories had minimal telomere length differences, but biogeographic clines had significant effect on telomere length variation. Plant telomere length increases closer to the equator, while telomere length decreases at higher altitude. Domestication was previously proposed to lengthen telomeres, but we found no consistent domestication effect on telomere lengths comparing crops and their wild progenitors. From our results we conclude plant telomere length contrasts with animal telomeres, in that life-history has minimal effect on length variation. Instead we propose the ecological adaptation hypothesis, where telomere length in plants is determined by selection from abiotic conditions, and suggest temperature is a major factor shaping telomere length in plants.

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.

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.

Seed ecophysiology is essential for understanding plant regeneration and developing effective conservation and ecological restoration strategies, yet knowledge remains scarce for most threatened Cerrado species. We characterised the germination niche, desiccation tolerance and storage behaviour of Xyris paradisiaca (Xyridaceae), an Endangered species endemic to Central Brazil, to assess whether its restricted distribution is associated with narrow germination requirements and to inform seed-based conservation. Germination was tested across light regimes, constant temperatures, decreasing water potentials and short-duration heat shocks, while thermal- and hydro-time models were used to quantify thermal and hydric thresholds. Seeds exhibited an absolute light requirement but broad abiotic tolerances. Germination remained >87% between 15 and 40 {degrees}C, with estimated cardinal temperatures of Tb = 8.94 {degrees}C, To = 33.13 {degrees}C and Tc = 45.53 {degrees}C. Germination remained similar to the control down to -0.6 MPa and exceeded 40% at -1.0 MPa, with a median base water potential of {Psi}b = -1.03 MPa. Germination also remained high after 1-min heat shocks up to 200 {degrees}C. Seeds were desiccation tolerant, with 100% germination after drying and a viability loss index of -0.04, and germination remained >90% after 24 months of ambient storage. Thus, the highly restricted distribution of X. paradisiaca is not driven by a narrow physiological germination niche. Its broad environmental tolerances, desiccation tolerance and favourable storage behaviour also highlight its unexplored potential for seed-based restoration, propagation and ex situ conservation.

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.

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.

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.

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.

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.

Distinct from plants which directly parasitise other plants (parasites), non-photosynthetic mycoheterophic plants (MHPs, previously known as achlorophyllous mycotrophs or saprophytes) depend upon fungal hyphae for their carbon and have been viewed as cheaters or parasites in their fungal partnerships rather than mutualistic as in most other plant-mycorrhizal interactions. Over 400 such species in 87 genera and 11 families are recorded which maintain this lifeform through their life-cycle, while an estimate 30,000 further species, mainly orchids, depend on this mode of life to establish themselves from seeds. In most cases the fungal partners and the ultimate carbon sources, usually autotrophic plants, are unknown. High fungal specificity is the norm in those MHPs researched. We describe a new MHP genus and species, Denisea fonensis that we discovered in the Simandou range of the Republic of Guinea, West Africa in October 2025 on the footprint of a mining project. While clearly placed in Dioscoreales (which has three MHP families), the features of Denisea are so unusual that it was not initially clear in which family to place it. Finally, with molecular data we placed Denisea in Thismiaceae, corroborated by morphology. The new discovery is all the more unexpected as this is the first record of Thismiaceae from West Africa. Until now the family has only been recorded in Africa from three species of the very different genus Oxygyne, of Cameroon and Central African Republic. We provisionally assess Denisea fonensis as Critically Endangered since less than ten individuals are recorded in only a few square metres at one site, where there are imminent threats from animals. We propose urgent measures to conserve this species to reduce the risk of global extinction that has eliminated other African MHP species such as Oxygyne triandra, and further research to determine the fungal and autotrophic plant partners of this remarkable plant genus.

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.

Understanding the genetic architecture of ecophysiological traits related to carbon gain and water loss is crucial to develop cultivars tolerant to abiotic stresses and that maintain productivity in the context of climate change. This can be achieved both through experiments in the greenhouse on potted plants under controlled conditions and in the field. However, the stability of genetic determinism of ecophysiological traits across environmental conditions has rarely been evaluated. We studied a grapevine diversity panel grown both in a greenhouse and in the field, to investigate the impact of growing conditions on leaf ecophysiological traits related to leaf morphology, fluorescence, gas exchange and water use efficiency. The correlation structure between traits was conserved within each environment even though, for most traits, cultivars ranking was not conserved between both environments. However, a set of cultivars, sharing the same functioning pattern in terms of gas exchange and fluorescence in both environments, was identified. Single- and multi-environment Genome-Wide Association Studies (GWAS), based on a dense genotyping matrix, were used to dissect the genetic architecture of leaf traits. The Quantitative Trait Loci (QTLs) identified through single-environment analyses differed substantially from those detected using multi-environment ones. Multi-environment approaches highlighted shared QTLs across environments for gas exchange and fluorescence traits. Candidate genes associated with carbon- and water-related functions were further identified in these genomic regions. Despite the weak correlation between greenhouse and field trait values, combining both environments enabled the identification of robust genetic regions associated with ecophysiological traits of interest.

Understanding floral visitor dynamics along elevational gradients is essential for assessing potential implications for pollen limitation and population genetic processes in fragmented populations of cliff-dwelling alpine endemics. Physoplexis comosa (Campanulaceae) is a chasmophytic paleoendemic restricted to vertical rock faces on dolomitic and limestone bedrock from 200 to 2,800 m a.s.l. The species exhibits a specific floral morphology with fused corolla apices, basal openings for nectar access, and secondary pollen presentation, yet its functional floral visitor assemblage has remained poorly characterized across the species' elevational range. In this pilot study, we tested a two-tier camera-monitoring setup combining action cameras for diurnal monitoring with Raspberry Pi-based systems equipped with IR illumination for combined diurnal and nocturnal recording. We conducted monitoring across five elevational sites (1,480 to 2,400 m a.s.l.) in the Eastern Alps. Recorded video material was downsampled and annotated using the Addax platform alongside manual cross-validation to record physical contact events. Preliminary results confirm the technical feasibility of continuous monitoring in steep terrain and suggest possible elevational variation in diurnal visitors, alongside nocturnal activity by Geometridae moths. Camera-based observations provided higher functional resolution and revealed a different visitor assemblage than previous eDNA metabarcoding trials. This proof-of-concept establishes a methodological foundation for forthcoming multi-day monitoring campaigns and complements ongoing studies of the mating system and population-genetic analyses.

The xylem safety-efficiency trade-off is a central paradigm in plant hydraulics, yet empirical support remains inconsistent. In this study, we highlight how this inconsistency may arise from variation in environmental and evolutionary constraints and from differences in observational scale. We analysed 38 diffuse-porous angiosperm and 24 conifer species grown in a common garden experiment to quantify the relationship between embolism resistance (P50) and specific hydraulic conductivity (Ks) across and within species, and its differences between three species groups differing in evolutionary ancestry and eco-hydrological strategy. When pooling over all species, we found evidence for a safety-efficiency trade-off both across and within species. However, this pattern was driven almost exclusively by the mesic broadleaved species. In contrast, the across-species pattern was suppressed in coniferous species due to the evolutionary constraints on efficiency posed by their tracheid-based wood anatomy, while there was no evidence for a trade-off on either level in hydrophilic broadleaved species, which evolved under conditions that pose reduced selective pressure on embolism resistance. Our results demonstrate that the safety-efficiency trade-off is not universal, but emerges only in specific ecological and evolutionary contexts, highlighting the importance of multi-scale analyses and constraint-based frameworks for interpreting plant hydraulic strategies.

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.

- 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.

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.

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.

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.

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.