Annals of Botany

1. Stomata and leaf traits are key regulators of plant water use efficiency and are expected to have changed in response to rising atmospheric CO2 concentrations and climate warming over the past centuries. However, long-term data documenting such changes are rare. 2. We leveraged herbarium collections to track changes in stomatal characteristics and leaf traits in 656 individuals of invasive Japanese knotweed and its hybrid Bohemian knotweed collected across their European range and spanning 160 years of invasive spread. 3. We found that several functional traits including stomatal density and maximum anatomical stomatal conductance did not show significant changes over time but that plants adjusted their stomatal size and shape over time, and that these changes were associated with increased atmospheric CO2 levels. Interestingly, Reynoutria japonica showed increases in stomatal size and stomatal elongation, while the hybrid R. x bohemica showed a reduction in stomatal size. Traits also varied systematically with climates of origin. Plants from warmer origins with higher evaporative demands during the growing season had thicker leaves, lower SLA, smaller stomata and higher stomatal density, indicating more conservative water-use strategies. Stomatal density and gas exchange capacity co-varied with leaf structural traits, and there was a trade-off between stomatal size and number. Overall, fast leaf economic traits were associated with slow physiological traits. 4. Our results suggest that stomatal anatomical plasticity may enhance climate resilience by maintaining a stable maximum gas exchange capacity across environmental gradients. Herbarium collections provide a unique resource for reconstructing plant responses to historical environmental changes and understanding intraspecific trait variation.

Background and AimsThe evolution of reproductive isolation between previously interbreeding populations is a fundamental driver of plant speciation. Within Amaryllidaceae, Nothoscordum represents an evolutionarily complex genus, characterized by an unusually high incidence of chromosomal rearrangements. During fieldwork, Nothoscordum montevidense and Nothoscordum bonariense were found growing in sympatry, along with individuals exhibiting intermediate morphological traits, suggesting a putative hybrid origin. To test this hypothesis, we employed an integrative approach to characterize the morphologically intermediate specimens and the two sympatric populations. Materials and methodsTo characterize the putative hybrids we have combined morphological, cytogenetic analyses (chromosome counts, CMA/DAPI banding, and FISH) and flow cytometry-based genome size estimation. Phylogenetic relationships and genomic structure were also investigated through Genotyping-by-Sequencing (GBS), complete chloroplast genome assembly, and comparative repetitive DNA analysis. We also performed species distribution modeling and phenological analyses of the putative parental species. Key ResultsMultiple lines of evidence confirm the hybrid origin of the studied plants. Cytogenetic analyses revealed specimens with 2n = 21 (1C {approx} 33 pg = 32.274 Mbp) and 2n = 25 (1C {approx} 37 pg = 36.186 Mbp), accompanied by meiotic irregularities consistent with interspecific hybridization. Chloroplast genome phylogeny identified N. montevidense (2n = 16, 1C {approx} 25 pg) as the maternal lineage, while GBS data confirmed N. bonariense (2n = 26, 1C {approx} 41 pg) as the paternal contributor and revealed evidence of subsequent backcrossing. Comparative analysis of repetitive DNA showed reduced 35S rDNA diversity in the hybrid, indicative of post-hybridization genomic restructuring. Despite the observed genomic complexity, no clear morphological differentiation was detected among hybrid individuals. Phenological analyses and species distribution models demonstrated broad overlap between parental species. ConclusionsOur findings highlight the role of hybridization in shaping genome architecture in cytogenetically labile plant lineages. Furthermore, our results underscore that morphological similarity can mask profound genomic complexity, reinforcing the value of integrative approaches to understand genera characterized by reticulate evolution and genomic instability.

Attracting specific pollinators can be favoured by natural selection to avoid reproductive interference between sympatric plant species. However, the ways in which fine differences in floral traits lead to the attraction of specific pollinators are diverse and unknown in many pollination interactions. We surveyed pollinators on three sympatric Aristolochia species (A. clematitis, A. pistolochia and A. rotunda) pollinated by Diptera to investigate if specific pollination occurs. To decipher if specific pollination may be mediated by different floral odours, we characterized the volatile organic compounds (VOCs) emitted by flowers and highlighted those VOCs electrophysiologically detected by pollinators in A. rotunda and A. pistolochia. Among the most abundant pollinators, Forcipomyia monilicornis was a specific pollinator of A. pistolochia while two Dasyhelea species were specific pollinators of A. clematitis. Forcipomyia aristolochiae and T. ruficeps were non-specific pollinators of A. rotunda, although they were more frequently found in A. rotunda flowers. The floral odours of A. rotunda and A. pistolochia differed significantly from each other and elicited specific electrophysiological responses in their respective pollinators. Although several pollinator species visit more than one Aristolochia species, those pollinators are preferentially found in one Aristolochia species. Selective attraction is likely mediated by specific VOCs.

Leaf shape varies widely across plant taxa and has repeatedly been shown to affect ecophysiology, interspecific interactions, and fitness. We used population genomics, genome wide association studies (GWAS), and comparative genomics to determine the genetic basis and evolutionary history of an uncharacterized Mendelian leaf shape polymorphism in Ipomoea hederacea. To do so, we assembled a reference genome and generated whole genome sequencing for 123 individuals from 55 populations. We identified a 117 kb indel that perfectly co-segregates with leaf shape by conducting a GWAS and assessing differences in coverage. Syntenic orthologs for genes on the indel were present in five other Ipomoea species with various leaf shapes, indicating the indel is newly arisen deletion in I. hederacea despite similar leaf shape phenotypes in the other species. More broadly, these results illustrate how a range of leaf shape phenotypes can be produced by distinct genetic mechanisms even in closely related species. Although none of the genes on the indel itself are known leaf shape candidates, there are multiple leaf shape candidate genes in close proximity that are involved in the auxin biosynthesis pathway. Additionally, the genes within the indel have gene functions that could affect other potentially ecologically relevant traits that have previously been shown to be associated with leaf shape in I. hederacea. Therefore, the pleiotropic effect of the indel polymorphism can have important implications for understanding the ecological mechanisms influencing a well-documented leaf shape latitudinal cline in I. hederacea. Significance StatementIpomoea hederacea has been used to investigate the ecological and evolutionary effects of leaf shape because of a well-documented latitudinal leaf shape cline governed by an uncharacterized Mendelian polymorphism in this species. We identified a 117 kb indel that perfectly co-segregates with leaf shape in I. hederacea. The indel appears to be a distinct genetic mechanism than those governing similar leaf shapes in other closely related Ipomoea species. Additionally, possible pleiotropic effects of the indel on other traits has important implications for understanding the ecology and evolution of leaf shape variation in I. hederacea. Therefore, both the results and the new genomic resources that we developed will help facilitate future work understanding the genetic, developmental, and ecological mechanisms governing leaf shape variation.

The tribe Spermacoceae (Rubiaceae) comprises a morphologically diverse assemblage of approximately 1,400 species distributed across the Neotropics, Africa, Asia, Australia, and Pacific region. It remains one of the most taxonomically intractable groups in the family, with generic limits repeatedly redefined for more than two centuries. Previous phylogenetic studies based on a limited number of plastid and nuclear markers left numerous relationships unresolved and provided sparse representation of Neotropical lineages. Here, we present the first phylogenomic study of the tribe based on plastome-scale data and expanded sampling of Neotropical taxa. We sampled 121 species representing 55 genera spanning all major clades and generated 123 new plastomes, including 25 species incorporated into a molecular phylogenetic framework for the first time. Maximum-likelihood and Bayesian analyses recovered a highly resolved and strongly supported phylogeny, with uncertainty restricted to a small number of deep backbone nodes. Pollen and seed micromorphology provided additional evidence for evaluating phylogenetic relationships. The resulting phylogenetic framework clarifies generic boundaries across several problematic lineages and supports multiple taxonomic changes. Pervasive homoplasy in seed and floral characters rendered several traditionally recognized genera non-monophyletic, warranting new combinations, including Edrastima oxycoccoides, Stenotis alexanderae, and S. prostrata, and a reassessment of taxa such as Terrellianthus serpyllaceus and Oldenlandia dusenii. We further identify genera requiring additional study and provide an updated key to the 82 recognized genera of Spermacoceae. Together, these results provide the most robust phylogenetic framework yet available for the tribe and establish a foundation for future systematic, biogeographic, and evolutionary research.

Background and AimsThe development of unisexual flowers relies on the tight coordination of flower organ identity and sex determination. The genus Quercus is typically considered strictly monoecious, bearing fully segregated male and female flowers within the same individual tree. However, several reports of atypical flowering across the genus challenge this canonical view, suggesting that flowering in oaks may be more flexible than traditionally assumed. In this work, the dynamics of flower development in Quercus orocantabrica were examined to correlate contrasting floral morphologies with divergent molecular profiles. MethodsThe flowering phenology of Q. orocantabrica trees was closely monitored over several individuals and years, together with a detailed floral morphological analysis of male, female and atypical flowers. Key floral homeotic gene homologues were identified, and their expression assayed in the development of different flowers. Key ResultsRecurrent and widespread hermaphroditic flowering was detected in several Q. orocantabrica trees, frequently associated with unseasonal flowering events. Gene expression analysis of male, female and hermaphroditic flowers revealed a sex-biased expression of Q. orocantabrica B- and C-class genes, with the B-class gene QoPI in particular being tightly associated with the presence of fully-developed stamens. In addition, the expression of the C-class gene QoSHP contrasted with reports in other Fagaceae, highlighting a potential functional divergence of the C/D-class lineage within the family. ConclusionsThe results here depicted indicate that the dynamics of floral sex identity in oaks are more plastic than traditionally assumed, supporting a reinterpretation of oak reproductive biology based on a versatile and resilient framework responsive to different developmental contexts.

Background and AimsComplex genomic histories driven by hybridization and polyploidy can shape key plant traits such as defense, stress tolerance, and toxicity, particularly in Amaran-thaceae, which includes crops such as quinoa and spinach. Within this family, white goosefoot (Chenopodium album) is both a widespread agricultural weed and a traditional food resource. However, its evolutionary history is complicated by discordant signals among genomic markers within the C. album complex, comprising diploid, tetraploid, and hexaploid taxa. Here, we tested whether reticulate evolution underlies this genome-wide discordance. MethodsUsing genome-scale phylogenomic data, we analysed 2,298 conserved nuclear loci (BUSCO genes) across 27 Amaranthaceae genomes. Both single- and multicopy gene families were included to capture signals of gene duplication, incomplete lineage sorting, and hybridization. Complementary phylogenomic approaches were used to evaluate whether the evolutionary history is best supported by strictly bifurcating relationships or by reticulate evolution. Key ResultsA consistent C. album lineage was recovered, comprising tetraploid and hexaploid C. album cytotypes together with C. suecicum, C. strictum, C. formosanum, C. acuminatum, and C. opulifolium. Phylogenetic discordance was concentrated within Chenopodium, particularly around the C. album and C. quinoa lineages. Models incorporating hybridization fit better than strictly bifurcating relationships, supporting at least two reticulation events. Hybridization signals were detected in 271 loci in tetraploid and 270 in hexaploid C. album, of which 232 were shared, indicating a shared hybrid origin rather than independent lineages. ConclusionsThe evolutionary history of the C. album lineage is best explained by reticulate processes involving hybridization and polyploidy. Conserved nuclear loci retain persistent signatures of these events, helping to resolve complex evolutionary histories in polyploid plant systems.

Background and aimsThis study combines morphological and environmental data to better understand a Brunfelsia (Solanaceae) species complex, aiming to clarify patterns of variation and identify ecological factors that shape morphotype boundaries. Such an approach provides a broader perspective on how organisms respond to environmental gradients and contributes to a more comprehensive understanding of biodiversity. MethodsWe analyzed 273 herbarium specimens for 13 morphological traits using univariate and ordination analyses, namely PCA and CVA. Climatic and edaphic variables were extracted for 147 specimens with georeferenced records. To assess habitat suitability and the ecological niche of each predefined morphotype, niche models under present conditions and niche overlap tests were conducted. A redundancy analysis (RDA) was applied to evaluate how environmental predictors explain variation in vegetative and floral traits. Finally, DAPC was used to estimate membership probabilities based on morphological and environmental data. Key ResultsTwo well-differentiated groups were recovered: the capitata-hydrangeiformis morphotype, allegedly composing a cline, and the ecologically and morphologically distinct "bahia" morphotype. Variation in floral traits was better explained by environmental predictors than variation in vegetative traits; moreover, floral traits were able to delineate morphotypes more robustly when plotted in isolation. However, when analyzing the results of ecological niche overlap, a significant ecological separation of the "bahia" morphotype from the others was observed. Therefore, key morphological characters for the taxonomy of Brunfelsia covary in part with environmental variables. ConclusionsThese findings support the recognition of "bahia" morphotype as a distinct species to be formally described. This integrative approach contributes to understanding diversification processes in biodiversity hotspots and highlights hidden taxonomic diversity within Brunfelsia, where many rare and narrow-endemic taxa lie.

PremiseDrought is intensifying in many regions under climate change, affecting plant growth, phenology, and resource allocation. Plants cope with water limitation through strategies such as escape, avoidance, and tolerance, but how these strategies are reflected in trait variation across natural drought gradients remains poorly understood. We examined how drought-related traits vary across a drought gradient in wild sunflower (Helianthus annuus L.). MethodsWe conducted a common-garden experiment using 19 wild sunflower populations collected across Southern California spanning a climatic moisture deficit gradient. We quantified growth, phenology, morphology, biomass allocation, and physiological traits and evaluated their relationships with climatic moisture deficit and soil variables using regression and correlation analyses. ResultsPopulations from drier environments exhibited coordinated shifts in life-history strategy, combining faster growth, earlier flowering, and increased reproductive output. These changes were coupled with reduced allocation to roots and structural tissues and diminished investment in traits that buffer plant water status, as reflected in declines in relative water content and higher (less negative) leaf water potentials. Ordinal modeling further indicated that populations from drier environments had a greater probability of severe wilting under drought stress. Together, these patterns are consistent with a drought escape strategy rather than drought avoidance or drought tolerance. ConclusionOur results suggest that increasing aridity can favor integrated drought escape strategies characterized by rapid growth and early reproduction. These coordinated shifts reveal trade-offs between fast life-history strategies and investment in traits associated with drought resistance, highlighting how environmental gradients shape adaptive trait syndromes in wild plant populations.

Background and AimsIn the plant kingdom, gene flow occurs through pollen and seed dispersal, shaping both within-population spatial genetic structure and among-population genetic differentiation. Anthropogenic land-use change can affect levels of gene flow by reducing pollinator abundance and altering pollen and seed dispersal pathways. Yet, how environmental features shape gene flow events in common herbaceous plants - a fundamental building block of many ecosystems - remains poorly understood. We address this gap by investigating how environmental context influences population genetic structure at regional and local scales in the red campion (Silene dioica). MethodsBy sampling 1,005 individuals from 29 populations across habitats ranging from semi-natural to strongly human-altered, we assessed whether population size and landscape composition influenced within-population genetic diversity and population genetic differentiation. At the local scale, we examined whether landscape composition affected fine-scale spatial genetic structure and pollen dispersal distances in a subset of six populations representing the two extremes of an anthropogenic gradient. Key ResultsNo effect of either demographic or landscape factors was found on levels of genetic diversity. We detected moderate levels of genetic differentiation among populations that matched an isolation-by-distance pattern, with high levels of population admixture, while landscape composition did not explain variation in population genetic differentiation. At the local scale, five of the six studied populations exhibited significant spatial genetic structuring, indicating distinct neighbourhoods at spatial scales less than 10 m. Paternity analyses based on 4,800 offspring further revealed predominantly short-distance pollen dispersal together with substantial immigration from external sources. Neither fine-scale genetic structure nor pollen dispersal distances differed between habitat types. ConclusionsAltogether, our results demonstrate a multimodal pattern of gene flow and a remarkable resilience of this common herbaceous species to anthropogenic habitat change, as substantial genetic connectivity is maintained despite insect-mediated pollen dispersal and gravity-driven seed dispersal.

O_LIThere is growing interest in developing perennial cereal crops for sustainable production, yet key differences in trait syndromes between annual and perennial grasses and their influence on environmental adaptation remain poorly understood. C_LIO_LIWe measured 25 traits in 16 annual and perennial Hordeum species (45 accessions), including barley, grown over three seasons in a common garden. Using a phylogenetic framework and repeated transitions between annual and perennial forms, we (i) identified traits distinguishing these life strategies and (ii) tested how they relate to climate at the accessions origins. C_LIO_LIWild and cultivated barley are distinguished within the Hordeum clade by high growth rates and large organs, which may have predisposed wild barley to domestication. Annual and perennial accessions differed in resource allocation: annuals had higher harvest index and leaf and grain nitrogen, while perennials produced carbon-rich tissues and sustained vegetative growth. Seasonal temperature variation shaped trait syndromes: annual traits aligned with temperature in the driest quarter, reflecting selection under terminal stress, while perennial traits correlated with temperature in the wettest quarter, the main growth phase shaping long-term performance and survival. C_LIO_LIWe provide important information on traits and climate adaptations underlying perennial persistence and annual resource strategies, relevant for developing perennial cereal crops. C_LI

Genomic variation within populations reflects both past and contemporary evolutionary processes. The genetic structure of the European peatland plant species is shaped by complex postglacial recolonization and recent habitat loss. Here, we investigate population genomic patterns in the vulnerable mire plant, round-leaved sundew (Drosera rotundifolia L.). Using ddRAD sequencing of 311 individuals from 38 populations across Europe, we detected significant genetic differentiation (Fst = 0.02-0.44, p < 0.05) and a consistent deficit of heterozygosity (Fis = 0.248) across populations. In contrast, sequencing of 10,449 bp of chloroplast DNA revealed extremely low variation, with only one SNP detected. Clustering analyses (Admixture and DAPC) both identified pronounced genetic structure comprising three major clusters (Western, Northern, and Eastern) that broadly correspond to European biogeographic regions. Genetic differentiation was partially explained by geographic distance (3.3%; p = 0.0001; Mantel test), while climatic variables, particularly temperature and precipitation, accounted for 2.9% of genomic variation (p < 0.0001; redundancy analysis). The observed pattern is consistent with polygenic responses to climatic gradients and is supported by 1,022 SNPs, distributed across 632 loci, that were significantly associated with environmental variables. Demographic reconstruction revealed distinct evolutionary trajectories among clusters, with the Western cluster showing a more recent expansion. Together, our results suggest that the genetic structure of D. rotundifolia is a result of postglacial recolonization, geographic isolation, and climate influence. We highlight the importance of conserving genetic diversity of D. rotundifolia across all three clusters and accounting for potential local adaptation and population vulnerability in future conservation strategies.

Shifts in flowering phenology are one of the most well studied plant responses to global climate change. Many studies have documented these shifts and their drivers, including some that describe altered patterns of co-flowering among taxonomically broad species within communities. In comparison, few analyses have examined systematic changes in co-flowering between closely related, interfertile species, where co-flowering can have unique evolutionary consequences. To address such shifts in co-flowering among close relatives, we investigate phenological responses to climate change and its effect on patterns of co-flowering over the past 124 years in 52 species of North America violets (Viola). This genus has many co-occurring species that reproductively interact via shared pollinators and hybridization. We use ~14,000 herbarium records along with environmental and species trait data to model the magnitude of recent flowering phenology shifts, environmental variables and/or species traits associated with these shifts, and resulting changes in co-flowering among species. While both the magnitude and direction of phenological shifts varied among Viola species, nearly half (25/52) show significant changes in flowering day. Regardless of whether flowering was advanced or delayed, flowering date was most consistently associated with local mean temperature. Of six species-level traits, geographical region also significantly predicted flowering shifts, consistent with environment and geography together explaining broad phenological responses across this group. These shifts have produced significant changes to pairwise patterns of co-flowering among species -- ranging from a 59 day increase in co-flowering to complete loss of co-flowering overlap. Sympatric pairs specifically have experienced both increases and decreases in co-flowering, with a geographic pattern of increased co-flowering occurring mainly in eastern US and decreased co-flowering common in western US. Because Viola species are generalist pollinated and already known to hybridize, these new co-flowering patterns could further undermine reproductive barriers among species in this genus.

Xylem and bark properties influence tree growth and drought resistance, yet their functional coordination and their environmental drivers remain unclear. We assessed xylem-bark coordination in branches of eight temperate woody Rosaceae species spanning different ecological preferences. We quantified xylem hydraulic efficiency and safety alongside bark traits governing permeability, hygroscopic water exchange, water storage, and anatomy, and evaluated phylogenetic signal and climatic associations. Bark water vapor conductance (Gbark) increased with maximum xylem hydraulic conductivity (Kh) and with xylem water potential at 50% loss of conductivity (P50), indicating species with more efficient but more embolism-vulnerable xylem developed more permeable bark. Species with higher Gbark showed reduced hygroscopic absorption time, consistent with faster rehydration from atmospheric water vapor. Both Gbark and P50 were phylogenetically conserved and covaried with climatic factors, namely air temperature, vapor pressure deficit (VPD), and isothermality. Species from warmer, high-VPD climates with greater diurnal temperature variability combined higher bark permeability with more vulnerable xylem, implying a shift from embolism avoidance to embolism tolerance strategies. Overall, xylem and bark hydraulics in Rosaceae evolved in concert along diurnal and annual gradients of evaporative demand, showing that drought resistance in woody angiosperms cannot be understood without considering bark traits alongside xylem function. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=104 SRC="FIGDIR/small/730605v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@14872c2org.highwire.dtl.DTLVardef@1662c8corg.highwire.dtl.DTLVardef@f6b4aaorg.highwire.dtl.DTLVardef@cf1aef_HPS_FORMAT_FIGEXP M_FIG C_FIG Caption: This study shows that xylem and bark in Rosaceae species form an integrated functional system in which xylem hydraulic safety, efficiency, and bark permeability are jointly tuned along diurnal and annual gradients of air temperature and evaporative demand.

Zingiberales includes eight morphologically distinctive families, but its family-level backbone has remained unstable, especially around Musaceae, Heliconiaceae, Lowiaceae, and Strelitziaceae. We analysed 1566 low-copy nuclear genes from 52 samples, representing all eight families and Pontederia crassipes as outgroup. Concatenated maximum likelihood and multispecies coalescent analyses recovered the same backbone: ((Zingiberaceae, Costaceae), (Cannaceae, Marantaceae)) is sister to (Musaceae, (Heliconiaceae, (Lowiaceae, Strelitziaceae))). Penalized-likelihood dating placed the sampled crown group in the Late Cretaceous, with several deep family-level divergences occurring on short internodes. Analysis of 1248 rerooted gene trees showed that conflict is concentrated on these deep branches and in several shallow clades. HyDe tests of empirical and simulated matrices, each including 62,475 triples, did not support widespread ancient hybridization among the major family-level lineages after filtering against the simulated null model. The nuclear data recover a stable Zingiberales backbone, and the long-standing instability of several deep nodes is best explained by rapid early divergence and extensive incomplete lineage sorting.

O_LIPhotoperiod is a stable seasonal signal. Although the photoperiodic flowering is well understood in short-day (SD) and long-day (LD) annual plants, regulatory mechanisms in perennials remain elusive. In a perennial woodland strawberry (Fragaria vesca L.), flowering is induced in SDs in autumn and plants flower following spring, while in plants with mutated FvTERMINAL FLOWER1 (FvTFL1), LDs induce flowering. C_LIO_LIWe investigated photoperiodic flowering of F. vesca through phenotypic and molecular characterization of transgenic lines and their crosses. We studied natural variation in flowering time and gene expression in European accessions, and explored their correlations with climatic, geographical and genetic origins. C_LIO_LIWe showed that FvGIGANTEA (FvGI) and FvCONSTANS (FvCO) activate FvFLOWERING LOCUS T1 (FvFT1) in LDs resulting in early flowering in fvtfl1 mutant, while in SD F. vesca, activation of FvTFL1 by FvFT1 reverses the photoperiodic requirement of flowering. In natural accessions, decreasing expression of FvFT1 and FvTFL1 towards colder climates in the east and north correlated with earlier flowering. C_LIO_LIWe define a photoperiodic flowering mechanism controlling floral transition of perennial F. vesca in autumn that differs from known mechanisms in annual and perennial plants. Our findings open new avenues to understand how perennial plants cope with changing seasons across climatic and geographical ranges. C_LI

Anthocyanin biosynthesis is a highly branched network with upstream mutations leading to the loss of many other branches beyond just anthocyanins like flavonols and isoflavonoids. Consequently, disentangling the specific effects of anthocyanins on plant fitness from those of other branches is difficult unless a mutant impacting the terminal step of anthocyanin biosynthesis can be found. We discovered the first Ipomoea hederacea plant (ivy-leaf morning glory) lacking anthocyanins due to a genetic deletion in the terminal step of the anthocyanin biosynthetic pathway. Anthocyanin loss follows a recessive, Mendelian inheritance pattern. Anthocyanin loss is perfectly correlated with a lack of expression of the anthocyanin biosynthesis gene BZ1. BZ1 is the gene most strongly and consistently differentially expressed between unpigmented and pigmented flowers in F2 siblings. A deletion of half of the BZ1 gene which includes the start codon is the genetic basis underlying a loss of BZ1 expression and, consequently, anthocyanins in unpigmented plants. The BZ1 deletion is rare and unique to the unpigmented line. Genome coverage analyses demonstrate that no other pigmented I. hederacea line (of 123 screened) has a loss-of-function mutation in BZ1. Furthermore, we confirm cosegregation of the BZ1 deletion with the unpigmented phenotype in F3 progeny from a cross where all unpigmented plants are homozygous for the BZ1 deletion. Taken together, we describe the genetic basis of a novel unpigmented mutant in I. hederacea, creating a potentially important model for studying the fitness effects of anthocyanin loss.

Most flowering plant species rely on insect pollinators for successful cross-pollination. While floral color, scent, and morphology help attract visitors, the primary motivation for visiting a flower is the nutritional reward - nectar and, especially, pollen. Pollen provides a complete set of macro- and micronutrients essential for adult and larval provisioning in many insect flower visitors. However, pollen nutrient composition (i.e., quality) varies widely among plant species, and different visitors may have distinct nutritional requirements. Whether these differences are linked to the selection of plants visited remains poorly understood. In this study, we investigated whether the nutrient composition of pollen is linked to the visitation patterns of two main alpine pollinator groups: bumblebees and hoverflies. We observed flower visits in the field and identified the origin of pollen collected by bumblebees and hoverflies via metabarcoding. In addition, we analyzed the nutrient profiles of alpine flowering plants from the same habitat, including amino acids, fatty acids, and sterols. We hypothesized that both bumblebees and hoverflies (i) differ in the plant spectrum they were observed on/collected from and visited for pollen collection, (ii) and that preferred nutrient profiles differ between the two pollinator groups. We additionally expected (iii) that, in the studied alpine plant communities, the nutrient composition of pollen of plant species collected by flower visitors is more strongly associated with pollinator identity than with plant phylogeny due to competition for pollinators. Our results revealed that alpine bumblebees and hoverflies were more similar in their pollen hosts and nutritional preferences than expected, but, for both groups, the network based on observed flower visits differed significantly from that based on pollen collection. Pollen fatty acids and amino acids content was positively correlated, and both bumblebees and hoverflies preferred pollen with low fatty and amino acid content. Pollen sterols did not differ between plants collected by pollinators and non-collected plants. Neither fatty acid, amino acid, nor sterol composition was linked to plant phylogeny. Our findings suggest that not only amino acid content, but also fatty acid content, plays a key role in shaping pollen-collection patterns of flower visitors in alpine ecosystems.

We tested whether seasonal plasticity in photosystem II (PSII) heat tolerance aligns with leaf phenology in tropical trees, comparing evergreen and deciduous species across the wettopostwet transition in a seasonally dry tropical forest of the central Western Ghats, India. This transition, preceding droughtinduced senescence, represents the key window to assess true unstressed thermal plasticity. PSII thermal traits: damage onset (T5), damage midpoint (T50), and temperature between damage onset to full loss (T95-T5), decline width (DW) were quantified in 27 cooccurring species during the wet (27.5{square}{degrees}C) and postwet (31.6{square}{degrees}C) periods. Contrary to phenologybased predictions, PSII plasticity was not structured by leaf habit or successional status. Both T5 (+1.7{square}{degrees}C) and T50 (+0.9{square}{degrees}C) increased significantly across seasons, but responses were speciesspecific, with evergreen and deciduous trees acclimating similarly. The preventionversusforbearance tradeoff (T5 - DW relationship) remained conserved, though leaf habits diverged under postwet conditions. Thermal safety margins based on T50 were large, but T5 revealed vulnerable latesuccessional evergreens (Saraca asoca, Ficus spp.) and Careya arborea. These results show that PSII thermotolerance regulation operates largely independently of droughtavoidance phenology, indicating species identity and not leaf habit drives plasticity in PSII thermal response in seasonally dry tropical forests.

O_LIHybridisation is frequently associated with plant invasions; however, its consequences for genome organisation and chromosome evolution remain poorly understood in invasive species. We investigated the extent of hybridisation in the invasive Carpobrotus edulis--acinaciformis hybrid complex and determined the cytogenomic contribution of parental species in hybrid accessions. C_LIO_LIWe combined whole-genome sequencing, population genomic analyses, genome size estimation, repeatome characterisation, chromosome counting and fluorescence in situ hybridisation to compare parental species and hybrid accessions from South Africa and the Mediterranean Basin. C_LIO_LIPopulation genomic analyses revealed widespread hybridisation and introgression, with most invasive accessions showing admixed ancestries. Pattersons D-statistic supported asymmetric allele sharing towards C. edulis. Hybrid accessions displayed genome sizes indistinguishable from C. edulis, whereas C. acinaciformis possessed significantly larger genomes. Repeatome analyses identified marked differences in repetitive DNA composition, particularly in satellite DNA abundance and chromosomal distribution. A newly identified satellite repeat (CarpoSat) showed contrasting chromosomal patterns between parental species, whereas hybrids resembled C. edulis satellite pattern. C_LIO_LIOur results demonstrate that Carpobrotus hybrid accessions are a swarm of later-generation hybrids and backcrosses showing a strong bias towards C. edulis, indicating asymmetric introgression. These findings highlight the value of integrating cytogenetic and genomic approaches to understand genome evolution in invasive hybrid complexes. C_LI