American Journal of Botany

Population genetic analysis of species of conservation concern provides information to devise management plans to effectively conserve the genetic variation of endangered species. One such endangered plant, Physaria globosa is a federally endangered species in the mustard family with a geographically restricted range that occurs in four disjunct locations in Indiana, Kentucky, and Tennessee (i.e., Highland Rim and Nashville Basin regions) and along the Wabash, Kentucky, and Cumberland Rivers. In this study, we sampled populations from throughout the range of P. globosa, genotyped them using 20 microsatellite loci, and assessed genetic diversity and structure within and among populations. The goals of the study were to understand: 1) levels of genetic diversity in P. globosa and whether populations show evidence of having experienced reductions in genetic diversity as the result of genetic bottlenecks, genetic drift, or inbreeding, 2) rangewide genetic diversity and structure in P. globosa and how genetic structure is affected by the disjunctions in the species range, and 3) implications for prioritization of in-situ and ex-situ conservation efforts. On average, P. globosa showed comparable levels of genetic diversity to other species of Physaria. However, some populations showed evidence of inbreeding, genetic bottlenecks, or decreases in genetic diversity, possibly due to anthropogenic or climate-related pressures and decreases in population size due to competition with invasive bush honeysuckle. Genetic variation was strongly structured into two main geographic groups, one in the northern part of the species range (KY and IN), and the other in the southern part of the species range (TN), but some populations likely originated via long-distance dispersal. We also found significant isolation by distance, likely due to both life history characteristics and physical barriers associated with the complex topological structure of the landscape occupied by P. globosa, limiting population connectivity. Given the strong genetic structure found in P. globosa, several populations should be protected and managed within each geographic region to conserve genetic variation. Ex situ conservation will also be important to protect genetic diversity, particularly for populations that are difficult to access and manage.

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

Premise of the StudyCarnivory has evolved repeatedly across the plant tree of life despite being a dramatic shift from typical plant nutrient acquisition strategies. It remains largely unclear whether the evolution of carnivory takes a similar genomic trajectory. Here, we explore the genomic consequences of carnivory in the pitcher plant genus Sarracenia. MethodsWe use a combination of Pacbio HiFi long-read sequencing and trio-binning to assemble chromosome-scale genome sequences for S. psittacina and S. rosea. We conduct comparative analyses with other asterid genomes to evaluate patterns of gene family expansion and contraction during the transition to carnivory. ResultsBoth Sarracenia genomes are large ([~]3.5 Gbp) and highly repetitive ([~]87% repeats) yet only contain [~]22,000 genes. This reduced gene content reflects widespread gene family contraction. In total, 3,654 gene families have contracted, including the complete loss of 934 gene families, while only 751 gene families have expanded. The gene losses are enriched for functions related to photosynthesis, including nuclear-encoded subunits of the NADH dehydrogenase (Ndh) complex, as well as immune-related genes. ConclusionsThese results indicate that the evolution of carnivory in Sarracenia is associated with widespread gene loss rather than extensive gene family expansion. The loss of genes involved in photosynthesis and immune response suggest the relaxation of selection on these functions, which may be partially supplanted by prey-derived nutrient acquisition and pitcher-associated microbiome. These chromosome-level assemblies will enable future comparative studies in plant evolution, while also serving as critical resources for the conservation of this ecologically significant lineage.

AimThe assembly of montane plant communities through time is underlain by historical and abiotic factors. However, the extent of evolutionary connectivity between ancient highland ecosystems and surrounding lowlands remains unclear. Here, we investigate the evolutionary connections between the campos rupestres, a hyperdiverse and fragmented montane vegetation complex in eastern South America, and lowland biomes surrounding it: savannas, rainforests, and seasonally dry tropical forests. LocationEastern South America. Time periodCenozoic. Major taxa studiedFlowering plants. MethodsUsing phylogenetic beta diversity analyses for 13 angiosperm clades, we assess the degree of lineage dissimilarity between campos rupestres subregions and adjacent biomes. We also apply generalized dissimilarity modeling to determine the role of climate, soil, and geographic distance in shaping spatial patterns of phylogenetic composition. ResultsOur results reveal high lineage permeability between campos rupestres and surrounding biomes, with lineage sharing largely reflecting biome adjacency. This pattern is mainly driven by shared climatic conditions, which are the strongest predictors of phylogenetic dissimilarity. Main conclusionsWe highlight the importance of lineage exchange between lowland and montane environments for the assembly of highland floras. By showing that lineage movements across biome boundaries have been common over time and spatial scales, our study challenges the idea that ancient Neotropical mountains are isolated sky-islands. Instead, we emphasize the dynamic nature of montane plant diversity and the pivotal role of climate in shaping evolutionary connections between highlands and lowlands.

PremiseHerbarium specimens are increasingly used to extract morphological traits for ecological and evolutionary studies, yet the effects of tissue desiccation on trait measurements remain poorly understood. Here, we tested whether higher tissue water content leads to greater measurement changes after herborization (H1) and whether fresh trait values can be reliably predicted from herbarium measurements (H2). MethodsWe evaluated the reliability of herbarium-based measurements by comparing fresh and dried traits of leaves, flowers, fleshy fruits, and seeds across 262 individuals representing 133 Neotropical Myrtaceae species. Phylogenetic least square models and machine-learning regressions were used to test H1 and H2. ResultsLeaves and flowers generally shrank after herborization, fruits size metrics tended to increase, and seeds were largely unaffected. Water content was significantly associated with the magnitude of herborization effects in flowers and some leaf and seed traits. Fresh trait values were accurately predicted from herbarium measurements. Prediction errors were lowest for leaf traits, followed by fruits, flowers, and seeds. DiscussionThese results partially support H1 and support H2, indicating that herbarium specimens can be reliably used for trait analyses when organ-specific responses are considered, providing a practical framework to account for potential desiccation bias in functional trait research.

Long-term field observations typically are the "gold-standard" for inferences of phenological sensitivities in montane systems but are spatially limited. Herbarium specimens provide broader spatial coverage, but their utility to accurately capture montane phenology remains poorly known. We compared flowering phenology of 45 species inferred from herbarium specimens with comparable data from nearly 50 years of direct observations at the Rocky Mountain Biological Laboratory. Estimates of flowering time and phenological sensitivity to snow density were consistent between herbarium specimens and observations, but observations revealed secondary flowering peaks. Herbarium specimens additionally yielded shallower estimates of phenological sensitivity to spring temperature than did field observations. Across co-occurring species, "early" flowering individuals inferred from herbarium specimens, rather than the mean response across all individuals, may better approximate community-level phenological responses to temperature changes. We conclude that herbarium specimens are reliable resources for closing gaps in understanding phenological variation along elevational gradients of montane systems.

The temperate rainforests of the Pacific Northwest of North America harbor many endemic taxa whose evolutionary histories have been shaped by major climatic and geologic events. The enigmatic taildropper slugs (genus Prophysaon) are one example, notable for their ability to autonomize their tails to escape predators. Despite extensive work uncovering the evolutionary history of individual lineages, relationships among the nine recognized species of Prophysaon remain poorly understood due to insufficient molecular data. To address this, we collected transcriptomes for six of the nine currently accepted species of Prophysaon. Using these data, we were able to resolve species relationships, calling into question the existing subgeneric classification based on morphology. We also detected undescribed phenotypic diversity within the P. andersonii--P. foliolatum species complex, with molecular data supporting the distinctness of two phenotypically distinct populations from Washington. Finally, our transcriptomic data suggest a moderate role of introgression in shaping the evolutionary history of Prophysaon. Here, we synonymize the subgenus Mimetarion with nominotypical Prophysaon. Future work should further investigate whether the undescribed diversity detected here represents species level differentiation.

Premise of studyUnder increasingly frequent pollinator-limited environments, plants need to rely on modes of reproductive assurance such as selfing and cloning. However, few studies investigate the interplay between selfing and cloning in plants that can do both. Here, we explore mechanisms determining the relative expression of selfing and cloning throughout the European distribution of the wild woodland strawberry (Fragaria vesca) under a pollinator-free environment. MethodsWe established an outdoor common garden with 121 woodland strawberry genotypes from across Europe and excluded them from pollinators. For each genotype, we recorded reproductive traits and performed hand-pollination treatments. Key ResultsWe found a weak trade-off between cloning and selfing, driven by increased seed and fruit provisioning rather than flower production. The capacity to autonomously self-fertilize was determined by the lateral proximity of the anthers to the pistils (lateral herkogamy), but not by early inbreeding depression. Genotypes sampled at lower latitudes and altitudes were better at self-fertilizing and had smaller petals. The propensity to clone increased towards the east, where genotypes also had smaller petals, particularly at higher latitudes. ConclusionAt the species level, we detected a trade-off between the propensity for clonal reproduction and the capacity for self-fertilization. At a continental scale, the capacity to self-fertilize varied along a north-south gradient, whereas clonal propensity varied along an east-west gradient. Our results suggest that these two modes of reproductive assurance may compensate for reduced pollinator attractiveness (smaller petals) in regions where each mode is most strongly expressed.

Ongoing climate change will negatively impact tree populations unless they are able to acclimate to the changes in their local environment. Effective planning for climate adaptation management requires an understanding of the current state of local adaptation and physiological performance to assess whether populations are at risk of local extinction, to determine if seed movement is appropriate, and to select appropriate seed sources if intervention is needed. We established a new reciprocal transplant experiment (ACE, Adaptation to Climate and Environment) across a latitudinal gradient in North America to examine the impacts of warming on three bur oak (Quercus macrocarpa) populations across much of the species range. We established common gardens in Minnesota, Illinois, and Oklahoma with seedlings grown from seeds collected within 50 km of each of those locations from a total of sixty maternal families. We aimed to 1) assess local adaptation in each of the populations using survival and size as fitness metrics, and 2) evaluate physiological responses to different environments along the latitudinal gradient. We found that northern populations are maladapted to hotter climates as evidenced by their low survival, growth, and photosynthetic rates in the warmest common garden. The southernmost population had the highest survival rate, growth rate, and fitness of the three populations in the southernmost garden, providing evidence for local adaptation to the warmest site. However, conditions in the middle garden resulted in the highest fitness and best physiological performance for all populations. Growth and survival were correlated in the middle garden but were decoupled in the northern and southern gardens. This decoupling is likely due to stress associated with more extreme climates at the ends of the gradient that led to greater resource allocation to survival than to growth. Our results suggest that southern seed sources may perform well in warmer conditions in the north brought on by climate change, which has important implications for managers assisting broadly ranged tree species in adapting to climate change.

Recent reviews of isolating barriers in plants conclude that prezygotic barriers play an outsized role in plant speciation; yet these conclusions derive overwhelmingly from studies of sympatric, perennial herbs in temperate zones, and at later stages of speciation. Trees possess several traits that are expected to influence barrier evolution, including prolonged generation times and reproduction, predominant outcrossing, and long-distance gene flow. We examined early-evolving post-pollination barriers between ecologically diverged, vegetatively distinct varieties of the tree species, Metrosideros polymorpha, that have a common floral morphology and highly overlapping flowering times. We performed controlled crosses between each of Hawaii Islands four varieties and maternal trees of the high-elevation variety and examined pollen-tube growth, fruit set, seed germination, and seedling phenotype. We then monitored survivorship, maturation rate, and fertility of F1 hybrids over [≥]8 years alongside parental controls and a fourth F1 genotype derived from companion studies. The four F1 crosses showed four contrasting patterns and strengths of predominantly postzygotic isolation, including high F1 mortality that manifested over several years. Results from this and other tree studies suggest that ecological speciation in trees follows the classical speciation model of early postzygotic barrier formation followed by reinforcement, whenever stable environments promote recurring hybridization.

1Microbial symbionts have the potential to contribute to host-plants ecological and evolutionary success, especially in plants adaptions to harsh environments, however their role has often been overlooked. Conversely, how host local adaptation (e.g., trait divergence across elevation) shapes the composition of associated microbial symbiont communities remains poorly understood. We explored how foliar endophytic fungi (FEF) abundance, richness, and community composition in three sympatric Monkeyflowers, an ecologically diverse group of flowering plants, change across elevation in the Sierra Nevada, CA, USA. We asked: Q1) Are there differences in leaf functional traits and FEF communities among sympatric Mimulus species populations at similar elevations? Q2) How do traits and FEF communities change across elevation within species? Q3) Are FEF richness, diversity and community composition correlated with leaf functional traits and/or elevation? Q4) How does FEF community composition differ with geographic distance within each species? We collected M. guttatus, M. nasutus, and M.laciniatus individuals from natural populations across the Sierra Nevada, measured leaf functional traits and used ITS sequencing to describe the leaf endophyte community. We found significant associations of FEF community composition with host species, and elevation, suggesting that these factors influence fungal community composition. Furthermore, FEF community dissimilarity was correlated with geographic distance indicating isolation by distance and limited dispersal of fungal endophytes. We detected the prevalence of Vishniacozyma victoriae, an endophyte found most commonly in Antarctica, across all Sierran Mimulus populations. The presence of V. victoriae could play a role in the adaptation of Mimulus to cold, high elevation environments. Our findings offer novel insights into the intricate interactions between fungal endophyte communities, plant traits, and elevation.

Reduced competition or facilitation between kin relative to nonkin can improve plant performance, particularly under resource-limited conditions. Understanding whether kin interactions differ between invasive and native species may provide insights into the mechanisms underlying the persistence and spread of invasive species, particularly for species that spread clonally. To explore this, we conducted a greenhouse experiment using the invasive Alternanthera philoxeroides and its native congener A. sessilis in China. For both species, we grew central plants without or with neighbors, and for the latter we had three intraspecific neighbor kinship treatments (kin only, nonkin only, and both kin and nonkin [mixed] neighbors). To test whether kinship effects are affected by resource limitation, we grew the plants under two watering conditions (well-watered and drought-stressed). Our findings revealed that at both the group (i.e., pot-level) and individual levels, invasive plants had a higher biomass production and experienced a less negative relative neighbor effect in kin groups than in nonkin groups, while these patterns were reversed in the native species. Although aboveground architecture of central plants did not differ significantly between kin and nonkin neighbors in either species, neighbor plants of the invasive species produced fewer nodes in kin groups than in nonkin groups, while the reverse was true for the native species. These patterns were not affected by the watering treatment. Together, these results indicate that while the native plants has stronger kin competition, the invasive species has reduced kin competition. Such reduced competition among kin in the invasive Alternanthera philoxeroides may enhance its population dominance and facilitate its spread.

Little is known about environmental drivers of opportunities for hybridization, but its phylogenetic distribution across species and areas is heterogeneous, suggesting that ecological traits may play an important role in concert with postzygotic isolation. Because plant-pollinator interactions are responsible for gene flow in most plant species, differences in the mosaic landscape of plant-pollinator interactions could explain why some plants are particularly prone to hybridization. Prezygotic isolation is mediated by sometimes complex pollen presentation; conversely, conserved pollination strategies would lead to evolutionary constraints on pollinator assemblage divergence in the speciation process and therefore predict higher opportunities for gene flow, although this hypothesis has yet to be tested. The plant taxonomic tribe Heuchereae (Saxifragaceae) is a well-characterized system for pollinator interactions and particularly for floral scent, the primary pollinator attractant in the group. Floral volatile organic compounds (VOCs) in this clade are hypervariable at the population level and are thought to be responsible for pollination selectivity, leading to divergent pollinator assemblages. Observing a contrast of hybridizing and non-hybridizing species, the levels of attractant divergence may therefore predict levels of hybridization. We investigated pollination biology in the plant genus Heuchera, notable for frequent interspecific gene flow compared to tribal relatives, asking whether high rates of hybridization may be associated with low interspecific divergence of VOCs and the pollinator assemblages they shape, using as our system the hybrid zone between H. americana var. americana and H. richardsonii in the midwestern USA. We optimized a closed-space collection and GC-MS (gas chromatography-mass spectrometry) protocol to characterize VOCs in Heuchera flowers. To identify floral visitation and effective pollinators we conducted pollination observations at 40 Heuchera populations over the span of two field seasons. GC-MS data from 89 Heuchera specimens representing 69 populations suggests that classes of VOCs, and to a large extent individual compounds, are shared within the hybrid complex while other Heuchera that are not thought to hybridize with these species have distinct species-specific compounds. Pollination observations and metabarcoding of pollinator pollen loads confirm shared effective pollinators in the hybrid zone and between adjacent parental populations. Attractant and visitation data considered together suggest that conservatism of pollinator interactions may be a typical feature associated with frequent hybridizers, perhaps arising from developmental or biochemical constraints on prezygotic isolation, and more broadly that the macroevolution of isolation mechanisms may be predictive of natural hybridization rate.

Montane plant populations are experiencing novel conditions due to climate change. Furthermore, climate change is causing increased climate perturbations, such as the 2012-2016 drought in the western US, remarkable in its aridity, longevity, and warmer temperatures. This drought provided an opportunity to understand how montane populations respond to extreme perturbations, including at range limits. We resurrected seeds of the endemic annual plant Erythranthe laciniata, collected in 2008 or earlier (before the drought) and in 2014 (the height of the drought), in a common garden experiment to understand how drought influenced evolution in contemporary field conditions. The study included nine populations across the species range, including range edges. Over 2,100 replicates were sown in three common gardens at natural populations at low, central, and high elevations. We recorded phenology and flower production to estimate lifetime fitness. This experiment took place in 2021, a year with low precipitation and high temperatures. We found higher fitness in the drought generation at the high garden, while both generations showed similar fitness at the central and low gardens. We detected climate adaptation at the low and high gardens, and rapidly evolved faster phenology at the high garden. Lifetime fitness was substantially lower at lower gardens overall, even for low-elevation populations. Low-elevation populations outperformed central populations at the central garden, suggesting adaptive mismatch. Together, these results indicate rapid contemporary adaptation that is beneficial at the leading edge of the species range. Nevertheless, low fitness at lower elevations may foreshadow range contraction under continued climate change.

Juncus bufonius L. s.l. is a species complex with several ploidy levels, for which species delimitation remains unclear due to a lack of reliable morphological characters and the paucity of molecular studies. To clarify taxonomic and geographic relationships in the complex, we combined genomic, cytometric and morphological data from a broad latitudinal range from England down to Spain. We collected morphometric and cytometric data from 31 populations, and genomic data were obtained through Hyb-Seq using the Angiosperm353 kit for a subset of individuals. These three datasets were combined to explore phylogenetic relationships, population structure, and the validity of four previously proposed morphospecies (J. bufonius s.str., a hexaploid; J. minutulus, a tetraploid; and J. ranarius and J. hybridus, both diploids). Sequencing supported the separation of diploids and polyploids as two distinct taxa, but morphometric characters used previously to describe morphospecies showed continuous variation with no diagnostic value, and were not congruent with genomic and cytometric data. Polyploids likely originated through allopolyploidisation from diploids and tetraploids. Phylogenetic lineages were extensively mixed geographically, both for diploid and polyploid taxa, which suggests repeated long-distance dispersal events for both diploids and polyploids, and no separation of taxa by geography. Splitting of diploids into J. ranarius and J. hybridus was not supported. We recommend J. ranarius be treated as a synonym of J. hybridus, and that tetraploids and hexaploids be grouped under J. bufonius. The observed geographical patterns are consistent with high rates of seed dispersal by migratory waterbirds.

Plant architecture, the spatial configuration of stems, branches, leaves, and inflorescences underpins essential physiological functions such as light capture, assimilate partitioning, flowering, and ultimately, yield. In cassava (Manihot esculenta), architectural traits such us plant height, branching level, and plant shape are agronomically important yet remain underexploited in breeding. Here, a large-scale analysis was conducted using phenotypic and genomic data from more than 14,000 cassava accessions evaluated across 34 field locations in Nigeria between 2010 and 2021, encompassing the national breeding programs of the National Root Crops Research Institute and the International Institute of Tropical Agriculture. The study aimed to dissect the genetic architecture, environmental stability, and breeding relevance of four key traits: plant full height, height to first branching, the branching level number (BranchlevelNum) and plant shape. Phenotypic analyses across breeding stages revealed consistent variation in plant height, branching height, and branching intensity, reflecting the cumulative effects of selection and evaluation across environments. Broad-sense heritability estimates ranged from 0.41 to 0.72, with BranchlevelNum and Cylindrical shape exhibiting strong genetic control and weak correlations with yield components, indicating their suitability for independent improvement. Genome-wide association analyses identified significant loci associated with BranchlevelNum, including a major region on chromosome 2 and an additional locus on chromosome 13, collectively explaining approximately 11% of the phenotypic variance. Candidate genes within these regions included regulators of meristem activity and hormone-related pathways, supporting a developmental basis for branching variation. Genomic prediction accuracy for BranchlevelNum reached 0.44, comparable to values reported for key agronomic traits in cassava. These results demonstrate that branching-related architectural traits are genetically tractable, largely independent of yield, and amenable to genomic selection. The findings support the integration of BranchlevelNum and plant shape into ideotype-driven breeding frameworks aimed at improving flowering efficiency, canopy structure, and field performance in cassava. Author SummaryCassava is a major food crop, and its plant shape plays an important role in how easily it can be grown, harvested, and improved through breeding. Traits such as plant height, branching, and canopy form affect flowering, seed production, and field management, yet they have received much less attention than yield or disease resistance. In this study, we examined plant architecture using field and genetic data from more than 14,000 cassava plants grown across Nigeria over twelve years. We focused on key traits describing plant height, branching level, and overall plant shape. We found that branching level is strongly controlled by genetics, remains stable across environments, and can be predicted accurately using genomic data. We also identified specific regions of the cassava genome linked to branching behavior. Our findings show that plant architecture can be improved using modern breeding tools without compromising yield. Incorporating branching traits into breeding programs can help develop cassava varieties that flower more reliably and perform better in farmers fields.

Understanding evolutionary relationships in hyperdiverse plant groups remains a major challenge in systematics. The orchid genus Bulbophyllum, the second largest genus of flowering plants, represents an exceptional example of phylogenetic and morphological complexity. Relationships, particularly within the species-rich Asian clade, have remained poorly resolved due to extensive morphological variation and limited resolution in previous phylogenetic studies. Here, we reconstructed phylogenetic relationships using 63 plastid genes from 355 specimens representing 322 species and 65 of the 97 recognised sections of Bulbophyllum. Our analyses confirmed that the genus comprises five major evolutionary lineages comprised of species predominantly from Australasia, Madagascar, Continental Africa, Neotropics, and Asia. We provide the first robust phylogenetic evidence for a dichotomous split within the Asian clade into two well-supported lineages: the Asian-Malesian clade and the Malesian-Papuasian clade, with the latter containing a strongly supported Papuasian subclade. Additionally, this study supports the monophyly of several currently recognised sections while clarifying relationships in previously problematic groups. This study provides the most comprehensive plastid-based phylogenomic framework for Bulbophyllum to date and establishes a foundation for future taxonomic revision and integrative analyses of diversification and trait evolution within this hyperdiverse genus.

Published studies have reported species-variance between profiles of Calvin-Benson cycle (CBC) intermediates, not only between C4 species and C3 species, but also within C3 species (Arrivault et al., 2019, Borghi et al. 2019). It was proposed that this variance reflects lineage-dependent changes in the balance between different reactions, or poising, of the CBC. These earlier studies investigated phylogenetically-unrelated C3 species. In the current study, CBC intermediates were profiled in five closely-related species from Solanum sect. lycopersicon subsect. Lycopersicum. The levels of individual CBC intermediates showed many significant differences. In a principal component analysis, whilst three species (Solanum lycopersicum, Solanum cheesmaniae, Solanum neorickii) overlapped, Solanum pimpinellifolium and especially Solanum pennellii grouped separately, and were at opposing ends of the distribution. When combined with published data, whilst the separation between Solanum species was retained, they formed a group that was separated from five other C3 species, as well as two C4 species. It is discussed that the observed variation in CBC metabolites profiles within Solanum, together with their separation from other C3 species, supports the idea that CBC evolution is shaped both by phylogenetic relatedness and lineage-specific adaptation. HighlightVariance of intermediate levels points to poising of the Calvin-Benson cycle varying between closely-related species in the tomato clade Solanum sect. lycopersicon subsect. Lycopersicum

BackgroundPlants are exposed to various environmental challenges. With ongoing climate change, droughts and insect outbreaks are expected to become more frequent. Thus, a better understanding is needed of how different plant species respond to such single and combined challenges. This study investigated common versus species-specific responses to environmental challenges in three perennial plant species of different growth forms and whether responses differ intraspecifically among accessions. Clones of different accessions of the herbaceous species Tanacetum vulgare, the woody vine Solanum dulcamara, and the tree Populus nigra were subjected to similar control, herbivory, drought, and combined (drought and herbivory) treatments for the same periods. After the exposure, concentrations of foliar phytohormones and various morphological traits were measured. ResultsAcross all species, several foliar phytohormones and one of ten morphological traits responded consistently to the environmental challenges. Jasmonoyl-isoleucine was induced by herbivory and the combined treatment, abscisic acid (ABA) by drought and the combined treatment, and indole acetic acid by the combined treatment in all species. Root mass remained unchanged in all species. However, structural equation models (SEMs) revealed a shared regulatory pathway across species in which ABA connected treatment and root mass, indicating a common hormonal response potentially linking challenges to growth responses. Despite these common patterns, species-specific responses were pronounced. In P. nigra, a unique induction of salicylic acid was found under the combined treatment, while aboveground mass and root-shoot ratio remained unaffected by any treatment, in contrast to the other two species. Species-specific SEMs further indicated distinct phytohormone-mediated pathways underlying morphological variation. Phenotypic plasticity reflected these species-specific patterns, with none of the phytohormones or morphological traits exhibiting uniform plasticity across species. Intraspecific variation further shaped responses, as phytohormone and morphological trait plasticity depended on accession, indicating substantial accession-specific plant responses. ConclusionsOur results indicate that some responses to comparable challenges may be conserved across species, while others are species-specific. The combined treatment elicited the most pronounced responses, and such complex responses may become more frequent under current global change. Our study highlights that comprehensive understanding of plant responses requires systematic comparisons at both interspecific and intraspecific scales.

Timing of flowering is shifting with climate change. Although climate-driven shifts in phenology sometimes affect seed production, whether changing phenology will scale up to affect population dynamics of long-lived plants remains largely unknown, particularly under changing precipitation. Understanding how phenology affects persistence and extinction risk is a pressing need given contemporary biodiversity loss. We combined nearly a decade of demographic censuses and a four-year phenological survey in a rainfall manipulation experiment to examine the effects of experimental drought and irrigation on flowering phenology, vital rates (e.g., survival and individual growth), and population growth in the perennial herb Lomatium utriculatum. We found that drought advanced flowering by 3.3 days on average, and that earlier-flowering plants produced more seeds regardless of treatment. However, both rainfall treatments reduced seed production compared to controls. We quantified the phenology-mediated and direct, non-phenological effects of rainfall manipulation on population growth rates using integral projection models and a life table response experiment. Drought and irrigation increased {lambda} through increased individual growth, but these effects were partially negated by treatment-driven declines in seed output. In contrast, changes to seed production resulting from shifting flowering times had negligible effects on population growth. Our results suggest that climate-driven phenological shifts may only marginally impact population dynamics in perennial plants and highlight that assessing phenologys consequences for persistence under climate change must also account for direct demographic effects of the climate driver(s) themselves. SignificanceWill changing flowering times under climate change increase extinction risk in plant populations? Despite well-documented earlier flowering and its influence on the number of offspring produced, how changing flowering times will affect population growth or decline is still mostly unknown. We study this in a perennial wildflower subject to changes in rainfall. While we found that drought meant earlier flowering and that, all else equal, early flowering meant more seeds, these effects only marginally affected population growth. Instead, population growth was influenced mostly by rainfall-driven changes to individual plant growth. While shifting flowering times remain an important indicator of climate change, assessing extirpation in plants requires considering flowering times as only one of many life cycle processes changing with climate.