Applications in Plant Sciences

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.

Orobanchaceae is the largest family of parasitic plants, encompassing a full spectrum of parasitic strategies, ranging from autotrophic to holoparasitic. Agalinis is a genus of facultative hemiparasites comprising about 70 species distributed throughout the Americas, including several endemic and rare taxa. Agalinis fasciculata, the beach false foxglove, is a widely distributed species across southeastern North America. Here, we use PacBio HiFi, Omni-C, and RNA-seq data to generate the first high-quality reference genome for the genus. The nuclear genome is 2.29 Gb in size, with most sequences anchored to 14 pseudochromosomes and an N50 of 162 Mb. BUSCO analyses indicate high completeness (98.4%). Structural genome annotation identified 34,133 protein-coding genes and 39,266 transcripts, most of which have at least one functional annotation. The plastid and mitochondrial genomes were also assembled. We further examined genetic diversity and demographic history in A. fasciculata, revealing low genome-wide heterozygosity and evidence of inbreeding. This reference genome is an important resource for understanding the evolutionary history of the genus and the evolutionary patterns of parasitism within Orobanchaceae. SignificanceThis high-quality genome is the first chromosome-level assembly for Agalinis, a hemiparasitic genus in the plant family Orobanchaceae. It improves the taxon sampling within Orobanchaceae, representing an important resource for investigating patterns of genome evolution in parasitic lineages. Furthermore, Agalinis has served as a focal genus for studies of the anatomy of haustorial development, and genome annotation incorporated RNA from multiple tissues, enabling the identification of genes expressed in different tissues, including roots and haustoria. This genome also serves as a reference for evolutionary studies of other Agalinis species, many of which are endemic and of conservation concern in North and South America. Overall, the beach false foxglove genome will support studies of the evolutionary history of Agalinis and genome evolution across Orobanchaceae.

Plant DNA metabarcoding enables the identification of plant taxa in mixed samples, with the trnL (UAA) intron and its P6 loop mini-barcode region performing as well as or better than other commonly used markers. Reliable metabarcoding requires high-quality reference databases, yet a regularly maintained trnL resource is currently lacking. Consequently, most studies use uncurated sequences downloaded directly from public repositories without essential validation. We address these gaps by providing guidance through a systematic comparison of three database curation tools - OBITools3/ecoPCR, RESCRIPt, and MetaCurator - to generate three trnL reference sequence databases and evaluate their classification performance across commonly sequenced trnL regions (CD, CH, and GH). Reference trnL sequences and taxonomy files were retrieved from public sequence repositories and curated using standardized filtering steps to reduce taxonomic errors, sequence ambiguity, and redundancy. Four simulated query datasets--two base sets and their mutated counterparts--were constructed to assess classification performance of the databases using the Naive Bayesian Classifier implemented in DADA2.- The evaluation showed that performance differed by trnL region: MetaCurator and RESCRIPt yielded higher and similar metrics for trnL CD; OBITools3/ecoPCR and RESCRIPt were comparable for trnL CH; and MetaCurator attained the highest performance for trnL GH region. All reference databases, taxonomy, and evaluation files are available at Zenodo (https://doi.org/10.5281/zenodo.17969450). The complete computational workflow and scripts are available on GitHub (https://github.com/oskuddar/trnL_DB). Although evaluation was focused on plant taxa in the United States, the resulting databases are suitable for use as global trnL reference databases.

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.

BackgroundIn recent years, the rise of AI-mediated technological assistance has impacted applied mycology. Various tools employ AI to analyze images of Macromycetes fruiting bodies for species identification. This trend has sparked widespread interest in online applications despite the potential risks associated with relying on AI-generated advice. MethodsWe conducted a comparative analysis of popular AI-based mushroom identification tools using over 100 original photographs of fungi fruiting bodies from nearly 60 species, taken in real-world conditions. Reference searches were conducted with mushroom names in five languages, including Latin species names. Functional test scores and an overall accuracy score were calculated for twelve selected AI applications to evaluate their general reliability. ResultsThe AI-based applications evaluated in the study were able to recognize only a portion of the provided mushroom images. Even the best-performing tools frequently failed to accurately identify fruiting bodies in real-world conditions. None of the tested applications consistently provided a single, correct species name. Instead, users were often presented with multiple options, among which the correct answer might have been found. ConclusionWhen addressing mycological queries, it is crucial to recognize the inherent risk of relying solely on AI-mediated resources for mushroom identification. Various limitations hinder their effectiveness in real-world environments. These tools should only be viewed as supplementary aids since they are inadequate for making definitive or safety-critical decisions.

O_LIRoot colonisation by endomycorrhizal fungi can indicate habitat condition. However, due to the significant time required to assess colonisation using traditional microscope techniques, studies of colonisation at large scales are impractical. AI-powered approaches may increase output and facilitate ecosystem assessments. C_LIO_LIWe trained our AI-powered tool MycorrhizaFinder (MFKew) on field roots from diverse ecosystems. It was trained to recognise a range of arbuscular and ericoid mycorrhizal fungal structures, and to differentiate dark septate endophytes common in field-sourced roots. C_LIO_LIHere we describe the semi-automated workflow from root processing and microscope slide scanning to model training and performance evaluation, proposing Macro F1 as the appropriate metric to be optimised. Without human supervision, Macro F1 currently stands at 66% for arbuscular and at 57% for ericoid mycorrhizal colonisation assessment. C_LIO_LIMFKew is user friendly, requires no programming skills and offers flexibility for advanced users who wish to further train the tool using their own labelled mycorrhizal root datasets, including images acquired from different devices or staining protocols. This adaptability allows users to customize the model for specific needs, making it optimal for ecologists and agronomists. Additionally, MFKew supports large-scale, repeatable, medium-throughput monitoring across ecosystems, enabling the assessment of mycorrhizal status and tracking changes over time. C_LI

Flow cytometry provides a reliable and fast method for estimating genome size and ploidy levels in plants. Until recently, most studies employed fresh tissues, which limits the use of the method with samples from remote areas or when an extremely high number of samples needs to be processed in a short time. Although there is growing evidence that silica-dried material can be used for ploidy estimation in some taxa, no flora-wide study has been available so far. Here, we provide methodological aspects of an unprecedented study exploring ploidy variation of non-apomictic angiosperms in the Eastern Alps. We have analysed ca. 45,000 silica-dried samples of 1135 species using flow cytometry with DAPI as stain. We were able to obtain ploidy level information from 1104 (97%) of species. The unsuccessful species included succulent plants of the family Crassulaceae (genera Jovibarba, Rhodiola, Sedum, Sempervivum), the achlorophyllous parasitic or mycoheterotrophic genera Orobanche and Hypopitis, and a handful of others. About 80% of samples were successfully analysed using a single universal protocol and leaf tissue, while in the remaining species the use of alternative tissues (such as petioles or flowers) and/or protocol modifications were needed (targeting composition of buffers, duration of fixation or staining time or use of alternative buffers). A total of 377 species (34%) included polyploid cytotypes and 179 (16%) species were ploidy-variable. As a community resource, we provide relative genome sizes and ploidy assignments of 1332 cytotypes retrieved from 1104 species along with methodological details (e.g. buffers, standards, analysed plant organs, histogram quality). We believe that this dataset will facilitate future research in particular species as well as in flora-wide investigations of ploidy level variation of the Central European flora in general. We are confident that novel cytotypes of many species will be discovered in other geographic areas, and we would be delighted if the present dataset could serve the botanical community for comparison.

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.

This paper presents a novel workflow leveraging Large Language Models (LLMs) to rapidly extract trait data from fungal species descriptions, addressing a significant bottleneck in ecological research. We developed and evaluated an LLM pipeline to extract morphological trait data from arbuscular mycorrhizal fungi, comparing performance against a manually curated dataset (TraitAM). Results demonstrate the potential of LLMs for automated trait data acquisition, though accuracy varies by trait and model, with systematic biases observed. This framework offers a blueprint for building trait databases across diverse taxa and domains, significantly accelerating ecological research and conservation efforts.

Root segmentation is a fundamental yet challenging task in image-based plant phenotyping. We present the first systematic comparison of Transformer and Convolutional Neural Network (ConvNet) architectures for root segmentation, evaluating 21 architectures across nine diverse datasets and comparing pre-trained models to training from scratch. Transformer-based models significantly outperform ConvNets for segmentation accuracy and root-diameter agreement. Pre-training significantly improves mean Dice from 0.623 to 0.666 (p = 3.3 x 10-10). We also find that Transformers benefit more from pre-training than ConvNets, with Dice improvements of +0.072 versus +0.022 (p = 3.7 x 10-4), supporting the hypothesis that fine-tuned Transformers transfer more effectively across large domain gaps. Among evaluated models, MobileSAM achieved the highest Dice score while maintaining computational efficiency. Dataset choice explained far more performance variance (70.9%) than model architecture (6.7%), suggesting that data curation matters more than model selection.

BackgroundPhylogenomic studies frequently reveal widespread gene tree discordance, primarily arising from incomplete lineage sorting (ILS) and hybridization and/or introgression. Disentangling these processes is especially challenging in rapidly radiating lineages. The genus Oryza, with its rapid diversification and multiple genome types, exemplifies this pervasive phylogenetic incongruence. We integrated multiple genomic datasets including whole-genome resequencing, transcriptomes, and published genomes from diploid Oryza species. Concatenation and multispecies coalescent analyses recovered a robust, congruent species tree, placing the FF and GG genome groups as a monophyletic basal clade, followed by successive divergence of the EE, CC, BB, and AA lineages, a topology differing from some prior hypotheses. ResultsTo assess the sources of discordance, we employed a suite of complementary phylogenomic methods. Quantifying introgression via Branch Lengths (QuIBL)-based model comparisons suggested that [~]74.17% of gene tree-species tree discordance is better explained by post-speciation introgression, whereas only [~]15.56% is consistent with ILS alone. Phylogenetic networks (PhyloNet) and allele-sharing statistics (D-statistics, f-branch) corroborated these results, indicating widespread historical introgression both within and between genome groups. Furthermore, genome-wide scans using the fdM statistic localized introgressed genomic regions, which showed reduced interspecific divergence and were enriched for genes involved in stress responses and metabolism. ConclusionsTaken together, our results demonstrate that historical introgression, not ILS, is the dominant force shaping phylogenetic discordance in diploid Oryza. The integrative phylogenomic framework implemented here, which quantifies the contributions of introgression versus ILS and maps the genomic footprint of gene flow, provides a replicable strategy for resolving complex evolutionary histories in other rapidly radiating lineages.

Understanding crop genetic diversity is essential for conservation and breeding, yet farmer-maintained germplasm remains largely underrepresented in genomic studies. Theobroma cacao L. has a complex domestication history and extensive global diversity, and cacao currently cultivated in Central America, particularly in Costa Rica, has been understudied compared to South American and Mexican cultivars despite cultural and historical importance. In this study, we investigate the genetic diversity of cacao from farmer-managed systems across Costa Rica to search for Criollo germplasm and identify and characterize any unique local genetic groups. Ninety-four trees were sampled from 17 farms across four regions of the country and sequenced using whole genome resequencing. Farmer materials were analyzed alongside 166 previously characterized reference accessions representing major cacao genetic groups. Population structure analyses, phylogenetic reconstruction, and network approaches revealed that Costa Rican cacao encompasses multiple known genetic groups, including Criollo-derived lineages, while also harboring locally distinct diversity not fully represented in current global reference collections. Analyses revealed close kinship between many accessions with no clear geographic patterns corresponding to the observed population differentiation, reflecting the effects of farmers in creating dominant patterns of gene flow through seed-saving, clonal propagation, and sharing genotypes among farms. Heterozygosity levels varied substantially among individuals, consistent with a mixture of highly inbred Criollo trees and more heterozygous, admixed genotypes. We find that farmer-managed cacao systems are reservoirs of genetic diversity, including possibly rare or historically important lineages, underscoring the value of these farming systems for effective conservation and management of genomic resources for cacao resilience and improvement.

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.

Integrating taxonomic data from various sources presents a significant challenge in the study of biodiversity research, due to non-standardized nomenclature and evolving species classifications. Discrepancies between major repositories like the Global Biodiversity Information Facility (GBIF) and the National Center for Biotechnology Information (NCBI), as well as citizen science platforms such as iNaturalist, lead to fragmented and sometimes inaccurate biological data. We present TaxonMatch, a tool designed to address these challenges. TaxonMatch aligns taxonomic names, resolves synonymy, and corrects typographical and structural inconsistencies across databases. We show how it can be used to build a common backbone arthropod taxonomy over NCBI, GBIF and iNaturalist, to find the closest molecular data to a given fossil, and to identify IUCN endangered species with molecular data. TaxonMatch provides a cohesive taxonomic framework and a consistent taxonomic backbone, and can be applied to any taxonomic source. The tool is available at https://github.com/MoultDB/TaxonMatch.

The combination of target capture sequencing (TCS) with low-coverage whole genome sequencing (lcWGS), an approach known as Hyb-Seq, has allowed the integration of natural history collections into the genomics revolution, transforming biodiversity research. To implement Hyb-Seq, a collection of genomic targets, often nuclear orthologs, is needed to design probes for TCS. In flowering plants, the universal Angiosperms353 probe set has been proven resolutive at multiple evolutionary scales, with caveats. Malpighiales is known to be one of the most challenging flowering plant orders to resolve. Within this order, the clusioid clade ([~]2.2K species, 94 genera, five families) is no exception. To resolve phylogenetic relationships in this recalcitrant clade, we design a custom probe set, the Clusioids626 kit, composed of 39,936 120-mer probes targeting 626 nuclear orthologs ([~]6.6M nucleotides). This probe set includes all Angiosperms353 targets and 273 clusioid-specific ones, carefully chosen taking copy-number, length evenness, and phylo-informativeness into account. We test our probe set on 70 accessions representing all families and tribes in the clusioid clade. On average, 50.4% of TCS reads mapped to our targets, recovering a median of [~]600 orthologs. Relationships for all clusioid families are fully resolved for our nuclear targets. Additionally, 105 plastid coding DNA sequences were retrieved from the lcWGS fraction. A strong cyto-nuclear conflict was detected. The Clusioids626 kit performs better than the universal Angiosperms353 enrichment panel alone. Our kit design workflow can be extended into other lineages for which a universal probe set exists but more resolution is needed.

Species are the fundamental analytical units of evolutionary processes; thus evidence-based species delimitation is a crucial step for understanding species radiations. However, the task of delimiting species is particularly challenging in the context of a syngameon--a group of distinct, but closely related species that have incomplete reproductive isolation and frequently hybridize in nature. This problem is further exacerbated by the presence of cryptic species--species that are phenotypically distinct, though difficult to distinguish with gross morphology alone. Heuchera subsect. Heuchera comprises both clear and cryptic species within a syngameon that has seen study from morphological, experimental, and phylogenetic aspects. This group has long been recognized for its taxonomic complexity, namely two recognized hybrid zones with extreme morphological variation and persistent non-monophyly among parental populations. Here, we reassess species limits within Heuchera subsect. Heuchera, focusing on the hybrid complex between H. americana and H. richardsonii and adjacent H. americana populations. We use a multipronged approach with deep population-level sampling to 1) assess the genetic structure of 655 individuals across the geographic range of the H. americana group to identify genetic lineages and 2) assess the phenotypic diagnosability of these lineages. Despite extensive admixture and gene tree conflict, we find multiple cohesive lineages with diagnosable phenotypes. We recognize five species and three varieties within the H. americana group, one new and four resurrected. Our results demonstrate that even highly reticulate syngameons can be partitioned into meaningful taxonomic units with multiple lines of evidence.

Genomic imprinting is a rare epigenetic phenomenon in plants and animals, defined by parent-of-origin specific gene expression. Its molecular mechanisms and evolutionary significance remain incompletely understood. In this study, we investigated whether genomic imprinting occurs in Scots pine and, by extension, in other conifers to gain insight into the evolutionary origins of imprinting. We performed reciprocal crosses to assess imprinting in seed embryos and applied a unique approach that used exome-capture data from the haploid, maternally inherited megagametophyte tissue to identify maternal alleles, thereby allowing us to infer paternal alleles in the embryos of the same seeds. Our findings show that maternally inherited haploid megagametophyte tissue offers an effective strategy for resolving parental alleles in offspring while simultaneously removing extensive paralogous variation from the dataset. This framework is broadly applicable to other conifer species and to taxa that possess comparable maternally derived haploid tissues. No evidence of genomic imprinting was detected. Although the limited overlap between the exome-capture and RNA-sequencing datasets and the stringent paralog filtering reduced the amount of analyzable data considerably, the absence of detectable imprinting may also reflect genuinely weak or absent imprinting signals in conifers. We identified several limitations in this preliminary study and outline recommendations for future work to overcome them, and additional research will be necessary to determine whether genomic imprinting occurs in conifers

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.

Abstract summaryThis report presents a preliminary bryoflora for Grand Staircase-Escalante National Monument (GSENM) in southern Utah. The inventory included over 1000 collections made across 40 localities (i.e. macrohabitat types) spanning two ecologically important gradients in bryophyte habitat: shade and moisture availability. At present, the growing checklist contains 117 taxa of liverworts and mosses including 27 families, 65 genera, 116 species, 9 varieties, and 1 subspecies. Noteworthy records include 49 putative taxa new for the state of Utah, and 2 undescribed species in the genera Grimmia and Schistidium. We propose 4 of these species be considered for addition to the recently revised bryoflora of North America. As expected for arid and semiarid environments, the bryophytes of GSENM are predominantly acrocarpous mosses (75%) followed by pleurocarpous mosses (16%), thalloid liverworts (7%), and leafy liverworts (2%). The most diverse families included xeric-soil acrocarpous mosses in the Pottiaceae (35%) and xeric-rock acrocarpous mosses in the Grimmiaceae (15%). Both xeric and mesic species were recovered in the Bryaceae (10% of species) while the pleurocarpous Amblystegiaceae included mesic and hydric species (7%). Most species in the bryoflora have broad global or disjunct distributions, but notably, the known distribution of 17 species appears limited in the United States, or globally, and warrant monitoring in GSENM. Using floristic habitat sampling across 19 macrohabitat types (combinations of 6 topography and 7 vegetation classes), mean site richness was 17.2 {+/-} 9 (SD) and ranged from 4 to 34 species. Six diversity hotspots supported [≥]30 species and were canyons with perennial or ephemeral streams dominated by mixed conifer, hardwood-riparian, riparian, or pinyon-juniper vegetation. High richness is likely supported by greater habitat diversity including xeric, mesic, and hydric conditions on variable substrates (e.g. rock, soil, biocrust, downed wood, seeps, and riparian aquatic/semi-aquatic habitat). Consequently, managing and monitoring diversity under future climate change and land-use alterations will necessitate a habitat-stratified approach that utilizes repeated floristic habitat sampling to document changes in site-level richness and to predict other candidate diversity hotspots on the basis of microhabitat-level diversity, which could be assessed by trained non-bryologists. Collection data are available to the public as georeferenced and photographed observations of half of the bryophyte collections on our iNaturalist.com project, Bryophytes of Grand Staircase Escalante, available for scientific, educational, or outreach activities. Observations are accessible to visitors (via the smartphone app) who wish to know what species have been found along popular trails in GSENM. Landscape-level richness may not reach that of the neighboring Grand Canyon National Park (>155 species), which supports a unique high-elevation bryophyte community sheltered in the mixed conifer and spruce-fir forests of the North Rims Kaibab Plateau. Future collecting by experts will inevitably uncover more species in this ecologically diverse monument important to conserving dryland bryophyte diversity and ecosystem function. This study will serve as a baseline for future research and long-term monitoring related to climate change impacts on dryland bryophytes including biocrust species. O_FIG O_LINKSMALLFIG WIDTH=158 HEIGHT=200 SRC="FIGDIR/small/708354v1_ufig1.gif" ALT="Figure 1"> View larger version (106K): org.highwire.dtl.DTLVardef@c50925org.highwire.dtl.DTLVardef@1db65c3org.highwire.dtl.DTLVardef@f60909org.highwire.dtl.DTLVardef@cb5244_HPS_FORMAT_FIGEXP M_FIG C_FIG Cover photos (by T. A. Clark): View of sandstone canyon wall along the Escalante River Trail taken during a July collection trip in 2015 (top) during which riparian bryophytes were collected by authro, T. A. Clark, (shown in photo) at a sandstone seep (bottom). Bureau of Land Managements National Landscape Conservation System Grant Cooperative Agreement #L14AC00275 issued to P.I. Lloyd R. Stark, UNLV O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=151 SRC="FIGDIR/small/708354v1_ufig2.gif" ALT="Figure 2"> View larger version (88K): org.highwire.dtl.DTLVardef@86109eorg.highwire.dtl.DTLVardef@1ad0efborg.highwire.dtl.DTLVardef@b2c239org.highwire.dtl.DTLVardef@1ed4a4a_HPS_FORMAT_FIGEXP M_FIG C_FIG Copyright 2020 by the authorsAll content contained herein is the property of the authors and all images the property of Theresa A. Clark and should not be used without permission except for education, in which case inclusion of the author/photographers name in citation or superimposed over any image(s) is requested.

Grapevine cluster architecture is a key selection target in breeding programs because it influences disease susceptibility, yield stability and juice quality. High-throughput phenotyping offers a rapid and non-destructive approach to capture biochemical and structural variation in these traits, yet the influence of plant organ reflectance and data partitioning strategies on trait prediction remains poorly understood. In this study, we evaluated how hyperspectral reflectance from different grapevine organs contributes to the prediction of cluster architecture and juice quality traits in two clonal populations of Riesling and Pinot. Using partial least squares regression (PLSR), we assessed the prediction accuracy of eight cluster architecture and six juice quality traits under two data partitioning strategies. Models based on cluster reflectance outperformed those using dry leaf reflectance for most traits, except for pH. Partitioning the dataset by cluster type increased trait variance and improved predictions for number of berries (R{superscript 2} = 0.53), berry diameter (R{superscript 2} = 0.79), and total acidity (R{superscript 2} = 0.48). Visible, red-edge and NIR spectra were most informative regions to predict the traits studied. Together, our results highlight the importance of organ-specific data and appropriate calibration strategies to improve phenomic models for the development of scalable proxies for grapevine improvement. HighlightSpectral phenomics reveals that prediction accuracy in grapevine depends on organ spectral signatures and traits, with cluster reflectance outperforming leaves, informing new phenotyping strategies for breeding improvement.