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Abstract Understanding the relationship between intraspecific trait variability (ITV) and its biotic and abiotic drivers is crucial for advancing population and community ecology. Despite its importance, there is a lack of guidance on how to effectively sample ITV and reduce bias in the resulting inferences. In this study, we explored how sample size affects the estimation of population‐level ITV, and how the distribution of sample sizes along an environmental gradient (i.e., sampling design) impacts the probabilities of committing Type I and II errors. We investigated Type I and II error probabilities using four simulated scenarios which varied sampling design and the strength of the ITV‐environment relationships. We also applied simulation scenarios to empirical data on populations of the small mammal,Peromyscus maniculatusacross gradients of latitude and temperature at sites in the National Ecological Observatory Network (NEON) in the continental United States. We found that larger sample sizes reduce error rates in the estimation of population‐level ITV for both in silico andPeromyscus maniculatuspopulations. Furthermore, the influence of sample size on detecting ITV‐environment relationships depends on how sample sizes and population‐level ITV are distributed along environmental gradients. High correlations between sample size and the environment result in greater Type I error, while weak ITV–environmental gradient relationships showed high Type II error probabilities. Therefore, having large sample sizes that are even across populations is the most robust sampling design for studying ITV‐environment relationships. These findings shed light on the complex interplay among sample size, sampling design, ITV, and environmental gradients. 

Soil microbiomes are heterogeneous, complex microbial communities. Metagenomic analysis is generating vast amounts of data, creating immense challenges in sequence assembly and analysis. Although advances in technology have resulted in the ability to easily collect large amounts of sequence data, soil samples containing thousands of unique taxa are often poorly characterized. These challenges reduce the usefulness of genome-resolved metagenomic (GRM) analysis seen in other fields of microbiology, such as the creation of high quality metagenomic assembled genomes and the adoption of genome scale modeling approaches. The absence of these resources restricts the scale of future research, limiting hypothesis generation and the predictive modeling of microbial communities. Creating publicly available databases of soil MAGs, similar to databases produced for other microbiomes, has the potential to transform scientific insights about soil microbiomes without requiring the computational resources and domain expertise for assembly and binning. 

Forecasting the impacts of changing climate on the phenology of plant populations is essential for anticipating and managing potential ecological disruptions to biotic communities. Herbarium specimens enable assessments of plant phenology across broad spatiotemporal scales. However, specimens are collected opportunistically, and it is unclear whether their collection dates – used as proxies of phenological stages – are closest to the onset, peak, or termination of a phenophase, or whether sampled individuals represent early, average, or late occurrences in their populations. Despite this, no studies have assessed whether these uncertainties limit the utility of herbarium specimens for estimating the onset and termination of a phenophase. Using simulated data mimicking such uncertainties, we evaluated the accuracy with which the onset and termination of population‐level phenological displays (in this case, of flowering) can be predicted from natural‐history collections data (controlling for biases in collector behavior), and how the duration, variability, and responsiveness to climate of the flowering period of a species and temporal collection biases influence model accuracy. Estimates of population‐level onset and termination were highly accurate for a wide range of simulated species' attributes, but accuracy declined among species with longer individual‐level flowering duration and when there were temporal biases in sample collection, as is common among the earliest and latest‐flowering species. The amount of data required to model population‐level phenological displays is not impractical to obtain; model accuracy declined by less than 1 day as sample sizes rose from 300 to 1000 specimens. Our analyses of simulated data indicate that, absent pervasive biases in collection and if the climate conditions that affect phenological timing are correctly identified, specimen data can predict the onset, termination, and duration of a population's flowering period with similar accuracy to estimates of median flowering time that are commonplace in the literature. 

Phenology varies widely over space and time because of its sensitivity to climate. However, whether phenological variation is primarily generated by rapid organismal responses (plasticity) or local adaptation remains unresolved. Here we used 1,038,027 herbarium specimens representing 1,605 species from the continental United States to measure flowering-time sensitivity to temperature over time (Stime) and space (Sspace). By comparing these estimates, we inferred how adaptation and plasticity historically influenced phenology along temperature gradients and how their contributions vary among species with different phenology and native climates and among ecoregions differing in species composition. Parameters Sspace and Stime were positively correlated (r = 0.87), of similar magnitude and more frequently consistent with plasticity than adaptation. Apparent plasticity and adaptation generated earlier flowering in spring, limited responsiveness in late summer and delayed flowering in autumn in response to temperature increases. Nonetheless, ecoregions differed in the relative contributions of adaptation and plasticity, from consistently greater importance of plasticity (for example, southeastern United States plains) to their nearly equal importance throughout the season (for example, Western Sierra Madre Piedmont). Our results support the hypothesis that plasticity is the primary driver of flowering-time variation along temperature gradients, with local adaptation having a widespread but comparatively limited role. 

The ecology of forest ecosystems depends on the composition of trees. Capturing fine-grained information on individual trees at broad scales provides a unique perspective on forest ecosystems, forest restoration, and responses to disturbance. Individual tree data at wide extents promises to increase the scale of forest analysis, biogeographic research, and ecosystem monitoring without losing details on individual species composition and abundance. Computer vision using deep neural networks can convert raw sensor data into predictions of individual canopy tree species through labeled data collected by field researchers. Using over 40,000 individual tree stems as training data, we create landscape-level species predictions for over 100 million individual trees across 24 sites in the National Ecological Observatory Network (NEON). Using hierarchical multi-temporal models fine-tuned for each geographic area, we produce open-source data available as 1 km²shapefiles with individual tree species prediction, as well as crown location, crown area, and height of 81 canopy tree species. Site-specific models had an average performance of 79% accuracy covering an average of 6 species per site, ranging from 3 to 15 species per site. All predictions are openly archived and have been uploaded to Google Earth Engine to benefit the ecology community and overlay with other remote sensing assets. We outline the potential utility and limitations of these data in ecology and computer vision research, as well as strategies for improving predictions using targeted data sampling. 

# Individual Tree Predictions for 100 million trees in the National Ecological Observatory Network Preprint: https://www.biorxiv.org/content/10.1101/2023.10.25.563626v1 ## Manuscript Abstract The ecology of forest ecosystems depends on the composition of trees. Capturing fine-grained information on individual trees at broad scales allows an unprecedented view of forest ecosystems, forest restoration and responses to disturbance. To create detailed maps of tree species, airborne remote sensing can cover areas containing millions of trees at high spatial resolution. Individual tree data at wide extents promises to increase the scale of forest analysis, biogeographic research, and ecosystem monitoring without losing details on individual species composition and abundance. Computer vision using deep neural networks can convert raw sensor data into predictions of individual tree species using ground truthed data collected by field researchers. Using over 40,000 individual tree stems as training data, we create landscape-level species predictions for over 100 million individual trees for 24 sites in the National Ecological Observatory Network. Using hierarchical multi-temporal models fine-tuned for each geographic area, we produce open-source data available as 1km^2 shapefiles with individual tree species prediction, as well as crown location, crown area and height of 81 canopy tree species. Site-specific models had an average performance of 79% accuracy covering an average of six species per site, ranging from 3 to 15 species. All predictions were uploaded to Google Earth Engine to benefit the ecology community and overlay with other remote sensing assets. These data can be used to study forest macro-ecology, functional ecology, and responses to anthropogenic change. ## Data Summary Each NEON site is a single zip archive with tree predictions for all available data. For site abbreviations see: https://www.neonscience.org/field-sites/explore-field-sites. For each site, there is a .zip and .csv. The .zip is a set 1km .shp tiles. The .csv is all trees in a single file. ## Prediction metadata *Geometry* A four pointed bounding box location in utm coordinates. *indiv_id* A unique crown identifier that combines the year, site and geoindex of the NEON airborne tile (e.g. 732000_4707000) is the utm coordinate of the top left of the tile.  *sci_name* The full latin name of predicted species aligned with NEON's taxonomic nomenclature.  *ens_score* The confidence score of the species prediction. This score is the output of the multi-temporal model for the ensemble hierarchical model.  *bleaf_taxa* Highest predicted category for the broadleaf submodel *bleaf_score* The confidence score for the broadleaf taxa submodel  *oak_taxa* Highest predicted category for the oak model  *dead_label* A two class alive/dead classification based on the RGB data. 0=Alive/1=Dead. *dead_score* The confidence score of the Alive/Dead prediction.  *site_id* The four letter code for the NEON site. See https://www.neonscience.org/field-sites/explore-field-sites for site locations. *conif_taxa* Highest predicted category for the conifer model *conif_score* The confidence score for the conifer taxa submodel *dom_taxa* Highest predicted category for the dominant taxa mode submodel *dom_score* The confidence score for the dominant taxa submodel ## Training data The crops.zip contains pre-cropped files. 369 band hyperspectral files are numpy arrays. RGB crops are .tif files. Naming format is __, for example. "NEON.PLA.D07.GRSM.00583_2022_RGB.tif" is RGB crop of the predicted crown of NEON data from Great Smoky Mountain National Park (GRSM), flown in 2022.Along with the crops are .csv files for various train-test split experiments for the manuscript. ### Crop metadata There are 30,042 individuals in the annotations.csv file. We keep all data, but we recommend a filtering step of atleast 20 records per species to reduce chance of taxonomic or data cleaning errors. This leaves 132 species. *score* This was the DeepForest crown score for the crop. *taxonID*For letter species code, see NEON plant taxonomy for scientific name: https://data.neonscience.org/taxonomic-lists *individual*unique individual identifier for a given field record and crown crop *siteID*The four letter code for the NEON site. See https://www.neonscience.org/field-sites/explore-field-sites for site locations. *plotID* NEON plot ID within the site. For more information on NEON sampling see: https://www.neonscience.org/data-samples/data-collection/observational-sampling/site-level-sampling-design *CHM_height* The LiDAR derived height for the field sampling point. *image_path* Relative pathname for the hyperspectral array, can be read by numpy.load -> format of 369 bands * Height * Weight *tile_year*  Flight year of the sensor data *RGB_image_path* Relative pathname for the RGB array, can be read by rasterio.open() # Code repository The predictions were made using the DeepTreeAttention repo: https://github.com/weecology/DeepTreeAttentionKey files include model definition for a [single year model](https://github.com/weecology/DeepTreeAttention/blob/main/src/models/Hang2020.py) and [Data preprocessing](https://github.com/weecology/DeepTreeAttention/blob/cae13f1e4271b5386e2379068f8239de3033ec40/src/utils.py#L59). 

ABSTRACT AimEcological theory suggests that dispersal limitation and selection by climatic factors influence bacterial community assembly at a continental scale, yet the conditions governing the relative importance of each process remains unclear. The carnivorous pitcher plantSarracenia purpureaprovides a model aquatic microecosystem to assess bacterial communities across the host plant's north–south range in North America. This study determined the relative influences of dispersal limitation and environmental selection on the assembly of bacterial communities inhabitingS. purpureapitchers at the continental scale. LocationEastern United States and Canada. Time Period2016. Major Taxa StudiedBacteria inhabitingS. purpureapitchers. MethodsPitcher morphology, fluid, inquilines and prey were measured, and pitcher fluid underwent DNA sequencing for bacterial community analysis. Null modelling of β‐diversity provided estimates for the contributions of selection and dispersal limitation to community assembly, complemented by an examination of spatial clustering of individuals. Phylogenetic and ecological associations of co‐occurrence network module bacteria was determined by assessing the phylogenetic diversity and habitat preferences of member taxa. ResultsDispersal limitation was evident from between‐site variation and spatial aggregation of individual bacterial taxa in theS. purpureapitcher system. Selection pressure was weak across the geographic range, yet network module analysis indicated environmental selection within subgroups. A group of aquatic bacteria held traits under selection in warmer, wetter climates, and midge abundance was associated with selection for traits held by a group of saprotrophs. Processes that increased pitcher fluid volume weakened selection in one module, possibly by supporting greater bacterial dispersal. ConclusionDispersal limitation governed bacterial community assembly inS. purpureapitchers at a continental scale (74% of between‐site comparisons) and was significantly greater than selection across the range. Network modules showed evidence for selection, demonstrating that multiple processes acted concurrently in bacterial community assembly at the continental scale. 

Conspecific negative density dependence (CNDD) promotes tree species diversity by reducing recruitment near conspecific adults due to biotic feedbacks from herbivores, pathogens, or competitors. While this process is well-described in tropical forests, tests of temperate tree species range from strong positive to strong negative density dependence. To explain this, several studies have suggested that tree species traits may help predict the strength and direction of density dependence: for example, ectomycorrhizal-associated tree species typically exhibit either positive or weaker negative conspecific density dependence. More generally, the strength of density dependence may be predictably related to other species-specific ecological attributes such as shade tolerance, or the relative local abundance of a species. To test the strength of density dependence and whether it affects seedling community diversity in a temperate forest, we tracked the survival of seedlings of three ectomycorrhizal-associated species experimentally planted beneath conspecific and heterospecific adults on the Prospect Hill tract of the Harvard Forest, in Massachusetts, USA. Experimental seedling survival was always lower under conspecific adults, which increased seedling community diversity in one of six treatments. We compared these results to evidence of CNDD from observed sapling survival patterns of 28 species over approximately 8 years in an adjacent 35-ha forest plot. We tested whether species-specific estimates of CNDD were associated with mycorrhizal association, shade tolerance, and local abundance. We found evidence of significant, negative conspecific density dependence (CNDD) in 23 of 28 species, and positive conspecific density dependence in two species. Contrary to our expectations, ectomycorrhizal-associated species generally exhibited stronger (e.g., more negative) CNDD than arbuscular mycorrhizal-associated species. CNDD was also stronger in more shade-tolerant species but was not associated with local abundance. Conspecific adult trees often have a negative influence on seedling survival in temperate forests, particularly for tree species with certain traits. Here we found strong experimental and observational evidence that ectomycorrhizal-associating species consistently exhibit CNDD. Moreover, similarities in the relative strength of density dependence from experiments and observations of sapling mortality suggest a mechanistic link between negative effects of conspecific adults on seedling and sapling survival and local tree species distributions.