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1. A UAV‐based high‐throughput phenotyping approach to assess time‐series nitrogen responses and identify trait‐associated genetic components in maize

   https://doi.org/10.1002/ppj2.20030  

   Rodene, Eric ; Xu, Gen ; Palali Delen, Semra ; Zhao, Xia ; Smith, Christine ; Ge, Yufeng ; Schnable, James ; Yang, Jinliang ( February 2022 , The Plant Phenome Journal)

   Advancements in the use of genome‐wide markers have provided unprecedented opportunities for dissecting the genetic components that control phenotypic trait variation. However, cost‐effectively characterizing agronomically important phenotypic traits on a large scale remains a bottleneck. Unmanned aerial vehicle (UAV)‐based high‐throughput phenotyping has recently become a prominent method, as it allows large numbers of plants to be analyzed in a time‐series manner. In this experiment, 233 inbred lines from the maize (Zea maysL.) diversity panel were grown in the field under different nitrogen treatments. Unmanned aerial vehicle images were collected during different plant developmental stages throughout the growing season. A workflow for extracting plot‐level images, filtering images to remove nonfoliage elements, and calculating canopy coverage and greenness ratings based on vegetation indices (VIs) was developed. After applying the workflow, about 100,000 plot‐level image clips were obtained for 12 different time points. High correlations were detected between VIs and ground truth physiological and yield‐related traits. The genome‐wide association study was performed, resulting inn = 29 unique genomic regions associated with image extracted traits from two or more of the 12 total time points. A candidate geneZm00001d031997, a maize homolog of theArabidopsis HCF244(high chlorophyll fluorescence 244), located underneath the leading single nucleotide polymorphisms of the canopy coverage associated signals were repeatedly detected under both nitrogen conditions. The plot‐level time‐series phenotypic data and the trait‐associated genes provide great opportunities to advance plant science and to facilitate plant breeding.

    

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2. Comparison of UAS-Based Structure-from-Motion and LiDAR for Structural Characterization of Short Broadacre Crops

   https://doi.org/10.3390/rs13193975  

   Zhang, Fei ; Hassanzadeh, Amirhossein ; Kikkert, Julie ; Pethybridge, Sarah Jane ; van Aardt, Jan ( October 2021 , Remote Sensing)

   The use of small unmanned aerial system (UAS)-based structure-from-motion (SfM; photogrammetry) and LiDAR point clouds has been widely discussed in the remote sensing community. Here, we compared multiple aspects of the SfM and the LiDAR point clouds, collected concurrently in five UAS flights experimental fields of a short crop (snap bean), in order to explore how well the SfM approach performs compared with LiDAR for crop phenotyping. The main methods include calculating the cloud-to-mesh distance (C2M) maps between the preprocessed point clouds, as well as computing a multiscale model-to-model cloud comparison (M3C2) distance maps between the derived digital elevation models (DEMs) and crop height models (CHMs). We also evaluated the crop height and the row width from the CHMs and compared them with field measurements for one of the data sets. Both SfM and LiDAR point clouds achieved an average RMSE of ~0.02 m for crop height and an average RMSE of ~0.05 m for row width. The qualitative and quantitative analyses provided proof that the SfM approach is comparable to LiDAR under the same UAS flight settings. However, its altimetric accuracy largely relied on the number and distribution of the ground control points. 

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3. Unoccupied aerial systems imagery for phenotyping in cotton, maize, soybean, and wheat breeding

   https://doi.org/10.1002/csc2.21028  

   Herr, Andrew W. ; Adak, Alper ; Carroll, Matthew E. ; Elango, Dinakaran ; Kar, Soumyashree ; Li, Changying ; Jones, Sarah E. ; Carter, Arron H. ; Murray, Seth C. ; Paterson, Andrew ; et al ( June 2023 , Crop Science)

   High‐throughput phenotyping (HTP) with unoccupied aerial systems (UAS), consisting of unoccupied aerial vehicles (UAV; or drones) and sensor(s), is an increasingly promising tool for plant breeders and researchers. Enthusiasm and opportunities from this technology for plant breeding are similar to the emergence of genomic tools ∼30 years ago, and genomic selection more recently. Unlike genomic tools, HTP provides a variety of strategies in implementation and utilization that generate big data on the dynamic nature of plant growth formed by temporal interactions between growth and environment. This review lays out strategies deployed across four major staple crop species: cotton (Gossypium hirsutumL.), maize (Zea maysL.), soybean (Glycine maxL.), and wheat (Triticum aestivumL.). Each crop highlighted in this review demonstrates how UAS‐collected data are employed to automate and improve estimation or prediction of objective phenotypic traits. Each crop section includes four major topics: (a) phenotyping of routine traits, (b) phenotyping of previously infeasible traits, (c) sample cases of UAS application in breeding, and (d) implementation of phenotypic and phenomic prediction and selection. While phenotyping of routine agronomic and productivity traits brings advantages in time and resource optimization, the most potentially beneficial application of UAS data is in collecting traits that were previously difficult or impossible to quantify, improving selection efficiency of important phenotypes. In brief, UAS sensor technology can be used for measuring abiotic stress, biotic stress, crop growth and development, as well as productivity. These applications and the potential implementation of machine learning strategies allow for improved prediction, selection, and efficiency within breeding programs, making UAS HTP a potentially indispensable asset.

    

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4. “Canopy fingerprints” for characterizing three-dimensional point cloud data of soybean canopies

   https://doi.org/10.3389/fpls.2023.1141153  

   Young, Therin J. ; Jubery, Talukder Z. ; Carley, Clayton N. ; Carroll, Matthew ; Sarkar, Soumik ; Singh, Asheesh K. ; Singh, Arti ; Ganapathysubramanian, Baskar ( March 2023 , Frontiers in Plant Science)

   Advances in imaging hardware allow high throughput capture of the detailed three-dimensional (3D) structure of plant canopies. The point cloud data is typically post-processed to extract coarse-scale geometric features (like volume, surface area, height, etc.) for downstream analysis. We extend feature extraction from 3D point cloud data to various additional features, which we denote as ‘canopy fingerprints’. This is motivated by the successful application of the fingerprint concept for molecular fingerprints in chemistry applications and acoustic fingerprints in sound engineering applications. We developed an end-to-end pipeline to generate canopy fingerprints of a three-dimensional point cloud of soybean [Glycine max(L.) Merr.] canopies grown in hill plots captured by a terrestrial laser scanner (TLS). The pipeline includes noise removal, registration, and plot extraction, followed by the canopy fingerprint generation. The canopy fingerprints are generated by splitting the data into multiple sub-canopy scale components and extracting sub-canopy scale geometric features. The generated canopy fingerprints are interpretable and can assist in identifying patterns in a database of canopies, querying similar canopies, or identifying canopies with a certain shape. The framework can be extended to other modalities (for instance, hyperspectral point clouds) and tuned to find the most informative fingerprint representation for downstream tasks. These canopy fingerprints can aid in the utilization of canopy traits at previously unutilized scales, and therefore have applications in plant breeding and resilient crop production.

    

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5. Florida mangrove saltmarsh reference surface soils

   https://doi.org/10.6073/pasta/0e08cbe07c84488cb7b9dd16669946d0  

   Lewis, David Bruce ( January 2021 , Environmental Data Initiative)

   Site description. This data package consists of data obtained from sampling surface soil (the 0-7.6 cm depth profile) in black mangrove (Avicennia germinans) dominated forest and black needlerush (Juncus roemerianus) saltmarsh along the Gulf of Mexico coastline in peninsular west-central Florida, USA. This location has a subtropical climate with mean daily temperatures ranging from 15.4 °C in January to 27.8 °C in August, and annual precipitation of 1336 mm. Precipitation falls as rain primarily between June and September. Tides are semi-diurnal, with 0.57 m median amplitudes during the year preceding sampling (U.S. NOAA National Ocean Service, Clearwater Beach, Florida, station 8726724). Sea-level rise is 4.0 ± 0.6 mm per year (1973-2020 trend, mean ± 95 % confidence interval, NOAA NOS Clearwater Beach station). The A. germinans mangrove zone is either adjacent to water or fringed on the seaward side by a narrow band of red mangrove (Rhizophora mangle). A near-monoculture of J. roemerianus is often adjacent to and immediately landward of the A. germinans zone. The transition from the mangrove to the J. roemerianus zone is variable in our study area. An abrupt edge between closed-canopy mangrove and J. roemerianus monoculture may extend for up to several hundred meters in some locations, while other stretches of ecotone present a gradual transition where smaller, widely spaced trees are interspersed into the herbaceous marsh. Juncus roemerianus then extends landward to a high marsh patchwork of succulent halophytes (including Salicornia bigellovi, Sesuvium sp., and Batis maritima), scattered dwarf mangrove, and salt pans, followed in turn by upland vegetation that includes Pinus sp. and Serenoa repens. Field design and sample collection. We established three study sites spaced at approximately 5 km intervals along the western coastline of the central Florida peninsula. The sites consisted of the Salt Springs (28.3298°, -82.7274°), Energy Marine Center (28.2903°, -82.7278°), and Green Key (28.2530°, -82.7496°) sites on the Gulf of Mexico coastline in Pasco County, Florida, USA. At each site, we established three plot pairs, each consisting of one saltmarsh plot and one mangrove plot. Plots were 50 m^2 in size. Plots pairs within a site were separated by 230-1070 m, and the mangrove and saltmarsh plots composing a pair were 70-170 m apart. All plot pairs consisted of directly adjacent patches of mangrove forest and J. roemerianus saltmarsh, with the mangrove forests exhibiting a closed canopy and a tree architecture (height 4-6 m, crown width 1.5-3 m). Mangrove plots were located at approximately the midpoint between the seaward edge (water-mangrove interface) and landward edge (mangrove-marsh interface) of the mangrove zone. Saltmarsh plots were located 20-25 m away from any mangrove trees and into the J. roemerianus zone (i.e., landward from the mangrove-marsh interface). Plot pairs were coarsely similar in geomorphic setting, as all were located on the Gulf of Mexico coastline, rather than within major sheltering formations like Tampa Bay, and all plot pairs fit the tide-dominated domain of the Woodroffe classification (Woodroffe, 2002, "Coasts: Form, Process and Evolution", Cambridge University Press), given their conspicuous semi-diurnal tides. There was nevertheless some geomorphic variation, as some plot pairs were directly open to the Gulf of Mexico while others sat behind keys and spits or along small tidal creeks. Our use of a plot-pair approach is intended to control for this geomorphic variation. Plot center elevations (cm above mean sea level, NAVD 88) were estimated by overlaying the plot locations determined with a global positioning system (Garmin GPS 60, Olathe, KS, USA) on a LiDAR-derived bare-earth digital elevation model (Dewberry, Inc., 2019). The digital elevation model had a vertical accuracy of ± 10 cm (95 % CI) and a horizontal accuracy of ± 116 cm (95 % CI). Soil samples were collected via coring at low tide in June 2011. From each plot, we collected a composite soil sample consisting of three discrete 5.1 cm diameter soil cores taken at equidistant points to 7.6 cm depth. Cores were taken by tapping a sleeve into the soil until its top was flush with the soil surface, sliding a hand under the core, and lifting it up. Cores were then capped and transferred on ice to our laboratory at the University of South Florida (Tampa, Florida, USA), where they were combined in plastic zipper bags, and homogenized by hand into plot-level composite samples on the day they were collected. A damp soil subsample was immediately taken from each composite sample to initiate 1 y incubations for determination of active C and N (see below). The remainder of each composite sample was then placed in a drying oven (60 °C) for 1 week with frequent mixing of the soil to prevent aggregation and liberate water. Organic wetland soils are sometimes dried at 70 °C, however high drying temperatures can volatilize non-water liquids and oxidize and decompose organic matter, so 50 °C is also a common drying temperature for organic soils (Gardner 1986, "Methods of Soil Analysis: Part 1", Soil Science Society of America); we accordingly chose 60 °C as a compromise between sufficient water removal and avoidance of non-water mass loss. Bulk density was determined as soil dry mass per core volume (adding back the dry mass equivalent of the damp subsample removed prior to drying). Dried subsamples were obtained for determination of soil organic matter (SOM), mineral texture composition, and extractable and total carbon (C) and nitrogen (N) within the following week. Sample analyses. A dried subsample was apportioned from each composite sample to determine SOM as mass loss on ignition at 550 °C for 4 h. After organic matter was removed from soil via ignition, mineral particle size composition was determined using a combination of wet sieving and density separation in 49 mM (3 %) sodium hexametaphosphate ((NaPO_3)_6) following procedures in Kettler et al. (2001, Soil Science Society of America Journal 65, 849-852). The percentage of dry soil mass composed of silt and clay particles (hereafter, fines) was calculated as the mass lost from dispersed mineral soil after sieving (0.053 mm mesh sieve). Fines could have been slightly underestimated if any clay particles were burned off during the preceding ignition of soil. An additional subsample was taken from each composite sample to determine extractable N and organic C concentrations via 0.5 M potassium sulfate (K_2SO_4) extractions. We combined soil and extractant (ratio of 1 g dry soil:5 mL extractant) in plastic bottles, reciprocally shook the slurry for 1 h at 120 rpm, and then gravity filtered it through Fisher G6 (1.6 μm pore size) glass fiber filters, followed by colorimetric detection of nitrite (NO_2^-) + nitrate (NO_3^-) and ammonium (NH_4^+) in the filtrate (Hood Nowotny et al., 2010,Soil Science Society of America Journal 74, 1018-1027) using a microplate spectrophotometer (Biotek Epoch, Winooski, VT, USA). Filtrate was also analyzed for dissolved organic C (referred to hereafter as extractable organic C) and total dissolved N via combustion and oxidation followed by detection of the evolved CO_2 and N oxide gases on a Formacs HT TOC/TN analyzer (Skalar, Breda, The Netherlands). Extractable organic N was then computed as total dissolved N in filtrate minus extractable mineral N (itself the sum of extractable NH_4-N and NO_2-N + NO_3-N). We determined soil total C and N from dried, milled subsamples subjected to elemental analysis (ECS 4010, Costech, Inc., Valencia, CA, USA) at the University of South Florida Stable Isotope Laboratory. Median concentration of inorganic C in unvegetated surface soil at our sites is 0.5 % of soil mass (Anderson, 2019, Univ. of South Florida M.S. thesis via methods in Wang et al., 2011, Environmental Monitoring and Assessment 174, 241-257). Inorganic C concentrations are likely even lower in our samples from under vegetation, where organic matter would dilute the contribution of inorganic C to soil mass. Nevertheless, the presence of a small inorganic C pool in our soils may be counted in the total C values we report. Extractable organic C is necessarily of organic C origin given the method (sparging with HCl) used in detection. Active C and N represent the fractions of organic C and N that are mineralizable by soil microorganisms under aerobic conditions in long-term soil incubations. To quantify active C and N, 60 g of field-moist soil were apportioned from each composite sample, placed in a filtration apparatus, and incubated in the dark at 25 °C and field capacity moisture for 365 d (as in Lewis et al., 2014, Ecosphere 5, art59). Moisture levels were maintained by frequently weighing incubated soil and wetting them up to target mass. Daily CO_2 flux was quantified on 29 occasions at 0.5-3 week intervals during the incubation period (with shorter intervals earlier in the incubation), and these per day flux rates were integrated over the 365 d period to compute an estimate of active C. Observations of per day flux were made by sealing samples overnight in airtight chambers fitted with septa and quantifying headspace CO_2 accumulation by injecting headspace samples (obtained through the septa via needle and syringe) into an infrared gas analyzer (PP Systems EGM 4, Amesbury, MA, USA). To estimate active N, each incubated sample was leached with a C and N free, 35 psu solution containing micronutrients (Nadelhoffer, 1990, Soil Science Society of America Journal 54, 411-415) on 19 occasions at increasing 1-6 week intervals during the 365 d incubation, and then extracted in 0.5 M K_2SO_4 at the end of the incubation in order to remove any residual mineral N. Active N was then quantified as the total mass of mineral N leached and extracted. Mineral N in leached and extracted solutions was detected as NH_4-N and NO_2-N + NO_3-N via colorimetry as above. This incubation technique precludes new C and N inputs and persistently leaches mineral N, forcing microorganisms to meet demand by mineralizing existing pools, and thereby directly assays the potential activity of soil organic C and N pools present at the time of soil sampling. Because this analysis commences with disrupting soil physical structure, it is biased toward higher estimates of active fractions. Calculations. Non-mobile C and N fractions were computed as total C and N concentrations minus the extractable and active fractions of each element. This data package reports surface-soil constituents (moisture, fines, SOM, and C and N pools and fractions) in both gravimetric units (mass constituent / mass soil) and areal units (mass constituent / soil surface area integrated through 7.6 cm soil depth, the depth of sampling). Areal concentrations were computed as X × D × 7.6, where X is the gravimetric concentration of a soil constituent, D is soil bulk density (g dry soil / cm^3), and 7.6 is the sampling depth in cm. 

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