<?xml version="1.0" encoding="UTF-8"?><article xml:lang="en" article-type="review-article"><front><journal-meta><journal-id journal-id-type="pmc-domain-id">69</journal-id><journal-id journal-id-type="pmc-domain">plntphys</journal-id><journal-title-group><journal-title>Plant Physiology</journal-title><abbrev-journal-title>Plant Physiol</abbrev-journal-title></journal-title-group><publisher><publisher-name>Oxford University Press</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmcid">PMC8561249</article-id><article-id pub-id-type="pmcaid">8561249</article-id><article-id pub-id-type="pmcaiid">8561249</article-id><article-id pub-id-type="pmid">34608963</article-id><article-id pub-id-type="doi">10.1093/plphys/kiab301</article-id><title-group><article-title>Resources for image-based high-throughput phenotyping in crops and data sharing challenges</article-title></title-group><contrib-group content-type="author"><contrib><name name-style="western"><surname>Danilevicz</surname><given-names initials="MF">Monica F</given-names></name><xref ref-type="aff" rid="kiab301-aff1">1</xref></contrib><contrib><name name-style="western"><surname>Bayer</surname><given-names initials="PE">Philipp E</given-names></name><xref ref-type="aff" rid="kiab301-aff1">1</xref></contrib><contrib><name name-style="western"><surname>Nestor</surname><given-names initials="BJ">Benjamin J</given-names></name><xref ref-type="aff" rid="kiab301-aff1">1</xref></contrib><contrib><name name-style="western"><surname>Bennamoun</surname><given-names initials="M">Mohammed</given-names></name><xref ref-type="aff" rid="kiab301-aff2">2</xref></contrib><contrib><name name-style="western"><surname>Edwards</surname><given-names initials="D">David</given-names></name><xref ref-type="aff" rid="kiab301-aff1">1</xref><xref rid="kiab301-cor1" ref-type="author-notes">✉</xref><xref rid="kiab301-FM1" ref-type="author-notes">†</xref></contrib></contrib-group><aff id="kiab301-aff1"><label>1</label>School of Biological Sciences and Institute of Agriculture, University of Western Australia, Perth, Western Australia 6009, Australia</aff><aff id="kiab301-aff2"><label>2</label>Department of Computer Science and Software Engineering, University of Western Australia, Perth, Western Australia 6009, Australia</aff><author-notes><fn id="kiab301-cor1"><label>✉</label><p>Author for communication: <email>dave.edwards@uwa.edu.au</email></p></fn><fn id="kiab301-FM1"><label>†</label><p>Senior author.</p></fn></author-notes><pub-date><day>28</day><month>6</month><year>2021</year></pub-date><volume>187</volume><issue>2</issue><fpage>699</fpage><page-range>699–715</page-range><pub-history><event event-type="pmc-release"><date><day>3</day><month>11</month><year>2021</year></date></event></pub-history><permissions><copyright-statement>© The Author(s) 2021. Published by Oxford University Press on behalf of American Society of Plant Biologists.</copyright-statement><license><license-p>This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs licence (<ext-link xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="https://creativecommons.org/licenses/by-nc-nd/4.0/" ext-link-type="uri">https://creativecommons.org/licenses/by-nc-nd/4.0/</ext-link>), which permits non-commercial reproduction and distribution of the work, in any medium, provided the original work is not altered or transformed in any way, and that the work is properly cited. For commercial re-use, please contact journals.permissions@oup.com</license-p></license></permissions><self-uri xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="kiab301.pdf" content-type="pmc-pdf"><?cloudpmc-path 0f62/8561249/186b561d578a/kiab301.pdf?><?cloudpmc-bucket app?><?size 638549?></self-uri><abstract id="abstract1"><title>Abstract</title><p>High-throughput phenotyping (HTP) platforms are capable of monitoring the phenotypic variation of plants through multiple types of sensors, such as red green and blue (RGB) cameras, hyperspectral sensors, and computed tomography, which can be associated with environmental and genotypic data. Because of the wide range of information provided, HTP datasets represent a valuable asset to characterize crop phenotypes. As HTP becomes widely employed with more tools and data being released, it is important that researchers are aware of these resources and how they can be applied to accelerate crop improvement. Researchers may exploit these datasets either for phenotype comparison or employ them as a benchmark to assess tool performance and to support the development of tools that are better at generalizing between different crops and environments. In this review, we describe the use of image-based HTP for yield prediction, root phenotyping, development of climate-resilient crops, detecting pathogen and pest infestation, and quantitative trait measurement. We emphasize the need for researchers to share phenotypic data, and offer a comprehensive list of available datasets to assist crop breeders and tool developers to leverage these resources in order to accelerate crop breeding.</p></abstract><abstract id="abstract2" abstract-type="teaser"><p>Various approaches are used to analyze high-throughput phenotyping data and tools can be developed and assessed using available image-based datasets.</p></abstract><custom-meta-group><custom-meta><meta-name>status</meta-name><meta-value>released</meta-value></custom-meta><custom-meta><meta-name>display-pdf</meta-name><meta-value>yes</meta-value></custom-meta><custom-meta><meta-name>is-olf</meta-name><meta-value>no</meta-value></custom-meta><custom-meta><meta-name>is-manuscript</meta-name><meta-value>no</meta-value></custom-meta><custom-meta><meta-name>is-preprint</meta-name><meta-value>no</meta-value></custom-meta><custom-meta><meta-name>is-journal-matter</meta-name><meta-value>no</meta-value></custom-meta><custom-meta><meta-name>is-scanned</meta-name><meta-value>no</meta-value></custom-meta><custom-meta><meta-name>is-retracted</meta-name><meta-value>no</meta-value></custom-meta></custom-meta-group></article-meta><notes notes-type="article-notes"><sec id="historyarticle-meta1" sec-type="history" disp-level="2"><p>Received 2020 Dec 4; Accepted 2021 May 26; Collection date 2021 Oct.</p></sec></notes></front><body><sec id="sec1" disp-level="1"><title>Introduction</title><p>Plant phenotypic variation is the result of the complex interplay between genetics and environmental conditions (<xref rid="kiab301-B23" ref-type="bibr">Boyer, 1982</xref>; <xref rid="kiab301-B57" ref-type="bibr">Ficke et al., 2018</xref>; <xref rid="kiab301-B61" ref-type="bibr">Frantzeskakis et al., 2020</xref>). Advances in genome sequencing have uncovered substantial genetic diversity within species (<xref rid="kiab301-B87" ref-type="bibr">Hirsch et al., 2014</xref>; <xref rid="kiab301-B74" ref-type="bibr">Golicz et al., 2016</xref>; <xref rid="kiab301-B269" ref-type="bibr">Zhao et al., 2018</xref>; <xref rid="kiab301-B93" ref-type="bibr">Hübner et al., 2019</xref>; <xref rid="kiab301-B222" ref-type="bibr">Song et al., 2020</xref>). However, the wealth of genetic information is rarely translated into gains for real-world agricultural crops (<xref rid="kiab301-B6" ref-type="bibr">Araus et al., 2018</xref>), partially due to the lack of phenotypic information associated with the genetic variation (<xref rid="kiab301-B65" ref-type="bibr">Furbank and Tester, 2011</xref>; <xref rid="kiab301-B149" ref-type="bibr">Mir et al., 2019</xref>). High-throughput phenotyping (HTP) has emerged to overcome the phenomics bottleneck. HTP platforms enable noninvasive data collection through several types of sensors that can be deployed in glasshouse facilities or field monitoring devices, including ground platforms to unmanned aerial vehicles (UAVs) and satellites (<xref rid="kiab301-B127" ref-type="bibr">Li et al., 2014</xref>; <xref rid="kiab301-B81" ref-type="bibr">Hank et al., 2015</xref>; <xref rid="kiab301-B109" ref-type="bibr">Kirchgessner et al., 2016</xref>; <xref rid="kiab301-B163" ref-type="bibr">Naito et al., 2017</xref>; <xref rid="kiab301-B40" ref-type="bibr">Danzi et al., 2019</xref>). These platforms can support the capture of temporal phenotypic variation for large populations across plant development, generating massive amounts of data. Systematic large-scale phenotyping platforms can be used for genetic dissection of targeted traits and assist the development of better performing varieties (<xref rid="kiab301-B128" ref-type="bibr">Li et al., 2018</xref>; <xref rid="kiab301-B149" ref-type="bibr">Mir et al., 2019</xref>). The increasing adoption of HTP platforms leads to a demand for new computer-based tools that can leverage these datasets and integrate-associated information (e.g. experimental conditions, weather measurements, and genotypic data) to extract meaningful insights regarding crop development and performance (<xref rid="kiab301-B227" ref-type="bibr">Tattaris et al., 2016</xref>; <xref rid="kiab301-B49" ref-type="bibr">van Eeuwijk et al., 2019</xref>). HTP data analysis is a nontrivial task, requiring a high level of expertise in computer science and plant development to understand the implications of phenotypic variation in the plant. Completeness of HTP metadata is crucial for plant physiologists to characterize the genetic and environmental conditions in which a phenotype occurs. Even though the majority of available datasets are lacking a clear description of conditions depicted, it is important that new datasets include metadata and methods to collect environmental data in their experimental design. Mathematical models, machine learning, and most recently deep learning models, can be used as guides to identify stress and predict crop performance under defined conditions (<xref rid="kiab301-B14" ref-type="bibr">Bai et al., 2016</xref>; <xref rid="kiab301-B9" ref-type="bibr">Atkinson et al., 2017a</xref>; <xref rid="kiab301-B103" ref-type="bibr">Joalland et al., 2017</xref>; <xref rid="kiab301-B153" ref-type="bibr">Moghadam et al., 2017</xref>; <xref rid="kiab301-B163" ref-type="bibr">Naito et al., 2017</xref>; <xref rid="kiab301-B56" ref-type="bibr">Fernandez-Gallego et al., 2018</xref>; <xref rid="kiab301-B187" ref-type="bibr">Prey et al., 2019</xref>; <xref rid="kiab301-B245" ref-type="bibr">Walter et al., 2019</xref>; <xref rid="kiab301-B47" ref-type="bibr">Ducournau et al., 2020</xref>; <xref rid="kiab301-B107" ref-type="bibr">Kerkech et al., 2020</xref>; <xref rid="kiab301-B206" ref-type="bibr">Selvaraj et al., 2020</xref>). Deep learning models have the advantage of automatically extracting features from the image by constructing increasingly abstract representations of the relationships within the dataset (<xref rid="kiab301-B122" ref-type="bibr">LeCun et al., 2015</xref>). In contrast, classic statistical approaches rely solely on the researcher to manually define the features before the analysis. Because deep learning models build the features based solely on the dataset, it usually requires large amounts of high-quality data to learn from these features to achieve high performance.</p><boxed-text id="kiab301-BOX1" position="float"><sec id="sec2" disp-level="2"><title>Advances</title><list list-type="bullet"><list-item><p>A broad diversity of sensors enables capturing and quantifying previously undetectable phenotypic traits. Combining the reflectance of different spectra allows for the detection of abiotic stress, such as nitrogen deficiency and frost damage.</p></list-item><list-item><p>• HTP has the potential to accelerate crop breeding, producing data that can be used to identify varieties with improved traits and higher performance, but there are technical challenges to overcome.</p></list-item><list-item><p>Deep learning models are effective in plant phenotyping tasks due to their capacity to leverage highly complex and multidimensional data, but their performance is dependent on the quality and diversity of the dataset.</p></list-item><list-item><p>A large effort is required to facilitate sharing high-quality phenotype datasets because they provide a key resource for developing tools for agronomic trait measurement and crop breeding.</p></list-item></list></sec></boxed-text><p>Developing a custom pipeline of software applications for processing HTP raw sensor data into traits, followed by its analysis, amounts to a major part of the cost to adopt HTP (<xref rid="kiab301-B193" ref-type="bibr">Reynolds et al., 2019</xref>). However, if the data analysis pipeline is being reutilized from a previous project, the cost of implementing the pipeline would drop to 10%–20% (<xref rid="kiab301-B193" ref-type="bibr">Reynolds et al., 2019</xref>), which means that being able to employ whole or part of a developed HTP analysis pipeline can decrease the costs for adopting HTP in research projects. Many challenges prevent the research community from efficiently reusing data processing tools and analysis pipelines. For example, the lack of interoperability between processing tools (image processing, weather data transformation) and analysis models (trait quantification, classification) due to the absence of standardized data processing methodology prevent the utilization of previously published analysis models (<xref rid="kiab301-B113" ref-type="bibr">Krajewski et al., 2015</xref>; <xref rid="kiab301-B99" ref-type="bibr">Janssen et al., 2017</xref>; <xref rid="kiab301-B260" ref-type="bibr">Yu et al., 2017</xref>; <xref rid="kiab301-B49" ref-type="bibr">van Eeuwijk et al., 2019</xref>). The inconsistency of data processing pipelines can be partially overcome by providing tools to standardize data input for target analysis models (<xref rid="kiab301-B27" ref-type="bibr">Busemeyer et al., 2013</xref>; <xref rid="kiab301-B260" ref-type="bibr">Yu et al., 2017</xref>; <xref rid="kiab301-B34" ref-type="bibr">Chopin et al., 2018</xref>; <xref rid="kiab301-B206" ref-type="bibr">Selvaraj et al., 2020</xref>); however, a robust solution requires standardizing syntax (formats) and semantics (definitions, ontology) of input/output files used by HTP data processing tools (<xref rid="kiab301-B99" ref-type="bibr">Janssen et al., 2017</xref>).</p><p>Data sharing is an important step for the advancement of crop breeding and the development of analysis pipelines (<xref rid="kiab301-B261" ref-type="bibr">Zamir, 2013</xref>; <xref rid="kiab301-B149" ref-type="bibr">Mir et al., 2019</xref>). The need to establish a repository to host raw phenotypic datasets with associated information has long been recognized (<xref rid="kiab301-B261" ref-type="bibr">Zamir, 2013</xref>; <xref rid="kiab301-B130" ref-type="bibr">Lobet, 2017</xref>). A centralized database with access to raw data and standardized metadata would increase discoverability and reutilization of the datasets, allowing researchers to reanalyze data using updated state-of-the-art tools, which may lead to the identification of novel and potentially interesting results (<xref rid="kiab301-B261" ref-type="bibr">Zamir, 2013</xref>). Even though some platforms have been developed to host selected datasets (<xref rid="kiab301-B76" ref-type="bibr">Granier et al., 2006</xref>; <xref rid="kiab301-B131" ref-type="bibr">Lobet et al., 2013</xref>; <xref rid="kiab301-B208" ref-type="bibr">Seren et al., 2017</xref>), the majority of datasets are insufficiently described, preventing plant researchers from properly analyzing phenotypic variations and leading to misinterpretation of results. The Minimum Information About a Plant Phenotyping Experiment (MIAPPE) initiative (<ext-link xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="https://www.miappe.org/" ext-link-type="uri">https://www.miappe.org/</ext-link>) provides a framework for phenotypic data sharing designed to standardize data publication with a controlled ontology vocabulary referencing multiple previously established ontologies (<xref rid="kiab301-B172" ref-type="bibr">Papoutsoglou et al., 2020</xref>). The MIAPPE guidelines are compatible with the Breeding Application Programming Interface (BrAPI), which aim to increase breeding datasets interoperability and provide easy access to breeding tools (<xref rid="kiab301-B205" ref-type="bibr">Selby et al., 2019</xref>; <xref rid="kiab301-B172" ref-type="bibr">Papoutsoglou et al., 2020</xref>). Adoption of these standardization guidelines for dataset description is a crucial step in transforming HTP datasets into data assets for plant researchers and breeders. Adequately described datasets can also be used to establish benchmark datasets (detailed in <xref rid="kiab301-BOX3" ref-type="boxed-text">Box 1</xref>). Benchmark datasets provide a standard to compare computer-based tools performance, helping uncover the tool limitations and strengths (<xref rid="kiab301-B261" ref-type="bibr">Zamir, 2013</xref>; <xref rid="kiab301-B147" ref-type="bibr">Minervini et al., 2016</xref>; <xref rid="kiab301-B130" ref-type="bibr">Lobet, 2017</xref>). Assessing tool performance will guide the user to apply the most effective methodology for their experimental design and data (<xref rid="kiab301-B130" ref-type="bibr">Lobet, 2017</xref>).
This review reports on previously published image-based HTP datasets with the aim of assisting the community to access and benefit from their development. The main contributions presented are (1) highlighting the challenges faced by researchers when reusing HTP datasets; (2) describing some of the criteria required when creating an effective benchmark dataset; and (3) presenting a collection of image-based HTP datasets available as a resource for researchers to improve model development and analysis.</p><boxed-text id="kiab301-BOX3" position="float"><?disp-level 2?><label>Box 1</label><caption><p>What is a benchmarking dataset?</p></caption><p>Benchmark dataset refers to a comprehensive data collection that represents real life data that a method or tool may encounter when performing the given task. Benchmark datasets are often employed as a standardized way to assess a new method’s performance, finding its strengths, and limitations (<xref rid="kiab301-B130" ref-type="bibr">Lobet, 2017</xref>). General requisites for benchmark datasets in most of the applications described in this study are: (1) intentional, the dataset must be designed to be employed on specific tasks; (2) relevant, the data should be coherent with the event it attempts to describe and have the limitations identified and clearly stated; (3) representative, meaning that the dataset covers most cases commonly encountered when performing a task within the defined scope (<xref rid="kiab301-B202" ref-type="bibr">Schaafsma and Vihinen, 2018</xref>), reporting any underrepresented classes; (4) sizable, the dataset must contain enough examples of each class or target to enable training machine learning and computer vision methods; (5) reliable, the data points must be experimentally obtained instead of artificially generated and annotations must be performed by plant experts (<xref rid="kiab301-B201" ref-type="bibr">Sasidharan Nair and Vihinen, 2013</xref>); and (6) descriptive, the dataset must have an extensive description of data collection methodology (sensors, UAV altitude), biological information (species, genotype, growth stage), and experimental conditions (temperature, soil, water availability). The importance of these criteria changes depending on the purpose of the dataset utilization. For computer tool developers, the first five criteria are probably more relevant as they can directly impact the performance and robustness of the new method. For plant physiology researchers, the sixth criteria is particularly important as it enables reutilization of the datasets to gain a deep understanding of the plant conditions, extract meaningful insights from plant phenotype analysis, and compare plant phenotype analyses with external phenotypic datasets.</p></boxed-text></sec><sec id="sec3" disp-level="1"><title>Applications of HTP</title><sec id="sec4" disp-level="2"><title>Improving crop productivity</title><p>A myriad of components contribute to yield, as plant performance is regulated by a combination of genetic factors (G), environmental factors (E), and the interaction between them (G × E; <xref rid="kiab301-B104" ref-type="bibr">Juliana et al., 2018</xref>; <xref rid="kiab301-B156" ref-type="bibr">Montesinos-López et al., 2018</xref>). Because of the high complexity that underlies plant performance, breeders have to submit potential varieties to extensive field testing to determine their potential yield (<xref rid="kiab301-B95" ref-type="bibr">Hunt et al., 2020</xref>). Field HTP can substantially accelerate the breeding process by allowing breeders to predict end-of-season traits, such as yield and biomass at early growth stages. Early yield prediction allows researchers to bypass plant growth time, a key limiting factor in crop breeding. In a soybean (<italic>Glycine max</italic>) study, 2,551 genotypes were grown in different locations, and it was observed that yield, plant maturity, and seed size can be predicted at an early stage using Cubist regression because it presented the best result in comparison to Partial Least Squares Regression, Random Forests, Artificial Neural Networks, and Support Vector Regression (<xref rid="kiab301-B258" ref-type="bibr">Yuan et al., 2019</xref>). Similar results were observed for wheat (<italic>Triticum aestivum</italic>), barley (<italic>Hordeum vulgare</italic>), and other soybean genotypes (<xref rid="kiab301-B14" ref-type="bibr">Bai et al., 2016</xref>; <xref rid="kiab301-B165" ref-type="bibr">Nevavuori et al., 2019</xref>). Although promising, the results are constrained to the conditions evaluated, since interannual weather variation, changes in agroecological zones, differences in farm management practices, sensor use and specifications, and other factors can cause instability in model accuracy.</p><p>The broad diversity of remote sensors enables capturing different aspects of the plant phenotype. Different combinations of RGB, multispectral, and thermal image data associated with weather and soil have been employed to train deep learning models for crop yield forecasting (<xref rid="kiab301-B242" ref-type="bibr">Vega et al., 2015</xref>; <xref rid="kiab301-B75" ref-type="bibr">Gracia-Romero et al., 2019</xref>; <xref rid="kiab301-B265" ref-type="bibr">Zhang et al., 2019a</xref>; <xref rid="kiab301-B134" ref-type="bibr">Maimaitijiang et al., 2020</xref>; <xref rid="kiab301-B214" ref-type="bibr">da Silva et al., 2020</xref>). The models can support differentiating crop performance in relation to irrigation regimes (<xref rid="kiab301-B75" ref-type="bibr">Gracia-Romero et al., 2019</xref>), quantify growth rate under nitrogen treatment (<xref rid="kiab301-B88" ref-type="bibr">Holman et al., 2016</xref>; <xref rid="kiab301-B7" ref-type="bibr">Arroyo et al., 2017</xref>; <xref rid="kiab301-B5" ref-type="bibr">Aranguren et al., 2020</xref>), estimate variation of wheat grain protein content (<xref rid="kiab301-B196" ref-type="bibr">Rodrigues et al., 2018</xref>; <xref rid="kiab301-B210" ref-type="bibr">Sharabiani et al., 2019</xref>), and monitor crop height variation during the season (<xref rid="kiab301-B271" ref-type="bibr">Ziliani et al., 2018</xref>). A systematic review on machine learning models for crop yield prediction was published by <xref rid="kiab301-B240" ref-type="bibr">van Klompenburg et al. (2020)</xref>, showing that deep learning models are increasing in popularity. The most used architectures were Convolutional Neural Networks (CNNs) and Long-Short Term Memory (LSTM). These architectures were created for different purposes: LSTM is designed specifically for sequence prediction tasks, while CNN’s structure is suited to extract features from complex image data. These architectures can benefit from transferred learning, in which pretrained weights obtained with different data can be implemented in the new model (<xref rid="kiab301-B86" ref-type="bibr">He et al., 2016</xref>) that allows rapid and high performance. Multimodal machine learning can be employed to analyze datasets with multiple data sources (rainfall, temperature, multispectral image, soil data), each data type is a modality that will be analyzed and combined to increase model performance (<xref rid="kiab301-B15" ref-type="bibr">Baltrušaitis et al., 2017</xref>). <xref rid="kiab301-B240" ref-type="bibr">van Klompenburg et al. (2020)</xref> observe in the review that temperature, rainfall, and soil type were the most used data types in machine learning models, but different feature combinations and the volume of data can directly impact the model performance and should be tested during development.</p><p>A few HTP datasets were recently released with the goal to improve yield prediction and more specifically support genotype to phenotype prediction. The Genomes to Field (G2F) datasets comprise genotype (single nucleotide polymorphism information), manual phenotype measurements, climatic data, soil information, inbred ear images, and UAV collected multispectral and hyperspectral images of several maize (<italic>Zea mays</italic>) varieties grown over multiple years (<xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 1</xref>; <xref rid="kiab301-B144" ref-type="bibr">McFarland et al., 2020</xref>). Detailed metadata are essential for understanding genotype to phenotype relationships in each season/environment. However, the G2F field trials were carried out in a single location, which limit the robustness of the traits identified. For sorghum (<italic>Sorghum bicolor</italic>) and wheat, the Transportation Energy Resources from Renewable Agriculture Phenotyping Reference Platform (TERRA-REF) database offers a comprehensive resource of sensor data (five thermal, spectral, and shape imaging sensors), phenotypic measurements, environmental and genomic data, including genome sequencing of 384 varieties and genotyping by sequencing of 768 varieties (<xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 1</xref>; <xref rid="kiab301-B121" ref-type="bibr">LeBauer et al., 2017</xref>; <xref rid="kiab301-B26" ref-type="bibr">Burnette et al., 2018</xref>). TERRA-REF has four sensing platforms to collect image-based phenotyping and agronomic traits from both controlled environment and field grown plants. TERRA-REF maintains a manuscript management section in their website where researchers willing to use the data can register their proposed manuscript to prevent overlap and encourage collaboration. Oftentimes, researchers will delay publishing datasets until their planned publications are completed. Nonetheless, the TERRA-REF approach to register publications enables early publishing of the data and allows other groups to explore different aspects of the dataset or collaborate. Federated learning is another strategy that can be used when the data must be protected due to privacy or security concerns, showing increasing use in medical research (<xref rid="kiab301-B123" ref-type="bibr">Lee et al., 2018</xref>; <xref rid="kiab301-B92" ref-type="bibr">Huang et al., 2019</xref>; <xref rid="kiab301-B194" ref-type="bibr">Rieke et al., 2020</xref>). Federated learning allows for training machine learning models collaboratively without exchanging the data, in this framework, each dataset owner institution downloads the model and trains it locally. The trained parameters from each institution are exported and aggregated, creating a model that benefits from previously inaccessible datasets while the data governance and accessibility remain in the control of the data owner (<xref rid="kiab301-B111" ref-type="bibr">Konečný et al., 2016</xref>).</p><p>Both G2F and TERRA-REF datasets present limitations regarding the types of environments represented, species grown, and the treatments that they were subjected to. Other similar phenotyping initiatives covering different locations and plant species (including noncrop plants) are needed to depict phenotypic variation. Nonetheless, the above datasets offer an extensive resource that can assist the identification of quantitative trait loci (QTLs) related to crop performance, develop tools for genotype to phenotype prediction based on the multidimensional dataset, and ultimately these could be used as benchmark datasets to assess tool performance. Moreover, smaller datasets for field trial experiments can be found at the Global Agricultural Research Data Innovation Acceleration Network and the International Maize and Wheat Improvement Center described in <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 1</xref>.</p><p>Grain yield in wheat is directly related to spike head population density, size, and maturity stage. The Annotated Crop Image Dataset (ACID) provides images with coordinates to identify wheat spikes under greenhouse conditions (<xref rid="kiab301-B185" ref-type="bibr">Pound et al., 2017b</xref>). ACID was designed for training novel tools for identifying the spike heads, and measuring individual head traits, but the tool could be further applied to new datasets and to link measured traits with genotypic variability. Limited metadata annotation in ACID prevents further exploration of the dataset itself for identification of yield-related traits because the genotypes and experimental conditions are not described. The global wheat head database compiles multiple RGB wheat images collected in the field, from several countries using different cameras (<xref rid="kiab301-B42" ref-type="bibr">David et al., 2020</xref>). The dataset was used in a challenge hosted on Kaggle (<ext-link xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="https://www.kaggle.com" ext-link-type="uri">https://www.kaggle.com</ext-link>) with the goal to benchmark wheat head detection approaches. Top solutions used object detection deep learning architectures (EfficientDet, Faster-RCNN, and Yolo-v3), with data augmentation techniques playing a major role for their success. Data augmentation is a computer vision technique to increase dataset size through a series of transformations, such as flipping or rotating the image (<xref rid="kiab301-B29" ref-type="bibr">Buslaev et al., 2020</xref>). It is important to note that with field images, a greater variability of conditions can occur such as genotype differences, head orientation, and mixed developmental stages, which can cause the object detection model to present performance instability such as mislabeling plant organs at a higher rate when the conditions differ from what was seen in the training data. The global wheat head dataset provides a valuable resource for developing and benchmarking tools due to the high variability of wheat genotypes and conditions represented. Similar datasets for different species can be developed collaboratively by annotating previously released HTP data (such as the G2F and TERRA-REF datasets). This can decrease the cost of producing a dataset and benefit from the described metadata.</p></sec><sec id="sec5" disp-level="2"><title>Developing crops tolerant to abiotic stress</title><p>The development of climate resilient crops must consider the effect of combined abiotic stresses occurring in the region (<xref rid="kiab301-B30" ref-type="bibr">Cammarano et al., 2019</xref>). As a result, datasets featuring combined abiotic stresses provide a resource to understand how their interaction impacts plant health and development. The Eschikon dataset (<xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 2</xref>) includes temporal images of beet (<italic>Beta vulgaris</italic>) under multiple independent and combined drought, nitrogen deficiency, and weed stresses (<xref rid="kiab301-B108" ref-type="bibr">Khanna et al., 2019</xref>). This dataset was employed to develop 3D representations of the plants from which the authors were able to extract canopy cover, height, and vegetation indices. These traits were used to classify stress in plants with 83%–93% accuracy depending on the stress measured. The dataset can be further explored to measure agronomic traits related to each stress and understand plant response, it can also be employed in further developing computer vision tools for stress classification (<xref rid="kiab301-B108" ref-type="bibr">Khanna et al., 2019</xref>). Plant researchers willing to predict the effects of climatic change in crop species will require the creation (or release) of more datasets in which the combined stresses are observed. These datasets must offer a detailed description of the environmental conditions and if possible, of the genetic data to enable accurate interpretation of the results. Ideally, the aggregated datasets must depict the diversity of agroecological zones including low latitude locations, which are currently underrepresented.</p><p>Crop water management is essential in regions currently facing or predicted to face water scarcity. Infrared thermography has been successfully implemented to assess water use by crops (<xref rid="kiab301-B166" ref-type="bibr">Nhamo et al., 2020</xref>), and for measuring genotype performance under salinity or water deficit stress (<xref rid="kiab301-B192" ref-type="bibr">Raza et al., 2014</xref>; <xref rid="kiab301-B115" ref-type="bibr">Kumar et al., 2017</xref>; <xref rid="kiab301-B230" ref-type="bibr">Thapa et al., 2018</xref>; <xref rid="kiab301-B90" ref-type="bibr">Hou et al., 2019</xref>; <xref rid="kiab301-B266" ref-type="bibr">Zhang et al., 2019b</xref>; <xref rid="kiab301-B116" ref-type="bibr">Kumar et al., 2020</xref>; <xref rid="kiab301-B140" ref-type="bibr">Masina et al., 2020</xref>). In cotton (<italic>Gossypium arboreum</italic>) monitored by infrared thermography, it was observed that yield, fiber length, and micronaire suffered reduction after canopy temperature exceeded a given threshold (<xref rid="kiab301-B38" ref-type="bibr">Conaty et al., 2015</xref>). Canopy temperature and evapotranspiration (ET) maps are used as a proxy for measuring the phenotypic response to both stresses as they influence stomatal conductance (<xref rid="kiab301-B58" ref-type="bibr">Fischer et al., 1998</xref>; <xref rid="kiab301-B216" ref-type="bibr">Sirault et al., 2009</xref>), and are observed from close range, at the aerial and spatial level. Remotely sensed thermal data collected by satellite platforms allow mapping water resource use through the prediction of ET maps (<xref rid="kiab301-B4" ref-type="bibr">Anderson et al., 2012</xref>). In 2018, a space station mission (ECOSTRESS) was launched to measure ET and identify plant stress (<xref rid="kiab301-B59" ref-type="bibr">Fisher et al., 2020</xref>). It provides a higher spatial and temporal resolution ratio (60 m with 1–5 d interval) in comparison to Landsat (&gt;60 m, 16-d interval) or MODIS (&gt;375 m, daily; <xref rid="kiab301-B4" ref-type="bibr">Anderson et al., 2012</xref>). The ECOSTRESS library provides satellite imagery associated with laboratory measurements of vegetation to help correlate the spectral patterns (<xref rid="kiab301-B146" ref-type="bibr">Meerdink et al., 2019</xref>; <xref rid="kiab301-B59" ref-type="bibr">Fisher et al., 2020</xref>). This dataset has been employed to assess plant species diversity in restoration areas, showing that sites with higher species diversity present lower temperatures (<xref rid="kiab301-B78" ref-type="bibr">Hamberg et al., 2020</xref>). For this review, we chose to focus on HTP images collected by aerial or ground devices since satellite images currently do not yet provide enough resolution to be used for assessing plants at the field level. However, satellite HTP imagery offers the potential to help understand abiotic stress at a large-scale (<xref rid="kiab301-B4" ref-type="bibr">Anderson et al., 2012</xref>; <xref rid="kiab301-B150" ref-type="bibr">Miralles et al., 2014</xref>), thus we have included links to satellite libraries (ECOSTRESS, Landsat and MODIS) in the resources in <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 2</xref>.</p><p>Besides infrared thermography, multispectral and hyperspectral sensors are also used in HTP. These sensors are capable of detecting physiological changes in the plant leaf composition (<xref rid="kiab301-B24" ref-type="bibr">Bruning et al., 2020</xref>). For example, decomposition of foliar hyperspectral signatures showed that C3 and C4 plants have divergent and well-defined patterns of reflectance (<xref rid="kiab301-B16" ref-type="bibr">Baranoski et al., 2016</xref>). Hyperspectral images were employed to quantitatively rank salt tolerance between four wheat varieties (dataset available in <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 2</xref>) using machine learning and dimensionality reduction. The authors observed that multiple trait measurements would be required to correctly assess the plants, whereas with hyperspectral images they were able to score them in a fast noninvasive way, dataset is described in <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 2</xref> (<xref rid="kiab301-B154" ref-type="bibr">Moghimi et al., 2018</xref>). Multispectral and hyperspectral images have also been employed to identify salt stress in sugarcane (<italic>Saccharum officinarum L.</italic>) and wheat irrigated with saline water (<xref rid="kiab301-B79" ref-type="bibr">Hamzeh et al., 2013</xref>; <xref rid="kiab301-B51" ref-type="bibr">El-Hendawy et al., 2019</xref>), acidic and heavy metal stress (<xref rid="kiab301-B126" ref-type="bibr">Liu et al., 2011</xref>; <xref rid="kiab301-B101" ref-type="bibr">Jin et al., 2013</xref>; <xref rid="kiab301-B129" ref-type="bibr">Li et al., 2015</xref>; <xref rid="kiab301-B264" ref-type="bibr">Zhang et al., 2017</xref>; <xref rid="kiab301-B247" ref-type="bibr">Wang et al., 2018a</xref>), nutrient deficiency (<xref rid="kiab301-B170" ref-type="bibr">Pacumbaba and Beyl, 2011</xref>), heat stress (<xref rid="kiab301-B70" ref-type="bibr">Gautam et al., 2015</xref>; <xref rid="kiab301-B233" ref-type="bibr">Trachsel et al., 2019</xref>), and frost (<xref rid="kiab301-B60" ref-type="bibr">Fitzgerald et al., 2019</xref>; <xref rid="kiab301-B168" ref-type="bibr">Nuttall et al., 2019</xref>; <xref rid="kiab301-B160" ref-type="bibr">Murphy et al., 2020</xref>).</p><p>Frost damage in wheat can have a major impact, as a single frost event can severely reduce quality and yield (<xref rid="kiab301-B21" ref-type="bibr">Boer et al., 1993</xref>; <xref rid="kiab301-B62" ref-type="bibr">Frederiks et al., 2012</xref>; <xref rid="kiab301-B136" ref-type="bibr">Martino and Abbate, 2019</xref>). Rapid detection of frost damage would enable growers to take management decisions to avoid losses. A study using hyperspectral images indicated that under controlled conditions, frosted and nonfrosted individuals, present significant spectral differences (<xref rid="kiab301-B160" ref-type="bibr">Murphy et al., 2020</xref>). Mixed results were observed when detecting frost under field conditions, indicating that more research is needed (<xref rid="kiab301-B60" ref-type="bibr">Fitzgerald et al., 2019</xref>; <xref rid="kiab301-B168" ref-type="bibr">Nuttall et al., 2019</xref>). The Frost nursery dataset provides multispectral images of several commercial wheat varieties grown in the field and were affected by frost at different developmental stages (<xref rid="kiab301-B2a" ref-type="bibr">AgReFed, 2019</xref>). This dataset includes final yield, leaf protein, and abundance of metabolites, which can be used to characterize the effect of frost (<xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 2</xref>). The association of hyperspectral data with physiological measurements may assist frost damage quantification, which can support crop breeders screening for more tolerant varieties. Hyperspectral imagery has the potential to capture traits related to the biochemical composition of target tissues. However, due to various technical factors, the recorded data are usually noisy with nonnegligible redundancy (<xref rid="kiab301-B152" ref-type="bibr">Mishra et al., 2019</xref>). Datasets including hyperspectral data may benefit from detailed description of the experimental conditions and sensors used, helping guide researchers how to better extract the information.</p><p>Platforms such as Quantitative Plant (<xref rid="kiab301-B131" ref-type="bibr">Lobet et al., 2013</xref>), Phenopsis (<xref rid="kiab301-B76" ref-type="bibr">Granier et al., 2006</xref>), and BrAPI (<xref rid="kiab301-B205" ref-type="bibr">Selby et al., 2019</xref>) are dedicated to assemble a wide range of phenotyping datasets that can be used to compare phenotypic response to stress within and between species. These platforms are focused on making phenotypic datasets more findable. Information and website links for other abiotic stress datasets are described in <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 2</xref>.</p></sec><sec id="sec6" disp-level="2"><title>Pathogen and pest detection in the field</title><p>Changes in environmental conditions are likely to shift pathogen and pest regional distributions (<xref rid="kiab301-B91" ref-type="bibr">Hovmøller et al., 2008</xref>; <xref rid="kiab301-B213" ref-type="bibr">Shaw and Osborne, 2011</xref>; <xref rid="kiab301-B17" ref-type="bibr">Bebber et al., 2013</xref>; <xref rid="kiab301-B68" ref-type="bibr">Garrett, 2013</xref>; <xref rid="kiab301-B135" ref-type="bibr">Mariette et al., 2016</xref>; <xref rid="kiab301-B217" ref-type="bibr">Skelsey et al., 2016</xref>). To provide suitable crop varieties and agricultural management recommendations for these new conditions, it is necessary to gain greater understanding of the ecological, phenotypic, and molecular basis of the interaction between plant and pathogens (<xref rid="kiab301-B217" ref-type="bibr">Skelsey et al., 2016</xref>). Pathogen identification and disease severity estimation are an important part of characterizing their distribution in the field (<xref rid="kiab301-B3" ref-type="bibr">Ali and Hodson, 2017</xref>). The detection and quantification of disease are usually performed by visually assessing crop symptoms, which may be subjected to bias and human error, besides being labor and time intensive. Various datasets have been released to assist the development of automated systems for disease identification and assessment (<xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 3</xref>). The Plant Village, BRACOL, RoCoLe, citrus (<italic>Citrus sp.</italic>), cassava (<italic>Manihot esculenta</italic>), and apple (<italic>Malus sp.</italic>) datasets offer close range annotated images of infected plant organs against a clean background, offering a resource for disease diagnosis and severity scoring in collected leaves (<xref rid="kiab301-B155" ref-type="bibr">Mohanty, 2016</xref>; <xref rid="kiab301-B8" ref-type="bibr">Arsenovic et al., 2019</xref>; <xref rid="kiab301-B35" ref-type="bibr">Chouhan et al., 2019</xref>; <xref rid="kiab301-B114" ref-type="bibr">Krohling, 2019</xref>; <xref rid="kiab301-B175" ref-type="bibr">Parraga-Alava et al., 2019</xref>; <xref rid="kiab301-B191" ref-type="bibr">Rauf et al., 2019</xref>; <xref rid="kiab301-B231" ref-type="bibr">Tian et al., 2019</xref>; <xref rid="kiab301-B164" ref-type="bibr">Nakatumba-Nabende et al., 2020</xref>; <xref rid="kiab301-B215" ref-type="bibr">Singh et al., 2020</xref>). Machine learning models using support vector machines, CNNs, and self-attention CNNs trained on similar datasets were published recently (<xref rid="kiab301-B1" ref-type="bibr">Abdu et al., 2020</xref>; <xref rid="kiab301-B52" ref-type="bibr">El Abidine et al., 2020</xref>; <xref rid="kiab301-B262" ref-type="bibr">Zeng and Li, 2020</xref>), some of which report increased efficiency when using segmented regions for pathogen identification (<xref rid="kiab301-B53" ref-type="bibr">Esgario et al., 2020</xref>; <xref rid="kiab301-B105" ref-type="bibr">Karlekar and Seal, 2020</xref>). A comprehensive review on machine learning for disease assessment in crops was published by <xref rid="kiab301-B83" ref-type="bibr">Hasan et al. (2020)</xref>.</p><p>Although disease detection models trained with the above datasets can be used in the field, the input samples have to be manually collected and imaged which can be time consuming. Hence, many researchers have focused on developing models that use UAV-collected images to accelerate disease detection (<xref rid="kiab301-B244" ref-type="bibr">Vergara-Diaz et al., 2015</xref>; <xref rid="kiab301-B225" ref-type="bibr">Sugiura et al., 2016</xref>; <xref rid="kiab301-B158" ref-type="bibr">Moriya et al., 2019</xref>; <xref rid="kiab301-B188" ref-type="bibr">Qiu et al., 2019</xref>; <xref rid="kiab301-B229" ref-type="bibr">Tetila et al., 2020</xref>; <xref rid="kiab301-B268" ref-type="bibr">Zhao et al., 2020</xref>). (<xref rid="kiab301-B139" ref-type="bibr">Marzougui et al., 2019</xref>) combined HTP images from greenhouse and field experiments to quantify Aphanomyces root rot resistance in lentils (<italic>Lens culinaris</italic>). The authors developed 12 normalized spectral indices that correlate with disease symptoms and severity, allowing breeders to objectively quantify genotype resistance. Another study used hyperspectral data and machine learning for early identification of charcoal rot disease in soybean, obtaining classification accuracy of 90% for plants 3 d after infection (<xref rid="kiab301-B162" ref-type="bibr">Nagasubramanian et al., 2018</xref>). These studies demonstrate the potential of image-based HTP to enable growers and breeders to automatically screen plants. To the best of our knowledge, there are no available datasets for disease-related tasks in the field which prevents the development and benchmarking of computer vision-based tools. Benchmark datasets created for this task requirements are shown in <xref rid="kiab301-BOX3" ref-type="boxed-text">Box 1</xref>, with specific image annotations depending on the target task (disease detection, identification, severity scoring, and lesion segmentation).</p><p>Field HTP is widely applied to the detection and quantification of pests. Rapid pest identification is important so growers can take action to control pest spread and limit damage to crops. A large benchmark dataset for insect pest detection was released containing 75,000 close range images of annotated pests belonging to 102 categories (see <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 3</xref> for a detailed description; <xref rid="kiab301-B253" ref-type="bibr">Wu et al., 2019</xref>). This benchmark dataset is a valuable resource for the development of crop monitoring and management approaches, allowing researchers to test model performance over a wide range of pests. This dataset can also be complemented with the mango (<italic>Mangifera indica</italic>) pest classification dataset, which has images of mango plants infected with 15 different categories of pests, with a large volume of augmented images to increase model robustness (<xref rid="kiab301-B118" ref-type="bibr">Kusrini et al., 2020a</xref>). Precise algorithms for the detection of pests can support assessing crop resistance by counting the pests, helping identify pest species in the field, and monitoring pest spread. Employing HTP datasets to measure plant–insect interactions can allow the use of RGB sensors to quantify leaf damage and defoliation (<xref rid="kiab301-B169" ref-type="bibr">O’Neal et al., 2002</xref>). Thermal infrared and hyperspectral images can also be used to capture physiological changes, such as stomata regulation (<xref rid="kiab301-B12" ref-type="bibr">Backoulou et al., 2011</xref>; <xref rid="kiab301-B161" ref-type="bibr">Nabity et al., 2013</xref>). Novel datasets targeting the plant–insect interactions should follow the guidelines proposed in <xref rid="kiab301-BOX3" ref-type="boxed-text">Box 1</xref> with special attention to providing detailed metadata (view MIAPPE project) and ground-truth measurements and labeling.</p><p>The development of navigation maps is particularly important for weed management systems, in which the map can be used for targeted herbicide application or by weed killing robots (<xref rid="kiab301-B221" ref-type="bibr">Somerville et al., 2019</xref>; <xref rid="kiab301-B69" ref-type="bibr">Gašparović et al., 2020</xref>; <xref rid="kiab301-B94" ref-type="bibr">Hunter et al., 2020</xref>; <xref rid="kiab301-B189" ref-type="bibr">Raja et al., 2020</xref>). Weed detection systems can reduce herbicide application by up to 60% in comparison to broadcast applications (<xref rid="kiab301-B221" ref-type="bibr">Somerville et al., 2019</xref>; <xref rid="kiab301-B94" ref-type="bibr">Hunter et al., 2020</xref>) and increase efficiency in organic production systems. A key challenge for implementing weed detection in the field using image-based HTP data is the difficulty in establishing robust computer vision-based models that can distinguish between crop and weed species under varying field conditions. To help overcome this challenge, many datasets have been released consisting of RGB and multispectral images of a wide variety of weed and crop species, some of which contain pixel level annotations to separate the weed from background (<xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 3</xref>; <xref rid="kiab301-B84" ref-type="bibr">Haug and Ostermann, 2015</xref>; <xref rid="kiab301-B46" ref-type="bibr">Dos Santos Ferreira, 2017</xref>; <xref rid="kiab301-B72" ref-type="bibr">Giselsson et al., 2017</xref>; <xref rid="kiab301-B198" ref-type="bibr">Sa et al., 2018</xref>; <xref rid="kiab301-B228" ref-type="bibr">Teimouri et al., 2018</xref>; <xref rid="kiab301-B218" ref-type="bibr">Skovsen et al., 2019</xref>; <xref rid="kiab301-B224" ref-type="bibr">Sudars et al., 2020</xref>). A few datasets feature images of weed seedlings, enabling the development of models that can detect weed infestation at an early stage. Studies using similar datasets employed computer vision and machine learning algorithms for weed detection, though these presented a high variability in the precision rate (69%–98%) depending on the crop field analyzed (<xref rid="kiab301-B246" ref-type="bibr">Wang et al., 2007</xref>; <xref rid="kiab301-B200" ref-type="bibr">dos Santos Ferreira et al., 2017</xref>; <xref rid="kiab301-B171" ref-type="bibr">Pallottino et al., 2018</xref>; <xref rid="kiab301-B237" ref-type="bibr">Umamaheswari et al., 2018</xref>; <xref rid="kiab301-B13" ref-type="bibr">Bah et al., 2019</xref>; <xref rid="kiab301-B176" ref-type="bibr">Partel et al., 2019</xref>). These results emphasize the need to produce more datasets with an increased variety of crop and weed species at different growth stages. Furthermore, the datasets need to reflect the management practices (e.g. sowing density) that the weed detection model would encounter in the field. Increasing model robustness to varied field conditions is essential to enable its adoption in agricultural management systems and allow plant researchers to quantify herbicide or other weed control practices efficiency.</p></sec><sec id="sec7" disp-level="2"><title>Root phenotyping</title><p>Root system architecture (RSA) greatly influences nutrient access, efficient water uptake, and plant tolerance to stress (<xref rid="kiab301-B138" ref-type="bibr">Mary et al., 2018</xref>; <xref rid="kiab301-B257" ref-type="bibr">York et al., 2018</xref>; <xref rid="kiab301-B141" ref-type="bibr">Mattupalli et al., 2019</xref>; <xref rid="kiab301-B28" ref-type="bibr">Busener et al., 2020</xref>; <xref rid="kiab301-B77" ref-type="bibr">Griffiths et al., 2020</xref>; <xref rid="kiab301-B145" ref-type="bibr">McKay Fletcher et al., 2020</xref>; <xref rid="kiab301-B207" ref-type="bibr">Seo et al., 2020</xref>). Increased efforts in breeding for desirable RSA traits can drive a breakthrough in crop productivity and resource efficiency (<xref rid="kiab301-B133" ref-type="bibr">Lynch, 2007</xref>). To leverage RSA potential in crop breeding, it is important that we improve current root phenotyping strategies.</p><p>Noninvasive RSA imaging is extremely challenging due to soil opacity. At the same time, soil replacements such as transparent gels or hydroponic mediums often lead to phenotypes that diverge substantially from the ones observed in regular soil (<xref rid="kiab301-B82" ref-type="bibr">Hargreaves et al., 2009</xref>; <xref rid="kiab301-B251" ref-type="bibr">Wojciechowski et al., 2009</xref>; <xref rid="kiab301-B36" ref-type="bibr">Clark et al., 2011</xref>; <xref rid="kiab301-B142" ref-type="bibr">Ma et al., 2019</xref>). A wide variety of sensors can be employed to acquire 2D or 3D images of plant root grown in the glasshouse, such as X-ray computed tomography, magnetic resonance imaging, positron emission tomography, and hyperspectral imaging (<xref rid="kiab301-B98" ref-type="bibr">Jahnke et al., 2009</xref>; <xref rid="kiab301-B67" ref-type="bibr">Garbout et al., 2012</xref>; <xref rid="kiab301-B157" ref-type="bibr">Mooney et al., 2012</xref>; <xref rid="kiab301-B48" ref-type="bibr">van Dusschoten et al., 2016</xref>; <xref rid="kiab301-B20" ref-type="bibr">Bodner et al., 2018</xref>). In <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 4</xref>, we list available RSA datasets with metadata at varied levels of detail including from plants grown in multiple types of media, such as gellan gum, soil, and hydroponics. In addition, a synthetic root system dataset is available. This large dataset was produced for tool calibration and modeling since it provides ground-truth of fibrous and tap-root images which help identify artifacts generated by the model when dealing with complex, overlapping root structures. The data were produced using ArchiSimple with three levels of noise, and the roots present varying degrees of complexity (<xref rid="kiab301-B132" ref-type="bibr">Lobet et al., 2017</xref>).</p><p>Field root phenotyping frequently requires the manual excavation of individual plants followed by imaging of the washed root crown system for quantitative trait analysis (<xref rid="kiab301-B234" ref-type="bibr">Trachsel et al., 2011</xref>; <xref rid="kiab301-B25" ref-type="bibr">Bucksch et al., 2014</xref>; <xref rid="kiab301-B37" ref-type="bibr">Colombi et al., 2015</xref>). Root crown datasets of multiple crop species are described in <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 4</xref>, some of which were produced with the aim to automatically quantify RSA traits using different tools. Noninvasive alternative approaches are not as commonly employed, but offer the potential to undertake a time-series analysis of crop development. These include electrical resistance tomography, electromagnetic inductance, and ground penetrating radar (<xref rid="kiab301-B43" ref-type="bibr">Diaz and Herrero, 1992</xref>; <xref rid="kiab301-B263" ref-type="bibr">Zenone et al., 2008</xref>; <xref rid="kiab301-B223" ref-type="bibr">Srayeddin and Doussan, 2009</xref>), which are used to characterize root water uptake of wheat and vine plants in the field (<xref rid="kiab301-B209" ref-type="bibr">Shanahan et al., 2015</xref>; <xref rid="kiab301-B250" ref-type="bibr">Whalley et al., 2017</xref>; <xref rid="kiab301-B138" ref-type="bibr">Mary et al., 2018</xref>).</p><p>Overall, image-based RSA phenotyping has many applications, such as linking RSA traits to micronutrient concentration and heritability (<xref rid="kiab301-B28" ref-type="bibr">Busener et al., 2020</xref>; <xref rid="kiab301-B145" ref-type="bibr">McKay Fletcher et al., 2020</xref>), the effect of dwarf genes in seedling roots (<xref rid="kiab301-B251" ref-type="bibr">Wojciechowski et al., 2009</xref>), changes in the root crown in response to disease (<xref rid="kiab301-B39" ref-type="bibr">Corona-Lopez et al., 2019</xref>; <xref rid="kiab301-B141" ref-type="bibr">Mattupalli et al., 2019</xref>), to investigate root plasticity (<xref rid="kiab301-B197" ref-type="bibr">Rosas et al., 2013</xref>), genetically driven root architecture differences (<xref rid="kiab301-B100" ref-type="bibr">Jiang et al., 2019</xref>), and QTL mapping of regions controlling RSA (<xref rid="kiab301-B232" ref-type="bibr">Topp et al., 2013</xref>). Most of the studies cited above use a combination of tools for RSA trait extraction (DIRT; <xref rid="kiab301-B41" ref-type="bibr">Das et al., 2015</xref>), RhizoVision (<xref rid="kiab301-B204" ref-type="bibr">Seethepalli and York, 2019</xref>), RSA-GiA (<xref rid="kiab301-B66" ref-type="bibr">Galkovskyi et al., 2012</xref>; <xref rid="kiab301-B232" ref-type="bibr">Topp et al., 2013</xref>), or Rootscape (<xref rid="kiab301-B195" ref-type="bibr">Ristova et al., 2013</xref>)) followed by statistical analysis (variations of ANOVA, three-parameter logistic function, PCA) or linear regression to test if the observed traits relate to environmental or genetic data. The wide range of approaches used reflects the diversity of input data formats. The sensors employed to collect RSA traits are very diverse and capture different aspects of the root. Hence, the decision for which feature extraction tool and analysis method to implement must be decided case by case. Even more important in this case is tool and data interoperability because it will allow researchers to explore the resources efficiently. Root image datasets from several major crop species can be downloaded from the Quantitative Plant platform (quantitative-plant.org/dataset) and Zenodo database(zenodo.org/).</p><p>The reconstruction of the data as 2D or 3D representations of the root system, and root segmentation from the medium usually assumes a high contrast between root and background, which is not always the case (<xref rid="kiab301-B11" ref-type="bibr">Atkinson et al., 2019</xref>). Machine and deep learning-based tools have been developed for root segmentation in 2D or 3D (<xref rid="kiab301-B97" ref-type="bibr">Iyer-Pascuzzi et al., 2010</xref>; <xref rid="kiab301-B25" ref-type="bibr">Bucksch et al., 2014</xref>; <xref rid="kiab301-B55" ref-type="bibr">Falk et al., 2020</xref>; <xref rid="kiab301-B255" ref-type="bibr">Yasrab et al., 2020a</xref>), including very thin (1–3 pixels) roots grown in visible medium (RootNet; <xref rid="kiab301-B256" ref-type="bibr">Yasrab et al., 2020b</xref>) and in soil (<xref rid="kiab301-B220" ref-type="bibr">Soltaninejad et al., 2020</xref>), while other tools aimed for RSA trait quantification (<xref rid="kiab301-B9" ref-type="bibr">Atkinson et al., 2017a</xref>; <xref rid="kiab301-B55" ref-type="bibr">Falk et al., 2020</xref>). Although there are many potential approaches to perform root segmentation, most are not suited for newer image data types. In addition, few tools are capable of linking observed RSA to genotypic information. Recently, deep learning models have been employed to attempt to bridge phenotype to genotype predictions (<xref rid="kiab301-B184" ref-type="bibr">Pound et al., 2017a</xref>; <xref rid="kiab301-B255" ref-type="bibr">Yasrab et al., 2020a</xref>) and can achieve similar results for QTL identification as user supervised methods (<xref rid="kiab301-B184" ref-type="bibr">Pound et al., 2017a</xref>). However, to effectively integrate high-throughput phenotype to genotype tools into the breeding process requires refined tools. These tools must be capable of dealing with phenotype and sensor variability and of aggregating experimental metadata into the analysis. The success in the development of such tools relies on the quality and size of the available datasets because these are the sole source of information for the deep learning model to adjust its internal parameters.</p></sec><sec id="sec8" disp-level="2"><title>Quantitative plant morphology</title><p>The description of plant morphological traits, for example, number of leaves, canopy cover, number of flowers, and seeds, provides a foundation to characterize plant phenotypic response, which is directly related to plant developmental stage, yield potential, and overall health (<xref rid="kiab301-B112" ref-type="bibr">Kouressy et al., 2008</xref>). The quantification of agronomic traits often relies on manual measurements, which are costly, labor-intensive, and prone to errors. Several approaches including neural networks and other machine learning models have been published to perform leaf counting, area estimation, folding and plant growth stage classification, stem–leaf segmentation, and seed counting (<xref rid="kiab301-B173" ref-type="bibr">Parmar et al., 2016</xref>; <xref rid="kiab301-B180" ref-type="bibr">Pereira et al., 2016</xref>; <xref rid="kiab301-B219" ref-type="bibr">Sodhi et al., 2017</xref>; <xref rid="kiab301-B228" ref-type="bibr">Teimouri et al., 2018</xref>; <xref rid="kiab301-B238" ref-type="bibr">Uzal et al., 2018</xref>; <xref rid="kiab301-B102" ref-type="bibr">Jin et al., 2019</xref>; <xref rid="kiab301-B190" ref-type="bibr">Rascio et al., 2020</xref>). Deep learning models are widely applied to image analysis due to the high complexity of the data and their potential for quantitative morphology lies partially in their capacity to segment the target object from the nontarget objects in the image. Hence, it is possible to measure the traits of the segmented object (number of seeds, color, fruit shape, fruit, or seed size). This measurement ability was shown in a study for fish morphology quantification that used Mask R-CNN for pixel-wise segmentation of the fish body followed by measurement of its morphological features (<xref rid="kiab301-B259" ref-type="bibr">Yu et al., 2020</xref>). A variety of trait phenotyping datasets have been released to develop pipelines for trait measurement, such as the hypocotyl dataset with images of <italic>A. thaliana</italic> seedlings for length determination (<xref rid="kiab301-B44" ref-type="bibr">Dobos et al., 2019</xref>), image time-series of <italic>A. thaliana</italic> growth that can be used to predict performance (<xref rid="kiab301-B226" ref-type="bibr">Taghavi Namin et al., 2018</xref>), and species identification datasets (<xref rid="kiab301-B117" ref-type="bibr">Kumar et al., 2012</xref>; Lee et al., <xref rid="kiab301-B125" ref-type="bibr">2015</xref>, <xref rid="kiab301-B124" ref-type="bibr">2017</xref>; <xref rid="kiab301-B63" ref-type="bibr">Fricker et al., 2019</xref>; <xref rid="kiab301-B270" ref-type="bibr">Zheng et al., 2019</xref>) as shown in <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 5</xref>.</p><p>PlantCV and Deep Plant Phenomics are the two platforms that offer packaged pretrained deep learning models to run as applications for phenotyping (<xref rid="kiab301-B54" ref-type="bibr">Fahlgren et al., 2015</xref>; <xref rid="kiab301-B235" ref-type="bibr">Ubbens and Stavness, 2017</xref>). However, tools for quantitative morphology analysis can only guarantee performance if under restricted image conditions and may require further image processing steps. Producing and sharing annotated datasets from a diverse set of species are the most efficient way to ensure new tools can be developed to incorporate them. The Plant Phenotyping Datasets (<xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 5</xref>) are the collection of annotated top-view images of <italic>A. thaliana</italic> and tobacco (<italic>Nicotiana tabacum</italic>) undergoing different treatments (<xref rid="kiab301-B147" ref-type="bibr">Minervini et al., 2016</xref>). It is a benchmark dataset (<xref rid="kiab301-BOX3" ref-type="boxed-text">Box 1</xref>), that was employed in the leaf segmentation and leaf counting challenges at the Computer Vision Problems in Plant Phenotyping conference, and propelled the development of tools for leaf segmentation and counting (<xref rid="kiab301-B2" ref-type="bibr">Aich and Stavness, 2017</xref>; <xref rid="kiab301-B45" ref-type="bibr">Dobrescu et al., 2017</xref>; <xref rid="kiab301-B73" ref-type="bibr">Giuffrida et al., 2018</xref>; <xref rid="kiab301-B186" ref-type="bibr">Praveen Kumar and Domnic, 2020</xref>), which can be later used for assessing plant growth and biomass. Other datasets focused on seed and fruit organs are available. Some datasets are useful to compare variance in seed morphological traits (<xref rid="kiab301-B47" ref-type="bibr">Ducournau et al., 2020</xref>), while others can be used for the development of computer vision tools for fruit counting and automatic quality assessment. In this category, there is a soybean image dataset to assess seed damage from mechanical and biological sources (<xref rid="kiab301-B179" ref-type="bibr">Pereira et al., 2019</xref>), a dataset for the identification of Indian basmati rice (<italic>Oryza sativa</italic>) seed varieties (<xref rid="kiab301-B211" ref-type="bibr">Sharma et al., 2020</xref>), sugar beet (<italic>Beta vulgaris</italic>) seed traits (<xref rid="kiab301-B47" ref-type="bibr">Ducournau et al., 2020</xref>), a cocoa bean (<italic>Theobroma cacao</italic>) dataset for quality assessment (<xref rid="kiab301-B199" ref-type="bibr">Santos et al., 2019</xref>), a banana (<italic>Musa sp.</italic>) tier abnormality classification (<xref rid="kiab301-B182" ref-type="bibr">Piedad, 2019</xref>), and hyperspectral images of different loose tea (<italic>Camellia sinensis</italic>; <xref rid="kiab301-B151" ref-type="bibr">Mishra, 2018</xref>; <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 5</xref>).</p><p>Determining leaf inclination and distribution on the plant is an important morphological trait, it impacts the plant spectral reflectance and is a mechanism to increase tolerance to abiotic stress, with impacts on leaf temperature, water loss, and drought tolerance (<xref rid="kiab301-B50" ref-type="bibr">Ehleringer and Comstock, 1987</xref>; <xref rid="kiab301-B64" ref-type="bibr">Fuchs, 1990</xref>; <xref rid="kiab301-B85" ref-type="bibr">He et al., 1996</xref>; <xref rid="kiab301-B249" ref-type="bibr">Werner et al., 1999</xref>). In common bean (<italic>Phaseolus vulgaris L.</italic>) the extent of leaf movement increases as the water availability drops, allowing the plants to maintain leaf temperature despite stomata closure (<xref rid="kiab301-B177" ref-type="bibr">Pastenes et al., 2005</xref>). A dataset for leaf angle estimation with ground-truth angles for 71 <italic>Eucalyptus</italic> species (<xref rid="kiab301-B183" ref-type="bibr">Pisek and Adamson, 2020</xref>) is described in <xref rid="sup1" ref-type="supplementary-material">Supplemental Data Set 5</xref>, it contains images of <italic>Eucalyptus</italic> canopies that can be used to estimate leaf angle distribution in trees. Automated pipelines for leaf angle extraction have been developed and tested for <italic>A. thaliana</italic>, beet, apple (<italic>Malus domestica</italic>), maize, and sorghum (<xref rid="kiab301-B159" ref-type="bibr">Müller-Linow et al., 2015</xref>; <xref rid="kiab301-B106" ref-type="bibr">Kenchanmane Raju et al., 2020</xref>), allowing researchers to track leaf angle variability and distribution over time. Identifying varieties with desired leaf angle distribution can assist breeders to select the varieties best adapted to specific environmental conditions, such as high planting densities where a narrow angle prevents the leaf from being shadowed by others (<xref rid="kiab301-B178" ref-type="bibr">Pepper et al., 1977</xref>; <xref rid="kiab301-B120" ref-type="bibr">Lambert and Johnson, 1978</xref>).</p><p>A multitask pipeline capable of phenotyping a comprehensive array of traits in different tissues would produce a snapshot that can be used to identify new QTLs. It was observed that genetic traits may contribute to different tissues causing multiple trait variance (<xref rid="kiab301-B128" ref-type="bibr">Li et al., 2018</xref>). This would provide a resource to detect QTLs and improve our understanding of the genetic basis of complex phenotypes (<xref rid="kiab301-B232" ref-type="bibr">Topp et al., 2013</xref>). Trait phenotyping can also be used for the construction of 3D representations of the plant structure (<xref rid="kiab301-B232" ref-type="bibr">Topp et al., 2013</xref>; <xref rid="kiab301-B239" ref-type="bibr">Vadez et al., 2015</xref>; <xref rid="kiab301-B48" ref-type="bibr">van Dusschoten et al., 2016</xref>; <xref rid="kiab301-B143" ref-type="bibr">McCormick et al., 2016</xref>; <xref rid="kiab301-B19" ref-type="bibr">Bengochea-Guevara et al., 2017</xref>; <xref rid="kiab301-B219" ref-type="bibr">Sodhi et al., 2017</xref>; <xref rid="kiab301-B241" ref-type="bibr">Vázquez-Arellano et al., 2018</xref>; <xref rid="kiab301-B248" ref-type="bibr">Wang et al., 2018b</xref>). This avoids loss of information caused by 2D compression and prevents the generation of artifacts that can occur due to lighting, occlusion, and overlaps.</p></sec></sec><sec id="sec9" disp-level="1"><title>Concluding remarks</title><p>HTP platforms and tools are revolutionizing the way we capture plant phenotypic variation, by allowing the quantification of agronomic traits, and the identification of genetic traits with potential for crop breeding. Publishing the collected phenotypic datasets and associated information would help drive the development of high-performance crops, allowing growers to more effectively monitor their crops and giving breeders the opportunity to explore research from a new perspective with updated tools. The research community must adhere to standardized practices for dataset release such as proposed by MIAPPE in order for the datasets to be explored and interpreted (see “Outstanding questions”). Because of the multiple types of data comprising a HTP dataset, it is important that the terms are clearly defined so researchers from different fields (computer science, remote sensing, and plant biology) can collaborate. In cases where data sharing is unfeasible due to privacy or security concerns, federative learning offers an opportunity to train machine learning algorithms collaboratively without exchanging data. A variety of mathematical and machine learning methods have recently been applied to address the bottleneck of phenotypic quantitative analysis. However, without established benchmark datasets, it is difficult to compare the performance of these approaches, imposing a barrier to improvements and our understanding of the limitations of techniques. It is also important that novel tools are intuitive and well documented, allowing domain experts with minimal programing background to benefit (<xref rid="kiab301-B110" ref-type="bibr">Klukas et al., 2014</xref>; <xref rid="kiab301-B235" ref-type="bibr">Ubbens and Stavness, 2017</xref>). Plant phenotyping is a rapidly evolving field with a growing community, it is important that we use this growth to establish structures such as public repositories and benchmarks to support the field so it may achieve its potential to accelerate crop breeding.</p>
<boxed-text id="kiab301-BOX2" position="float"><sec id="sec10" disp-level="2"><title>Outstanding questions</title><list list-type="bullet"><list-item><p>What is the best approach to solve the high variability in HTP data collection and processing methodology? Should we define standard methodologies for these tasks or develop tools to detect variance?</p></list-item><list-item><p>How can we collate sufficient benchmark datasets to evaluate tool performance? Are the current benchmarks capable of exposing limitations of the tools?</p></list-item><list-item><p>How should authors be encouraged to release datasets with their publications, similar to what is required when publishing the results from analysis of genomic datasets? What structures are needed to support the release and maintenance of these datasets?</p></list-item><list-item><p>How can we increase data interoperability to integrate datasets from multiple sources (genomic, environmental data)? What is the minimum metadata needed to ensure that?</p></list-item></list></sec></boxed-text>
</sec><sec id="sec11" disp-level="1"><title>Supplemental data</title><p>The following materials are available in the online version of this article.</p><p><bold><xref rid="sup1" ref-type="supplementary-material">Supplementary Data Set 1</xref></bold>. Available image-based HTP datasets for crop yield prediction.</p><p><bold><xref rid="sup1" ref-type="supplementary-material">Supplementary Data Set 2</xref></bold>. Available image-based HTP datasets for abiotic stress phenotyping</p><p><bold><xref rid="sup1" ref-type="supplementary-material">Supplementary Data Set 3</xref></bold>. Available image-based HTP datasets for disease and pest detection</p><p><bold><xref rid="sup1" ref-type="supplementary-material">Supplementary Data Set 4</xref></bold>. Root phenotyping datasets</p><p><bold><xref rid="sup1" ref-type="supplementary-material">Supplementary Data Set 5</xref></bold>. Other miscellaneous databases that may be useful for applications not discussed in this review.</p></sec><sec id="sec12" disp-level="1"><title>Funding</title><p>This work was supported by the Australian Government through the Australian Research Council (Projects DP200100762, DP1601004497, and LP140100537) and the Grains Research and Development Corporation (Projects 9177539 and 9177591) Benjamin J. Nestor is supported by a university postgraduate award at The University of Western Australia. Monica F. Danilevicz and Philipp E. Bayer received support from the Forrest Research Foundation. Monica F. Danilevicz and Benjamin J. Nestor are further supported by the Research Training Program scholarships.</p><p><italic>Conflict of interest statement</italic>. The authors declare no competing interests.</p></sec><sec id="sec13" disp-level="1"><title>Supplementary Material</title><supplementary-material id="sup1" position="float"><?disp-level 2?><label>kiab301_Supplementary_Data</label><media xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="kiab301_Supplementary_Data.zip" mimetype="application" mime-subtype="zip"><?cloudpmc-path 0f62/8561249/e77636d9bb03/kiab301_Supplementary_Data.zip?><?cloudpmc-bucket app?><?size 46696?><caption><p>Click here for additional data file.</p></caption></media></supplementary-material></sec><sec id="sec14" disp-level="1"><title> </title><p>M.F.D. wrote the manuscript with input and edits from P.E.B., M.B., B.J.N., and D.E.</p><p>The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (<ext-link xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="https://academic.oup.com/plphys/pages/general-instructions" ext-link-type="uri">https://academic.oup.com/plphys/pages/general-instructions</ext-link>) is: David Edwards (<email>dave.edwards@uwa.edu.au</email>).</p></sec><sec id="ref1" sec-type="ref-list" disp-level="1"><title>References</title><sec id="ref1_sec2" disp-level="2"><ref-list><ref id="kiab301-B1"><mixed-citation><named-content content-type="citation-string">Abdu AM, Mokji MM, Sheikh UU (2020) Automatic vegetable disease identification approach using individual lesion features. Comput Electron Agric
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