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Plant phenotyping methods.

植物形質を測っただけの研究ではなく、フェノタイピング手法の開発・検証・実質的利用・ベンチマーク・方法レビューとの関連性が見つかった論文を中心に表示します。

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10 papers · code / dataset availability confirmedLatest completed run · 2016-01-01 – 2026-09-15
Code / dataset availability confirmedCrossref · checked 5 Sept 2026
Published23 Dec 2025PlantsCited by 0 · OpenAlex ↗

Real-Time Callus Instance Segmentation in Plant Tissue Culture Using Successive Generations of YOLO Architectures

Yunus Egi · Tülay Oter · Mortaza Hajyzadeh · Muammer Catak

LentilLaboratory / benchtopLeafTissueSegmentation

Callus induction is a complex procedure in plant organ, cell, and tissue culture that underpins processes such as metabolite production, regeneration, and genetic transformation. It is important to monitor callus formation alongside subjective evaluations, which require labor-intensive care. In this research, the first curated lentil (Lens culinaris) callus dataset for instance segmentation was experimentally generated using three genotypes as one data set: Firat-87, Cagil, and Tigris. Leaf explants were cultured on MS medium fortified with different concentrations of gross regulators of BA and NAA to induce callus formation. Three biologically relevant stages, the leaf stage, the green callus, and the necrosis callus, were produced. During this process, 122 high-resolution images were obtained, resulting in 1185 total annotations across them. The dataset was evaluated across four successive generations (v5/7/8/11) of YOLO deep learning models under identical conditions using mAP, Dice coefficient, Precision, Recall, and IoU, together with efficiency metrics including parameter counts, FLOPs, and inference speed. The results show that anchor-based variants (YOLOv5/7) relied on predefined priors and showed limited boundary precision, whereas anchor-free designs (YOLOv8/11) used decoupled heads and direct center/boundary regression that provided clear advantages for callus structures. YOLOv8 reached the highest instance segmentation precision with mAP50@0.855, while it matched the accuracy with greater efficiency and achieved real-time inference with 166 FPS.

Reproduction assets foundThe paper's lentil callus image dataset with annotations (122 images, 1185 annotations) is publicly available on Roboflow Universe per the Data Availability Statement. The YOLOv5 GitHub link and Ultralytics docs are generic third-party libraries, not authors' analysis code, and the FAO link is a cited reference, so all
Dataset · publicThe dataset used in this study, including annotated images for callus detection, is publicly available and can be accessed at Roboflow Universe: https://universe.roboflow.com/yunus-7v2b5/callus-hug7d , accessed on 13 September 2025. This repository contains all images and annotations generated and analyzed during the current study.Open asset ↗Roboflow Universe · callus-hug7dlines:314-345
Code / dataset availability confirmedOpenAlex · Crossref · checked 14 Sept 2026
Published9 Nov 2025The Plant Phenome JournalCited by 0 · OpenAlex ↗

UAV‐based high‐throughput phenotyping for crop growth analysis and seed yield prediction in a nested association mapping population of lentils

Sandesh Neupane · Kirstin E. Bett · Steven J. Shirtliffe

LentilAerial / UAVField / plotWhole plant / canopy / plot / fieldMorphology / geometry measurementGrowth / time-series analysisYield / biomass estimationArchitecture / morphology / geometryGrowth / development / phenologyPlant / canopy height

Abstract Unoccupied aerial vehicle (UAV)‐based high‐throughput phenotyping provides scalable and cost‐effective access to phenotypic information for crop improvement, yet its application in minor crops such as lentil ( Lens culinaris Medik.) remains limited. This study applied UAV‐derived canopy traits and crop growth regression modeling to a nested association mapping population developed from CDC Redberry crossed with 32 diverse founder lines. UAV imagery collected across four site‐years was used to capture canopy height, crop area, and crop volume per plot basis at multiple time points. Crop growth regression models were fitted to derive crop growth parameters, maximum canopy size, growth rate, and cumulative growth anchored to phenological stages. These static and time‐series traits were evaluated for seed yield prediction using partial least squares regression with a 70:20:10 data split and 10‐fold cross‐validation. Static traits such as maximum crop volume and maximum crop area were consistently associated with yield. Dynamic trait‐based models improved prediction accuracy and identified the swollen pod stage (R5–R6) as the most informative forecasting window. External validation using an independent trial confirmed the generalizability of the approach. This study presents a UAV phenotyping framework that supports crop growth dissection and early yield prediction and downstream trait discovery in lentil.

Reproduction assets foundThe paper's data availability statement points to a public KnowPulse experiment page hosting the study's UAV-derived phenotyping and yield data; no author analysis code repository is stated.
Dataset · publicof S). We thank Dr. Ana Vargas at the Crop Development Center, U of S for generously providing yield data from the independent field trial. C O N F L I C T O F I N T E R E S T S TAT E M E N T The authors declare no conflicts of interest. DATA AVA I L A B I L I T Y S TAT E M E N T The data supporting this study are available at: https://knowpulse.usask.ca/experiment/AGILE-NAM-UAV-growth-modelling or from the authors upon request. O RC I D SandeshNeupane https://orcid.org/0000-0003-3679-1046 KirstinE. Bett https://orcid.org/0000-0001-7959-6959 SteveJ. Shirtliffe https://orcid.org/0000-0002-3603-7417 R E F E R E N C E S Araus, J. L., Kefauver, S. C., Zaman-Allah, M., Olsen, M. S., & Cairns, J.Open asset ↗KnowPulse · AGILE-NAM-UAV-growth-modellingpdf-raw-page:15 lines:1-90
Code / dataset availability confirmedOpenAlex · checked 14 Sept 2026
Published19 Aug 2025The Plant Phenome JournalCited by 1 · OpenAlex ↗

Dissecting lentil crop growth in contrasting environments using digital imaging and genome‐wide association studies

Derek Wright · Sandesh Neupane · Steven J. Shirtliffe · Tania Gioia · Giuseppina Logozzo · Stefania Marzario · Karsten M. E. Nielsen · Kirstin E. Bett

LentilAerial / UAVField / plotRGB / grayscaleMultispectral / hyperspectralWhole plant / canopy / plot / fieldMorphology / geometry measurementGrowth / time-series analysisArchitecture / morphology / geometryGrowth / development / phenology

Abstract The development of high‐throughput phenotyping platforms to capture time‐series data on large, diverse populations holds promise for crop researchers and breeders investigating growth‐related traits. We used imagery from unoccupied aerial vehicles (UAVs) with red/green/blue (RGB) and multispectral cameras flown over multiple site‐years in Saskatchewan, Canada, and Metaponto, Italy, to gather data for crop height, area, and volume in a lentil diversity panel (324 genotypes). The temporal nature of the UAV image‐derived data enabled the modeling of growth curves for volume, height, and area, something that would be impractical under traditional phenotyping procedures in such a large population grown in multiple environments. A principal component analysis and hierarchical clustering revealed differential growth patterns across contrasting environments, with large variations in temperature and photoperiod, within our lentil diversity panel. Combining this analysis with genome‐wide genotyping data, we identified markers, from an exome capture array (267,845 single nucleotide polymorphisms), associated with crop growth that could be used for marker‐assisted selection. Our study demonstrates the potential for UAV‐based imaging to obtain large‐scale time‐series data across multiple environments to model growth curves and investigate genotype‐by‐environment interactions. In addition, we can now use phenotypic traits that were once impractical to collect and derive novel phenotypes to improve our understanding of crop growth and the genetics underlying adaptation in lentil, approaches that will be useful for both researchers and breeders.

Reproduction assets foundThe paper's UAV-derived lentil growth phenotypes are publicly available on KnowPulse, and the authors' full analysis code/workflow is public on GitHub with a rendered vignette. Both are explicitly stated in the data availability statement and methods.
Dataset · publiciluppo e di Innovazione in Agricoltura) in Metaponto, Italy. Special thanks to Laura Jardine for help with editing. C O N F L I C T O F I N T E R E S T S TAT E M E N T The authors declare no conflicts of interest. DATA AVA I L A B I L I T Y S TAT E M E N T The data that support the findings of this study are available online at https://knowpulse.usask.ca/research-experiment/AGILE-UAV and https://github.com/derekmichaelwright/AGILE_LDP_UAV or from the authors upon request. O RC I D DerekM. Wright https://orcid.org/0000-0002-9639-7596 SandeshNeupane https://orcid.org/0000-0003-3679-1046 Tania Gioia https://orcid.org/0000-0001-8980-3034 Giuseppina Logozzo https://orcid.org/0000-0002-7951-2425 SOpen asset ↗knowpulse.usask.ca · AGILE-UAVpdf-raw-page:11 lines:1-84
Code · publical user- calculated traits as described in Figure 2. G × E analysis was done with “lme4” using linear mixed models (Bates et al., 2015). Principal component analysis (PCA) and hierarchical k-means clustering were performed using the “FactoMineR” R package (Lê et al., 2008). The source code for all data analyses is available at: https://derekmichaelwright.github.io/AGILE_LDP_UAV/LDP_UAV_Vignette.html.25782703, 2025, 1, Downloaded from https://acsess.onlinelibrary.wiley.com/doi/10.1002/ppj2.70040, Wiley Online Library on [20/08/2025]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on WileyOpen asset ↗derekmichaelwright.github.io/AGILE_LDP_UAV · LDP_UAV_Vignettepdf-raw-page:3 lines:1-106
Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Published12 Dec 2024Data in briefCited by 3 · OpenAlex ↗

Lentil plant disease and quality assessment: A detailed dataset of high-resolution images for deep learning research.

Mahamud E, Assaduzzaman M, Sharmin S.

LentilField / plotWhole plant / canopy / plot / fieldStress / disease detectionDisease symptoms / severity

The Lentil, a vital legume globally cultivated, faces significant challenges from diseases like ascochyta blight, lentil rust, and powdery mildew. Ensuring optimal harvest timing and effectively discerning healthy and diseased lentil plants are crucial for maintaining crop quality and economic viability, particularly in regions such as Bangladesh. This paper introduces a comprehensive dataset comprising high-resolution images of lentil plants gathered meticulously over four months from diverse locations across Bangladesh, under expert supervision. The dataset aims to support the development of machine-learning models for precise disease detection and quality assessment in lentil cultivation. Potential applications include enhancing the accuracy of quality evaluation, and improving packaging processes, thereby enhancing overall lentil production efficiency. Agricultural researchers can utilize this dataset to advance applications of computer vision and deep learning in managing crop diseases and enhancing yield outcomes. The dataset's creation involved collaboration with domain experts to ensure its relevance and reliability for agricultural research. By leveraging this dataset, researchers can explore innovative approaches to tackle challenges in lentil farming, contributing to sustainable agricultural practices and food security. Moreover, the dataset serves as a valuable resource for training and testing machine learning algorithms tailored to agricultural settings, facilitating advancements in automated agricultural technologies. Ultimately, this initiative aims to empower stakeholders in the lentil industry with tools to mitigate disease impact and optimize production practices, paving the way for more resilient and efficient agricultural systems globally .

Reproduction assets foundThe paper is a Data in Brief article describing a lentil plant disease image dataset (1,898 original and 4,550 augmented images across four classes) deposited publicly on Mendeley Data with DOI 10.17632/7vb77bz2st.1. This is a paper-specific, publicly accessible image dataset directly reproducing the paper's phenotypic
Dataset · publicnd research stations in Barisal, Bangladesh Coordinates: 1. Farm A: 22.7083° N, 90.3653° E 2. Farm B: 22.6715° N, 90.3252° E 3. Research Station C: 22.7032° N, 90.3863° E Zone: Barisal] Country: [Bangladesh] Data accessibility Repository name: [Mendeley Data] Data identification number: 10.17632/7vb77bz2st.1 Direct URL to data: https://data.mendeley.com/datasets/7vb77bz2st/1 Related research article [None] How the dataset helps in packaging 1. [ Quality Assessment : Automates the assessment of lentil quality, ensuring only high-quality products are packaged. 2. Sorting and Grading : Aids in developing algorithms for sorting lentils based on size, color, and disease presence, enhancing efficiOpen asset ↗Mendeley Data · 10.17632/7vb77bz2st.1lines:1-55
Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Published7 Dec 2022Foods (Basel, Switzerland)Cited by 9 · OpenAlex ↗

Characterization of a Collection of Colored Lentil Genetic Resources Using a Novel Computer Vision Approach.

Del Coco M, Laddomada B, Romano G, Carcagnì P, Kumar S, Kumar S, Leo M.

LentilRGB / grayscaleSeed / grainClassificationMorphology / geometry measurementFruit / seed / panicle traits

The lentil ( Lens culinaris Medik.) is one of the major pulse crops cultivated worldwide. However, in the last decades, lentil cultivation has decreased in many areas surrounding Mediterranean countries due to low yields, new lifestyles, and changed eating habits. Thus, many landraces and local varieties have disappeared, while local farmers are the only custodians of the treasure of lentil genetic resources. Recently, the lentil has been rediscovered to meet the needs of more sustainable agriculture and food systems. Here, we proposed an image analysis approach that, besides being a rapid and non-destructive method, can characterize seed size grading and seed coat morphology. The results indicated that image analysis can give much more detailed and precise descriptions of grain size and shape characteristics than can be practically achieved by manual quality assessment. Lentil size measurements combined with seed coat descriptors and the color attributes of the grains allowed us to develop an algorithm that was able to identify 64 red lentil genotypes collected at ICARDA with an accuracy approaching 98% for seed size grading and close to 93% for the classification of seed coat morphology.

Reproduction assets foundThe paper's lentil seed image dataset (64 images of ICARDA genotypes used for the computer vision phenotyping pipeline) is explicitly stated to be publicly available on the authors' GitHub repository. The Data Availability Statement only offers other data upon request, but the image dataset itself has a public URL.
Dataset · publicpaigns, and we found that its repositioning contained a non-negligible error for the purposes of the evaluation process here described. This means that the images in some cases showed a different scaling factor that was handled by the algorithms. In Figure 1 , the acquisition setup is shown. The dataset is publicly available at https://github.com/beppe2hd/unconstrainedLentils (accessed on 10 November 2022). 2.1. Plant Materials In the present study, we analyzed the grains of 64 lentil genotypes received by ICARDA in Lebanon, including 48 varieties released in 19 different countries between 1984 and 2018, 9 germplasm accessions, and 7 elite breeding lines developed at ICARDA in Lebanon ( TablOpen asset ↗https://github.com/beppe2hd/unconstrainedLentilslines:31-42
Code / dataset availability confirmedEurope PMC · checked 8 Sept 2026
Published3 Nov 2022Frontiers in plant scienceCited by 8 · OpenAlex ↗

Climate change conditions the selection of rust-resistant candidate wild lentil populations for in situ conservation.

Civantos-Gómez I, Rubio Teso ML, Galeano J, Rubiales D, Iriondo JM, García-Algarra J.

LentilField / plotStress / disease detectionDisease symptoms / severity

Crop Wild Relatives (CWR) are a valuable source of genetic diversity that can be transferred to commercial crops, so their conservation will become a priority in the face of climate change. Bizarrely, in situ conserved CWR populations and the traits one might wish to preserve in them are themselves vulnerable to climate change. In this study, we used a quantitative machine learning predictive approach to project the resistance of CWR populations of lentils to a common disease, lentil rust, caused by fungus Uromyces viciae-fabae . Resistance is measured through a proxy quantitative value, DSr (Disease Severity relative), quite complex and expensive to get. Therefore, machine learning is a convenient tool to predict this magnitude using a well-curated georeferenced calibration set. Previous works have provided a binary outcome (resistant vs. non-resistant), but that approach is not fine enough to answer three practical questions: which variables are key to predict rust resistance, which CWR populations are resistant to rust under current environmental conditions, and which of them are likely to keep this trait under different climate change scenarios. We first predict rust resistance in present time for crop wild relatives that grow up inside protected areas. Then, we use the same models under future climate IPCC (Intergovernmental Panel on Climate Change) scenarios to predict future DSr values. Populations that are rust-resistant by now and under future conditions are optimal candidates for further evaluation and in situ conservation of this valuable trait. We have found that rust-resistance variation as a result of climate change is not uniform across the geographic scope of the study (the Mediterranean basin), and that candidate populations share some interesting common environmental conditions.

Reproduction assets foundThe paper's data availability statement points to a public Zenodo deposit containing the study's datasets (lentil CWR distribution/calibration data and DSr-related analysis data). Supplementary material is also available via the Frontiers article page, but the Zenodo deposit is the explicit, actionable paper-specific资产
Dataset · publicPublicly available datasets were analyzed in this study. This data can be found here: https://zenodo.org/record/6883274 .Open asset ↗Zenodo · 6883274lines:494-522
Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
Published13 Nov 2020Plant phenomics (Washington, D.C.)Cited by 16 · OpenAlex ↗

Generalized Linear Model with Elastic Net Regularization and Convolutional Neural Network for Evaluating Aphanomyces Root Rot Severity in Lentil.

Marzougui A, Ma Y, McGee RJ, Khot LR, Sankaran S.

LentilRGB / grayscaleRootClassificationDisease symptoms / severity

Phenomics technologies allow quantitative assessment of phenotypes across a larger number of plant genotypes compared to traditional phenotyping approaches. The utilization of such technologies has enabled the generation of multidimensional plant traits creating big datasets. However, to harness the power of phenomics technologies, more sophisticated data analysis methods are required. In this study, Aphanomyces root rot (ARR) resistance in 547 lentil accessions and lines was evaluated using Red-Green-Blue (RGB) images of roots. We created a dataset of 6,460 root images that were annotated by a plant breeder based on the disease severity. Two approaches, generalized linear model with elastic net regularization (EN) and convolutional neural network (CNN), were developed to classify disease resistance categories into three classes: resistant, partially resistant, and susceptible. The results indicated that the selected image features using EN models were able to classify three disease categories with an accuracy of up to 0.91 ± 0.004 (0.96 ± 0.005 resistant, 0.82 ± 0.009 partially resistant, and 0.92 ± 0.007 susceptible) compared to CNN with an accuracy of about 0.84 ± 0.009 (0.96 ± 0.008 resistant, 0.68 ± 0.026 partially resistant, and 0.83 ± 0.015 susceptible). The resistant class was accurately detected using both classification methods. However, partially resistant class was challenging to detect as the features (data) of the partially resistant class often overlapped with those of resistant and susceptible classes. Collectively, the findings provided insights on the use of phenomics techniques and machine learning approaches to provide quantitative measures of ARR resistance in lentil.

Reproduction assets foundThe paper's Data Availability statement points to a Zenodo deposit (DOI 10.5281/zenodo.4018168) containing the paper-specific lentil root rot image dataset (6,460 annotated RGB root images) used for the EN and CNN phenotyping analyses. No separate author analysis code URL is given; the R project URL is a generic tool,
Dataset · publicData available at: https://doi.org/10.5281/zenodo.4018168 .Open asset ↗zenodo · 10.5281/zenodo.4018168lines:121-134
Code / dataset availability confirmedCrossref · Europe PMC · checked 9 Sept 2026
Published10 Apr 2020Plant MethodsCited by 32 · OpenAlex ↗

The BELT and phenoSEED platforms: shape and colour phenotyping of seed samples.

Keith Halcro · Kaitlin McNabb · Ashley Lockinger · Didier Socquet-Juglard · Kirstin E. Bett · Scott D. Noble

LentilRGB / grayscaleSeed / grainMorphology / geometry measurementPigment / colour / senescenceFruit / seed / panicle traits

Abstract Background Quantitative and qualitative assessment of visual and morphological traits of seed is slow and imprecise with potential for bias to be introduced when gathered with handheld tools. Colour, size and shape traits can be acquired from properly calibrated seed images. New automated tools were requested to improve data acquisition efficacy with an emphasis on developing research workflows. Results A portable imaging system (BELT) supported by image acquisition and analysis software (phenoSEED) was created for small-seed optical analysis. Lentil ( Lens culinaris L.) phenotyping was used as the primary test case. Seeds were loaded into the system and all seeds in a sample were automatically individually imaged to acquire top and side views as they passed through an imaging chamber. A Python analysis script applied a colour calibration and extracted quantifiable traits of seed colour, size and shape. Extraction of lentil seed coat patterning was implemented to further describe the seed coat. The use of this device was forecasted to eliminate operator biases, increase the rate of acquisition of traits, and capture qualitative information about traits that have been historically analyzed by eye. Conclusions Increased precision and higher rates of data acquisition compared to traditional techniques will help to extract larger datasets and explore more research questions. The system presented is available as an open-source project for academic and non-commercial use.

Reproduction assets foundThe paper explicitly states that the phenoSEED analysis script is publicly available on GitLab and that the BELT-captured seed image datasets are available on the first author's Figshare page. Both are paper-specific, public, and actionable.
Code · publicAt the time of publication, a version of the processing script is available at https://gitlab.com/usask-speclab/phenoseed .Open asset ↗usask-speclab/phenoseedlines:145-153
Dataset · publicThe image datasets captured by BELT analysed for the sample study are available from https://figshare.com/authors/Keith_Halcro/8363580 . The phenoSEED script is available from https://gitlab.com/usask-speclab/phenoseed .Open asset ↗lines:173-192
Code / dataset availability confirmedEurope PMC · bioRxiv · checked 15 Sept 2026
Published31 Oct 2019bioRxivCited by 1 · OpenAlex ↗

The BELT and phenoSEED platforms: shape andcolour phenotyping of seed samples

Halcro K, McNabb K, Lockinger A, Socquet-Juglard D, Bett KE, Noble SD.

LentilRGB / grayscaleSeed / grainMorphology / geometry measurementCalibration / preprocessingPigment / colour / senescenceFruit / seed / panicle traits

Background Seed analysis is currently a bottleneck in phenotypic analysis of seeds. Measurements are slow and imprecise with potential for bias to be introduced when gathered manually. New acquisition tools were requested to improve phenotyping efficacy with an emphasis on obtaining colour information. Results A portable imaging system (BELT) supported by image acquisition and analysis software (phenoSEED) was created for small-seed optical analysis. Lentil ( Lens culinaris L.) phenotyping was used as the primary test case. Seeds were loaded into the system and all seeds in a sample were automatically and individually imaged to acquire top and side views as they passed through an imaging chamber. A Python analysis script applied a colour calibration and extracted quantifiable traits of seed colour, size and shape. Extraction of lentil seed coat patterning was implemented to further describe the seed coat. The use of this device was forecasted to eliminate operator biases, increase the rate of acquisition of traits, and capture qualitative information about traits that have been historically analyzed by eye. Conclusions Increased precision and higher rates of data acquisition compared to traditional techniques will help breeders to develop more productive cultivars. The system presented is available as an open-source project for academic and non-commercial use.

Reproduction assets foundThe paper's phenoSEED image-analysis script (used for seed shape, size, colour, and clustering phenotyping) is explicitly stated to be publicly available on the authors' GitLab repository. The BELT image datasets themselves are only available on request, so they do not qualify as public assets.
Code · publicde and hard- ware plans available for academic and non-commercial use. It is hoped that this will support the collection of more easily cross-comparable data and encourage other research groups to contribute to further de- velopment of the project. At the time of publica- tion, a version of the processing script is available at https://gitlab.com/usask-speclab/phenoseed. Further information on a comprehensive hardware and soft- ware bundle will be made available as it is packaged for distribution. Methods BELT System Design and Description BELT (Figure 1) was designed around a 150 mm wide conveyor with at white, low-gloss belt (Mini-Mover Conveyors, Volcano CA) mounted on an audio-visual carOpen asset ↗usask-speclab/phenoseedpdf-raw-page:8 lines:1-99
Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Published9 Oct 2017Frontiers in plant scienceCited by 20 · OpenAlex ↗

A Biosensor-Based Leaf Punch Assay for Glutamine Correlates to Symbiotic Nitrogen Fixation Measurements in Legumes to Permit Rapid Screening of Rhizobia Inoculants under Controlled Conditions.

Thilakarathna MS, Moroz N, Raizada MN.

CowpeaLentilSoybeanGrowth chamberLeafPhysiological trait estimation

Legumes are protein sources for billions of humans and livestock. These traits are enabled by symbiotic nitrogen fixation (SNF), whereby root nodule-inhabiting rhizobia bacteria convert atmospheric nitrogen (N) into usable N. Unfortunately, SNF rates in legume crops suffer from undiagnosed incompatible/suboptimal interactions between crop varieties and rhizobia strains. There are opportunities to test much large numbers of rhizobia strains if cost/labor-effective diagnostic tests become available which may especially benefit researchers in developing countries. Inside root nodules, fixed N from rhizobia is assimilated into amino acids including glutamine (Gln) for export to shoots as the major fraction (amide-exporting legumes) or as the minor fraction (ureide-exporting legumes). Here, we have developed a new leaf punch based technique to screen rhizobia inoculants for SNF activity following inoculation of both amide exporting and ureide exporting legumes. The assay is based on measuring Gln output using the GlnLux biosensor, which consists of Escherichia coli cells auxotrophic for Gln and expressing a constitutive lux operon. Subsistence farmer varieties of an amide exporter (lentil) and two ureide exporters (cowpea and soybean) were inoculated with different strains of rhizobia under controlled conditions, then extracts of single leaf punches were incubated with GlnLux cells, and light-output was measured using a 96-well luminometer. In the absence of external N and under controlled conditions, the results from the leaf punch assay correlated with 15 N-based measurements, shoot N percentage, and shoot total fixed N in all three crops. The technology is rapid, inexpensive, high-throughput, requires minimum technical expertise and very little tissue, and hence is relatively non-destructive. We compared and contrasted the benefits and limitations of this novel diagnostic assay to methods.

Reproduction assets foundThe article's Supplementary Material (Figures S1–S4) contains the paper's own phenotyping measurements (GlnLux luminometer outputs, Gln standard curves, nodulation counts, root/shoot morphometric data, and root images) and is publicly available at the Frontiers supplementary-material URL stated in the text. No author分析
Supplement · publicewan) and Alice Chang (University of British Columbia) for 15 N analysis. Footnotes Funding. This research was supported by CIFSRF grant 107791 to MR from the International Food Development Centre (IDRC, Ottawa) and Global Affairs Canada. Supplementary Material The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fpls.2017.01714/full#supplementary-material FIGURE S1 Luminescence measurement of Gln standards using the GlnLux 96-well luminometer bioassay to demonstrate the linearity of the assay. Luminescence was measured using a concentration gradient of pure Gln standards (0, 125 × 10 -8 , 25 × 10 -7 , 5 × 10 -6 , and 1 × 10 -5 M) uOpen asset ↗lines:155-178