Evaluation of 3D seed structure and cellular traits in-situ using X-ray microscopy

Abstract Phenotyping methods for seed morphology are mostly limited to two-dimensional imaging or manual measures. In this study, we present a novel seed phenotyping approach utilizing lab-based X-ray microscopy (XRM) to characterize 3D seed morphology, internal structures, and cellular analysis fro...

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Main Authors: Marcus Griffiths, Barsanti Gautam, Clara Lebow, Keith Duncan, Xinxin Ding, Pubudu Handakumbura, John C. Sedbrook, Christopher N. Topp
Format: Article
Language:English
Published: Nature Portfolio 2025-02-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-025-88482-7
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author Marcus Griffiths
Barsanti Gautam
Clara Lebow
Keith Duncan
Xinxin Ding
Pubudu Handakumbura
John C. Sedbrook
Christopher N. Topp
author_facet Marcus Griffiths
Barsanti Gautam
Clara Lebow
Keith Duncan
Xinxin Ding
Pubudu Handakumbura
John C. Sedbrook
Christopher N. Topp
author_sort Marcus Griffiths
collection DOAJ
description Abstract Phenotyping methods for seed morphology are mostly limited to two-dimensional imaging or manual measures. In this study, we present a novel seed phenotyping approach utilizing lab-based X-ray microscopy (XRM) to characterize 3D seed morphology, internal structures, and cellular analysis from a single scan. Seeds of pennycress (Thlaspi arvense L.) an oilseed cover crop, were scanned and segmented using a machine learning model. Seed morphological analysis and a coat thickness map was applied to compare seed volumes of four genotypes. Notably, the 3D seed volume measurement alone was not enough to reveal differences in seed coat between the genotypes. Applying a seed coat thickness map revealed that the Large-Golden genotype had a thinner seed coat compared to wildtype despite a greater seed coat volume. Cellular analysis showed that cotyledon cell size was a driving factor for larger seeds. XRM was compared to traditional seed morphology traits and positive correlations were observed. Between Large-Golden and wildtype, differences could only be resolved by XRM highlighting the limitations of 2D seed area measures. These results demonstrate that XRM can provide quantitative measures extracted from seeds for enhancing our understanding of seed structure, development, and facilitate breeding efforts for enhanced seed quality and crop performance.
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issn 2045-2322
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spelling doaj-art-6d59fc3498f442f98129a4e084330b492025-02-09T12:30:41ZengNature PortfolioScientific Reports2045-23222025-02-0115111110.1038/s41598-025-88482-7Evaluation of 3D seed structure and cellular traits in-situ using X-ray microscopyMarcus Griffiths0Barsanti Gautam1Clara Lebow2Keith Duncan3Xinxin Ding4Pubudu Handakumbura5John C. Sedbrook6Christopher N. Topp7Donald Danforth Plant Science CenterSchool of Biological Sciences, Illinois State UniversityDonald Danforth Plant Science CenterDonald Danforth Plant Science CenterPacific Northwest National LaboratoryPacific Northwest National LaboratorySchool of Biological Sciences, Illinois State UniversityDonald Danforth Plant Science CenterAbstract Phenotyping methods for seed morphology are mostly limited to two-dimensional imaging or manual measures. In this study, we present a novel seed phenotyping approach utilizing lab-based X-ray microscopy (XRM) to characterize 3D seed morphology, internal structures, and cellular analysis from a single scan. Seeds of pennycress (Thlaspi arvense L.) an oilseed cover crop, were scanned and segmented using a machine learning model. Seed morphological analysis and a coat thickness map was applied to compare seed volumes of four genotypes. Notably, the 3D seed volume measurement alone was not enough to reveal differences in seed coat between the genotypes. Applying a seed coat thickness map revealed that the Large-Golden genotype had a thinner seed coat compared to wildtype despite a greater seed coat volume. Cellular analysis showed that cotyledon cell size was a driving factor for larger seeds. XRM was compared to traditional seed morphology traits and positive correlations were observed. Between Large-Golden and wildtype, differences could only be resolved by XRM highlighting the limitations of 2D seed area measures. These results demonstrate that XRM can provide quantitative measures extracted from seeds for enhancing our understanding of seed structure, development, and facilitate breeding efforts for enhanced seed quality and crop performance.https://doi.org/10.1038/s41598-025-88482-7
spellingShingle Marcus Griffiths
Barsanti Gautam
Clara Lebow
Keith Duncan
Xinxin Ding
Pubudu Handakumbura
John C. Sedbrook
Christopher N. Topp
Evaluation of 3D seed structure and cellular traits in-situ using X-ray microscopy
Scientific Reports
title Evaluation of 3D seed structure and cellular traits in-situ using X-ray microscopy
title_full Evaluation of 3D seed structure and cellular traits in-situ using X-ray microscopy
title_fullStr Evaluation of 3D seed structure and cellular traits in-situ using X-ray microscopy
title_full_unstemmed Evaluation of 3D seed structure and cellular traits in-situ using X-ray microscopy
title_short Evaluation of 3D seed structure and cellular traits in-situ using X-ray microscopy
title_sort evaluation of 3d seed structure and cellular traits in situ using x ray microscopy
url https://doi.org/10.1038/s41598-025-88482-7
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