Publication details

PAT: An Image Analysis Tool for Automated Scoring of Pollen in Alexander-Stained Anthers.

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Authors

VOLKAVA Darya ŘÍHA Karel RAXWAL Vivek Kumar

Year of publication 2026
Type Peer-reviewed scientific article
Magazine / Source Journal of Experimental Botany
MU Faculty or unit

Central European Institute of Technology

Citation
web https://academic.oup.com/jxb/advance-article/doi/10.1093/jxb/erag436/8785351
Doi https://doi.org/10.1093/jxb/erag436
Keywords Alexander staining; CAP-D2; Cellpose; Condensin; Deep learning; Meiotic exit; Pollen viability; SMG7
Attached files
Description Quantitative pollen viability analysis is a critical but labor-intensive step in plant reproductive biology. Existing deep-learning Segment Anything Models (SAM) fail to reliably segment viable pollen in Alexander-stained anthers. To address this, we fine-tuned an existing Cellpose-SAM model for pollen segmentation. We integrated it into PAT (Pollen Analysis Tool), a cross-platform desktop application. PAT features instance segmentation with interactive quality control, an in-app model retraining module, and publication-ready statistical outputs. We deployed PAT in an EMS suppressor screen of semi-sterile Arabidopsis smg7-6 mutants, enabling efficient candidate prioritization for whole-genome sequencing and mapping of the candidate mutation. This screen led to the identification of a point mutation in CAP-D2 (capd2-2), a Condensin I subunit, that rescues the smg7-6 meiotic phenotype. Notably, mutation in a Condensin II subunits (CAP-D3 and CAP-H2) does not confer rescue. Further characterization suggests the capd2-2 allele is hypomorphic, showing no defects in vegetative growth, chromocenter compaction, or transposable element silencing. Collectively, we demonstrate that accessible AI tools have the potential to bridge gaps in plant phenotyping and accelerate the pace of biological discovery.
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