# Overcoming difficulties in segmentation of hyperspectral plant images with small projection areas using machine learning

**Authors:** Eva Neuwirthová, Jiří Chuchlík, Miroslav Pikl, Zuzana Lhotáková, Ivan Kashkan, Klára Panzarová, Jan Stejskal, Jana Albrechtová, Milan Lstibůrek, Jaroslav Čepl

PMC · DOI: 10.1038/s41598-025-31952-9 · Scientific Reports · 2026-01-30

## TL;DR

This study uses hyperspectral imaging and machine learning to segment and evaluate conifer seedlings, enabling non-destructive physiological assessment.

## Contribution

A novel hyperspectral image processing pipeline using K-means and random forest models for conifer seedling segmentation and classification.

## Key findings

- K-means algorithm derived 23 hyperspectral centroids classified into ten biologically distinct groups.
- Random forest model effectively differentiated Scots pine seedlings based on origin during stress and recovery.
- Hyperspectral imaging and machine learning show promise for non-destructive evaluation of conifer physiology.

## Abstract

Segmentation of hyperspectral image data is a well-established technique in remote sensing. While it is commonly applied to individual field crops, its use for individual trees is less prevalent. Conifers are crucial in forestry, and assessing physiological status, or genetic diversity is required for effective early-age treatment in nurseries and hyperspectral imaging (HSI) combined with high-throughput phenotyping (HTP) offers faster and non-destructive evaluation. NDVI-based thresholding is sufficient for detection of leaves with large projection areas, but needles of conifers present challenges due to spatial resolution constraints and increased proportion of border pixels. This study monitored the offspring of three locally adapted Scots pine (Pinus sylvestris L.) populations, representing distinct upland and lowland ecotypes. This study presents a hyperspectral image processing pipeline for segmenting and isolating individual Scots pine seedlings. Using a K-means algorithm, 23 hyperspectral centroids were successfully derived and subsequently classified into ten biologically distinct groups. Random forest classification model effectively differentiated Scots pine seedlings based on origin during water stress and recovery periods. This study highlights the potential of hyperspectral imaging and machine learning in evaluating the physiological state of conifer seedlings, demonstrating promising applications in forest tree physiology research and tree breeding.

## Full-text entities

- **Diseases:** HSI (MESH:C564543), drought (MESH:C536747), dehydration (MESH:D003681), SRWC (MESH:D005242)
- **Chemicals:** QL24010275 (-), chlorophyll (MESH:D002734), Water (MESH:D014867)
- **Species:** Pinus subgen. Pinus (diploxylon pines, subgenus) [taxon 139271], Arabidopsis thaliana (mouse-ear cress, species) [taxon 3702], Fagus sylvatica (European beech, species) [taxon 28930], Pinus taeda (loblolly pine, species) [taxon 3352], Conifers [taxon 3312], Picea abies (Norway spruce, species) [taxon 3329], Sorghum bicolor (broomcorn, species) [taxon 4558], Pinus sylvestris (Scotch pine, species) [taxon 3349]

## Full text

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## Figures

2 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12864965/full.md

## References

5 references — full list in the complete paper: https://tomesphere.com/paper/PMC12864965/full.md

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Source: https://tomesphere.com/paper/PMC12864965