Scale-dependent variation among destructive and non-destructive chlorophyll estimation methods across crop species

Abstract

Abstract Chlorophyll estimation is fundamental in plant physiology, crop management, and ecological studies; however, destructive and non-destructive methods are often interpreted interchangeably despite differing measurement principles. The present study compared four chlorophyll estimation approaches—two non-destructive (SPAD meter and GreenSeeker) and two destructive (80% acetone and DMSO extraction)—across eight crop species under uniform field conditions. Significant interspecific variation was observed for all methods. Correlation and regression analyses revealed generally weak relationships among methods, particularly between leaf-level (SPAD, solvent extraction) and canopy-level (GreenSeeker) measurements, reflecting scale-dependent behavior and methodological differences. Moderate associations were observed between SPAD and acetone-extracted chlorophyll for certain traits, whereas GreenSeeker showed poor agreement with solvent-based estimates. Differences between DMSO and acetone extraction further highlighted solvent-specific extraction efficiency. The results demonstrate that chlorophyll estimation methods are not directly interchangeable and should be selected based on study objectives, biological scale, and leaf anatomical characteristics. Species-specific calibration and integration of canopy structural parameters are required to improve cross-method interpretability.

Plant phenotyping relevance

複数の葉・キャノピーのクロロフィル推定法を作物種間で比較し、相関、回帰、スケール依存性、互換性を評価しており、植物表現型測定法の技術的検証が中心である。

Code and data availability

The preprint declares that the datasets generated in this chlorophyll-method comparison study (SPAD, GreenSeeker, acetone and DMSO measurements across eight crop species) are publicly deposited in Figshare under DOI 10.6084/m9.figshare.31817989. This is a paper-specific, publicly actionable phenotype dataset. No author