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Association of leaf spectral variation with functional genetic variants

bioRxiv · 25 Sept 2025 · 10.1101/2023.10.03.560760

Abstract

The application of in-field and aerial spectroscopy to assess functional and phylogenetic variation in plants has led to novel ecological insights and supports global assessments of plant biodiversity. Understanding how plant genetic variation influences reflectance spectra will help harness this potential for biodiversity monitoring and improve understanding of why plants differ in functional responses to environmental change. Here, we use a well-resolved genetic mapping population derived from Multi-parent Advanced Generation Inter-cross (MAGIC) lines of Nicotiana attenuata to associate genetic differences with differences in leaf spectra between plants in a field experiment in their natural environment. We analyzed the leaf reflectance spectra using a hand-held spectroradiometer (350-2500 nm) on 616 fully genotyped plants of N. attenuata grown in a randomized block design. We tested three approaches to conducting Genome-Wide Association Studies on spectral variants. We introduce a new Hierarchical Spectral Clustering with Parallel Analysis (HSC-PA) method. This method efficiently captured the variation in our high-dimensional dataset and allowed us to discover a novel association, between a locus on chromosome 1 and the 734-1143 nm spectral range, spanning the red-edge and near-infrared regions that are sensitive to leaf structure and photosynthetic activity. This locus contains a candidate gene annotated as carbonic anhydrase, an enzyme involved in CO2 hydration and regulation of photosynthetic efficiency, suggesting a physiological link between variation in leaf optical properties and carbon assimilation. In contrast, an approach treating single wavelengths as phenotypes identified the same associations as HSC-PA, but without the statistical power to pinpoint significant associations. An index-based approach, which reduces complex spectra to a few dimensionless variables, detected two significant associations for ARDSI_Cw (a water-content-related index) with loci on chromosome 1 near genes annotated as a Zeta toxin domain-containing protein, and an Exocyst subunit Exo70 family protein. While these findings are biologically plausible, they represent a very narrow subset of the spectral variation captured by HSC-PA. The HSC-PA approach supports a comprehensive understanding of the genetic determinants of leaf spectral variation which is data-driven but human-interpretable, and lays a robust foundation for future research in linking plant genetics with biodiversity monitoring, large-scale ecological assessment and remote-sensing applications.

Plant phenotyping relevance

葉の反射スペクトルを植物表現型として取得し、高次元スペクトルを解析する新規HSC-PA手法を導入・評価しており、植物フェノタイピング手法が研究の中心的な技術的貢献である。

abstractWe analyzed the leaf reflectance spectra using a hand-held spectroradiometer (350-2500 nm) on 616 fully genotyped plants of N. attenuata
abstractWe introduce a new Hierarchical Spectral Clustering with Parallel Analysis (HSC-PA) method.
abstractThe HSC-PA approach supports a comprehensive understanding of the genetic determinants of leaf spectral variation

Code and data availability

The supplied blocks describe leaf spectral phenotyping (FieldSpec 4, 350–2500 nm) of 616 Nicotiana attenuata MAGIC plants and HSC-PA-GWAS analysis, but contain no data or code availability statement, no public deposit of spectra or genotypes, and no author code URL. The only URL mentioned (N. attenuata Data Hub) is a引用

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