A Comparative Study on the Identification of Xanthium sibiricum Patrin ex Widder and Xanthium italicum Moretti Based on Three Microscopy Technology.

Abstract

Xanthium sibiricum Patrin ex Widder and Xanthium italicum Moretti are morphologically similar fructus that are frequently misidentified. Xanthium italicum Moretti may possess inherent toxicity, and its adulteration of genuine medicinal materials poses a threat to clinical drug safety. Macroscopic observation and three microscopic techniques including stereo microscope, optical microscope, and 3D X-ray microscope were used for morphological identification of Xanthium sibiricum Patr ex Widder and Xanthium italicum Moretti in this study. 3D X-ray microscopy was applied as a novel tool for non-destructive, high-resolution discrimination of the two taxa. Intact fructus (n = 30 per species) were first screened macroscopically, then examined by stereo microscopy, optical microscopy, and 3D X-ray microscopy (0.3, 0.7, 1.5, 3.5, 18.06, 20.01 μm voxel size, Zeiss Xradia 520 Versa). The results showed that stereo microscopy, optical microscopy, and 3D X-ray microscopy collectively confirm the same conclusion from three distinct physical perspectives: surface topography, internal two-dimensional structure, and internal three-dimensional density distribution. The two Xanthium species differ significantly in burr spine morphology, fructus size and shape, the architecture and distribution of non-glandular and glandular trichomes, cotyledon conformation, and seed-coat cell patterning. In particular, 3D X-ray microscopy clearly resolves internal cotyledon spatial configurations and involucral cavity architectures, which furnishes critical endomorphic characters for taxonomic diagnosis. 3D X-ray microscopy provides unprecedented volumetric contrast of surface spines and internal seed architecture, permitting confident, non-destructive species identification. This study provides a basis for the safe clinical use of Xanthium sibiricum Patrin ex Widder. The frontier of 3D X-ray microscopy in plant systematics offers a novel, rapid, accurate and non-destructive protocol for the discrimination of morphologically elusive species.

Plant phenotyping relevance

3D X線顕微鏡を含む複数の画像計測法を用いて果実・種子の形態形質を抽出し、近縁2分類群の非破壊識別プロトコルとして比較・検証しており、植物フェノタイピング手法が中心である。

Code and data availability

The paper's phenotyping data (microscopy images, 3D X-ray scans, measurements) are not publicly deposited; the Data Availability Statement says data are available only on request from the corresponding author. Supporting Information contains only sample metadata and scanning parameters, not the underlying image/phenome

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