Oxidative cleavage of carotenoids leads to dialdehydes (diapocarotenoids, DIALs) in addition to the widely known apocarotenoids. DIALs are biologically active compounds that presumably impact human health and play different roles in plant development and carotenoid metabolism. However, detection of DIALs in plants is challenging due to their instability, low abundance, and poor ionization efficiency in mass spectrometry. Here, we developed a solid-phase extraction and derivatization protocol coupled with ultrahigh performance liquid chromatography−mass spectrometry for quantitative profiling of DIALs. Our method significantly enhances the sensitivity of DIAL detection with a detection limit of 0.05 pg/mg of dried food materials, allowing unambiguous profiling of 30 endogenous DIALs with C
5
to C
24
from vegetables. Our work provides a new and efficient approach for determining the content of DIALs from various complex matrices, paving the way for uncovering the functions of DIALs in human health and plant growth and development.
Carotenoid-derived regulatory metabolites and hormones are generally known to arise through the oxidative cleavage of a single double bond in the carotenoid backbone, which yields mono-carbonyl products called apocarotenoids. However, the extended conjugated double bond system of these pigments predestines them also to repeated cleavage forming dialdehyde products, diapocarotenoids, which have been less investigated due to their instability and low abundance. Recently, we reported on the short diapocarotenoid anchorene as an endogenous Arabidopsis metabolite and specific signaling molecule that promotes anchor root formation. In this work, we investigated the biological activity of a synthetic isomer of anchorene, iso-anchorene, which can be derived from repeated carotenoid cleavage. We show that iso-anchorene is a growth inhibitor that specifically inhibits primary root growth by reducing cell division rates in the root apical meristem. Using auxin efflux transporter marker lines, we also show that the effect of iso-anchorene on primary root growth involves the modulation of auxin homeostasis. Moreover, by using liquid chromatography-mass spectrometry analysis, we demonstrate that iso-anchorene is a natural Arabidopsis metabolite. Chemical inhibition of carotenoid biosynthesis led to a significant decrease in the iso-anchorene level, indicating that it originates from this metabolic pathway. Taken together, our results reveal a novel carotenoid-derived regulatory metabolite with a specific biological function that affects root growth, manifesting the biological importance of diapocarotenoids.
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