Significance: Roots produce hundreds to thousands of small molecules with unknown functions. We targeted the apocarotenoid pathway, which has been linked to numerous developmental processes in Arabidopsis , for a sensitized chemical genetic screen to identify regulators of root development. β-Cyclocitral, a small molecule derived from β-carotene, was identified as a regulator of root stem cell behavior in Arabidopsis as well as in rice and tomato. β-Cyclocitral promotes root stem cell divisions to enhance root growth and branching. In rice, β-cyclocitral enhanced both root and shoot growth during salt stress, which has important implications for agriculture.

Arabidopsis is anchored by a carotenoid-derived metabolite.

Verticillium wilt, caused by the soil-borne fungus Verticillium dahliae, is one of the most devastating diseases in cotton (Gossypium spp.). Lignin in the cell wall forms a physical barrier to inhibit pathogen invasion, and defense-induced lignification reinforces secondary cell wall to prevent pathogens from further spreading. Cinnamyl alcohol dehydrogenases (CADs) catalyze the production of three main monolignols, p-coumaryl- (H), coniferyl- (G), and sinapyl-alcohols (S), which are the fundamental blocks of lignin. Here, we identified CAD genes in G. hirsutum, analyzed their expression profiles in cotton leaf, stem, and root from different developmental stages, and selected GhCAD35, GhCAD45, and GhCAD43, which were consistently induced by V. dahliae inoculation in G. hirsutum cultivars resistant or susceptible to V. dahliae. On the basis of confirmation of the in vitro enzymatic activity of the three proteins in generation of the three monolignols, we used virus-induced gene silencing (VIGS) to investigate the effects of silencing of GhCAD35, GhCAD45, or GhCAD43 on resistance to V. dahliae as well as on deposition and the composition of lignin. Silencing each of the three CADs impaired the defense-induced lignification and salicylic acid biosynthesis in stem, and compromised resistance to V. dahliae. Moreover, our study showed that silencing the three GhCADs severely affected the biosynthesis of S-lignin, leading to a decrease of the syringyl/guaiacyl (S/G) ratio. Heterogeneous overexpression of GhCAD35, GhCAD45, or GhCAD43 in Arabidopsis enhanced disease resistance. Taken together, our study demonstrates a role of the three GhCADs in defense-induced lignin biosynthesis and resistance to V. dahliae in G. hirsutum.

Roots are important plant organs for the uptake of water and nutrient elements. Plant root development is finely regulated by endogenous signals and environmental cues, which shapes the root system architecture to optimize the plant growth and adapt to the rhizospheric environments. Carotenoids are precursors of plant hormones strigolactones (SLs) and ABA, as well as multiple bioactive molecules. Numerous studies have demonstrated SLs and ABA as essential regulators of plant root growth and development. In addition, a lot carotenoid-derived bioactive metabolites are recently identified as plant root growth regulators, such as anchorene, β-cyclocitral, retinal and zaxinone. However, our knowledge on how these metabolites affect the root architecture to cope with various stressors and how they interact with each other during these processes is still quite limited. In the present review, we will briefly introduce the biosynthesis of carotenoid-derived root regulators and elaborate their biological functions on root development and architecture, focusing on their contribution to the rhizospheric environmental adaption of plants.

Summary: 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.