SummaryPlants and certain bacteria use a non-mevalonate alternative route for the biosynthesis of many isoprenoids, including carotenoids. This route has been discovered only recently and has been designated the deoxyxylulose phosphate pathway or methylerythritol phosphate (MEP) pathway. We report here that colonisation of roots from wheat, maize, rice and barley by the arbuscular mycorrhizal fungal symbiont Glomus intraradices involves strong induction of transcript levels of two of the pivotal enzymes of the MEP pathway, 1-deoxy-D-xylulose 5-phosphate synthase (DXS) and 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR). This induction is temporarily and spatially correlated with speci®c and concomitant accumulation of two classes of apocarotenoids, namely glycosylated C 13 cyclohexenone derivatives and mycorradicin (C 14 ) conjugates, the latter being a major component of the long-knowǹ yellow pigment'. A total of six cyclohexenone derivatives were characterised from mycorrhizal wheat and maize roots. Furthermore, the acyclic structure of mycorradicin described previously only from maize has been identi®ed from mycorrhizal wheat roots after alkaline treatment of an`apocarotenoid complex' of yellow root constituents. We propose a hypothetical scheme for biogenesis of both types of apocarotenoids from a common oxocarotenoid (xanthophyll) precursor. This is the ®rst report demonstrating (i) that the plastidic MEP pathway is active in plant roots and (ii) that it can be induced by a fungus.

We investigated the accumulation of reactive oxygen species (ROS) in arbuscular mycorrhizal (AM) roots from Medicago truncatula, Zea mays and Nicotiana tabacum using three independent staining techniques. Colonized root cortical cells and the symbiotic fungal partner were observed to be involved in the production of ROS. Extraradical hyphae and spores from Glomus intraradices accumulated small levels of ROS within their cell wall and produced ROS within the cytoplasm in response to stress. Within AM roots, we observed a certain correlation of arbuscular senescence and H2O2 accumulation after staining by diaminobenzidine (DAB) and a more general accumulation of ROS close to fungal structures when using dihydrorhodamine 123 (DHR 123) for staining. According to electron microscopical analysis of AM roots from Z. mays after staining by CeCl3, intracellular accumulation of H2O2 was observed in the plant cytoplasm close to intact and collapsing fungal structures, whereas intercellular H2O2 was located on the surface of fungal hyphae. These characteristics of ROS accumulation in AM roots suggest similarities to ROS accumulation during the senescence of legume root nodules.

Contents Summary 22 Introduction 23  Mycorradicin 23  Cyclohexenone derivatives 24  Occurrence of apocarotenoids 24 Biosynthesis of apocarotenoids 25 Root plastids 27 Significance of carotenoid metabolism 29 Acknowledgements 32 References  32 Summary Plant root‐colonizing arbuscular mycorrhizal (AM) fungi activate the methylerythritol phosphate pathway, carotenoid biosynthesis and oxidative carotenoid cleavage in roots, leading to C13 and C14 apocarotenoids, that is, cyclohexenone and mycorradicin derivatives. Mycorradicin causes the characteristic yellow coloration of many AM roots accumulating within a complex mixture of unknown components. The accumulating C13 cyclohexenones exhibit various ring substitutions and different glycosyl moieties. Transcript levels of the first two enzymes of the MEP pathway, 1‐deoxy‐d‐xylulose 5‐phosphate synthase and 1‐deoxy‐d‐xylulose 5‐phosphate reductoisomerase, and of the carotenoid pathway, phytoene desaturase and ζ‐carotene desaturase, along with a carotenoid‐cleaving dioxygenase, are markedly increased in AM roots. This correlates with proliferation and reorganization of root plastids. These results allow at this point only speculation about the significance of apocarotenoid accumulation: participation in the production of signaling molecules and control of fungal colonization or protection against soil‐borne pathogens; protection of root cells against oxidative damage of membranes by reactive oxygen species; and promotion of the symbiotic interactions between plant roots and AM fungi.

Colonization of root cortical cells by arbuscular mycorrhizal fungi leads to marked cytological changes of plastids and mitochondria. Plastids in particular are forming tubular extensions partially connecting individual organelles in a network-like way. These cytological changes correspond to an increased need for plastid and mitochondrial products during establishment and functioning of the symbiosis. The analysis of metabolite and transcript levels in mycorrhizal and nonmycorrhizal roots from Medicago truncatula revealed concomitant changes regarding a number of metabolic pathways. Our results indicate the activation of the mitochondrial tricarboxylic acid cycle and of plastid biosynthetic pathways producing fatty acids, amino acids, and apocarotenoids. These observations provide a general overview of structural and metabolic changes of plastids and mitochondria during colonization of root cortical cells by arbuscular mycorrhizal fungi.