orobanchol and 5-deoxystrigol

CYP722C from cotton, a homolog of the enzyme involved in orobanchol synthesis in cowpea and tomato, catalyzes the conversion of carlactonoic acid to 5-deoxystrigol. Strigolactones (SLs) are important phytohormones with roles in the regulation of plant growth and development. These compounds also function as signaling molecules in the rhizosphere by interacting with beneficial arbuscular mycorrhizal fungi and harmful root parasitic plants. Canonical SLs, such as 5-deoxystrigol (5DS), consist of a tricyclic lactone ring (ABC-ring) connected to a methylbutenolide (D-ring). Although it is known that 5DS biosynthesis begins with carlactonoic acid (CLA) derived from β-carotene, the enzyme that catalyzes the conversion of CLA remains elusive. Recently, we identified cytochrome P450 (CYP) CYP722C as the enzyme that catalyzes direct conversion of CLA to orobanchol in cowpea and tomato (Wakabayashi et al., Sci Adv 5:eaax9067, 2019). Orobanchol has a different C-ring configuration from that of 5DS. The present study aimed to characterize the homologous gene, designated GaCYP722C, from cotton (Gossypium arboreum) to examine whether this gene is involved in 5DS biosynthesis. Expression of GaCYP722C was upregulated under phosphate starvation, which is an SL-producing condition. Recombinant GaCYP722C was expressed in a baculovirus-insect cell expression system and was found to catalyze the conversion of CLA to 5DS but not to 4-deoxyorobanchol. These results strongly suggest that GaCYP722C from cotton is a 5DS synthase and that CYP722C is the crucial CYP subfamily involved in the generation of canonical SLs, irrespective of the different C-ring configurations.

Topics: beta Carotene; Cytochrome P-450 Enzyme System; Gossypium; Heterocyclic Compounds, 3-Ring; Lactones; Mass Spectrometry; Phosphates; Plant Growth Regulators; Plant Proteins

Significant losses in sorghum biomass and grain yield occur in sub-Saharan Africa owing to infection by the root-parasitic weed Striga hermonthica (Del.) Benth. One strategy to avoid these losses is to adopt resistant crop varieties. For further delineation of the role of germination stimulants in resistance, we conducted a field experiment employing six sorghum genotypes, in eastern Sudan, and in parallel analysed the strigolactone levels in the root exudates of these genotypes under controlled conditions in Wageningen.. The root exudates of these genotypes displayed large differences in strigolactone composition and Striga-germination-inducing activity. Korokollow, Fakimustahi and Wadfahel exuded the highest amounts of 5-deoxystrigol. Fakimustahi was by far the highest sorgomol producer, and Wadbaco and SRN39 produced the highest amount of orobanchol. The concentration of 5-deoxystrigol in the root exudate showed a significant positive correlation with in vitro Striga germination and was positively associated with Striga infection in the field experiments, whereas orobanchol was negatively associated with Striga infection in the field experiments.. For the first time a close association is reported between strigolactone levels analysed under laboratory conditions and Striga infection in the field in sorghum genotypes. These genotypes may be used for further study of this resistance mechanism and for the introgression of the low germination trait in other sorghum varieties to breed for a strigolactone composition with low stimulant activity. The use of such improved varieties in combination with other Striga management tools could possibly alleviate the current Striga problem on the African continent. © 2016 Society of Chemical Industry.

Topics: Antibiosis; Lactones; Netherlands; Plant Growth Regulators; Plant Roots; Plant Weeds; Sorghum; Striga; Sudan

A major strigolactone produced by the model legume Medicago truncatula (barrel medic) has been tentatively identified as a didehydro-orobanchol isomer. In this study, a putative didehydro-orobanchol isomer was isolated from root exudates collected from barrel medic grown hydroponically under phosphate-starved conditions. The structure and absolute configurations of this strigolactone, named medicaol, were determined by a combination of spectroscopic analysis and spectral comparison with 4-deoxymedicaol which was synthesized using solvolysis and rearrangement of hydroxymethylhexahydroindenone to tetrahydroazulenone as a key step. Medicaol has a seven-membered cycloheptadiene in the A ring instead of a typical six-membered cyclohexene. Medicaol and 4-deoxymedicaol showed activity comparable to their corresponding six-membered A ring relatives, orobanchol and 4-deoxyorobanchol (ent-2'-epi-5-deoxystrigol), in inducing hyphal branching of germinating spores of an arbuscular mycorrhizal fungus Gigaspora margarita. Plausible biosynthetic pathways from 4-deoxyorobanchol to medicaol are also proposed.

Topics: Fabaceae; Glomeromycota; Heterocyclic Compounds, 4 or More Rings; Lactones; Medicago truncatula; Mycorrhizae; Phosphates; Plant Growth Regulators; Plant Roots; Symbiosis

Strigolactones (SLs) are a class of phytohormones and rhizosphere signaling compounds with high structural diversity. Three enzymes, carotenoid isomerase DWARF27 and carotenoid cleavage dioxygenases CCD7 and CCD8, were previously shown to convert all-trans-β-carotene to carlactone (CL), the SL precursor. However, how CL is metabolized to SLs has remained elusive. Here, by reconstituting the SL biosynthetic pathway in Nicotiana benthamiana, we show that a rice homolog of Arabidopsis More Axillary Growth 1 (MAX1), encodes a cytochrome P450 CYP711 subfamily member that acts as a CL oxidase to stereoselectively convert CL into ent-2'-epi-5-deoxystrigol (B-C lactone ring formation), the presumed precursor of rice SLs. A protein encoded by a second rice MAX1 homolog then catalyzes the conversion of ent-2'-epi-5-deoxystrigol to orobanchol. We therefore report that two members of CYP711 enzymes can catalyze two distinct steps in SL biosynthesis, identifying the first enzymes involved in B-C ring closure and a subsequent structural diversification step of SLs.

Topics: Arabidopsis; Arabidopsis Proteins; beta Carotene; Biocatalysis; Dioxygenases; Gene Expression Regulation, Plant; Lactones; Metabolic Networks and Pathways; Models, Molecular; Molecular Docking Simulation; Nicotiana; Oryza; Plant Growth Regulators; Plants, Genetically Modified; Sequence Homology, Amino Acid

Major strigolactones (SLs) produced by rice (Oryza sativa L. cv. Nipponbare) and tobacco (Nicotiana tabacum L. cv. Michinoku No. 1) were purified and their stereochemical structures were determined by comparing with optically pure synthetic standards for their NMR and CD data and retention times and mass fragmentations in ESI-LC/MS and GC-MS. SLs purified from root exudates of rice plants were orobanchol, orobanchyl acetate, and ent-2'-epi-5-deoxystrigol. In addition to these SLs, 7-oxoorobanchyl acetate and the putative three methoxy-5-deoxystrigol isomers were detected by LC-MS/MS. The production of 7-oxoorobanchyl acetate seemed to occur in the early growth stage, as it was detected only in the root exudates collected during the first week of incubation. The root exudates of tobacco contained at least 11 SLs, including solanacol, solanacyl acetate, orobanchol, ent-2'-epi-orobanchol, orobanchyl acetate, ent-2'-epi-orobanchyl acetate, 5-deoxystrigol, ent-2'-epi-5-deoxystrigol, and three isomers of putative didehydro-orobanchol whose structures remain to be clarified. Furthermore, two sorgolactone isomers but not sorgolactone were detected as minor SLs by LC-MS/MS analysis. It is intriguing to note that rice plants produced only orobanchol-type SLs, derived from ent-2'-epi-5-deoxystrigol, but both orobanchol-type and strigol-type SLs, derived from 5-deoxystrigol were detected in tobacco plants.

Topics: Chromatography, Liquid; Lactones; Mass Spectrometry; Nicotiana; Oryza; Plant Exudates; Plant Roots; Reproducibility of Results; Stereoisomerism