Temporal accumulation of pigments during colour transformation from white to red in Combretum indicum (L.) DeFilipps (syn. Quisqualis indica L.) flowers.

This study focuses on the identification of major anthocyanin following its temporal accumulation in colour changing flowers of Combretum indicum (L.) DeFilipps (syn. Quisqualis indica L.). Separation and identification of pigments governing changes in floral colour were performed using HPLC-DAD. Comparison of chromatographic runs with retention time and UV-Vis spectra of authentic standards determined cyanidin 3-O-glucoside as the major anthocyanin accumulating in the petals. Acid hydrolysis of anthocyanin extracts further confirmed cyanidin as the major anthocyanidin in floral tissue. Light microscopic studies revealed gradual accumulation of pigments in the epidermal and hypodermal cell layers of petals. Antioxidant potentials of floral extracts in ethanol, methanol, water and ethyl acetate were determined by DPPH assay where methanolic extracts showed highest free-radical scavenging capacity, and petals of red stage showed maximum activity. Antioxidative potentials measured in terms of FRAP and ABTS also indicated similar results showing highest activity in the red stage.

Characterization of an alcohol acetyltransferase GcAAT responsible for the production of antifungal volatile esters in endophytic Geotrichum candidum PF005.

Alcohol acetyltransferases (AATs) are a group of enzymes that catalyze the formation of esters from different alcohols and acetyl-CoA. However, these enzymes are not well characterized with regard to synthesis of antifungal compounds. The present study aims to investigate the AAT enzyme from Geotrichum candidum PF005, an endophytic yeast-like fungus that emits fruity scented antifungal volatiles, primarily comprising of acetate esters. After PCR-based cloning of the GcAAT gene, the encoded enzyme was characterized structurally through in silico methods and functionally via heterologous expression in Saccharomyces cerevisiae. In native host, the single copy GcAAT gene exhibited induced expression upon supplementation with metabolic precursors, like L-leucine (Leu) or α-ketoisocaproate (α-KIC). Docking studies using the modelled structure of GcAAT revealed differential but favourable binding interactions for three alcohol substrates (i.e., isoamyl alcohol, isobutyl alcohol and 2-phenylethanol) and the co-substrate acetyl-CoA. Binding sites for both substrate and co-substrate are found to be located inside a tunnel identified in the structure, wherein the H208 of the acetyltransferase conserved motif HXXXD was found at a hydrogen bond distance from the substrate. Functional complementation of GcAAT in S. cerevisiae AAT knockout strain caused 32% decrease in dry biomass weight of the test phytopathogenic fungus, Rhizoctonia solani as compared to the control (AAT knockout strain with empty plasmid) after 72 h of incubation due to the emitted volatiles. When the transformed yeast cells were fed with Leu and α-KIC, the relative abundance of the isoamyl acetate ester increased by 21% and 48%, respectively as compared to the control (without precursor). Further analysis documented that volatiles from α-KIC fed GcAAT transformant exhibited 58% higher antifungal activity against the test fungus R. solani than the control, engendered by increased oxidative stress that led to distorted mycelial morphology and increased hyphal branching. Together, the augmented antifungal effect displayed by the GcAAT expressing S. cerevisiae AAT knockout strain is clearly attributable to the acetate esters, especially isoamyl acetate, which are inherently produced in endophytic G. candidum PF005 as antifungal volatiles.

Revealing floral metabolite network in tuberose that underpins scent volatiles synthesis, storage and emission.

KEY MESSAGE: The role of central carbon metabolism in the synthesis and emission of scent volatiles in tuberose flowers was revealed through measurement of changes in transcripts and metabolites levels. Tuberose or Agave amica (Medikus) Thiede & Govaerts is a widely cultivated ornamental plant in several subtropical countries. Little is known about metabolite networking involved in biosynthesis of specialized metabolites utilizing primary metabolites. In this study, metabolite profiling and gene expression analyses were carried out from six stages of maturation throughout floral lifespan. Multivariate analysis indicated distinction between early and late maturation stages. Further, the roles of sugars viz. sucrose, glucose and fructose in synthesis, glycosylation and emission of floral scent volatiles were studied. Transcript levels of an ABC G family transporter (picked up from the floral transcriptome) was in synchronization with terpene volatiles emission during the anthesis stage. A diversion from phenylpropanoid/benzenoid to flavonoid metabolism was observed as flowers mature. Further, it was suggested that this metabolic shift could be mediated by isoforms of 4-Coumarate-CoA ligase along with Myb308 transcription factor. Maximum glycosylation of floral scent volatiles was shown to occur at the late mature stage when emission declined, facilitating both storage and export from the floral tissues. Thus, this study provides an insight into floral scent volatiles synthesis, storage and emission by measuring changes at transcripts and metabolites levels in tuberose throughout floral lifespan.

Glandular trichomes of the flowers and leaves in Millingtonia hortensis (Bignoniaceae).

MAIN CONCLUSION: Three types of the glandular trichomes are developed on the flowers and leaves of Millingtonia hortensis. Morphology, cell ultrastructure and content of the volatile compounds are specific to each trichome type. The aim of this study was to characterize the structural and histochemical features of the glandular trichomes (GTs) of two types localized on the different flower parts and leaves in Millingtonia hortensis, as well as to identify the composition of the internal pool of metabolites. The peltate GTs are most common; they are founded on peduncle, calyx, ovary, and leaves. GTs consist of 12-24-cell disk-shaped head and a single-celled neck. The capitate GTs are located on corolla tube and have four to eight-cell head, single-celled neck and a wide multicellular stalk. A series of histochemical reactions and fluorescent microscopy revealed the various substances in the chemical composition of GTs. Acid polysaccharides are predominately identified in the capitate trichomes of the corolla tube and peltate trichomes of calyx, terpenes present in larger quantity in the trichomes of the corolla tube and ovary, whilst phenolic substances prevail in the trichomes of the calyx and ovary. GTs of each type are characterized by specific ultrastructural traits. Smooth endoplasmic reticulum (SER) and leucoplasts prevail in the peltate trichomes of peduncle, calyx and ovary; Golgi apparatus is the common organelle in the capitate trichomes of the corolla tube and peltate trichomes of calyx; the huge aggregates of the RER cisterns there are in cytoplasm of all leaf trichomes. Synthesized secretion accumulates in the subcuticular cavity of all GTs except the leaf peltate trichomes. In the trichomes of the leaves secretion is stored in the thick upper cell wall with the wide cutinized layer. For the first time content of the internal pool of metabolites from the flowers and leaves was identified by GC-MS. Seventeen compounds, including alcohols, fatty acid derivatives, monoterpenes, sesquiterpenes, and benzenoids were identified. 1-octen 3-ol, 3-carene, methyl salicylate, p-hydroxybenzeneethanol and 1-hydroxy-2,4-di-tertbutyl-benzene were the main compounds of the flower scent. We consider GTs of the reproductive organs in M. hortensis synthesizing acid polysaccharides and volatile compounds as secretory structures attracting of pollinators, whereas the leaf peltate trichomes accumulating predominately non-volatile phenols, protect young vegetative shoots against small herbivorous insects and pathogens.

Specialized metabolites contributing to colour and scent volatiles in Uvaria hamiltonii flowers.

The present study focuses on the emitted and endogenous scent profiles of Uvaria hamiltonii flowers. Among the 34 compounds identified, sesquiterpenoids were found to dominate the floral volatiles composition. Profiles from endogenous scent volatiles showed higher number of compounds than the emitted ones. The anthocyanin pigment responsible for the flower colour was also explored. It was found that a single anthocyanin compound, cyanidin-3-O-glucoside, was principally responsible for petal colour. Total phenolic content was evaluated and antioxidant capacities were studied with the help of DPPH, FRAP and ABTS assays. The total phenolic content and the antioxidant capacity were higher in methanolic extract as compared to aqueous, petroleum ether and ethyl acetate extracts of U. hamiltonii flowers.