Angeli, A. | Search bioRxiv

bioRxiv2023

Generation and characterisation of an Arabidopsis thaliana f3h/fls1/ans triple mutant that accumulates eriodictyol derivatives

BackgroundFlavonoids are plant specialised metabolites, which derive from phenylalanine and acetate metabolism. They possess a variety of beneficial characteristics for plants and humans. Several modification steps in the synthesis of tricyclic flavonoids cause for the amazing diversity of flavonoids in plants. The 2-oxoglutarate-dependent dioxygenases (2-ODDs) flavanone 3-hydroxylase (F3H, synonym FHT), flavonol synthase (FLS) and anthocyanidin synthase (ANS, synonym leucoanthocyanidin dioxigenase (LDOX)), catalyse oxidative modifications to the central C ring. They are highly similar and have been shown to catalyse, at least in part, each others reactions. FLS and ANS have been identified as bifunctional enzymes in many species, including Arabidopsis thaliana, stressing the capability of plants to bypass missing or mutated reaction steps on the way to flavonoid production. However, little is known about such bypass reactions and the flavonoid composition of plants lacking all three central flavonoid 2-ODDs. ResultsTo address this issue, we generated a f3h/fls1/ans mutant, as well as the corresponding double mutants and investigated the flavonoid composition of this mutant collection. The f3h/fls1/ans mutant was further characterised at the genomic level by analysis of a nanopore DNA sequencing generated genome sequence assembly and at the transcriptomic level by RNA-Seq analysis. The mutant collection established, including the novel double mutants f3h/fls1 and f3h/ans, was used to validate and analyse the multifunctionalities of F3H, FLS1, and ANS in planta. Metabolite analyses revealed the accumulation of eriodictyol and additional glycosylated derivatives in mutants carrying the f3h mutant allele, resulting from the conversion of naringenin to eriodictyol by flavonoid 3-hydroxylase (F3H) activity. ConclusionsWe describe the in planta multifunctionality of the three central flavonoid 2-ODDs from A. thaliana and identify a bypass in the f3h/fls1/ans triple mutant that leads to the formation of eriodictyol derivatives. As (homo-)eriodictyols are known as bitter taste maskers, the annotated eriodictyol (derivatives) and in particular the observations made on their in planta production, could provide valuable insights for creating of novel food supplements.

plant biology↗

bioRxiv2021

Reaching to inhibit a prepotent response: a wearable 3-axis accelerometer kinematic analysis

The present work explores the distinctive contribution of motor planning and control to human reaching movements. In particular, the movements were triggered by the selection of a prepotent response (Dominant) or, instead, by the inhibition of the prepotent response, that required the selection of an alternative one (Non-dominant). To this aim, we adapted a Go/No-Go task to investigate both the dominant and non-dominant movements of a cohort of 19 adults, utilizing kinematic measures to discriminate between the planning and control components of the two actions. To sample such measures, a low-cost, easy to use, 3-axis wrist worn accelerometer was put to good use to obtain raw acceleration data and to compute and break down its velocity components. The values obtained with such task indicate that with the inhibition of a prepotent response, the selection and execution of the alternative one yields both a longer reaction time and movement duration. Moreover, the peak velocity occurred later in time with respect to the dominant response, revealing that participants tended to indulge more in motor planning rather than in adjusting their movement along the way. Finally, comparing such results to the findings obtained by other means in literature, we discuss the feasibility of an accelerometer-based analysis to disentangle distinctive cognitive mechanisms of human movements.

bioinformatics↗