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Generation and characterisation of an Arabidopsis thaliana f3h/fls1/ans triple mutant that accumulates eriodictyol derivatives
Get PDFBackground: Flavonoids 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 dioxygenase (LDOX)), catalyse oxidative modifications to the central C ring. They are highly similar and have been shown to catalyse, at least in part, each other's 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. Results: To 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 (F3'H) activity. Conclusions: We 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 the creation of novel food supplements
The effect of different medium compositions and LAB strains on fermentation Volatile Organic Compounds (VOCs) analysed by Proton Transfer Reaction-Time of Flight-Mass Spectrometry (PTR-ToF-MS)
Get PDFLactic acid bacteria (LAB) fermentation is a viable approach for producing plant-based flavour compounds; however, little is understood about the impact of different LAB strains and medium compositions on the production of volatile organic compounds (VOCs). This study investigated the impact of the addition of individual amino acids (AAs) (L-leucine, L-isoleucine, L-phenylalanine, L-glutamic acid, L-aspartic acid, L-threonine, or L-methionine) to a defined medium (DM) on the generation of VOCs (after 0, 7, and 14 days) by one of three LAB strains (Levilactobacillus brevis WLP672 (LB672), Lactiplantibacillus plantarum LP100 (LP100), and Pediococcus pentosaceus PP100 (PP100)), using proton transfer reaction-time of flight-mass spectrometry (PTR-ToF-MS). The concentration of m/z 45.031 (t.i. acetaldehyde) was significantly (p < 0.05) higher after 7 days of fermentation by LP100 in the DM supplemented with threonine compared to all other media fermented by all three strains. The concentrations of m/z 49.012 (t.i. methanethiol) and m/z 95.000 (t.i. dimethyl disulfide) were significantly (p < 0.05) higher after 7 days of fermentation by either LP100, PP100, or LB672 in the DM supplemented with methionine compared to all other media. Information on the role of individual AAs on VOCs generation by different LAB strains will help to guide flavour development from the fermentation of plant-based substrate
Elucidation of arsenic detoxification mechanism in Marchantia polymorpha: the role of ACR3
Get PDFThe arsenic-specific ACR3 transporter plays pivotal roles in As detoxification in yeast and a group of ancient tracheophytes, the ferns. Despite putative ACR3 genes being present in the genomes of bryophytes, whether they have the same relevance also in this lineage is currently unknown. In this study, we characterized the MpACR3 gene from the bryophyte Marchantia polymorpha L. through a multiplicity of functional approaches ranging from phylogenetic reconstruction, expression analysis, loss- and gain-of-function as well as genetic complementation with an MpACR3 gene tagged with a fluorescent protein. Genetic complementation demonstrates that MpACR3 plays a pivotal role in As tolerance in M. polymorpha, with loss-of-function Mpacr3 mutants being hypersensitive and MpACR3 overexpressors more tolerant to As. Additionally, MpACR3 activity regulates intracellular As concentration, affects its speciation and controls the levels of intracellular oxidative stress. The MpACR3::3xCitrine appears to localize at the plasma membrane and possibly in other endomembrane systems. Taken together, these results demonstrate the pivotal function of ACR3 detoxification in both sister lineages of land plants, indicating that it was present in the common ancestor to all embryophytes. We propose that Mpacr3 mutants could be used in developing countries as low-cost and low-technology visual bioindicators to detect As pollution in wate
Isotopic, mycotoxin, and pesticide analysis for organic authentication along the production chain of wheat-derived products
Get PDFWheat-based products are staples in diets worldwide. Organic food frauds continuously threaten consumer trust in the agri-food system. A multi-method approach was conducted for the organic authentication and safety assessment of pasta and bakery products along their production chain. Bulk and Compound-Specific (CS) Isotope Ratio Mass Spectrometry (IRMS) suggested the δ15Nbulk, δ15Nleucine and δ15Nproline as promising organic markers, with CS able to distinguish between pairs which bulk analysis could not. Processing significantly affected the values of δ15Nleucine, δ13Cproline and δ13Cleucine. Multi-mycotoxin analysis (HT-2, T-2, DON, ZEN, OTA, AFB1) revealed higher contamination in conventional than organic samples, while both milling and baking significantly reduced mycotoxin content. Lastly, from the evaluation of 400 residues, isopyrazam was present at the highest concentration (0.12 mg/kg) in conventional wheat, exhibiting a 0.12 Processing Factor (PF), while tebuconazole levels remained unchanged in pasta production (90 °C) and reduced below LOQ in biscuits and crackers (180–250 °C
Importanza del monitoraggio nella gestione della flavescenza
No full textIn un’area viticola collinare del Trentino meridionale caratterizzata da elevate criticità ambientali si è svolta un’indagine pluriennale sulla diffusione delle fitoplasmosi della vite e degli insetti vettori. Il monitoraggio nelle diverse fasi della stagione e la sensibilizzazione dei viticoltori sono fondamentali per una corretta gestione di questa fitopati
Light-assisted room temperature ammonia gas sensor based on porphyrin-coated V2O5 nanosheets
No full textThis study explores the gas-sensing capabilities of light-activated porphyrin-coated V2O5 nanosheets for ammonia detection. The nanosheets were synthesized via a hydrothermal method, heat treatment, and solution dispersion. They were then coated with thin layers of Zn-porphyrin and Cu-porphyrin derivatives. The gas-sensing performance of the porphyrin/V2O5 sensor was evaluated under dark and visible light conditions. The combination of materials significantly improves the sensor's performance compared to pure V2O5 nanosheets sensor. The study also investigated selectivity, temporal stability, and proposed a mechanism for ammonia detection based on the interaction between photo-excited charge carriers and adsorbed gas molecules. With such performance, compact size, cost-effectiveness, and low energy consumption, this sensor holds substantial promise for monitoring freshness and spoilage along food supply chain