Effects of the <i>Agrobacterium rhizogenes rolC</i> Gene Insertion on Secondary Metabolites Profile and In Vitro Biological Activity of <i>Acmella oleracea</i> (L.) R.K. Jansen

This study investigates the transformation of <i>Acmella oleracea</i> with the <i>Agrobacterium rhizogenes rolC</i> gene and evaluates its impact on phytochemical composition and biological activity. A total of 480 plant nodes were subjected to <i>Agrobacterium</i>...

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Main Authors: Priscilla Paola Bettini, Martina Imbesi, Patrizia Bogani, Valentina Maggini, Filippo Firenzuoli, Fabio Firenzuoli, Domenico Trombetta, Antonella Smeriglio
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Language:English
Published: MDPI AG 2025-05-01
Series:Plants
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Online Access:https://www.mdpi.com/2223-7747/14/9/1373
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author Priscilla Paola Bettini
Martina Imbesi
Patrizia Bogani
Valentina Maggini
Filippo Firenzuoli
Fabio Firenzuoli
Domenico Trombetta
Antonella Smeriglio
author_facet Priscilla Paola Bettini
Martina Imbesi
Patrizia Bogani
Valentina Maggini
Filippo Firenzuoli
Fabio Firenzuoli
Domenico Trombetta
Antonella Smeriglio
author_sort Priscilla Paola Bettini
collection DOAJ
description This study investigates the transformation of <i>Acmella oleracea</i> with the <i>Agrobacterium rhizogenes rolC</i> gene and evaluates its impact on phytochemical composition and biological activity. A total of 480 plant nodes were subjected to <i>Agrobacterium</i>−mediated transformation, leading to the regeneration of 35 putative transgenic plants. Molecular analysis confirmed the presence of the <i>rolC</i> transgene in 17 clones, of which four (C123, C127, C129, and C132) exhibited <i>rolC</i> mRNA expression. Phytochemical profiling of hydroalcoholic extracts of aerial parts (AP) and roots (R) revealed significant differences (<i>p</i> ≤ 0.05) between transgenic and non-transgenic plants (CTR). Compared to non−transgenic plants, transgenic AP exhibited lower total phenolic content but retained or increased flavonoid concentrations, particularly flavan−3−ols, whereas R extracts consistently showed reduced secondary metabolite levels. LC−DAD−ESI−MS analysis identified a diverse metabolite profile, with AP being notably rich in flavonoids (48.65%) and alkylamides (32.43%), including spilanthol. Functional assessments across antioxidant and anti−inflammatory assays demonstrated that R extracts exhibited stronger bioactivity compared to AP extracts, as indicated by lower IC<sub>50</sub> values (0.004–2.18 mg/mL for R vs. 0.007–7.24 mg/mL for AP). However, iron−chelating capacity was higher in AP extracts, correlating with flavonoid concentration. Hierarchical clustering confirmed that transgenic lines C123 and C127 most closely resembled the control, while C129 and C132 displayed distinct metabolic profiles. These findings highlight <i>rolC</i>’s role in modulating secondary metabolite synthesis, influencing both the phytochemical composition and functional properties of <i>A. oleracea</i> extracts.
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spelling doaj-art-e6b87c6e9640479ca001de76e01aaf352025-08-20T02:31:03ZengMDPI AGPlants2223-77472025-05-01149137310.3390/plants14091373Effects of the <i>Agrobacterium rhizogenes rolC</i> Gene Insertion on Secondary Metabolites Profile and In Vitro Biological Activity of <i>Acmella oleracea</i> (L.) R.K. JansenPriscilla Paola Bettini0Martina Imbesi1Patrizia Bogani2Valentina Maggini3Filippo Firenzuoli4Fabio Firenzuoli5Domenico Trombetta6Antonella Smeriglio7Department of Biology, University of Florence, Via Madonna del Piano 6, Sesto F.no, 50019 Florence, ItalyDepartment of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Viale Ferdinando Stagno d’Alcontres 31, 98166 Messina, ItalyDepartment of Biology, University of Florence, Via Madonna del Piano 6, Sesto F.no, 50019 Florence, ItalyDepartment of Biology, University of Florence, Via Madonna del Piano 6, Sesto F.no, 50019 Florence, ItalyCERFIT, Research and Innovation Center in Phytotherapy and Integrated Medicine, Careggi University Hospital, Largo Giovanni Alessandro Brambilla 3, 50134 Florence, ItalyCERFIT, Research and Innovation Center in Phytotherapy and Integrated Medicine, Careggi University Hospital, Largo Giovanni Alessandro Brambilla 3, 50134 Florence, ItalyDepartment of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Viale Ferdinando Stagno d’Alcontres 31, 98166 Messina, ItalyDepartment of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Viale Ferdinando Stagno d’Alcontres 31, 98166 Messina, ItalyThis study investigates the transformation of <i>Acmella oleracea</i> with the <i>Agrobacterium rhizogenes rolC</i> gene and evaluates its impact on phytochemical composition and biological activity. A total of 480 plant nodes were subjected to <i>Agrobacterium</i>−mediated transformation, leading to the regeneration of 35 putative transgenic plants. Molecular analysis confirmed the presence of the <i>rolC</i> transgene in 17 clones, of which four (C123, C127, C129, and C132) exhibited <i>rolC</i> mRNA expression. Phytochemical profiling of hydroalcoholic extracts of aerial parts (AP) and roots (R) revealed significant differences (<i>p</i> ≤ 0.05) between transgenic and non-transgenic plants (CTR). Compared to non−transgenic plants, transgenic AP exhibited lower total phenolic content but retained or increased flavonoid concentrations, particularly flavan−3−ols, whereas R extracts consistently showed reduced secondary metabolite levels. LC−DAD−ESI−MS analysis identified a diverse metabolite profile, with AP being notably rich in flavonoids (48.65%) and alkylamides (32.43%), including spilanthol. Functional assessments across antioxidant and anti−inflammatory assays demonstrated that R extracts exhibited stronger bioactivity compared to AP extracts, as indicated by lower IC<sub>50</sub> values (0.004–2.18 mg/mL for R vs. 0.007–7.24 mg/mL for AP). However, iron−chelating capacity was higher in AP extracts, correlating with flavonoid concentration. Hierarchical clustering confirmed that transgenic lines C123 and C127 most closely resembled the control, while C129 and C132 displayed distinct metabolic profiles. These findings highlight <i>rolC</i>’s role in modulating secondary metabolite synthesis, influencing both the phytochemical composition and functional properties of <i>A. oleracea</i> extracts.https://www.mdpi.com/2223-7747/14/9/1373<i>Acmella oleracea</i> (L.) R.K. Jansenaerial partsrootin vitro plant culture<i>Agrobacterium rhizogenes</i><i>rolC</i> gene
spellingShingle Priscilla Paola Bettini
Martina Imbesi
Patrizia Bogani
Valentina Maggini
Filippo Firenzuoli
Fabio Firenzuoli
Domenico Trombetta
Antonella Smeriglio
Effects of the <i>Agrobacterium rhizogenes rolC</i> Gene Insertion on Secondary Metabolites Profile and In Vitro Biological Activity of <i>Acmella oleracea</i> (L.) R.K. Jansen
Plants
<i>Acmella oleracea</i> (L.) R.K. Jansen
aerial parts
root
in vitro plant culture
<i>Agrobacterium rhizogenes</i>
<i>rolC</i> gene
title Effects of the <i>Agrobacterium rhizogenes rolC</i> Gene Insertion on Secondary Metabolites Profile and In Vitro Biological Activity of <i>Acmella oleracea</i> (L.) R.K. Jansen
title_full Effects of the <i>Agrobacterium rhizogenes rolC</i> Gene Insertion on Secondary Metabolites Profile and In Vitro Biological Activity of <i>Acmella oleracea</i> (L.) R.K. Jansen
title_fullStr Effects of the <i>Agrobacterium rhizogenes rolC</i> Gene Insertion on Secondary Metabolites Profile and In Vitro Biological Activity of <i>Acmella oleracea</i> (L.) R.K. Jansen
title_full_unstemmed Effects of the <i>Agrobacterium rhizogenes rolC</i> Gene Insertion on Secondary Metabolites Profile and In Vitro Biological Activity of <i>Acmella oleracea</i> (L.) R.K. Jansen
title_short Effects of the <i>Agrobacterium rhizogenes rolC</i> Gene Insertion on Secondary Metabolites Profile and In Vitro Biological Activity of <i>Acmella oleracea</i> (L.) R.K. Jansen
title_sort effects of the i agrobacterium rhizogenes rolc i gene insertion on secondary metabolites profile and in vitro biological activity of i acmella oleracea i l r k jansen
topic <i>Acmella oleracea</i> (L.) R.K. Jansen
aerial parts
root
in vitro plant culture
<i>Agrobacterium rhizogenes</i>
<i>rolC</i> gene
url https://www.mdpi.com/2223-7747/14/9/1373
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