An UHPLC-HRMS-Based Untargeted Metabolomics Approach to Explore the Effects of Bacterial Endophyte Co-Culture on <i>Alkanna tinctoria</i> (L.) Tausch Cell Suspension Metabolome

Colonization of plant tissues by bacterial endophytes might lead to qualitative and quantitative changes in secondary metabolites (SMs). In this work, in vitro co-culture experiments were performed using cell suspensions of the medicinal plant <i>Alkanna tinctoria</i> and eight of its ba...

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Main Authors: Elodie Bossard, Adrien Cousy, Antonio Grondin, Nikolaos Tsafantakis, Angélique Rat, Nektarios Aligiannis, Anne Willems, Laetitia Cattuzzato, Thien Nguyen, Nikolas Fokialakis
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Language:English
Published: MDPI AG 2025-07-01
Series:Microorganisms
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Online Access:https://www.mdpi.com/2076-2607/13/7/1601
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author Elodie Bossard
Adrien Cousy
Antonio Grondin
Nikolaos Tsafantakis
Angélique Rat
Nektarios Aligiannis
Anne Willems
Laetitia Cattuzzato
Thien Nguyen
Nikolas Fokialakis
author_facet Elodie Bossard
Adrien Cousy
Antonio Grondin
Nikolaos Tsafantakis
Angélique Rat
Nektarios Aligiannis
Anne Willems
Laetitia Cattuzzato
Thien Nguyen
Nikolas Fokialakis
author_sort Elodie Bossard
collection DOAJ
description Colonization of plant tissues by bacterial endophytes might lead to qualitative and quantitative changes in secondary metabolites (SMs). In this work, in vitro co-culture experiments were performed using cell suspensions of the medicinal plant <i>Alkanna tinctoria</i> and eight of its bacterial endophytes. An untargeted metabolomics approach using Ultra-High-Performance Liquid Chromatography High-Resolution Mass Spectrometry (UHPLC-HRMS) was employed to investigate plant–microbe interactions. Hierarchical clustering analysis and principal component analysis highlighted significant modifications of specific regulation patterns in SM production, caused by bacterial endophytes. The annotation step lead to the identification of 32 stimulated compounds in <i>A. tinctoria</i> cell suspensions. Among them, 3′-hydroxy-14-hydroxyshikonofuran H (<b>5</b>), 8′-decarboxy-rosmarinic acid (<b>18</b>), 8‴-decarboxy-salvianolic B (<b>23</b>), 8″-8‴-didecarboxy-salvianolic acid B (<b>26</b>) were putatively identified for the first time. Our findings highlight that employing selected microbial inoculants under controlled conditions can be an effective strategy for enhancing or stimulating the production of specific high-value metabolites.
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spelling doaj-art-02e15620fd0946eaa2b4dc30d7d3cf312025-08-20T03:35:38ZengMDPI AGMicroorganisms2076-26072025-07-01137160110.3390/microorganisms13071601An UHPLC-HRMS-Based Untargeted Metabolomics Approach to Explore the Effects of Bacterial Endophyte Co-Culture on <i>Alkanna tinctoria</i> (L.) Tausch Cell Suspension MetabolomeElodie Bossard0Adrien Cousy1Antonio Grondin2Nikolaos Tsafantakis3Angélique Rat4Nektarios Aligiannis5Anne Willems6Laetitia Cattuzzato7Thien Nguyen8Nikolas Fokialakis9Department of Pharmacy, National and Kapodistrian University of Athens, 15771 Athens, GreecePierre Fabre Dermo-Cosmetic & Personal Care (PFDC&PC), 31100 Toulouse, FranceHerbal Products Laboratory, Green Mission Department, Pierre Fabre Research Institute, 31035 Toulouse, FranceDepartment of Pharmacy, National and Kapodistrian University of Athens, 15771 Athens, GreeceLaboratory of Microbiology, Faculty of Sciences, Ghent University, 9000 Ghent, BelgiumDepartment of Pharmacy, National and Kapodistrian University of Athens, 15771 Athens, GreeceLaboratory of Microbiology, Faculty of Sciences, Ghent University, 9000 Ghent, BelgiumPierre Fabre Dermo-Cosmetic & Personal Care (PFDC&PC), 31100 Toulouse, FrancePierre Fabre Dermo-Cosmetic & Personal Care (PFDC&PC), 31100 Toulouse, FranceDepartment of Pharmacy, National and Kapodistrian University of Athens, 15771 Athens, GreeceColonization of plant tissues by bacterial endophytes might lead to qualitative and quantitative changes in secondary metabolites (SMs). In this work, in vitro co-culture experiments were performed using cell suspensions of the medicinal plant <i>Alkanna tinctoria</i> and eight of its bacterial endophytes. An untargeted metabolomics approach using Ultra-High-Performance Liquid Chromatography High-Resolution Mass Spectrometry (UHPLC-HRMS) was employed to investigate plant–microbe interactions. Hierarchical clustering analysis and principal component analysis highlighted significant modifications of specific regulation patterns in SM production, caused by bacterial endophytes. The annotation step lead to the identification of 32 stimulated compounds in <i>A. tinctoria</i> cell suspensions. Among them, 3′-hydroxy-14-hydroxyshikonofuran H (<b>5</b>), 8′-decarboxy-rosmarinic acid (<b>18</b>), 8‴-decarboxy-salvianolic B (<b>23</b>), 8″-8‴-didecarboxy-salvianolic acid B (<b>26</b>) were putatively identified for the first time. Our findings highlight that employing selected microbial inoculants under controlled conditions can be an effective strategy for enhancing or stimulating the production of specific high-value metabolites.https://www.mdpi.com/2076-2607/13/7/1601bacterial endophytesplant–endophyte relationshipsplant cell cultureco-culturehydroxynaphthoquinones
spellingShingle Elodie Bossard
Adrien Cousy
Antonio Grondin
Nikolaos Tsafantakis
Angélique Rat
Nektarios Aligiannis
Anne Willems
Laetitia Cattuzzato
Thien Nguyen
Nikolas Fokialakis
An UHPLC-HRMS-Based Untargeted Metabolomics Approach to Explore the Effects of Bacterial Endophyte Co-Culture on <i>Alkanna tinctoria</i> (L.) Tausch Cell Suspension Metabolome
Microorganisms
bacterial endophytes
plant–endophyte relationships
plant cell culture
co-culture
hydroxynaphthoquinones
title An UHPLC-HRMS-Based Untargeted Metabolomics Approach to Explore the Effects of Bacterial Endophyte Co-Culture on <i>Alkanna tinctoria</i> (L.) Tausch Cell Suspension Metabolome
title_full An UHPLC-HRMS-Based Untargeted Metabolomics Approach to Explore the Effects of Bacterial Endophyte Co-Culture on <i>Alkanna tinctoria</i> (L.) Tausch Cell Suspension Metabolome
title_fullStr An UHPLC-HRMS-Based Untargeted Metabolomics Approach to Explore the Effects of Bacterial Endophyte Co-Culture on <i>Alkanna tinctoria</i> (L.) Tausch Cell Suspension Metabolome
title_full_unstemmed An UHPLC-HRMS-Based Untargeted Metabolomics Approach to Explore the Effects of Bacterial Endophyte Co-Culture on <i>Alkanna tinctoria</i> (L.) Tausch Cell Suspension Metabolome
title_short An UHPLC-HRMS-Based Untargeted Metabolomics Approach to Explore the Effects of Bacterial Endophyte Co-Culture on <i>Alkanna tinctoria</i> (L.) Tausch Cell Suspension Metabolome
title_sort uhplc hrms based untargeted metabolomics approach to explore the effects of bacterial endophyte co culture on i alkanna tinctoria i l tausch cell suspension metabolome
topic bacterial endophytes
plant–endophyte relationships
plant cell culture
co-culture
hydroxynaphthoquinones
url https://www.mdpi.com/2076-2607/13/7/1601
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