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Competition between FLS and DFR regulates the distribution of flavonols and proanthocyanidins in Rubus chingii Hu

Rubus chingii Hu is a berry plant of the genus Rubus of the Rosaceae family, which has high nutritional and medicinal value and is rich in flavonoids. Flavonol synthase (FLS) and dihydroflavonol 4-reductase (DFR) compete for the common substrate dihydroflavonols to regulate the metabolic flux of fla...

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Autores principales: Lei, Ting, Huang, Jun, Ruan, Haixiang, Qian, Wei, Fang, Zhou, Gu, Chunyang, Zhang, Niuniu, Liang, Yaxuan, Wang, Ziyun, Gao, Liping, Wang, Yunsheng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10031046/
https://www.ncbi.nlm.nih.gov/pubmed/36968391
http://dx.doi.org/10.3389/fpls.2023.1134993
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author Lei, Ting
Huang, Jun
Ruan, Haixiang
Qian, Wei
Fang, Zhou
Gu, Chunyang
Zhang, Niuniu
Liang, Yaxuan
Wang, Ziyun
Gao, Liping
Wang, Yunsheng
author_facet Lei, Ting
Huang, Jun
Ruan, Haixiang
Qian, Wei
Fang, Zhou
Gu, Chunyang
Zhang, Niuniu
Liang, Yaxuan
Wang, Ziyun
Gao, Liping
Wang, Yunsheng
author_sort Lei, Ting
collection PubMed
description Rubus chingii Hu is a berry plant of the genus Rubus of the Rosaceae family, which has high nutritional and medicinal value and is rich in flavonoids. Flavonol synthase (FLS) and dihydroflavonol 4-reductase (DFR) compete for the common substrate dihydroflavonols to regulate the metabolic flux of flavonoids. However, the competition between FLS and DFR based on enzyme is rarely reported. Here, we isolated and identified two FLS genes (RcFLS1 and RcFLS2) and one DFR gene (RcDFR) from Rubus chingii Hu. RcFLSs and RcDFR were highly expressed in stems, leaves, and flowers, although the flavonol accumulation in these organs was significantly higher than that of proanthocyanidins (PAs). The recombinant RcFLSs demonstrated bifunctional activities via hydroxylation and desaturation at the C-3α position having a lower Michaelis constant (Km) for dihydroflavonols than RcDFR. We also found that a low concentration of flavonols could significantly inhibit RcDFR activity. To investigate the competitive relationship between RcFLSs and RcDFR, we used a prokaryotic expression system (E. coli) to co-express these proteins. The transgenic cells expressing recombinant proteins were incubated with substrates, and the reaction products were analyzed. Furthermore, two transient expression systems (tobacco leaves and strawberry fruits) and a stable genetic system (Arabidopsis thaliana) were used to co-express these proteins in vivo. The results showed that RcFLS1 was dominant in the competition with RcDFR. Our results demonstrated that the competition between FLS and DFR regulated the metabolic flux distribution of flavonols and PAs, which will be of great significance for the molecular breeding of Rubus plants.
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spelling pubmed-100310462023-03-23 Competition between FLS and DFR regulates the distribution of flavonols and proanthocyanidins in Rubus chingii Hu Lei, Ting Huang, Jun Ruan, Haixiang Qian, Wei Fang, Zhou Gu, Chunyang Zhang, Niuniu Liang, Yaxuan Wang, Ziyun Gao, Liping Wang, Yunsheng Front Plant Sci Plant Science Rubus chingii Hu is a berry plant of the genus Rubus of the Rosaceae family, which has high nutritional and medicinal value and is rich in flavonoids. Flavonol synthase (FLS) and dihydroflavonol 4-reductase (DFR) compete for the common substrate dihydroflavonols to regulate the metabolic flux of flavonoids. However, the competition between FLS and DFR based on enzyme is rarely reported. Here, we isolated and identified two FLS genes (RcFLS1 and RcFLS2) and one DFR gene (RcDFR) from Rubus chingii Hu. RcFLSs and RcDFR were highly expressed in stems, leaves, and flowers, although the flavonol accumulation in these organs was significantly higher than that of proanthocyanidins (PAs). The recombinant RcFLSs demonstrated bifunctional activities via hydroxylation and desaturation at the C-3α position having a lower Michaelis constant (Km) for dihydroflavonols than RcDFR. We also found that a low concentration of flavonols could significantly inhibit RcDFR activity. To investigate the competitive relationship between RcFLSs and RcDFR, we used a prokaryotic expression system (E. coli) to co-express these proteins. The transgenic cells expressing recombinant proteins were incubated with substrates, and the reaction products were analyzed. Furthermore, two transient expression systems (tobacco leaves and strawberry fruits) and a stable genetic system (Arabidopsis thaliana) were used to co-express these proteins in vivo. The results showed that RcFLS1 was dominant in the competition with RcDFR. Our results demonstrated that the competition between FLS and DFR regulated the metabolic flux distribution of flavonols and PAs, which will be of great significance for the molecular breeding of Rubus plants. Frontiers Media S.A. 2023-03-08 /pmc/articles/PMC10031046/ /pubmed/36968391 http://dx.doi.org/10.3389/fpls.2023.1134993 Text en Copyright © 2023 Lei, Huang, Ruan, Qian, Fang, Gu, Zhang, Liang, Wang, Gao and Wang https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Lei, Ting
Huang, Jun
Ruan, Haixiang
Qian, Wei
Fang, Zhou
Gu, Chunyang
Zhang, Niuniu
Liang, Yaxuan
Wang, Ziyun
Gao, Liping
Wang, Yunsheng
Competition between FLS and DFR regulates the distribution of flavonols and proanthocyanidins in Rubus chingii Hu
title Competition between FLS and DFR regulates the distribution of flavonols and proanthocyanidins in Rubus chingii Hu
title_full Competition between FLS and DFR regulates the distribution of flavonols and proanthocyanidins in Rubus chingii Hu
title_fullStr Competition between FLS and DFR regulates the distribution of flavonols and proanthocyanidins in Rubus chingii Hu
title_full_unstemmed Competition between FLS and DFR regulates the distribution of flavonols and proanthocyanidins in Rubus chingii Hu
title_short Competition between FLS and DFR regulates the distribution of flavonols and proanthocyanidins in Rubus chingii Hu
title_sort competition between fls and dfr regulates the distribution of flavonols and proanthocyanidins in rubus chingii hu
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10031046/
https://www.ncbi.nlm.nih.gov/pubmed/36968391
http://dx.doi.org/10.3389/fpls.2023.1134993
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