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Comprehensive analysis of putative dihydroflavonol 4-reductase gene family in tea plant

One identified dihydroflavonol 4-reductases (DFR) encoding gene (named as CsDFRa herein) and five putative DFRs (named as CsDFRb1, CsDFRb2, CsDFRb3, CsDFRc and CsDFRd) in tea (Camellia sinensis) have been widely discussed in recent papers concerning multi-omics data. However, except for CsDFRa, thei...

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Autores principales: Mei, Xin, Zhou, Caibi, Zhang, Wenting, Rothenberg, Dylan O’Neill, Wan, Shihua, Zhang, Lingyun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6932780/
https://www.ncbi.nlm.nih.gov/pubmed/31877197
http://dx.doi.org/10.1371/journal.pone.0227225
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author Mei, Xin
Zhou, Caibi
Zhang, Wenting
Rothenberg, Dylan O’Neill
Wan, Shihua
Zhang, Lingyun
author_facet Mei, Xin
Zhou, Caibi
Zhang, Wenting
Rothenberg, Dylan O’Neill
Wan, Shihua
Zhang, Lingyun
author_sort Mei, Xin
collection PubMed
description One identified dihydroflavonol 4-reductases (DFR) encoding gene (named as CsDFRa herein) and five putative DFRs (named as CsDFRb1, CsDFRb2, CsDFRb3, CsDFRc and CsDFRd) in tea (Camellia sinensis) have been widely discussed in recent papers concerning multi-omics data. However, except for CsDFRa, their function and biochemical characteristics are not clear. This study aims to compare all putative CsDFRs and preliminarily evaluate their function. We investigated the sequences of genes (coding and promoter regions) and predicted structures of proteins encoded, and determined the activities of heterologously expressed CsDFRs under various conditions. The results showed that the sequences of five putative CsDFRs were quite different from CsDFRa, and had lower expression levels as well. The five putative CsDFRs could not catalyze three dihydroflavonol substrates. The functional CsDFRa had the strongest affinity with dihydroquercetin, and performed best at pH around 7 and 35°C but was not stable at lower pHs or higher temperatures. Single amino acid mutation at position 141 modified the preference of CsDFRa for dihydroquercetin and dihydromyricetin, and also weakened its stability. These data suggest that only CsDFRa works in the pathway for generating anthocyanidins and catechins. This study provides new insights into the function of CsDFRs and may assist to develop new strategies to manipulate the composition of tea flavonoids in the future.
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spelling pubmed-69327802020-01-07 Comprehensive analysis of putative dihydroflavonol 4-reductase gene family in tea plant Mei, Xin Zhou, Caibi Zhang, Wenting Rothenberg, Dylan O’Neill Wan, Shihua Zhang, Lingyun PLoS One Research Article One identified dihydroflavonol 4-reductases (DFR) encoding gene (named as CsDFRa herein) and five putative DFRs (named as CsDFRb1, CsDFRb2, CsDFRb3, CsDFRc and CsDFRd) in tea (Camellia sinensis) have been widely discussed in recent papers concerning multi-omics data. However, except for CsDFRa, their function and biochemical characteristics are not clear. This study aims to compare all putative CsDFRs and preliminarily evaluate their function. We investigated the sequences of genes (coding and promoter regions) and predicted structures of proteins encoded, and determined the activities of heterologously expressed CsDFRs under various conditions. The results showed that the sequences of five putative CsDFRs were quite different from CsDFRa, and had lower expression levels as well. The five putative CsDFRs could not catalyze three dihydroflavonol substrates. The functional CsDFRa had the strongest affinity with dihydroquercetin, and performed best at pH around 7 and 35°C but was not stable at lower pHs or higher temperatures. Single amino acid mutation at position 141 modified the preference of CsDFRa for dihydroquercetin and dihydromyricetin, and also weakened its stability. These data suggest that only CsDFRa works in the pathway for generating anthocyanidins and catechins. This study provides new insights into the function of CsDFRs and may assist to develop new strategies to manipulate the composition of tea flavonoids in the future. Public Library of Science 2019-12-26 /pmc/articles/PMC6932780/ /pubmed/31877197 http://dx.doi.org/10.1371/journal.pone.0227225 Text en © 2019 Mei et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Mei, Xin
Zhou, Caibi
Zhang, Wenting
Rothenberg, Dylan O’Neill
Wan, Shihua
Zhang, Lingyun
Comprehensive analysis of putative dihydroflavonol 4-reductase gene family in tea plant
title Comprehensive analysis of putative dihydroflavonol 4-reductase gene family in tea plant
title_full Comprehensive analysis of putative dihydroflavonol 4-reductase gene family in tea plant
title_fullStr Comprehensive analysis of putative dihydroflavonol 4-reductase gene family in tea plant
title_full_unstemmed Comprehensive analysis of putative dihydroflavonol 4-reductase gene family in tea plant
title_short Comprehensive analysis of putative dihydroflavonol 4-reductase gene family in tea plant
title_sort comprehensive analysis of putative dihydroflavonol 4-reductase gene family in tea plant
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6932780/
https://www.ncbi.nlm.nih.gov/pubmed/31877197
http://dx.doi.org/10.1371/journal.pone.0227225
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