Identification of chalcone isomerase gene family in Astragalus mongholicus revealed genes regulating isoflavone synthesis

Astragalus mongholicus (AM) is the original plant of the famous traditional Chinese medicine Astragali Radix, and its isoflavones are important bioactive substances with wide-ranging medicinal values. The chalcone isomerase (CHI) serves a pivotal function in flavonoid synthesis. However, the CHI gen...

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Main Authors: Zhen Wang, Panpan Wang, Xinxin Wang, Lingyang Kong, Jianhao Wu, Weichao Ren, Xiubo Liu, Wei Ma
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Plant Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fpls.2025.1612434/full
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author Zhen Wang
Zhen Wang
Panpan Wang
Xinxin Wang
Lingyang Kong
Jianhao Wu
Weichao Ren
Xiubo Liu
Wei Ma
author_facet Zhen Wang
Zhen Wang
Panpan Wang
Xinxin Wang
Lingyang Kong
Jianhao Wu
Weichao Ren
Xiubo Liu
Wei Ma
author_sort Zhen Wang
collection DOAJ
description Astragalus mongholicus (AM) is the original plant of the famous traditional Chinese medicine Astragali Radix, and its isoflavones are important bioactive substances with wide-ranging medicinal values. The chalcone isomerase (CHI) serves a pivotal function in flavonoid synthesis. However, the CHI gene family in AM has not yet been characterized and systematically analyzed. The present study identified a number of eight AmCHIs distributed on five chromosomes and classified them into four types. The evolutionary relationships, conserved motifs, gene structures, and cis-elements of AmCHIs are discussed. The transcriptome revealed the expression profiles of the AmCHI genes in roots, stems and leaves. In order to characterize AmCHIs function, recombinant proteins AmCHI3, AmCHI4 and AmCHI5 were expressed in Escherichia coli, and the enzyme activity assay showed that the typeI AmCHI4 could only catalyze naringenin chalcone to chalcone, and the typeII AmCHI3 catalyzed the conversion of naringenin chalcone to chalcone and of isoliquiritigenin to Liquiritigenin. Type III AmCHI5 lacked catalytic activity. In addition, gene suppression of AmCHI3 was carried out by using antisense oligodeoxynucleotides (AsODN). Transient gene silencing of AmCHI3 decreased the contents of calycosin, calycosin-7-glucoside and formononetin. indicating that AmCHI3 gene has a positive regulatory effect on the synthesis of isoflavonoids in AM. These results provide the data support for future elucidation of the regulatory mechanism of CHI in isoflavone biosynthesis.
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spelling doaj-art-5c9e2e00641e4c92af1baf38d216e7f22025-08-20T04:03:22ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2025-08-011610.3389/fpls.2025.16124341612434Identification of chalcone isomerase gene family in Astragalus mongholicus revealed genes regulating isoflavone synthesisZhen Wang0Zhen Wang1Panpan Wang2Xinxin Wang3Lingyang Kong4Jianhao Wu5Weichao Ren6Xiubo Liu7Wei Ma8Pharmacy of College, Heilongjiang University of Chinese Medicine, Harbin, ChinaKey Laboratory of Xinjiang Phytomedicine Resource and Utilization, Ministry of Education, School of Pharmacy, Shihezi University, Shihezi, ChinaPharmacy of College, Heilongjiang University of Chinese Medicine, Harbin, ChinaPharmacy of College, Heilongjiang University of Chinese Medicine, Harbin, ChinaPharmacy of College, Heilongjiang University of Chinese Medicine, Harbin, ChinaPharmacy of College, Heilongjiang University of Chinese Medicine, Harbin, ChinaPharmacy of College, Heilongjiang University of Chinese Medicine, Harbin, ChinaCollege of Jiamusi, Heilongjiang University of Chinese Medicine, Jiamusi, ChinaPharmacy of College, Heilongjiang University of Chinese Medicine, Harbin, ChinaAstragalus mongholicus (AM) is the original plant of the famous traditional Chinese medicine Astragali Radix, and its isoflavones are important bioactive substances with wide-ranging medicinal values. The chalcone isomerase (CHI) serves a pivotal function in flavonoid synthesis. However, the CHI gene family in AM has not yet been characterized and systematically analyzed. The present study identified a number of eight AmCHIs distributed on five chromosomes and classified them into four types. The evolutionary relationships, conserved motifs, gene structures, and cis-elements of AmCHIs are discussed. The transcriptome revealed the expression profiles of the AmCHI genes in roots, stems and leaves. In order to characterize AmCHIs function, recombinant proteins AmCHI3, AmCHI4 and AmCHI5 were expressed in Escherichia coli, and the enzyme activity assay showed that the typeI AmCHI4 could only catalyze naringenin chalcone to chalcone, and the typeII AmCHI3 catalyzed the conversion of naringenin chalcone to chalcone and of isoliquiritigenin to Liquiritigenin. Type III AmCHI5 lacked catalytic activity. In addition, gene suppression of AmCHI3 was carried out by using antisense oligodeoxynucleotides (AsODN). Transient gene silencing of AmCHI3 decreased the contents of calycosin, calycosin-7-glucoside and formononetin. indicating that AmCHI3 gene has a positive regulatory effect on the synthesis of isoflavonoids in AM. These results provide the data support for future elucidation of the regulatory mechanism of CHI in isoflavone biosynthesis.https://www.frontiersin.org/articles/10.3389/fpls.2025.1612434/fullAstragalus mongholicuschalcone isomeraseexpression patternprokaryotic expressionAsODN
spellingShingle Zhen Wang
Zhen Wang
Panpan Wang
Xinxin Wang
Lingyang Kong
Jianhao Wu
Weichao Ren
Xiubo Liu
Wei Ma
Identification of chalcone isomerase gene family in Astragalus mongholicus revealed genes regulating isoflavone synthesis
Frontiers in Plant Science
Astragalus mongholicus
chalcone isomerase
expression pattern
prokaryotic expression
AsODN
title Identification of chalcone isomerase gene family in Astragalus mongholicus revealed genes regulating isoflavone synthesis
title_full Identification of chalcone isomerase gene family in Astragalus mongholicus revealed genes regulating isoflavone synthesis
title_fullStr Identification of chalcone isomerase gene family in Astragalus mongholicus revealed genes regulating isoflavone synthesis
title_full_unstemmed Identification of chalcone isomerase gene family in Astragalus mongholicus revealed genes regulating isoflavone synthesis
title_short Identification of chalcone isomerase gene family in Astragalus mongholicus revealed genes regulating isoflavone synthesis
title_sort identification of chalcone isomerase gene family in astragalus mongholicus revealed genes regulating isoflavone synthesis
topic Astragalus mongholicus
chalcone isomerase
expression pattern
prokaryotic expression
AsODN
url https://www.frontiersin.org/articles/10.3389/fpls.2025.1612434/full
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