首页分享 Nature Plants :通过抑制衰老调节因子EPHEMERAL1延长日本牵牛花寿命的化学方法

Nature Plants :通过抑制衰老调节因子EPHEMERAL1延长日本牵牛花寿命的化学方法

来源：花匠小妙招时间：2026-02-16 22:03

AbstractPetal senescence in flowering plants is a type of programmed cell death with highly regulated onset and progression. A NAM/ATAF1,2/CUC2 transcription factor, EPHEMERAL1 (EPH1), has been identified as a key regulator of petal senescence in Japanese morning glory (Ipomoea nil). Here we used a novel chemical approach to delay petal senescence in Japanese morning glory by inhibiting the DNA-binding activity of EPH1.

。NAM/ATAF1,2/CUC2转录因子EPHEMERAL1（EPH1）已被确定为日本牵牛花（Ipomoea nil）花瓣衰老的关键调节因子。在这里，我们使用了一种新的化学方法，通过抑制EPH1的DNA结合活性来延缓日本牵牛花的花瓣衰老。

A cell-free high-throughput screening system and subsequent bioassays found two tetrafluorophthalimide-based compounds, Everlastin1 and Everlastin2, that inhibited the EPH1–DNA interaction and delayed petal senescence. The inhibitory mechanism was due to the suppression of EPH1 dimerization. RNA-sequencing analysis revealed that the chemical treatment strongly suppressed the expression of programmed cell death- and autophagy-related genes.

无细胞高通量筛选系统和随后的生物测定发现了两种基于四氟邻苯二甲酰亚胺的化合物，Everlastin1和Everlastin2，它们抑制了EPH1-DNA的相互作用并延缓了花瓣的衰老。。RNA测序分析表明，化学处理强烈抑制了程序性细胞死亡和自噬相关基因的表达。

These results suggest that a chemical approach targeting a transcription factor can regulate petal senescence..

这些结果表明，靶向转录因子的化学方法可以调节花瓣衰老。。

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Fig. 1: Establishment of a high-throughput assay to detect the EPH1–DNA interaction and screening of EPH1 inhibitors.Fig. 2: Screening of analogues of the compounds obtained from the first screening.Fig. 3: Effects of Everlastin1 and Everlastin2 on the DNA interaction and dimerization of EPH1.Fig. 4: ChIP and transcriptome analysis of Everlastin1- and Everlastin2-treated petals.Fig.

图1：建立高通量测定法以检测EPH1-DNA相互作用和筛选EPH1抑制剂。图2：从第一次筛选获得的化合物类似物的筛选。图3:Everlastin1和Everlastin2对EPH1的DNA相互作用和二聚化的影响。图4:Everlastin1和Everlastin2处理的花瓣的ChIP和转录组分析。图。

5: Effects of Everlastin1 and Everlastin2 on petal senescence..

5： Everlastin1和Everlastin2对花瓣衰老的影响。。

Data availability

数据可用性

All data generated or analysed in this study are included in this published article and its supplementary information files. Raw RNA-seq data are available at DDBJ Sequence Read Archive (https://www.ddbj.nig.ac.jp/dra/index-e.html, with accession numbers DRR437987–DRR437998, DRR545318–DRR545320). The I.

本研究中生成或分析的所有数据均包含在本文及其补充信息文件中。原始RNA-seq数据可在DDBJ序列读取存档中获得(https://www.ddbj.nig.ac.jp/dra/index-e.html，登录号为DRR437987–DRR437998，DRR545318–DRR545320）。I。

nil genome (RefSeq assembly accession: GCF_001879475.1) was used for the sequencing analysis. Source data are provided with this paper..

nil基因组（RefSeq组装登录号：GCF001879475.1）用于测序分析。本文提供了源数据。。

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Download referencesAcknowledgementsWe thank the Applied Protein Research Laboratory at Ehime University for protein analysis, the Drug Discovery Initiative at the University of Tokyo for providing the chemical libraries and the National BioResource Project ‘Morning glory’, Japan, for providing seeds of I.

下载参考文献致谢我们感谢爱媛大学应用蛋白质研究实验室进行蛋白质分析，东京大学药物发现计划提供化学文库，日本国家生物资源项目“牵牛花”提供I的种子。

nil. We also thank M. Nakayama (National Agriculture and Food Research Organization) for valuable discussions about chemical structures, and K. Maruyama (Ehime University) for assistance with in vitro interaction assays. This work was supported by a grant from a commissioned project study on ‘Development of post-harvest technology in cut flowers’ (JP15650941 to K.S.) from the Ministry of Agriculture, Forestry and Fisheries, Japan.; the Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research) from Japan Agency for Medical Research and Development (AMED) under grant numbers JP21am0101086 (support number 1590 to K.S.) and JP21am0101077 (support number 1144 to K.S.); and a Grant-in-Aid for Scientific Research on Innovative Areas (JP16H06579 to T.S.), Grants-in-Aid for Scientific Research (21K05589 to K.S., 19H03218 to T.S.

零。我们还感谢M.Nakayama（国家农业和食品研究组织）对化学结构的宝贵讨论，以及K.Maruyama（爱媛大学）对体外相互作用测定的帮助。这项工作得到了日本农林水产省委托进行的“切花采后技术开发”项目研究（JP15650941至K.S.）的资助。；日本医学研究与发展署（AMED）支持药物发现和生命科学研究的平台项目（支持创新药物发现和生命科学研究的基础），资助号为JP21am0101086（K.S.的支持号1590）和JP21am0101077（K.S.的支持号1144）；以及创新领域科学研究补助金（T.S.的JP16H06579），科学研究补助金（K.S.的21K05589，T.S.的19H03218）。

and 19K05815 to A.N.) from the Japan Society for the Promotion of Science.Author informationAuthors and AffiliationsInstitute of Vegetable and Floriculture Science, National Agriculture and Food Research Organization (NARO), Tsukuba, JapanKenichi ShibuyaProteo-Science Center (PROS), Ehime University, Matsuyama, JapanAkira Nozawa, Chikako Takahashi & Tatsuya SawasakiAuthorsKenichi ShibuyaView author publicationsYou can also search for this author in.

和日本科学促进会的19K05815）。作者信息作者和所属机构国家农业和食品研究组织（NARO）蔬菜和花卉科学研究所，筑波，日本爱媛大学，松山，日本野崎骏，高桥千代子和SawasakiAuthorsKenichi ShibuyaView作者出版物你也可以在中搜索这位作者。

PubMed Google ScholarAkira NozawaView author publicationsYou can also search for this author in

PubMed Google ScholarAkira NozawaView作者出版物您也可以在

PubMed Google ScholarChikako TakahashiView author publicationsYou can also search for this author in

PubMed Google ScholarChikako TakahashiView作者出版物您也可以在

PubMed Google ScholarTatsuya SawasakiView author publicationsYou can also search for this author in

PubMed Google ScholarTatsuya SawasakiView作者出版物您也可以在

PubMed Google ScholarContributionsK.S., A.N. and T.S. designed experiments. K.S. and A.N. performed chemical screening. A.N., C.T. and K.S. performed in vitro interaction assays. K.S. performed the experiments with Japanese morning glory plants and RNA-seq analysis. K.S., A.N.

PubMed谷歌学术贡献SK。S、，A.N.和T.S.设计的实验。K、 S.和A.N.进行了化学筛选。A、 N.，C.T.和K.S.进行了体外相互作用测定。K、美国用日本牵牛花植物和RNA-seq分析进行了实验。K、。

and T.S. analysed and interpreted data. K.S. and A.N. wrote the manuscript, and all authors contributed to editing.Corresponding authorsCorrespondence to.

。K、 S.和A.N.撰写了手稿，所有作者都为编辑做出了贡献。通讯作者通讯。

Kenichi Shibuya or Akira Nozawa.Ethics declarations

涉谷健一或野泽明。道德宣言

Competing interests

相互竞争的利益

The authors declare no competing interests.

作者声明没有利益冲突。

Peer review

同行评审

Peer review information

同行评审信息

Nature Plants thanks Junping Gao and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Nature Plants感谢高俊平和另一位匿名审稿人为这项工作的同行评审做出的贡献。

Additional informationPublisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.Extended dataExtended Data Fig. 1 Chemical structures of G1-1 and its analogues.The inhibition assay of EPH1–DNA interaction activity was performed in the presence of G1-1 analogues at a final concentration of 5 µM.Extended Data Fig.

Additional informationPublisher的注释Springer Nature在已发布的地图和机构隶属关系中的管辖权主张方面保持中立。扩展数据扩展数据图1 G1-1及其类似物的化学结构。EPH1-DNA相互作用活性的抑制测定是在最终浓度为5µM的G1-1类似物存在下进行的。扩展数据图。

2 Inhibition rate of EPH1–DNA interaction activity for fluorine-substituted compounds of G1-5 and G1-6.a, b, Chemical structures (a) and the inhibition rate of EPH1–DNA interaction activity (b) for fluorine-substituted compounds of G1-5. c, d, Chemical structures (c) and the inhibition rate of EPH1–DNA interaction activity (d) for fluorine-substituted compounds of G1-6.

。c、 d，化学结构（c）和EPH1-DNA相互作用活性（d）对G1-6氟取代化合物的抑制率。

The inhibition assays were performed in the presence of compounds at a final concentration of 10 µM. Values are means ± s.e.m. of n = 3 independent experiments (b, d). Asterisks indicate significant differences. **** P < 0.0001; one-way ANOVA with Dunnett’s test compared with G1-5 (b) or G1-6 (d).Source dataExtended Data Fig.

在终浓度为10μM的化合物存在下进行抑制测定。。星号表示显著差异。****P<0.0001；与G1-5（b）或G1-6（d）相比，Dunnett检验的单因素方差分析。源数据扩展数据图。

3 Effects of Everlastin1 and Everlastin2 on the DNA interaction of NAC TFs other than EPH1.a, Alignment of EPH1 (GenBank Accession No. AB849126), Arabidopsis ORE1 (NM_123323), and Lilium LhNAP (LC807006) polypeptide sequences. Identical amino acids are shown in reverse type. The overline indicates the conserved DNA-binding domain.

3 Everlastin1和Everlastin2对除EPH1.a以外的NAC-TF的DNA相互作用的影响，EPH1（GenBank登录号AB849126），拟南芥ORE1（NMU 123323）和百合LhNAP（LC807006）多肽序列的比对。。上划线表示保守的DNA结合结构域。

b, Phylogenic tree of EPH1, Arabidopsis ORE1 and Lilium LhNAP proteins. Sequence alignment with ClustalW2 and phylogenic tree analysis with the neighbor-joining method were performed using Geneious Prime software (Biomatters, Auckland, New Zealand). c, Interaction assay for NAC TFs and DNA. Interactions between DNA and full-length ORE1 or LhNAP in the presence of Everlasti.

b、。使用Geneious Prime软件（Biomatters，奥克兰，新西兰）进行与ClustalW2的序列比对和与邻居连接方法的系统发育树分析。c、 NAC TFs和DNA的相互作用测定。在Everlasti存在下，DNA与全长ORE1或LhNAP之间的相互作用。

Nat. Plants (2024). https://doi.org/10.1038/s41477-024-01767-zDownload citationReceived: 14 January 2023Accepted: 19 July 2024Published: 29 August 2024DOI: https://doi.org/10.1038/s41477-024-01767-zShare this articleAnyone you share the following link with will be able to read this content:Get shareable linkSorry, a shareable link is not currently available for this article.Copy to clipboard.

《自然植物》（2024）。https://doi.org/10.1038/s41477-024-01767-zDownload引文接收日期：2023年1月14日接收日期：2024年7月19日发布日期：2024年8月29日OI：https://doi.org/10.1038/s41477-024-01767-zShare本文与您共享以下链接的任何人都可以阅读此内容：获取可共享链接对不起，本文目前没有可共享的链接。复制到剪贴板。

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