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一种用于监测植物乙烯反应的强大合成报告基因。

a robust synthetic reporter for monitoring ethylene responses in plants.

作者信息

Fernandez-Moreno Josefina-Patricia, Fenech Mario, Yaschenko Anna E, Zhao Chengsong, Neubauer Matthew, Davis Hannah N, Marchi Alex J, Concannon Raine, Keren-Keiserman Alexandra, Reuveni Moshe, Levitsky Victor, Oshchepkov Dmitry, Dolgikh Vladislav, Goldshmidt Alexander, Ascencio-Ibáñez José T, Zemlyanskaya Elena, Alonso Jose M, Stepanova Anna N

机构信息

Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC 27695, USA.

Department of Vegetables and Field Crops, Institute of Plant Science, Agricultural Research Organization, The Volcani Center, 7528809 Rishon Lezion, Israel.

出版信息

bioRxiv. 2025 May 28:2025.05.23.655144. doi: 10.1101/2025.05.23.655144.

Abstract

Ethylene is a gaseous plant hormone that controls a wide array of physiologically relevant processes, including plant responses to biotic and abiotic stress, and induces ripening in climacteric fruits. To monitor ethylene in plants, analytical methods, phenotypic assays, gene expression analysis, and transcriptional or translational reporters are typically employed. In the model plant Arabidopsis, two ethylene-sensitive synthetic transcriptional reporters have been described, and . These reporters harbor a different type, arrangement, and number of homotypic -elements in their promoters and thus may recruit the ethylene master regulator EIN3 in the context of alternative transcriptional complexes. Accordingly, the patterns of GUS activity in these transgenic lines differ and neither of them encompasses all plant tissues even in the presence of saturating levels of exogenous ethylene. Herein, we set out to develop and test a more sensitive version of the ethylene-inducible promoter that we refer to as (abbreviated as ). leverages a tandem of ten non-identical, natural copies of a novel, dual, everted, 11bp-long EIN3-binding site, . We show that in Arabidopsis, outperforms its predecessors in terms of its ethylene sensitivity, having the capacity to monitor endogenous levels of ethylene and displaying more ubiquitous expression in response to the exogenous hormone. We demonstrate that the promoter is also functional in tomato, opening new avenues to manipulating ethylene-regulated processes, such as ripening and senescence, in crops.

摘要

乙烯是一种气态植物激素,它控制着一系列与生理相关的过程,包括植物对生物和非生物胁迫的反应,并诱导跃变型果实成熟。为了监测植物中的乙烯,通常采用分析方法、表型分析、基因表达分析以及转录或翻译报告基因。在模式植物拟南芥中,已经描述了两种乙烯敏感型合成转录报告基因,即[具体名称1]和[具体名称2]。这些报告基因在其启动子中具有不同类型、排列和数量的同型[具体元件名称]元件,因此可能在不同的转录复合物中招募乙烯主调控因子EIN3。相应地,这些转基因系中GUS活性的模式不同,即使在外源乙烯达到饱和水平的情况下,它们也都不能涵盖所有植物组织。在此,我们着手开发并测试一种更敏感的乙烯诱导型启动子版本,我们将其称为[具体名称3](简称为[缩写名称])。[具体名称3]利用了串联的十个不同的、天然的新型双反向11bp长的EIN3结合位点[具体序列]的拷贝。我们表明,在拟南芥中,[具体名称3]在乙烯敏感性方面优于其前身,能够监测乙烯的内源水平,并在外源激素作用下表现出更广泛的表达。我们证明,[具体名称]3启动子在番茄中也具有功能,为调控作物中乙烯调节的过程,如成熟和衰老,开辟了新途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7017/12154813/3d3bd686b7b0/nihpp-2025.05.23.655144v1-f0001.jpg

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