Fast and simple fluorometric measurement of phloem loading exposes auxin-dependent regulation of Arabidopsis sucrose transporter AtSUC2.

IF 6.2 1区 生物学 Q1 PLANT SCIENCES The Plant Journal Pub Date : 2024-11-01 DOI:10.1111/tpj.17110
Yunjuan Ren, Ziyu Zhang, Diana Zhanakhmetova, Wenhui Li, Shaolin Chen, Tomáš Werner, Johannes Liesche
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Abstract

The rate of sucrose export from leaves is a major factor in balancing whole-plant carbon and energy partitioning. A comprehensive study of its dynamics and relationship to photosynthesis, sink demand, and other relevant processes is hampered by the shortcomings of current methods for measuring sucrose phloem loading. We utilize the ability of sucrose transporter proteins, known as SUCs or SUTs, to specifically transport the fluorescent molecule esculin in a novel assay to measure phloem loading rates. Esculin was administered to source leaves and its fluorescence in the leaf extract was measured after 1 or 2 h. Dicot plants with an active phloem loading strategy showed an export-dependent reduction of esculin fluorescence. Relative leaf esculin export rates correlated with leaf export rates of isotopic carbon and phloem exudate sucrose levels. We used esculin experiments to examine the effects of phytohormones on phloem loading in Arabidopsis, showing, for example, that auxin induces phloem loading while cytokinin reduces it. Transcriptional regulation of AtSUC2 by AUXIN RESPONSE FACTOR1 (ARF1) corroborated the link between auxin signaling and phloem loading. Unlike established methods, the esculin assay is rapid and does not require specialized equipment. Potential applications and limitations of the esculin assay are discussed.

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对韧皮部负荷进行快速、简单的荧光测量,揭示拟南芥蔗糖转运体 AtSUC2 受植物生长素调控的情况。
叶片的蔗糖输出率是平衡整个植物碳和能量分配的一个主要因素。由于目前测量蔗糖韧皮部负载的方法存在缺陷,因此无法全面研究蔗糖韧皮部负载的动态及其与光合作用、吸收汇需求和其他相关过程的关系。我们利用蔗糖转运蛋白(又称 SUC 或 SUT)特异性转运荧光分子 esculin 的能力,采用一种新的测定方法来测量韧皮部负载率。双子叶植物具有活跃的韧皮部负载策略,表现出依赖于输出的埃斯库林荧光减少。叶片的相对esculin输出率与叶片的同位素碳输出率和韧皮部渗出的蔗糖水平相关。我们利用esculin实验研究了植物激素对拟南芥韧皮部负载的影响,结果显示,例如,辅助素会诱导韧皮部负载,而细胞分裂素则会降低韧皮部负载。AUXIN RESPONSE FACTOR1(ARF1)对 AtSUC2 的转录调控证实了叶绿素信号与韧皮部负载之间的联系。与既有方法不同的是,esculin 分析法快速且不需要专门设备。本文讨论了埃斯库灵测定法的潜在应用和局限性。
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来源期刊
The Plant Journal
The Plant Journal 生物-植物科学
CiteScore
13.10
自引率
4.20%
发文量
415
审稿时长
2.3 months
期刊介绍: Publishing the best original research papers in all key areas of modern plant biology from the world"s leading laboratories, The Plant Journal provides a dynamic forum for this ever growing international research community. Plant science research is now at the forefront of research in the biological sciences, with breakthroughs in our understanding of fundamental processes in plants matching those in other organisms. The impact of molecular genetics and the availability of model and crop species can be seen in all aspects of plant biology. For publication in The Plant Journal the research must provide a highly significant new contribution to our understanding of plants and be of general interest to the plant science community.
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