Deciphering signal transduction processes is crucial for understanding how plants sense and respond to environmental changes. Various chemical compounds function as central messengers within deeply intertwined signaling networks. How such compounds act in concert remains to be elucidated. We have developed dual-reporting transcriptionally linked genetically encoded fluorescent indicators (2-in-1-GEFIs) for multiparametric in vivo analyses of the phytohormone abscisic acid (ABA), Ca²⁺, protons (H⁺), chloride (anions), the glutathione redox potential, and H₂O₂. Simultaneous analyses of two signaling compounds in Arabidopsis (Arabidopsis thaliana) roots revealed that ABA treatment and uptake did not trigger rapid cytosolic Ca²⁺ or H⁺ dynamics. Glutamate, ATP, Arabidopsis PLANT ELICITOR PEPTIDE, and glutathione disulfide (GSSG) treatments induced rapid spatiotemporally overlapping cytosolic Ca²⁺, H⁺, and anion dynamics, but except for GSSG, only weakly affected the cytosolic redox state. Overall, 2-in-1-GEFIs enable complementary, high-resolution in vivo analyses of signaling compound dynamics and facilitate an advanced understanding of the spatiotemporal coordination of signal transduction processes in Arabidopsis.

破译信号转导过程对于理解植物如何感知和响应环境变化至关重要。各种化合物在紧密交织的信号网络中充当中心信使。这些化合物如何协同作用仍有待阐明。我们开发了双报告转录连锁基因编码荧光指示剂 (2-in-1-GEFI)，用于植物激素脱落酸 (ABA)、Ca ²⁺ 、质子 (H ⁺ )、氯离子（阴离子）、谷胱甘肽氧化还原电位和H ₂ O ₂ 。对拟南芥 ( Arabidopsis thaliana ) 根中两种信号化合物的同时分析表明，ABA 处理和摄取不会触发快速的胞质 Ca ²⁺或 H ⁺动力学。谷氨酸、ATP、拟南芥植物诱导肽和二硫化谷胱甘肽 (GSSG) 处理诱导快速时空重叠的胞质 Ca ²⁺ 、 H ⁺和阴离子动力学，但除 GSSG 外，仅对胞质氧化还原状态产生微弱影响。总体而言，二合一GEFI能够对信号化合物动力学进行互补的高分辨率体内分析，并促进对拟南芥中信号转导过程的时空协调的深入理解。