Characterization of the cytokinin sensor TCSv2 in arabidopsis and tomato
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Plant Methods Open Access
METHODOLOGY
Characterization of the cytokinin sensor TCSv2 in arabidopsis and tomato Evyatar Steiner1, Alon Israeli1, Rupali Gupta2, Ido Shwartz1, Ido Nir1,5, Meirav Leibman‑Markus2, Lior Tal3,6, Mika Farber1, Ziva Amsalem3, Naomi Ori1, Bruno Müller4,7 and Maya Bar2*
Abstract Background: Hormones are crucial to plant life and development. Being able to follow the plants hormonal response to various stimuli and throughout developmental processes is an important and increasingly widespread tool. The phytohormone cytokinin (CK) has crucial roles in the regulation of plant growth and development. Results: Here we describe a version of the CK sensor Two Component signaling Sensor (TCS), referred to as TCSv2. TCSv2 has a different arrangement of binding motifs when compared to previous TCS versions, resulting in increased sensitivity in some examined tissues. Here, we examine the CK responsiveness and distribution pattern of TCSv2 in arabidopsis and tomato. Conclusions: The increased sensitivity and reported expression pattern of TCSv2 make it an ideal TCS version to study CK response in particular hosts, such as tomato, and particular tissues, such as leaves and flowers. Keywords: Cytokinin, Imaging, Two-component sensor (TCS), TCSv2, Tomato, Arabidopsis Background Cytokinins (CKs) are a class of adenine-derived plant hormones that control multiple processes throughout the plant life cycle. They provide positional information for growth and patterning, and integrate biotic and abiotic cues from the environment. Notable examples are meristem maintenance in both the shoot apical meristem (SAM) and root apical meristem (RAM), cell division and cell differentiation. CK is also involved in regulating traits that affect yield and fruit quality. The roles of CK in plant growth and development have been reviewed extensively [1–6]. Cytokinin signalling is mediated via a two-component multistep phosphorelay cascade. As the final step, type-B response regulators (RRs) activate transcription in response to phosphorelay signalling activity, while type-A *Correspondence: [email protected] 2 Department of Plant Pathology and Weed Research, Plant Protection Institute, Agricultural Research Organization, The Volcani Center, 7505101 Rishon LeZion, Israel Full list of author information is available at the end of the article
response regulators are rapidly induced by CK via Type-B RRs, and, in turn, repress signalling via a negative-feedback loop [6–10]. There are many CK derivatives, and methods for the detection of a large number of them have emerged in recent years [11–20]. However, it is often difficult to know for certain which of these derivatives represent active CKs, and not all of them are detectable. In parallel to the advances made in hormone substance detection, efforts have also been invested in the detection of CK signalling via transcriptional sensors that mark the site of CK-derived response within a specific tissue or organ. Whereas specific genes and promoters involved in the C
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