The plant hormones referred to as cytokinins are chemical signals that control numerous developmental processes throughout the plant life cycle, including gametogenesis, root meristem specification, vascular development, shoot and root growth, meristem homeostasis, senescence, and more. In addition, they mediate responses to environmental cues such as light, stress, and nutrient conditions. The core mechanistics of cytokinin metabolism and signaling have been elucidated, but more layers of regulation, additional functions, and interactions with other signals are continuously discovered and described. In this chapter, we recapitulate the highlights of over 100 years of cytokinin research covering its isolation, the elucidation of phosphorelay signaling, and how cytokinin functions in various developmental contexts including its interaction with other pathways. Additionally, given cytokinin's paracrine signaling mechanism, we postulate that cellular exporters for cytokinins exist.

Cytokinin Synthesis, Signaling, and Function—Advances and New Insights

Cytokinin is a multifaceted plant hormone that plays major roles not only in diverse plant growth and development processes, but also stress responses. We summarize knowledge of the roles of its metabolism, transport, and signalling in responses to changes in levels of both macronutrients (nitrogen, phosphorus, potassium, sulphur) and micronutrients (boron, iron, silicon, selenium). We comment on cytokinin’s effects on plants’ xenobiotic resistance, and its interactions with light, temperature, drought, and salinity signals. Further, we have compiled a list of abiotic stress-related genes and demonstrate that their expression patterns overlap with those of cytokinin metabolism and signalling genes.

https://www.mdpi.com/1422-0067/19/8/2450/pdf

Cytokinin at the Crossroads of Abiotic Stress Signalling Pathways

Woody perennials enter seasonal dormancy to avoid unfavorable environmental conditions. Plant hormones are the critical mediators regulating this complex process, which is subject to the influence of many internal and external factors. Over the last two decades, our knowledge of hormone-mediated dormancy has increased considerably, primarily due to advancements in molecular biology, omics, and bioinformatics. These advancements have enabled the elucidation of several aspects of hormonal regulation associated with bud dormancy in various deciduous tree species. Plant hormones interact with each other extensively in a context-dependent manner. The dormancy-associated MADS (DAM) transcription factors appear to enable hormones and other internal signals associated with the transition between different phases of bud dormancy. These proteins likely hold a great potential in deciphering the underlying mechanisms of dormancy initiation, maintenance, and release. In this review, a recent understanding of the roles of plant hormones, their cross talks, and their potential interactions with DAM proteins during dormancy is discussed.

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Hormonal Orchestration of Bud Dormancy Cycle in Deciduous Woody Perennials.

The signal molecules melatonin and ethylene play key roles in abiotic stress tolerance. The interplay between melatonin and ethylene in regulating salt tolerance and the underlying molecular mechanism of this interplay remain unclear. Here, we found that both melatonin and 1-aminocyclopropane-1-carboxylic acid (ACC, a precursor of ethylene) enhanced the tolerance of grapevine to NaCl; additionally, ethylene participated in melatonin-induced salt tolerance. Further experiments indicated that exogenous treatment and endogenous induction of melatonin increased the ACC content and ethylene production in grapevine and tobacco plants, respectively. The expression of MYB108A and ACS1, which function as a transcription factor and a key gene involved in ethylene production, respectively, was strongly induced by melatonin treatment. Additionally, MYB108A directly bound to the promoter of ACS1 and activated its transcription. MYB108A expression promoted ACC synthesis and ethylene production by activating ACS1 expression in response to melatonin treatment. The suppression of MYB108A expression partially limited the effect of melatonin on the induction of ethylene production and reduced melatonin-induced salt tolerance. Collectively, melatonin promotes ethylene biosynthesis and salt tolerance through the regulation of ACS1 by MYB108A.

/pdf/melatonin-enhances-salt-tolerance-by-promoting-myb108a-47zr2bgxpe.pdf

Melatonin enhances salt tolerance by promoting MYB108A -mediated ethylene biosynthesis in grapevines

Sub-optimal growing conditions have a major effect on plants; therefore, large efforts are devoted to maximizing the availability of agricultural inputs to crops. To increase the sustainable use of non-renewable inputs, attention is currently given to the study of plants under non-optimal conditions. In this work, we investigated the impact of sub-optimal macrocations availability and light intensity in two lettuce varieties that differ for the accumulation of secondary metabolites (i.e., 'Red Salanova' and 'Green Salanova'). Photosynthesis-related measurements and untargeted metabolomics were used to identify responses and pathways involved in stress resilience. The pigmented ('Red') and the non-pigmented ('Green Salanova') lettuce exhibited distinctive responses to sub-optimal conditions. The cultivar specific metabolomic signatures comprised a broad modulation of metabolism, including secondary metabolites, phytohormones, and membrane lipids signaling cascade. Several stress-related metabolites were altered by either treatment, including polyamines (and other nitrogen-containing compounds), phenylpropanoids, and lipids. The metabolomics and physiological response to macrocations availability and light intensity also implies that the effects of low-input sustainable farming systems should be evaluated considering a range of cultivar-specific positive and disadvantageous metabolic effects in addition to yield and other socio-economic parameters.

The Metabolic Reprogramming Induced by Sub-Optimal Nutritional and Light Inputs in Soilless Cultivated Green and Red Butterhead Lettuce.

Integrating important environmental signals with intrinsic developmental programmes is a crucial adaptive requirement for plant growth, survival, and reproduction. Key environmental cues include changes in several light variables, while important intrinsic (and highly interactive) regulators of many developmental processes include the phytohormones cytokinins (CKs) and ethylene. Here, we discuss the latest discoveries regarding the molecular mechanisms mediating CK/ethylene crosstalk at diverse levels of biosynthetic and metabolic pathways and their complex interactions with light. Furthermore, we summarize evidence indicating that multiple hormonal and light signals are integrated in the multistep phosphorelay (MSP) pathway, a backbone signalling pathway in plants. Inter alia, there are strong overlaps in subcellular localizations and functional similarities in components of these pathways, including receptors and various downstream agents. We highlight recent research demonstrating the importance of CK/ethylene/light crosstalk in selected aspects of plant development, particularly seed germination and early seedling development. The findings clearly demonstrate the crucial integration of plant responses to phytohormones and adaptive responses to environmental cues. Finally, we tentatively identify key future challenges to refine our understanding of the molecular mechanisms mediating crosstalk between light and hormonal signals, and their integration during plant life cycles.

/pdf/illuminating-light-cytokinin-and-ethylene-signalling-4fcmu3qfmo.pdf

Illuminating light, cytokinin, and ethylene signalling crosstalk in plant development

In plants, the multistep phosphorelay (MSP) pathway mediates a range of regulatory processes, including those activated by cytokinins. The cross talk between cytokinin response and light has been known for a long time. However, the molecular mechanism underlying the interaction between light and cytokinin signaling remains elusive. In the screen for upstream regulators we identified a LONG PALE HYPOCOTYL (LPH) gene whose activity is indispensable for spatiotemporally correct expression of CYTOKININ INDEPENDENT1 (CKI1), encoding the constitutively active sensor His kinase that activates MSP signaling. lph is a new allele of HEME OXYGENASE1 (HY1) that encodes the key protein in the biosynthesis of phytochromobilin, a cofactor of photoconvertible phytochromes. Our analysis confirmed the light-dependent regulation of the CKI1 expression pattern. We show that CKI1 expression is under the control of phytochrome A (phyA), functioning as a dual (both positive and negative) regulator of CKI1 expression, presumably via the phyA-regulated transcription factors (TF) PHYTOCHROME INTERACTING FACTOR3 and CIRCADIAN CLOCK ASSOCIATED1. Changes in CKI1 expression observed in lph/hy1-7 and phy mutants correlate with misregulation of MSP signaling, changed cytokinin sensitivity, and developmental aberrations that were previously shown to be associated with cytokinin and/or CKI1 action. Besides that, we demonstrate a novel role of phyA-dependent CKI1 expression in the hypocotyl elongation and hook development during skotomorphogenesis. Based on these results, we propose that the light-dependent regulation of CKI1 provides a plausible mechanistic link underlying the well-known interaction between light- and cytokinin-controlled plant development.

http://www.plantphysiol.org/content/plantphysiol/174/1/387.full.pdf

Light Controls Cytokinin Signaling via Transcriptional Regulation of Constitutively Active Sensor Histidine Kinase CKI1

Tightly controlled spatiotemporal specificity of gene expression is intrinsic to developmental and adaptation responses of living systems throughout the kingdoms. Forward genetic screens employing well-characterized reporter lines can be used to identify as yet unknown genetic factors driving specific aspects of individual regulatory pathways. However, such screens are demanding with respect to data acquisition and analysis from thousands of mutant lines. Here, we describe a method that allows screening of a mutagenized GUS reporter line in Arabidopsis using an automated microscopy imaging system as a tool for rapid and efficient identification of mutants with modified expression profile for a gene of interest.

Automated microscopy in forward genetic screening of Arabidopsis.

Seedling de-etiolation is one of the key stages of the plant life cycle, characterized by a strong rearrangement of the plant development and metabolism. The conversion of dark accumulated protochlorophyllide to chlorophyll in etioplasts of de-etiolating plants is taking place in order of ns to µs after seedlings illumination, leading to detectable increase of chlorophyll levels in order of minutes after de-etiolation initiation. The highly complex chlorophyll biosynthesis integrates number of regulatory events including light and hormonal signaling, thus making de-etiolation an ideal model to study the underlying molecular mechanisms. Here we introduce the iReenCAM, a novel tool designed for non-invasive fluorescence-based quantitation of early stages of chlorophyll biosynthesis during de-etiolation with high spatial and temporal resolution. iReenCAM comprises customized HW configuration and optimized SW packages, allowing synchronized automated measurement and analysis of the acquired fluorescence image data. Using the system and carefully optimized protocol, we show tight correlation between the iReenCAM monitored fluorescence and HPLC measured chlorophyll accumulation during first 4h of seedling de-etiolation in wild type Arabidopsis and mutants with disturbed chlorophyll biosynthesis. Using the approach, we demonstrate negative effect of exogenously applied cytokinins and ethylene on chlorophyll biosynthesis during early de-etiolation. Accordingly, we identify type-B response regulators, the cytokinin-responsive transcriptional activators ARR1 and ARR12 as negative regulators of early chlorophyll biosynthesis, while contrasting response was observed in case of EIN2 and EIN3, the components of canonical ethylene signaling cascade. Knowing that, we propose the use of iReenCAM as a new phenotyping tool, suitable for quantitative and robust characterization of the highly dynamic response of seedling de-etiolation.

https://www.frontiersin.org/articles/10.3389/fpls.2023.1093292/pdf

iReenCAM: automated imaging system for kinetic analysis of photosynthetic pigment biosynthesis at high spatiotemporal resolution during early deetiolation

Tereza Dobisová

Papers

Illuminating light, cytokinin, and ethylene signalling crosstalk in plant development

Light Controls Cytokinin Signaling via Transcriptional Regulation of Constitutively Active Sensor Histidine Kinase CKI1

Automated microscopy in forward genetic screening of Arabidopsis.

iReenCAM: automated imaging system for kinetic analysis of photosynthetic pigment biosynthesis at high spatiotemporal resolution during early deetiolation