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Phototropin (Phot), a blue light‐sensing LOV domain protein, mediates blue light responses and is evolutionarily conserved across the green lineage. Klebsormidium nitens, a green terrestrial alga, presents a valuable opportunity to study adaptive responses from aquatic to land habitat transitions. We determined the crystal structure of Klebsormidium nitens Phot LOV1 domain (KnLOV1) in the dark and engineered different mutations (R60K, Q122N, and D33N) to modulate the lifetime of the photorecovery cycle. We observed unusual, slow recovery kinetics in the wild‐type KnLOV1 domain (τ = 41 ± 3 min) compared to different mutants (R60K: τ = 2.0 ± 0.1 min, Q122N: τ = 1.7 ± 0.1 min, D33N: τ = 9.6 ± 0.1 min). Crystal structures of wild‐type KnLOV1 and mutants revealed subtle but critical changes near the protein chromophore that is responsible for modulating protein dark recovery time. Our findings shed light on the unique structural and biochemical characteristics of the newly studied KnLOV1 and its evolutionary importance for phototropin‐mediated physiology. [ABSTRACT FROM AUTHOR]

In Chlamydomonas, the directly light‐gated, plasma membrane‐localized cation channels channelrhodopsins ChR1 and ChR2 are the primary photoreceptors for phototaxis. Their targeting and abundance is essential for optimal movement responses. However, our knowledge how Chlamydomonas achieves this is still at its infancy. Here we show that ChR1 internalization occurs via light‐stimulated endocytosis. Prior or during endocytosis ChR1 is modified and forms high molecular mass complexes. These are the solely detectable ChR1 forms in extracellular vesicles and their abundance therein dynamically changes upon illumination. The ChR1‐containing extracellular vesicles are secreted via the plasma membrane and/or the ciliary base. In line with this, ciliogenesis mutants exhibit increased ChR1 degradation rates. Further, we establish involvement of the cysteine protease CEP1, a member of the papain‐type C1A subfamily. ΔCEP1‐knockout strains lack light‐induced ChR1 degradation, whereas ChR2 degradation was unaffected. Low light stimulates CEP1 expression, which is regulated via phototropin, a SPA1 E3 ubiquitin ligase and cyclic AMP. Further, mutant and inhibitor analyses revealed involvement of the small GTPase ARL11 and SUMOylation in ChR1 targeting to the eyespot and cilia. Our study thus defines the degradation pathway of this central photoreceptor of Chlamydomonas and identifies novel elements involved in its homoeostasis and targeting. Summary Statement: How Chlamydomonas targets and controls the level of its main photoreceptor for photomovement responses is largely unknown. Here we show involvement of light‐stimulated endocytosis, cilia, a small GTPase, and a phototropin E3 ubiquitin ligase‐ and cAMP‐controlled cysteine protease in these processes. [ABSTRACT FROM AUTHOR]

Light is a critical environmental signal that is perceived by various photoreceptors and is of great significance in the photosynthetic growth of algae and plants. The phytochrome‐lacking model green alga Chlamydomonas reinhardtii possesses heme oxygenase (HMOX1) and phycocyanobilin ferredoxin oxidoreductase (PCYA1) to synthesize the linear tetrapyrrole bilin from heme in the chloroplast. The hmox1 mutant has photosynthetic growth deficiency and accumulation of photosystem I proteins such as LHCA1 is severely inhibited, and these defects could be rescued by exogenous bilin feeding in a blue light‐dependent manner. To investigate the contribution of the typical blue/ultraviolet light photoreceptors PHOT, aCRY, pCRY, and UVR8 in the process of bilin and blue light‐dependent recovery of LHCA1 protein in hmox1, we generated double mutants of these photoreceptors in hmox1, as well as a triple mutant of phot uvr8 hmox1, to analyze the LHCA1 protein abundance in these mutants. Results clearly showed that PHOT, CRYs, and UVR8 do not participate in this process. In addition, transcriptome profiling analysis of the hmox1 and its genetically complemented strain ho1C2 during dark‐to‐blue light transition revealed a total of 269 blue light‐responsive genes independent of bilin (|fold change| ≥ 2). RNA‐seq also identified a set of 249 differentially expressed genes that are dependent on both blue light and bilin. These findings provide valuable insights for elucidating the role of bilin in mediating blue light signaling pathways in Chlamydomonas. [ABSTRACT FROM AUTHOR]

Phototropin (phot) is a blue light (BL) receptor and thermosensor that mediates chloroplast movements in plants. Liverworts, as early‐diverging plant species, have a single copy of PHOT gene, and the phot protein in each liverwort activates the signaling pathway adapted to its specific growing environment. In this study, we functionally compared phot from two different liverworts species: Apopellia endiviifolia (Aephot) and Marchantia polymorpha (Mpphot). The BL‐dependent photochemical activity of Aephot was similar to that of Mpphot, whereas the thermochemical activity of Aephot was lower than that of Mpphot. Therefore, the phot‐mediated signaling pathways of the two plant species may differ more in response to temperature than to BL. Furthermore, we analyzed the functional compatibility of Aephot and Mpphot in chloroplast movements by transiently expressing AePHOT or MpPHOT. The transient expression of AePHOT did not mediate chloroplast movement in M. polymorpha, showing the incompatibility of Aephot with the signaling pathway of M. polymorpha. By contrast, the transient expression of MpPHOT mediated chloroplast movement in A. endiviifolia, indicating the compatibility of Mpphot with the signaling pathway of A. endiviifolia. Our findings reveal both functional similarities and differences between Aephot and Mpphot proteins from the closely related liverworts. [ABSTRACT FROM AUTHOR]

Many ferns thrive even in low-light niches such as under an angiosperm forest canopy. However, the shade adaptation strategy of ferns is not well understood. Phytochrome 3/neochrome (phy3/neo) is an unconventional photoreceptor, found in the fern Adiantum capillus-veneris , that controls both red and blue light-dependent phototropism and chloroplast photorelocation, which are considered to improve photosynthetic efficiency in ferns. Here we show that phy3/neo localizes not only at the plasma membrane but also in the nucleus. Since both phototropism and chloroplast photorelocation are mediated by membrane-associated phototropin photoreceptors, we speculated that nucleus-localized phy3/neo possesses a previously undescribed biological function. We reveal that phy3/neo directly interacts with Adiantum cryptochrome 3 (cry3) in the nucleus. Plant cryptochromes are blue light receptors that transcriptionally regulate photomorphogenesis; therefore, phy3/neo may function via cry3 to synchronize light-mediated development with phototropism and chloroplast photorelocation to promote fern growth under low-light conditions. Furthermore, we demonstrate that phy3/neo regulates the expression of the Cyclin-like gene AcCyc1 and promotes prothallium expansion growth. These findings provide insight into the shade adaptation strategy of ferns and suggest that phy3/neo plays a substantial role in the survival and growth of ferns during the tiny gametophytic stage under low-light conditions, such as those on the forest floor. [ABSTRACT FROM AUTHOR]

Summary: Leaves are the main photosynthetic organs in plants, and their anatomy is optimized for light interception and gas exchange. Although each species has a characteristic leaf anatomy, which depends on the genotype, leaves also show a large degree of developmental plasticity. Light and temperature regulate leaf development from primordia differentiation to late stages of blade expansion. While the molecular mechanisms of light and temperature signaling have been mostly studied in seedlings, in the latest years, research has focused on leaf development. Here, I will describe the latest work carried out in the environmental regulation of Arabidopsis leaf development, comparing signaling mechanisms between leaves and seedlings, highlighting the new discoveries, and pointing out the most exciting open questions. [ABSTRACT FROM AUTHOR]

Oriented movement (phototaxis) is an efficient way to optimize light‐driven processes and to avoid photodamage for motile algae. In Chlamydomonas the receptors for phototaxis are the channelrhodopsins ChR1 and ChR2. Both are directly light‐gated, plasma membrane‐localized cation channels. To optimally adjust its overall light‐dependent responses, Chlamydomonas must tightly control the ChRs cellular abundance and integrate their activities into its general photoprotective network. How this is achieved is largely unknown. Here we show that the ChR1 protein level decreases upon illumination in a light‐intensity and quality‐dependent manner, whereas it is stable in prolonged darkness. Analysis of knockout strains of six major photoreceptors absorbing in the blue–violet range, which is most effective in evoking ChR1 degradation, revealed that only phototropin (PHOT) is involved. Notably, ChR2 degradation was normal in a ΔPHOT strain. Further, our results indicate that a COP1‐SPA1 E3 ubiquitin ligase, the transcription factor Hy5 as well as changes in the cellular redox poise and cyclic nucleotide levels are additional components involved in this light acclimation response of Chlamydomonas. Our data highlight the presence of an adaptive framework connecting phototaxis with general photoprotective mechanisms via the use of overlapping signaling components already at the level of the primary photoreceptor. Summary Statement: How Chlamydomonas controls the level of ChR1, the main photoreceptor for its movement responses, is largely unknown. Here we show that regulatory elements involved in photoprotective gene expression are parts of this cascade(s), supporting an integrating link between phototaxis and photoprotection. [ABSTRACT FROM AUTHOR]

SUMMARY: Directional movements impact the ability of plants to respond and adjust their growth accordingly to the prevailing light environment. The plasma‐membrane associated protein, ROOT PHOTOTROPISM 2 (RPT2) is a key signalling component involved in chloroplast accumulation movement, leaf positioning, and phototropism, all of which are regulated redundantly by the ultraviolet/blue light‐activated AGC kinases phototropin 1 and 2 (phot1 and phot2). We recently demonstrated that members of the NON‐PHOTOTROPIC HYPOCOTYL 3 (NPH3)/RPT2‐like (NRL) family in Arabidopsis thaliana, including RPT2, are directly phosphorylated by phot1. However, whether RPT2 is a substrate for phot2, and the biological significance of phot phosphorylation of RPT2 remains to be determined. Here, we show that RPT2 is phosphorylated by both phot1 and phot2 at a conserved serine residue (S591) within the C‐terminal region of the protein. Blue light triggered the association of 14‐3‐3 proteins with RPT2 consistent with S591 acting as a 14‐3‐3 binding site. Mutation of S591 had no effect on the plasma membrane localization of RPT2 but reduced its functionality for leaf positioning and phototropism. Moreover, our findings indicate that S591 phosphorylation within the C‐terminus of RPT2 is required for chloroplast accumulation movement to low level blue light. Taken together, these findings further highlight the importance of the C‐terminal region of NRL proteins and how its phosphorylation contributes to phot receptor signalling in plants. Significance Statement: ROOT PHOTOTROPISM 2 (RPT2) is a key signalling component involved in chloroplast accumulation movement, leaf positioning, and phototropism, which are regulated by phototropin receptor kinases (phot1 and phot2). Here we demonstrate that RPT2 is phosphorylated by phot1 and phot2 at S591 within the C‐terminus and phosphorylation of this residue plays a role in establishing phototropism, leaf positioning and is functionally required for RPT2 to mediate chloroplast movement to low levels of blue light. [ABSTRACT FROM AUTHOR]

Plants recognize the direction of a light source and exhibit phototropic responses. Physiological studies have predicted that differences in the light intensity received by the cells on the irradiated and shaded sides of a coleoptile or hypocotyl cause differences in the amounts of photoproduct. This hypothetical photoproduct appears to regulate a signaling pathway that controls cell elongation in which cells under lower light intensity elongate more than those under higher light intensity. This results in a bending growth toward a light source and has been proposed as the photoproduct-gradient model of phototropism. In this review, we summarize recent findings on the photosensory adaptation mechanisms involving a blue-light photoreceptor, phototropin1 (phot1), ROOT PHOTOTROPISM2, NONPHOTOTROPIC HYPOCOTYL3 (NPH3), and another photoreceptor family, the phytochromes. The current evidence demonstrates that, in addition to the transition of the phot1–NPH3 photoreceptor complexes to their active state, the presence of a certain population of the phot1–NPH3 complexes showing a steady state, even in a light environment, is essential for recognition of the light source direction in phototropism. This is consistent with the photoproduct-gradient model, and a dissociation state of the phot1–NPH3 complex would be considered an entity of the hypothetical photoproduct in this model. [ABSTRACT FROM AUTHOR]

Summary: Green plants are equipped with photoreceptors that are capable of sensing radiation in the ultraviolet‐to‐blue and the red‐to‐far‐red parts of the light spectrum. However, plant cells are not particularly sensitive to green light (GL), and light which lies within this part of the spectrum does not efficiently trigger the opening of stomatal pores. Here, we discuss the current knowledge of stomatal responses to light, which are either provoked via photosynthetically active radiation or by specific blue light (BL) signaling pathways. The limited impact of GL on stomatal movements provides a unique option to use this light quality to control optogenetic tools. Recently, several of these tools have been optimized for use in plant biological research, either to control gene expression, or to provoke ion fluxes. Initial studies with the BL‐activated potassium channel BLINK1 showed that this tool can speed up stomatal movements. Moreover, the GL‐sensitive anion channel GtACR1 can induce stomatal closure, even at conditions that provoke stomatal opening in wild‐type plants. Given that crop plants in controlled‐environment agriculture and horticulture are often cultivated with artificial light sources (i.e. a combination of blue and red light from light‐emitting diodes), GL signals can be used as a remote‐control signal that controls stomatal transpiration and water consumption. [ABSTRACT FROM AUTHOR]

Abstract Background Many regulatory circuits in plants contain steps of targeted proteolysis, with the ubiquitin proteasome system (UPS) as the mediator of these proteolytic events. In order to decrease ubiquitin-dependent proteolysis, we inducibly expressed a ubiquitin variant with Arg at position 48 instead of Lys (ubK48R). This variant acts as an inhibitor of proteolysis via the UPS, and allowed us to uncover processes that are particularly sensitive to UPS perturbation. Results Expression of ubK48R during germination leads to seedling death. We analyzed the seedling transcriptome, proteome and metabolome 24 h post ubK48R induction and confirmed defects in chloroplast development. We found that mutations in single genes can suppress seedling lethality, indicating that a single process in seedlings is critically sensitive to decreased performance of the UPS. Suppressor mutations in phototropin 2 (PHOT2) suggest that a contribution of PHOT2 to chloroplast protection is compromised by proteolysis inhibition. Conclusions Overall, the results reveal protein turnover as an integral part of a signal transduction chain that protects chloroplasts during development.

Photosensory proteins known as photoreceptors (PHRs) are crucial for delineating light environments in synchronization with other environmental cues and regulating their physiological variables in plants. However, this has not been well studied in the Brassica genus, which includes several important agricultural and horticultural crops. Herein, we identified five major PHR gene families—phytochrome (PHY), cryptochrome (CRY), phototropin (PHOT), F-box containing flavin binding proteins (ZTL/FKF1/LKP2), and UV RESISTANCE LOCUS 8 (UVR8)—genomic scales and classified them into subfamilies based on their phylogenetic clustering with Arabidopsis homologues. The molecular evolution characteristics of Brassica PHR members indicated indirect expansion and lost one to six gene copies at subfamily levels. The segmental duplication was possibly the driving force of the evolution and amplification of Brassica PHRs. Gene replication retention and gene loss events of CRY, PHY, and PHOT members found in diploid progenitors were highly conserved in their tetraploid hybrids. However, hybridization events were attributed to quantitative changes in UVR8 and ZTL/FKF1/LKP2 members. All PHR members underwent purifying selection. In addition, the transcript expression profiles of PHR genes in different tissue and in response to exogenous ABA, and abiotic stress conditions suggested their multiple biological significance. This study is helpful in understanding the molecular evolution characteristics of Brassica PHRs and lays the foundation for their functional characterization. [ABSTRACT FROM AUTHOR]

Chloroplasts accumulate on the cell surface under weak light conditions to efficiently capture light but avoid strong light to minimize photodamage. The blue light receptor phototropin regulates the chloroplast movement in various plant species. In Arabidopsis thaliana, phototropin mediates the light-induced chloroplast movement and positioning via specialized actin filaments on the chloroplasts, chloroplast-actin filaments. KINESIN-LIKE PROTEIN FOR ACTIN-BASED CHLOROPLAST MOVEMENT (KAC) and CHLOROPLAST UNUSUAL POSITIONING 1 (CHUP1) are pivotal for chloroplast-actin-based chloroplast movement and positioning in land plants. However, the mechanisms by which KAC and CHUP1 regulate chloroplast movement and positioning remain unclear. In this study, we characterized KAC and CHUP1 orthologs in the liverwort Marchantia polymorpha, MpKAC and MpCHUP1, respectively. Their knockout mutants, Mpkack° and Mpchup1k°, impaired the light-induced chloroplast movement. Although Mpchup1k° showed mild chloroplast aggregation, Mpkack° displayed severe chloroplast aggregation, suggesting the greater contribution of MpKAC to the chloroplast anchorage to the plasma membrane. Analysis of the subcellular localization of the functional MpKAC-Citrine indicated that MpKAC-Citrine formed a punctate structure on the plasma membrane. Structure-function analysis of MpKAC revealed that a deletion of the conserved C-terminal domain abrogates the targeting to the plasma membrane and its function. A deletion of the N-terminal motor domain retained the plasma membrane targeting but abrogates the formation of punctate structure and showed severe defect in the light-induced chloroplast movement. Our findings suggest that the formation of the punctate structure on the plasma membrane of MpKAC is essential for chloroplast movement., (© The Author(s) 2024. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site–for further information please contact journals.permissions@oup.com.)

SUMMARY: Plant cells alter the intracellular positions of chloroplasts to ensure efficient photosynthesis, a process controlled by the blue light receptor phototropin. Chloroplasts migrate toward weak light (accumulation response) and move away from excess light (avoidance response). Chloroplasts are encircled by the endoplasmic reticulum (ER), which forms a complex network throughout the cytoplasm. To ensure rapid chloroplast relocation, the ER must alter its structure in conjunction with chloroplast relocation movement, but little is known about the underlying mechanism. Here, we searched for interactors of phototropin in the liverwort Marchantia polymorpha and identified a RETICULON (RTN) family protein; RTN proteins play central roles in ER tubule formation and ER network maintenance by stabilizing the curvature of ER membranes in eukaryotic cells. Marchantia polymorpha RTN1 (MpRTN1) is localized to ER tubules and the rims of ER sheets, which is consistent with the localization of RTNs in other plants and heterotrophs. The Mprtn1 mutant showed an increased ER tubule diameter, pointing to a role for MpRTN1 in ER membrane constriction. Furthermore, Mprtn1 showed a delayed chloroplast avoidance response but a normal chloroplast accumulation response. The live cell imaging of ER dynamics revealed that ER restructuring was impaired in Mprtn1 during the chloroplast avoidance response. These results suggest that during the chloroplast avoidance response, MpRTN1 restructures the ER network and facilitates chloroplast movement via an interaction with phototropin. Our findings provide evidence that plant cells respond to fluctuating environmental conditions by controlling the movements of multiple organelles in a synchronized manner. Significance Statement: Although chloroplasts are encircled by the endoplasmic reticulum (ER), they move rapidly within a cell in response to light to ensure efficient photosynthesis, but the regulatory mechanism of the ER structure during chloroplast movement remains unknown. Here, we reveal that in Marchantia polymorpha, the blue light receptor phototropin, which mediates chloroplast movement, interacts with an ER membrane‐bending protein reticulon (MpRTN1), and that MpRTN1 restructures the ER network and facilitates phototropin‐mediated chloroplast movement. [ABSTRACT FROM AUTHOR]

Background: Many regulatory circuits in plants contain steps of targeted proteolysis, with the ubiquitin proteasome system (UPS) as the mediator of these proteolytic events. In order to decrease ubiquitin-dependent proteolysis, we inducibly expressed a ubiquitin variant with Arg at position 48 instead of Lys (ubK48R). This variant acts as an inhibitor of proteolysis via the UPS, and allowed us to uncover processes that are particularly sensitive to UPS perturbation. Results: Expression of ubK48R during germination leads to seedling death. We analyzed the seedling transcriptome, proteome and metabolome 24 h post ubK48R induction and confirmed defects in chloroplast development. We found that mutations in single genes can suppress seedling lethality, indicating that a single process in seedlings is critically sensitive to decreased performance of the UPS. Suppressor mutations in phototropin 2 (PHOT2) suggest that a contribution of PHOT2 to chloroplast protection is compromised by proteolysis inhibition. Conclusions: Overall, the results reveal protein turnover as an integral part of a signal transduction chain that protects chloroplasts during development. [ABSTRACT FROM AUTHOR]

Photosensor proteins are important not only because of their biological functions but also because of their applications in optogenetics. To understand the molecular mechanism behind their biological functions and consequently seek possible applications to optogenetics, the dynamics of their intermolecular interaction (for example, association/dissociation reaction and conformational changes) upon photoexcitation need to be elucidated. Although it has been difficult to trace such reactions in the time domain using traditional spectroscopic techniques, the time-resolved diffusion method based on the transient grating technique has been demonstrated to possess a significant advantage in detecting such spectrally silent dynamics in a time-resolved manner. In this paper, the principle and studies on blue light sensor proteins, phototropins, are described. Reaction kinetics of dimerization, dissociation reactions, and conformational changes were measured, and reaction schemes were determined. This method can be employed to study protein reactions from the viewpoint of diffusion and to elucidate the reaction schemes and kinetics that cannot be detected by other spectroscopic methods. [ABSTRACT FROM AUTHOR]

Plants growing in dense vegetation stands need to flexibly position their photosynthetic organs to ensure optimal light capture in a competitive environment. They do so through a suite of developmental responses referred to as the shade avoidance syndrome. Belowground, root development is also adjusted in response to aboveground neighbour proximity. Canopies are dynamic and complex environments with heterogenous light cues in the far-red, red, blue and UV spectrum, which can be perceived with photoreceptors by spatially separated plant tissues. Molecular regulation of plant architecture adjustment via PHYTOCHROME-INTERACTING FACTOR (PIF) transcription factors and growth-related hormones such as auxin, gibberellic acid, brassinosteroids and abscisic acid were historically studied without much attention to spatial or tissue-specific context. Recent developments and technologies have, however, sparked strong interest in spatially explicit understanding of shade avoidance regulation. Other environmental factors such as temperature and nutrient availability interact with the molecular shade avoidance regulation network, often depending on the spatial location of the signals, and the responding organs. Here, we aim to review recent advances in how plants respond to heterogenous light cues and integrate these with other environmental signals., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

This document is a profile of Martina Legris, a postdoctoral researcher at the Center for Integrative Genomics at the University of Lausanne in Switzerland. Martina's interest in plant science was inspired by her upbringing in a family of veterinarians and farmers, as well as her experiences playing with plants and animals in nature. She decided to pursue a career in research after participating in scientific meetings and outreach activities, where she met inspiring scientists and learned about their research. Martina is motivated by her love for her work and the sense of duty to contribute to society. She considers her mentors and female colleagues with children to be role models, and she appreciates the opportunity to choose her research topics and the variety of tasks in her everyday life. Martina's research focuses on plant development in response to light and temperature signals from the environment. She has published papers on the convergence of light and temperature signaling pathways in plants and the regulation of meristem activity. Martina's favorite plant is Ceiba speciosa, a large tree native to South America, and she also has a fondness for Arabidopsis thaliana, a tiny plant that has allowed her to uncover exciting aspects of plant biology. [Extracted from the article]

Hypocotyl phototropism is redundantly mediated by phot1 and phot2, two blue light receptor phototropins, under the intensity of blue light>1 μmol m−2 s−1. As light intensity increases, phot1 inhibits the phot2-mediated response. To date, only Arabidopsis Root Phototropism2 (RPT2) has been shown to participate in phot1-mediated inhibition of phototropism. To dissect the signaling network that underlies phot1-mediated inhibition, we carried out a yeast two-hybrid (Y2H) screening assay for RPT2 interacting proteins and identified J-domain protein required for chloroplast accumulation response 1 (JAC1). The interaction between JAC1 and RPT2 was verified by bimolecular fluorescence complementation and Co-IP assays. JAC1 is expressed mainly in cotyledons and hypocotyls. Like RPT2 , JAC1 can be induced by blue light, suggesting that it may function similarly to RPT2 in the inhibition of phototropism. Genetic analysis showed that jac1 mutation significantly enhanced the hypocotyl bending of phot1 mutants towards intermediate-intensity blue light, and this effect was inhibited by the constitutive expression of JAC1 in the phot1 jac1 mutant. The phot1 rpt 2 double mutant also exhibited enhanced phototropism compared with the phot1 mutant. Taken together, our data clearly demonstrate that JAC1 cooperates with RPT2 to negatively regulate hypocotyl phototropism in plants and may act either downstream of or in parallel with phot1. • JAC1 interacts with RPT2. • JAC1 is expressed mainly in cotyledons and hypocotyls, similarly with RPT2 induced by blue light. • Mutations in JAC1 or RPT2 caused an enhanced hypocotyl bending towards intermediate-intensity blue light in phot1 mutant. • JAC1 acts downstream of RPT2 to inhibit phot2-mediated phototropism. [ABSTRACT FROM AUTHOR]

To efficiently use light for photosynthesis, chloroplasts move to the appropriate location according to ambient light conditions. Chloroplasts relocate towards dimly lit cell parts to maximise light capture (the accumulation response), whereas they escape from the strong light to minimise photodamage (the avoidance response). In most plant species, blue light is the most effective to induce chloroplast movements. Since the first report of light-induced chloroplast movement in 1856, many researchers have tried to elucidate the molecular mechanism underlying chloroplast movement including the photoreceptor system, the signal transduction and the motility system. However, the molecular mechanism remained unknown until a molecular genetic analysis using Arabidopsis thaliana as a model plant has been applied in the analysis of chloroplast movement. In this article, we review physiological and molecular analyses that have studied signal transduction of light-induced chloroplast movement. [ABSTRACT FROM AUTHOR]

Phototropism represents a simple physiological mechanism—differential growth across the growing organ of a plant—to respond to gradients of light and maximize photosynthetic light capture (in aerial tissues) and water/nutrient acquisition (in roots). The phototropin blue light receptors, phot1 and phot2, have been identified as the essential sensors for phototropism. Additionally, several downstream signal/response components have been identified, including the phot-interacting proteins NON-PHOTOTROPIC HYPOCOTYL 3 (NPH3) and PHYTOCHROME SUBSTRATE 4 (PKS4). While the structural and photochemical properties of the phots are quite well understood, much less is known about how the phots signal through downstream regulators. Recent advances have, however, provided some intriguing clues. It appears that inactive receptor phot1 is found dispersed in a monomeric form at the plasma membrane in darkness. Upon light absorption dimerizes and clusters in sterol-rich microdomains where it is signal active. Additional studies showed that the phot-regulated phosphorylation status of both NPH3 and PKS4 is linked to phototropic responsiveness. While PKS4 can function as both a positive (in low light) and a negative (in high light) regulator of phototropism, NPH3 appears to function solely as a key positive regulator. Ultimately, it is the subcellular localization of NPH3 that appears crucial, an aspect regulated by its phosphorylation status. While phot1 activation promotes dephosphorylation of NPH3 and its movement from the plasma membrane to cytoplasmic foci, phot2 appears to modulate relocalization back to the plasma membrane. Together these findings are beginning to illuminate the complex biochemical and cellular events, involved in adaptively modifying phototropic responsiveness under a wide varying range of light conditions. [ABSTRACT FROM AUTHOR]

Premise: While essential for photosynthesis, excess light can damage plants. We investigated how growth light conditions affect two photoprotective strategies, chloroplast movement and nonphotochemical quenching (NPQ), as well as electron transport rates (ETR), and the relative importance of these processes in the short‐term stress tolerance of Arabidopsis thaliana. Methods: We grew wild‐type (WT) and mutant plants with impaired chloroplast movement (phot1, phot2, phot1 phot2, chup1) or NPQ (npq1) at low (160 μmol photons m−2 s−1) or intermediate light (400 μmol photons m−2 s−1) before quantifying transmission changes due to chloroplast movement, NPQ, ETR, and the ability to recover from a short‐term high‐light treatment. Results: Plants with impaired chloroplast avoidance movement (phot2, phot1 phot2, chup1) did not recover as well from a short‐term high light treatment as the WT or npq1 and phot1 mutants. Plants grown at intermediate light recovered more completely from the same stress treatment regardless of their genotype and despite reduced degrees of transmission changes due to chloroplast movement. This result was due in part to all genotypes having up to a 2‐fold increase in ETRmax and a slight increase in NPQmax. Conclusions: Growth light conditions affect which mechanisms are important in dealing with short‐term high‐light stress. The chloroplast avoidance response is important for low‐light‐grown plants, while increases in ETRmax and NPQmax allow plants grown at intermediate light intensities to avoid being damaged. [ABSTRACT FROM AUTHOR]

Abstract The blue‐light photoreceptor kinase phototropin (phot) mediates chloroplast movement in response to light and temperature. Phot predominantly localizes at the plasma membrane, but also resides in the cytosol and the chloroplast periphery. Although the phot localized to the chloroplast periphery is thought to mediate chloroplast movement, the localization mechanism is unknown. In this study, we found that chloroplast movement does not occur in 0‐day‐old gemma cells of the liverwort Marchantia polymorpha but that the movement is induced in 1‐day‐old gemmaling cells. Along with this physiological change, the subcellular localization of phot also changed: In 0‐day‐old gemma cells, phot localized at the plasma membrane and the cytosol, but in 1‐day‐old gemmaling cells, the phot disappeared from the cytosol and appeared at the chloroplast periphery. When the relocalization was tracked using a photoconvertible fluorescent protein, the cytosolic phot relocated to the plasma membrane, and the plasma membrane‐resident phot relocated to the chloroplast periphery. The blue‐light‐dependent activation of phot kinase activity enhanced this relocalization. Mutated phot deficient in blue‐light reception or kinase activity had a severely reduced ability to localize at the chloroplast periphery. These findings suggest that photoactivated phot localizes at the chloroplast periphery to initiate chloroplast movement.

Senescence is the final stage of plant development, affecting individual organs or the whole organism, and it can be induced by several environmental factors, including shading or darkness. Although inevitable, senescence is a complex and tightly regulated process, ensuring optimal remobilization of nutrients and cellular components from senescing organs. Photoreceptors such as phytochromes and cryptochromes are known to participate in the process of senescence, but the involvement of phototropins has not been studied to date. We investigated the role of these blue light photoreceptors in the senescence of individually darkened Arabidopsis thaliana leaves. We compared several physiological and molecular senescence markers in darkened leaves of wild-type plants and phototropin mutants (phot1, phot2, and phot1phot2). In general, all the symptoms of senescence (lower photochemical activity of photosystem II, photosynthetic pigment degradation, down-regulation of photosynthetic genes, and up-regulation of senescence-associated genes) were less pronounced in phot1phot2, as compared to the wild type, and some also in one of the single mutants, indicating delayed senescence. This points to different mechanisms of phototropin operation in the regulation of senescence-associated processes, either with both photoreceptors acting redundantly, or only one of them, phot1, playing a dominant role.

The ability to enhance photosynthetic capacity remains a recognized bottleneck to improving plant productivity. Phototropin blue light receptors (phot1 and phot2) optimize photosynthetic efficiency in Arabidopsis thaliana by coordinatingmultiple light-capturing processes. In this study, we explore the potential of using protein engineering to improve photoreceptor performance and thereby plant growth. We demonstrate that targeted mutagenesis can decrease or increase the photocycle lifetime of Arabidopsis phototropins in vitro and show that these variants can be used to reduce or extend the duration of photoreceptor activation in planta. Our findings show that slowing the phototropin photocycle enhanced several light-capturing responses, while accelerating it reduced phototropin's sensitivity for chloroplast accumulation movement. Moreover, plants engineered to have a slow-photocycling variant of phot1 or phot2 displayed increased biomass production under low-light conditions as a consequence of their improved sensitivity. Together, these findings demonstrate the feasibility of engineering photoreceptors to manipulate plant growth and offer additional opportunities to enhance photosynthetic competence, particularly under suboptimal light regimes. [ABSTRACT FROM AUTHOR]

The haploid gametophyte generation of ferns is an excellent experimental material for cell biology studies because of its simple structure and high sensitivity to light. Each step of the developmental process, such as cell growth, cell cycle and the direction of cell division, is controlled, step by step, by light, unlike what happens in complex seed plant tissues. To perform analyses at the cell or organelle level, we have developed special tools, instruments and techniques, such as a cuvette suitable for repeated centrifugation in particular directions, microbeam irradiators for partial cell irradiation and single-cell ligation technique to create enucleated cells. Some of our main discoveries are as follows: (1) changes in the intracellular position of the nucleus in long protonemal cells by centrifugation revealed that the nuclear position or a factor(s) that is/are co-centrifuged with the nucleus is important for the decision regarding the place of the formation of preprophase bands and the timing of their disappearance, which determines the position where the new cell wall attaches to the mother cell wall; (2) even within a single cell, various phenomena could be induced by blue or red light, with the localization of the blue or red light receptors being different depending on the phenomenon; (3) de novo mRNA synthesis is not involved in the signal transduction pathways underlying light-induced chloroplast movements. In this review article, various microscopic techniques, in addition to the results of physiology studies in fern gametophytes, are described. [ABSTRACT FROM AUTHOR]

Although plant responses to artificial lighting spectra often produce abnormal morphogenesis and reduced productivity, no quantification method to determine how plants perceive and respond to light has been available. Our objective in this study was to test whether a plant’s spectral perception can be quantified using the light absorption of its major photoreceptors, phytochrome, cryptochrome, and phototropin. We developed an artificial solar lamp and three different light sources, based on a high-pressure sodium lamp, a fluorescent lamp, and red and blue light-emitting diodes, whose absorption by photoreceptors was equal to that of the standard solar spectrum. Cucumber plants grown under the artificial solar and developed light sources showed normal photomorphogenesis and were indistinguishable from each other. Plants grown under unmodified commercial light sources had abnormal photomorphogenesis that made them short and small. The photosynthetic rate was higher under the unmodified light sources; however, dry masses were highest under the artificial solar and modified light sources, indicating that the cucumber plants are optimized to the solar spectrum. Our results clearly demonstrate that the spectral perceptions of plants can be quantified using the light absorption of their photoreceptors, not visual color or spectra. We expect that our findings will contribute to a better understanding of plant perceptions of and responses to light quality, and improve the productivity of plants cultivated under artificial light.

Phototropins (phot1 and phot2) are blue-light receptors that control cotyledon flattening and positioning under strong light; however, their functional redundancy restricts our understanding of the specific roles of phot2. To identify the factors responsible for phot2-dependent cotyledon flattening and growth, we screened for light-insensitive mutants among mutagenized phot1 mutants in Arabidopsis thaliana. The double mutant phot1lea1 (leaf expansion associated 1), which is defective in cotyledon flattening and positioning but not the phototropic response was selected. This mutant phenotype could be alleviated by constitutively expressing MORE AXILLARY GROWTH 2 (MAX2), indicating that LEA1 was allelic to MAX2. The max2 mutants (max2-2 and max2-3) are defective in cotyledon flattening, which is similar to that of the phot1 phot2 mutants. Moreover, the amounts of MAX2 transcripts are inhibited in leaves of phot1 mutant. However, the additional disruption of PHOT1 gene in max2-2 or max2-3 did not affect their phenotype, including MAX2-mediated inhibition of hypocotyl elongation. By contrast, phototropins-mediated hypocotyl phototropism was not regulated by MAX2. Together, these results suggest that cotyledon flattening was mediated by both phototropins and MAX2 signaling, but the relationship between two pathways need further study.

Phototropins (phot1 and phot2) are blue-light receptors that control cotyledon flattening and positioning under strong light; however, their functional redundancy restricts our understanding of the specific roles of phot2. To identify the factors responsible for phot2-dependent cotyledon flattening and growth, we screened for light-insensitive mutants among mutagenized phot1 mutants in Arabidopsis thaliana. The double mutant phot1 lea1 (leaf expansion associated 1), which is defective in cotyledon flattening and positioning but not the phototropic response was selected. This mutant phenotype could be alleviated by constitutively expressing MORE AXILLARY GROWTH 2 (MAX2), indicating that LEA1 was allelic to MAX2. The max2 mutants (max2-2 and max2-3) are defective in cotyledon flattening, which is similar to that of the phot1 phot2 mutants. Moreover, the amounts of MAX2 transcripts are inhibited in leaves of phot1 mutant. However, the additional disruption of PHOT1 gene in max2-2 or max2-3 did not affect their phenotype, including MAX2-mediated inhibition of hypocotyl elongation. By contrast, phototropins-mediated hypocotyl phototropism was not regulated by MAX2. Together, these results suggest that cotyledon flattening was mediated by both phototropins and MAX2 signaling, but the relationship between two pathways need further study. [ABSTRACT FROM AUTHOR]

Background: Phototropism is the response a plant exhibits when it is faced with a directional blue light stimulus. Though a seemingly simple differential cell elongation response within a responding tissue that results in organ curvature, phototropism is regulated through a complex set of signal perception and transduction events that move from the plasma membrane to the nucleus. In nature phototropism is one of several plant responses that have evolved to optimize photosynthesis and growth.Objective: In the present work we will review the state of the field with respect to the molecules and mechanisms associated with phototropism in land plants.Methods: A systematic literature search was done to identify relevant advances in the field. Though we tried to focus on literature within the past decade (1998-present), we have discussed and cited older literature where appropriate because of context or its significant impact to the present field. Several previous review articles are also cited where appropriate and readers should seek those out.Results: A total of 199 articles are cited that fulfill the criteria listed above.Conclusions: Though important numerous and significant advances have been made in our understanding of the molecular, biochemical, cell biological and physiologic mechanisms underlying phototropism in land plants over the past decade, there are many remaining unanswered questions. The future is indeed bright for researchers in the field and we look forward to the next decade worth of discoveries. [ABSTRACT FROM AUTHOR]

Low light (LL) and high light (HL)-acclimated plants of A. thaliana were exposed to blue (BB) or red (RR) light or to a mixture of blue and red light (BR) of incrementally increasing intensities. The light response of photosystem II was measured by pulse amplitude-modulated chlorophyll fluorescence and that of photosystem I by near infrared difference spectroscopy. The LL but not HL leaves exhibited blue light-specific responses which were assigned to relocation of chloroplasts from the dark to the light-avoidance arrangement. Blue light (BB and BR) decreased the minimum fluorescence (F0′) more than RR light. This extra reduction of the F0′ was stronger than theoretically predicted for F0′ quenching by energy dissipation but actual measurement and theory agreed in RR treatments. The extra F0′ reduction was assigned to decreased light absorption of chloroplasts in the avoidance position. A maximum reduction of 30% was calculated. Increasing intensities of blue light affected the fluorescence parameters NPQ and qP to a lesser degree than red light. After correcting for the optical effects of chloroplast relocation, the NPQ responded similarly to blue and red light. The same correction method diminished the color-specific variations in qP but did not abolish it; thus strongly indicating the presence of another blue light effect which also moderates excitation pressure in PSII but cannot be ascribed to absorption variations. Only after RR exposure, a post-illumination overshoot of F0′ and fast oxidation of PSI electron acceptors occurred, thus, suggesting an electron flow from stromal reductants to the plastoquinone pool. [ABSTRACT FROM AUTHOR]

Plants grow in constantly changing environments, including highly variable light intensities. Sunlight provides the energy that drives photosynthesis and is thus of the utmost importance for plant growth and the generation of oxygen, which the majority of life on Earth depends on. However, exposure to either insufficient or excess levels of light can have detrimental effects and cause light stress. Whereas exposure to insufficient light limits photosynthetic activity, resulting in ‘energy starvation’, exposure to excess light can damage the photosynthetic apparatus. Furthermore, strong sunlight is associated with high levels of potentially damaging UV-B radiation. Different classes of photoreceptors play important roles in coping with the negative aspects of sunlight, for which specific mechanisms are emerging that are reviewed here. [ABSTRACT FROM AUTHOR]

Plants are sessile and require diverse strategies to adapt to fluctuations in the surrounding light conditions. Consequently, the photorelocation movement of chloroplasts is essential to prevent damages that are induced by intense light (avoidance response) and to ensure efficient photosynthetic activities under weak light conditions (accumulation response). The mechanisms that underlie chloroplast movements have been revealed through analysis of the behavior of individual chloroplasts and it has been found that these organelles can move in any direction without turning. This implies that any part of the chloroplast periphery can function as the leading or trailing edge during movement. This ability is mediated by a special structure, which consists of short actin filaments that are polymerized at the leading edge of moving chloroplasts and are specifically localized in the space between the chloroplast and the plasma membrane, and is called chloroplast-actin. In addition, several of the genes that encode proteins that are involved in chloroplast-actin polymerization or maintenance have been identified. In this Review, we discuss the mechanisms that regulate chloroplast movements through polymerization of the chloroplast-actin and propose a model for actin-driven chloroplast photorelocation movement. [ABSTRACT FROM AUTHOR]

[Objective] In this study, a full-length complementary DNA (cDNA) sequence of a novel blue photoreceptor phololropin (PHOT), designated Haephot, was cloned, and the bioinformatics from the green alga Haematococcus pluvialis was analyzed. [Method] The homology cloning and rapid-amplification of cDNA ends (RACE) strategies were applied to obtain the sequence ol Haephot, and its sequence was analyzed. [Result] The cloned cDNA o[ Haephot contained an open reading frame (ORF) of2139 bp encoding a protein of 712 amino acids, and the deduced protein had a calculated molecular mass of 78.73 kD with an estimated isoelectric point ( pi) of 8. 66. Multiple sequence alignment analysis revealed that HaePHOT was homologs to known PHOTs, e. g., Arabidopsis thaliana (58%) and Chlamydomonas reinhardtii (68%). Conserved domains analysis showed that the typical catalytic motifs of the PHOT were highly conserved in the protein sequences of HaePHOT. The phylogenetic analysis of the PHOTs implied that HaePHOT had higher similarity to the PHOTs from high plants and green algae, which indicated that they share a common ancestor origin. [Conclusion]The first identified algal phot gene from green alga Haematococcus pluvialis was obtained, which facilitated the expression and functional investigations of PHOT-mediated signaling pathways involved in astaxanlhin biosynthesis regulation. [ABSTRACT FROM AUTHOR]

Light-induced chloroplast movement is found in most plant species, including algae and land plants. In land plants with multiple small chloroplasts, under weak light conditions, the chloroplasts move towards the light and accumulate on the periclinal cell walls to efficiently perceive light for photosynthesis (the accumulation response). Under strong light conditions, chloroplasts escape from light to avoid photodamage (the avoidance response). In most plant species, blue light induces chloroplast movement, and phototropin receptor kinases are the blue light receptors. Molecular mechanisms for photoreceptors, signal transduction and chloroplast motility systems are being studied using the model plant Arabidopsis thaliana. However, to further understand the molecular mechanisms and evolutionary history of chloroplast movement in green plants, analyses using other plant systems are required. Here, we review recent works on chloroplast movement in green algae, liverwort, mosses and ferns that provide new insights on chloroplast movement. [ABSTRACT FROM AUTHOR]

Ca2+ signaling is an important biological process that enable to perceive and communicate information in the cell. Our current understanding of the signaling system suggests that plants and animals have certain differences in signal-sensing mechanisms. The Ca2+-mediated CBL-CIPK module has emerged as a major sensor responder network for Ca2+ signaling and has been speculated to be involved in plant terrestrial life adaptation. This module has previously been reported in Archaeplastids, Chromalveolates, and Excavates. In our experimental analysis of Chlamydomonas reinhardtii CBLs, we proved that the CrCBL1 protein interacts with Phototropin and Channelrhodopsin, and the expression of CrCBLs is modulated by light. Further analysis using chlorophyte and streptophyte algal sequences allowed us to identify the differences that have evolved in CBL and CIPK proteins since plants have progressed from aquatic to terrestrial habitats. Moreover, an investigation of Klebsormidium CBL and CIPK genes led us to know that they are abiotic stress stimuli-responsive, indicating that their role was defined very early during terrestrial adaptations. Structure-based prediction and Ca2+-binding assays indicated that the KnCBL1 protein in Klebsormidium showed a typical Ca2+-binding pocket. In summary, the results of this study suggest that these stress-responsive proteins enable crosstalk between Ca2+ and light signaling pathways very early during plant adaptation from aquatic to terrestrial habitats. [ABSTRACT FROM AUTHOR]

Nitrite plays an important role in the nitrogen metabolism of most cells, including Chlamydomonas reinhardtii. We have shown that vegetative cells of C. reinhardtii are attracted by nitrite. The Nia1nit2 mutant with defects in genes encoding the nitrate reductase and regulatory protein NIT2 respectively was found to exhibit normal chemotaxis to nitrite. The data suggest that chemotaxis events appear to be specific and independent of those involved in nitrate assimilation. Unlike vegetative cells and noncompetent pregametes, mature gametes did not show chemotaxis to nitrite. Just like gamete formation, the change in chemotaxis mode is controlled by the sequential action of two environmental cues, removal of nitrogen from the medium and light. Comparative analysis of wild-type and RNAi strains with reduced level of phototropin has indicated that switch-off of chemotaxis towards nitrite is dependent on phototropin. The studies revealed individual elements of the phototropin-dependent signal transduction pathway involved in the blue-light-controlled change in chemotaxis mode of C. reinhardtii during gamete formation: three protein kinases, one operating against signal flux and two that promote signal transduction. We have proposed a working model for the signaling pathway by which blue light controls chemotaxis towards attractants, which are nitrogen sources, during pregamete-to-gamete conversion of C. reinhardtii.