Nyctinastic Leaf Research Articles

An outward-rectifying plant K+ channel SPORK2 exhibits temperature-sensitive ion-transport activity

Temperature sensing is critical for the survival of living organisms.1,2 Thermosensitive transient receptor-potential (TRP) cation channels function as thermosensors in mammals.2,3,4,5,6 In contrast to animals, landplants lack TRP genes.7,8,9 Previous patch-clamp studies in plant cells suggested the presence of ion channels whose activities are related to temperature, implying the presence of TRP-like channels.10,11,12,13,14 However, the molecular entities of such temperature-sensitive ion channels were still unknown in land plants. In this study, we observed that the unique rainfall-induced leaf-folding movement of the legumetree Samanea saman15 was temperature-sensitive by using a rainfall-mimicking assay. Chilling-induced leaf folding in S.saman was shown to be related to the swelling of the motor cells16,17 at the base of the leaflet. This swelling suggested involvement of temperature-sensitive inactivation of K+ currents, independent of fluctuations in ion channel gene expression in motor cells. These findings led us to examine the temperature sensitivity of an outward-rectifying K+ channel, SPORK2, which was reported as an ion channel responsible for the nyctinastic (circadian-rhythmic) leaf movement of S.saman.18 We alsodiscovered that SPORK2 exhibits temperature-sensitive K+ transport activity in the Xenopus oocyte expression system. Using chimeric channels, we showed that two domains of SPORK2 regulated the temperaturesensitivity. Furthermore, heterologously expressed SPORK2 in Arabidopsis guard cells induced temperature-dependent stomatal closure. Therefore, SPORK2 is an ion channel in land plantswith temperature-sensitive ion-transport activity that functions similarly to mammalian TRP channels. Our current findings advance the molecular understanding of temperature-sensing mechanisms in plants.

Pulvinus and petiole comparative anatomy in a species of Mimosa with slow nyctinastic leaf movement

Pulvinus and petiole comparative anatomy in a species of Mimosa with slow nyctinastic leaf movement

Morphology, anatomy and sleep movements of Ludwigia sedoides

The diurnal motion of higher plants, responding to the alternation of day and night, known as nyctinastic movements or “sleep movements”, has been discussed frequently. We present the first description of the circadian rhythm of the water plant Ludwigia sedoides (Humb. & Bonpl.) H.Hara of the family Onagraceae, furthermore its morphology and anatomy. Our results indicate that the plant’s movements are endogenous, although environmental factors certainly have an influence. The majority of plants with nyctinastic leaf movements have a pulvinus, as the crucial part of the plant enabling this movement. Although the basal section of the L. sedoides petiole is not swollen, the tissue functions similarly to a pulvinus. It consists of a central conducting tissue with thick-walled cells, which is surrounded by thin-walled motor cells that can undergo visible shrinking and swelling. Thus, the tissue functionally corresponds to a pulvinus. Examinations of cellular processes, like measurements of the turgor pressure in the petiole, need to be evaluated in future studies.

Heterologous expression of flowering locus T promotes flowering but does not affect diurnal movement in the legume Lotus japonicus.

Flowering locus T (FT) is known to promote flowering in response to photoperiodic conditions and has recently been shown to contribute to other phenomenon, such as diurnal stomatal movement. In legumes, FTs are classified into three subtypes, though the role of each subtype is not well defined. It has been reported that when FT of Lotus japonicus (LjFT) is heterologously expressed in Arabidopsis, LjFT functions as a mobile florigen to promote flowering, similar to Arabidopsis FT (AtFT). In this study, we expressed AtFT in L. japonicus using the SUC2 promoter and showed that heterologous expression of AtFT was able to promote flowering in the plant. We also showed that AtFT expression does not affect stomatal closing nor nyctinastic leaf movement. These findings contribute to our understanding of flower development and have potential application to breeding or plant biotechnology.

Multidimensional Gene Regulatory Landscape of Motor Organ Pulvinus in the Model Legume Medicago truncatula.

Nyctinastic leaf movement of Fabaceae is driven by the tiny motor organ pulvinus located at the base of the leaf or leaflet. Despite the increased understanding of the essential role of ELONGATED PETIOLULE1 (ELP1)/PETIOLE LIKE PULVINUS (PLP) orthologs in determining pulvinus identity in legumes, key regulatory components and molecular mechanisms underlying this movement remain largely unclear. Here, we used WT pulvinus and the equivalent tissue in the elp1 mutant to carry out transcriptome and proteome experiments. The omics data indicated that there are multiple cell biological processes altered at the gene expression and protein abundance level during the pulvinus development. In addition, comparative analysis of different leaf tissues provided clues to illuminate the possible common primordium between pulvinus and petiole, as well as the function of ELP1. Furthermore, the auxin pathway, cell wall composition and chloroplast distribution were altered in elp1 mutants, verifying their important roles in pulvinus development. This study provides a comprehensive insight into the motor organ of the model legume Medicago truncatula and further supplies a rich dataset to facilitate the identification of novel players involved in nyctinastic movement.

The Conserved and Specific Roles of the LUX ARRHYTHMO in Circadian Clock and Nodulation.

LUX ARRHYTHMO (LUX) plays a key role in circadian rhythms and flowering. Here, we identified the MtLUX gene which is the putative ortholog of LUX in Medicago truncatula. The roles of MtLUX, in both the nodulation belowground and leaf movement aboveground, were investigated by characterizing a loss-of-function mtlux mutant. MtLUX was required for the control of flowering time under both long-day and short-day conditions. Further investigations showed that the early flowering in the mtlux mutant was correlated with the elevated expression level of the MtFTa1 gene but in a CO-like independent manner. MtLUX played a conserved role in the regulatory interactions with MtLHY, MtTOC1, and MtPRR genes, which is similar to those in other species. Meanwhile, the unexpected functions of MtLUX were revealed in nodule formation and nyctinastic leaf movement, probably through the indirect regulation in MtLHY. Its participation in nodulation is of interest in the context of functional conservation and the neo-functionalization of the products of LUX orthologs.

Cover Image: Nyctinastic leaf movement in legume

Diurnal leaf movement, named nyctinasty, has been of great interest to researchers since Darwin's time. In this issue, Kong et al. (pp. 1880–1894) report that leaf movement is orchestrated by clock gene MtLHY in a legume species Medicago truncatula, which is also influenced by the availability of nitrogen produced by the nodules in an MtLHY dependent manner.

The nodulation and nyctinastic leaf movement is orchestrated by clock gene LHY in Medicago truncatula

As sessile organisms, plants perceive, respond, and adapt to the environmental changes for optimal growth and survival. The plant growth and fitness are enhanced by circadian clocks through coordination of numerous biological events. In legume species, nitrogen-fixing root nodules were developed as the plant organs specialized for symbiotic transfer of nitrogen between microsymbiont and host. Here, we report that the endogenous circadian rhythm in nodules is regulated by MtLHY in legume species Medicago truncatula. Loss of function of MtLHY leads to a reduction in the number of nodules formed, resulting in a diminished ability to assimilate nitrogen. The operation of the 24-h rhythm in shoot is further influenced by the availability of nitrogen produced by the nodules, leading to the irregulated nyctinastic leaf movement and reduced biomass in mtlhy mutants. These data shed new light on the roles of MtLHY in the orchestration of circadian oscillator in nodules and shoots, which provides a mechanistic link between nodulation, nitrogen assimilation, and clock function.

Ion Channels Regulate Nyctinastic Leaf Opening in Samanea saman

The circadian leaf opening and closing (nyctinasty) of Fabaceae has attracted scientists' attention since the era of Charles Darwin. Nyctinastic movement is triggered by the alternate swelling and shrinking of motor cells at the base of the leaf. This, in turn, is facilitated by changing osmotic pressures brought about by ion flow through anion and potassium ion channels. However, key regulatory ion channels and molecular mechanisms remain largely unknown. Here, we identify three key ion channels in mimosoid tree Samanea saman: the slow-type anion channels, SsSLAH1 and SsSLAH3, and the Shaker-type potassium channel, SPORK2. We show that cell-specific circadian expression of SsSLAH1 plays a key role in nyctinastic leaf opening. In addition, SsSLAH1 co-expressed with SsSLAH3 in flexor (abaxial) motor cells promoted leaf opening. We confirm the importance of SLAH1 in leaf movement using SLAH1-impaired Glycine max. Identification of this "master player" advances our molecular understanding of nyctinasty.

Enantiodifferential Approach for the Target Protein Detection of the Jasmonate Glucoside That Controls the Leaf Closure of Samanea saman.

The synthetic photoaffinity probe designed to mimic bioactive molecules is one of the powerful tools for the identification of the target protein in living organisms. However, nonspecific interaction between the probe and nontargets would cause a misleading result in many cases of the photoaffinity labeling. In this chapter, we describe an enantiodifferential approach as a reliable method for the detection of the specific target protein of the bioactive natural product, jasmonate glucoside, a chemical factor that controls the nyctinastic leaf movement of the leguminous plants.

Identification and characterization of petiolule-like pulvinus mutants with abolished nyctinastic leaf movement in the model legume Medicago truncatula.

Leaves of many plant species open during the day and fold at night. Diurnal leaf movement, named nyctinasty, has been of great interest to researchers since Darwin's time. Nyctinastic leaf movement is generated by the pulvinus, which is a specialized motor organ located at the base of leaf and leaflet. The molecular basis and functional reason behind nyctinasty are unknown.In a forward screening of a retrotransposon-tagged mutant population of Medicago truncatula, four petiolule-like pulvinus (plp) mutant lines with defects in leaf movement were identified and characterized.Loss of function of PLP results in the change of pulvini to petiolules. PLP is specifically expressed in the pulvinus, as demonstrated by quantitative reverse-transcription polymerase chain reaction analysis, expression analysis of a PLP promoter-β-glucuronidase construct in transgenic plants and in situ hybridization. Microarray analysis revealed that the expression levels of many genes were altered in the mutant during the day and at night. Crosses between the plp mutant and several leaf pattern mutants showed that the developmental mechanisms of pulvini and leaf patterns are likely independent.Our results demonstrated that PLP plays a crucial role in the determination of pulvinus development. Leaf movement generated by pulvini may have an impact on plant vegetative growth.

12-Hydroxyjasmonic Acid Glucoside Is a COI1-JAZ-Independent Activator of Leaf-Closing Movement in Samanea saman

Jasmonates are ubiquitously occurring plant growth regulators with high structural diversity that mediate numerous developmental processes and stress responses. We have recently identified 12-O-β-D-glucopyranosyljasmonic acid as the bioactive metabolite, leaf-closing factor (LCF), which induced nyctinastic leaf closure of Samanea saman. We demonstrate that leaf closure of isolated Samanea pinnae is induced upon stereospecific recognition of (-)-LCF, but not by its enantiomer, (+)-ent-LCF, and that the nonglucosylated derivative, (-)-12-hydroxyjasmonic acid also displays weak activity. Similarly, rapid and cell type-specific shrinkage of extensor motor cell protoplasts was selectively initiated upon treatment with (-)-LCF, whereas flexor motor cell protoplasts did not respond. In these bioassays related to leaf movement, all other jasmonates tested were inactive, including jasmonic acid (JA) and the potent derivates JA-isoleucine and coronatine. By contrast, (-)-LCF and (-)-12-hydroxyjasmonic acid were completely inactive with respect to activation of typical JA responses, such as induction of JA-responsive genes LOX2 and OPCL1 in Arabidopsis (Arabidopsis thaliana) or accumulation of plant volatile organic compounds in S. saman and lima bean (Phaseolus lunatus), generally considered to be mediated by JA-isoleucine in a COI1-dependent fashion. Furthermore, application of selective inhibitors indicated that leaf movement in S. saman is mediated by rapid potassium fluxes initiated by opening of potassium-permeable channels. Collectively, our data point to the existence of at least two separate JA signaling pathways in S. saman and that 12-O-β-D-glucopyranosyljasmonic acid exerts its leaf-closing activity through a mechanism independent of the COI1-JAZ module.

ChemInform Abstract: Chemistry and Biology of Plant Leaf Movements

The leaves of Mimosa pudica L. are well known for their rapid movement when touched. Recently, we were able to isolate an excitatory substance in small quantities from this plant, which consists of three different components (potassium L-malate, magnesium trans-aconitate, and dimethylammonium salt). Many plants close their leaves in the evening, as if to sleep, and open them early in the morning (nyctinastic leaf movement). This circadian rhythm is known to be controlled by the biological clock of such plants. Extensive studies on other nyctinastic plants led to the isolation of a variety of leaf-opening substances (LOSs) and leaf-closing substances (LCSs). Based on our experiments on these bioactive substances, we found that the circadian rhythmic leaf movement of these plants is initiated by the regulated balance of LOSs and LCSs. The balance of concentration between the two leaf-movement factors (LMFs) is inversed during the day. The glycoside-type LMF is hydrolyzed with β-glucosidase, the activity of which is regulated by the biological clock. The circadian rhythm observed in the leaf movement is introduced by activation of β-glucosidase regulated by the biological clock.

Chemical Basis of Plant Leaf Movement

Nyctinastic plants open and close leaves with a circadian rhythm. Here we discuss chemical aspects of the mechanism of nyctinastic leaf movement. Nyctinastic plants from five different genera are known to contain species-specific leaf-opening and leaf-closing factors. The relative concentrations of leaf-closing and leaf-opening factors of the nyctinastic plant Phyllanthus urinaria change circadianly, suggesting that nyctinastic movement is regulated by two classes of circadianly regulated factors with opposing functions. A closing and an opening factor of Albizzia, when linked to a fluorescent dye, both specifically labeled motor cells of pluvini. A membrane fraction of pluvini contains proteins of 210 and 180 kDa that bind to a leaf-opening factor of Cassia mimosoides. The molecular identification of these proteins is underway.

Endogenous factors involved in the regulation of movement and "memory" in plants

Abstract The bioorganic basis of plant movement in two plant systems is described in this article: the circadian rhythmic leaf movement known as nyctinasty and trap movement in the Venus flytrap. The bioactive substances responsible for plant movement, the chemical mechanism of the rhythm, and studies on the key protein controlling nyctinasty are presented. The nyctinastic leaf movement is induced by a pair of leaf-movement factors, and one of each pair is a glucoside. There are two key proteins that are involved in the control of nyctinasty. One is β-glucosidase: a biological clock regulates the activity of β-glucosidase, which deactivates the glucoside-type leaf-movement factor, controlling the balance in the concentrations of the leaf-closing and -opening factors. The other is the specific receptor for each leaf-movement factor: the genuine target cell for each leaf-movement factor is confirmed to be a motor cell from leaflet pulvini, and the specific receptors that regulate the turgor of motor cells are localized in the membrane fraction. The article also discusses the isolation of the "memory" substance from the Venus flytrap and presents a mechanism for this action.

Enantio-differential approach to identify the target cell for glucosyl jasmonate-type leaf-closing factor, by using fluorescence-labeled probe compounds

Potassium β- d-glucopyranosyl 12-hydroxyjasmonate ( 1) is a leaf-closing factor of Albizzia plants that induces nyctinastic leaf closure. In this paper, we synthesized probe 3 and its congener 4 by using a pair of enantiomerically pure methyl jasmonate that was prepared by using optical resolution, and carried out fluorescence studies using 3 and 4 to identify the target cell of 1. The probe 3 bound to the motor cells of two Albizzia plants, whereas it could not bind to the motor cells of plants belonging to other genus. On the other hand, probe 4 did not bind to the motor cell at all. These results suggested that a specific receptor for 1 is involved in the motor cell of Albizzia plants.

Direct observation of the target cell for jasmonate-type leaf-closing factor: genus-specific binding of leaf-movement factors to the plant motor cell

We report the synthesis of the novel fluorescence-labeled jasmonate glycoside 2 based on β- d-glucopyranosyl 12-hydroxyjasmonate 1 , which is a leaf-closing substance of Albizzia julibrissin Durazz. The fluorescence study using 2 revealed that the target cell for 1 is a motor cell. Probe 2 bound to the motor cells of two plants belonging to genus Albizzia. This result suggested that a receptor for 2 , which is common among genus Albizzia would be involved in the nyctinastic leaf movement.

Bioorganic studies on plant movement, from natural products to its receptor

The chemical aspects of the circadian leaf movement known as "nyctinasty" are discussed in this paper. Each of the nyctinastic plants of five different genera so far examined contained a pair of factors, one of which induced leaf closure and another induced leaf opening. The relative contents of the closing and opening factors changed correlating with the nyctinastic leaf movement. The use of fluorescence-labeled and photoaffinity-labeled factors revealed that the factors bind to specific cells, the motor cells, present in the pulvini, and that the membrane fraction of the motor cells contained two proteins of 210 and 180 kDa, which can bind to the factors.

Concise synthesis of glyconoamidines as affinity ligands for the purification of β-glucosidase involved in control of some biological events including plant leaf movement

Glycosidases are involved in deactivation or storage of some endogenous bioactive substances through biologically intriguing processes. For example, nyctinastic leaf movement is controlled by a biological clock through the regulation of β-glucosidase activity. Ganem’s glyconoamidine ( 1 ) is used as a micromolar inhibitor of glucosidase in biochemical studies and would be useful as an affinity ligand for purification of glycosidase. However, its use for the specific inhibition of glucosidase which is highly specific to a glycoside with voluntary aglycon is seriously restricted because no universal method for the synthesis of N-alkylated glyconoamidine has been reported. Here, we report a concise synthesis of N-alkylated Ganem’s glyconoamidine with voluntary aglycon using a non-protected sugar derivative.

The plasma membrane aquaporin NtAQP1 is a key component of the leaf unfolding mechanism in tobacco

Epinastic leaf movement of tobacco is based on differential growth of the upper and lower leaf surface and is distinct from the motor organ-driven mechanism of nyctinastic leaf movement of, for example, mimosa species. The epinastic leaf movement of tobacco is observed not only under diurnal light regimes but also in continuous light, indicating a control by light and the circadian clock. As the transport of water across membranes by aquaporins is an important component of rapid plant cell elongation, the role of the tobacco aquaporin Nt aquaporin (AQP)1 in the epinastic response was studied in detail. In planta NtAQP1-luciferase (LUC) activity studies, Northern and Western blot analyses demonstrated a diurnal and circadian oscillation in the expression of this plasma membrane intrinsic protein (PIP)1-type aquaporin in leaf petioles, exhibiting peaks of expression coinciding with leaf unfolding. Cellular water permeability of protoplasts isolated from leaf petioles was found to be high in the morning, i.e. during the unfolding reaction, and low in the evening. Moreover, diurnal epinastic leaf movement was shown to be reduced in transgenic tobacco lines with an impaired expression of NtAQP1. It is concluded that the cyclic expression of PIP1-aquaporin represents an important component of the leaf movement mechanism.