RESUMEN
Quiescence is a crucial survival attribute in which cell division is repressed in a reversible manner. Although quiescence has long been viewed as an inactive state, recent studies have shown that it is an actively monitored process that is influenced by environmental stimuli. Here, we provide a perspective of the quiescent state and discuss how this process is tuned by energy, nutrient and oxygen status, and the pathways that sense and transmit these signals. We not only highlight the governance of canonical regulators and signalling mechanisms that respond to changes in nutrient and energy status, but also consider the central significance of mitochondrial functions and cues as key regulators of nuclear gene expression. Furthermore, we discuss how reactive oxygen species and the associated redox processes, which are intrinsically linked to energy carbohydrate metabolism, also play a key role in the orchestration of quiescence.
Asunto(s)
Plantas , Transducción de Señal , Plantas/genética , Plantas/metabolismo , División Celular , Metabolismo de los Hidratos de CarbonoRESUMEN
Quiescence is an essential property of meristematic cells, which restrains the cell cycle while retaining the capacity to divide. This crucial process not only facilitates life-long tissue homeostasis and regenerative capacity but also provides protection against adverse environmental conditions, enabling cells to conserve the proliferative potency while minimising DNA damage. As a survival attribute, quiescence is inherently regulated by the products of aerobic life, in particular reactive oxygen species (ROS) and the redox (reduction/ oxidation) mechanisms that plant have evolved to channel these into pervasive signals. Adaptive responses allow quiescent cells to compensate for reduced oxygen tension (hypoxia) in a reversible manner, while the regulated production of the superoxide anion (.O2-) facilitates cell division and the maintenance of stem cells. Here we discuss the role of ROS and redox reactions in the control of the quiescent state in plant meristems, and how this process is integrated with cellular energy and hormone biochemistry. We consider the pathways that sense and transmit redox signals with a focus on the central significance of redox regulation in the mitochondria and nucleus, which is a major regulator of quiescence in meristems. We discuss recent studies that suggest ROS are a critical component of the feedback loops that control stem cell identity and fate and suggest that the ROS/hypoxia interface is an important "outside/ in" positional cue for plant cells, particularly in meristems.
RESUMEN
Yield losses due to nutrient deficiency are estimated as the primary cause of the yield gap worldwide. Understanding how plant roots perceive external nutrient status and elaborate morphological adaptations in response to it is necessary to develop reliable strategies to increase crop yield. In the last decade, reactive oxygen species (ROS) were shown to be key players of the mechanisms underlying root responses to nutrient limitation. ROS contribute in multiple ways to shape the root system in response to nutritional cues, both as direct effectors acting on cell wall architecture and as second messengers in signalling pathways. Here, we review the mutual interconnections existing between perception and signalling of the most common forms of the major macronutrients (nitrogen, phosphorus and potassium), and ROS in shaping plant root system architecture. We discuss recent advances in dissecting the integration of these elements and their impact on morphological traits of the root system, highlighting the functional ductility of ROS and enzymes implied in ROS metabolism, such as class III peroxidases.
Asunto(s)
Fósforo , Raíces de Plantas , Raíces de Plantas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Fósforo/metabolismo , Fenotipo , Nutrientes , Nitrógeno/metabolismoRESUMEN
Oxygen and reactive oxygen species (ROS) have been co-opted during evolution into the regulation of plant growth, development, and differentiation. ROS and oxidative signals arising from metabolism or phytohormone-mediated processes control almost every aspect of plant development from seed and bud dormancy, liberation of meristematic cells from the quiescent state, root and shoot growth, and architecture, to flowering and seed production. Moreover, the phytochrome and phytohormone-dependent transmissions of ROS waves are central to the systemic whole plant signaling pathways that integrate root and shoot growth. The sensing of oxygen availability through the PROTEOLYSIS 6 (PRT6) N-degron pathway functions alongside ROS production and signaling but how these pathways interact in developing organs remains poorly understood. Considerable progress has been made in our understanding of the nature of hydrogen peroxide sensors and the role of thiol-dependent signaling networks in the transmission of ROS signals. Reduction/oxidation (redox) changes in the glutathione (GSH) pool, glutaredoxins (GRXs), and thioredoxins (TRXs) are important in the control of growth mediated by phytohormone pathways. Although, it is clear that the redox states of proteins involved in plant growth and development are controlled by the NAD(P)H thioredoxin reductase (NTR)/TRX and reduced GSH/GRX systems of the cytosol, chloroplasts, mitochondria, and nucleus, we have only scratched the surface of this multilayered control and how redox-regulated processes interact with other cell signaling systems.
Asunto(s)
Oxígeno/metabolismo , Desarrollo de la Planta/fisiología , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Grapevine (Vitis vinifera L.) displays wide plasticity to climate; however, the physiology of dormancy along a seasonal continuum is poorly understood. Here we investigated the apparent disconnect between dormancy and the underlying respiratory physiology and transcriptome of grapevine buds, from bud set in summer to bud burst in spring. The establishment of dormancy in summer was pronounced and reproducible; however, this was coupled with little or no change in physiology, indicated by respiration, hydration, and tissue oxygen tension. The release of dormancy was biphasic; the depth of dormancy declined substantially by mid-autumn, while the subsequent decline towards spring was moderate. Observed changes in physiology failed to explain the first phase of dormancy decline, in particular. Transcriptome data contrasting development from summer through to spring also indicated that dormancy was poorly reflected by metabolic quiescence during summer and autumn. Gene Ontology and enrichment data revealed the prevailing influence of abscisic acid (ABA)-related gene expression during the transition from summer to autumn, and promoter motif analysis suggested that photoperiod may play an important role in regulating ABA functions during the establishment of dormancy. Transcriptomic data from later transitions reinforced the importance of oxidation and hypoxia as physiological cues to regulate the maintenance of quiescence and resumption of growth. Collectively these data reveal a novel disconnect between growth and metabolic quiescence in grapevine following bud set, which requires further experimentation to explain the phenology and dormancy relationships.
Asunto(s)
Latencia en las Plantas , Vitis , Ácido Abscísico/metabolismo , Regulación de la Expresión Génica de las Plantas , Fotoperiodo , Latencia en las Plantas/genética , Estaciones del Año , Vitis/metabolismoRESUMEN
Plant growth is mediated by cell proliferation and expansion. Both processes are controlled by a network of endogenous factors such as phytohormones, reactive oxygen species (ROS), sugars, and other signals, which influence gene expression and post-translational regulation of proteins. Stress resilience requires rapid and appropriate responses in plant growth and development as well as defence. Regulation of ROS accumulation in different cellular compartments influences growth responses to abiotic and biotic stresses. While ROS are essential for growth, they are also implicated in the stress-induced cessation of growth and, in some cases, programmed cell death. It is widely accepted that redox post-translational modifications of key proteins determine the growth changes and cell fate responses to stress, but the molecular pathways and factors involved remain poorly characterized. Here we discuss ROS as a signalling molecule, the mechanisms of ROS-dependent regulation that influence protein-protein interactions, protein function, and turnover, together with the relocation of key proteins to different intracellular compartments in a manner that can alter cell fate. Understanding how the redox interactome responds to stress-induced increases in ROS may provide a road map to tailoring the dynamic ROS interactions that determine growth and cell fate in order to enhance stress resilience.
Asunto(s)
Desarrollo de la Planta , Reguladores del Crecimiento de las Plantas , Plantas , Especies Reactivas de Oxígeno , Estrés FisiológicoRESUMEN
Drought stress is the main limiting factor for global soybean growth and production. Genetic improvement for water and nutrient uptake efficiency is critical to advance tolerance and enable more sustainable and resilient production, underpinning yield growth. The identification of quantitative traits and genes related to water and nutrient uptake will enhance our understanding of the mechanisms of drought tolerance in soybean. This review summarizes drought stress in the context of the physiological traits that enable effective acclimation, with a particular focus on roots. Genes controlling root system architecture play an important role in water and nutrient availability, and therefore important targets for breeding strategies to improve drought tolerance. This review highlights the candidate genes that have been identified as regulators of important root traits and responses to water stress. Progress in our understanding of the function of particular genes, including GmACX1, GmMS and GmPEPCK are discussed in the context of developing a system-based platform for genetic improvement of drought tolerance in soybean.
Asunto(s)
Fabaceae , Glycine max , Sequías , Raíces de Plantas/genética , Glycine max/genética , AguaRESUMEN
The physiological constraints on bud burst in woody perennials, including vascular development and oxygenation, remain unresolved. Both light and tissue oxygen status have emerged as important cues for vascular development in other systems; however, grapevine buds have only a facultative light requirement, and data on the tissue oxygen status have been confounded by the spatial variability within the bud. Here, we analysed apoplastic development at early stages of grapevine bud burst and combined molecular modelling with histochemical techniques to determine the pore size of cell walls in grapevine buds. The data demonstrate that quiescent grapevine buds were impermeable to apoplastic dyes (acid fuchsin and eosin Y) until after bud burst was established. The molecular exclusion size was calculated to be 2.1 nm, which would exclude most macromolecules except simple sugars and phytohormones until after bud burst. We used micro-computed tomography to demonstrate that tissue oxygen partial pressure data correlated well with structural heterogeneity of the bud and differences in tissue density, confirming that the primary bud complex becomes rapidly and preferentially oxygenated during bud burst. Taken together, our results reveal that the apoplastic porosity is highly regulated during the early stages of bud burst, suggesting a role for vascular development in the initial, rapid oxygenation of the primary bud complex.
Asunto(s)
Bencenosulfonatos/metabolismo , Eosina Amarillenta-(YS)/metabolismo , Luz , Oxígeno , Vitis/metabolismo , Transporte Biológico , Poro Nuclear/metabolismo , Vitis/crecimiento & desarrollo , Microtomografía por Rayos XRESUMEN
The superior agronomic and human nutritional properties of grain legumes (pulses) make them an ideal foundation for future sustainable agriculture. Legume-based farming is particularly important in Africa, where small-scale agricultural systems dominate the food production landscape. Legumes provide an inexpensive source of protein and nutrients to African households as well as natural fertilization for the soil. Although the consumption of traditionally grown legumes has started to decline, the production of soybeans (Glycine max Merr.) is spreading fast, especially across southern Africa. Predictions of future land-use allocation and production show that the soybean is poised to dominate future production across Africa. Land use models project an expansion of harvest area, whereas crop models project possible yield increases. Moreover, a seed change in farming strategy is underway. This is being driven largely by the combined cash crop value of products such as oils and the high nutritional benefits of soybean as an animal feed. Intensification of soybean production has the potential to reduce the dependence of Africa on soybean imports. However, a successful "soybean bonanza" across Africa necessitates an intensive research, development, extension, and policy agenda to ensure that soybean genetic improvements and production technology meet future demands for sustainable production.
Asunto(s)
Producción de Cultivos , Grano Comestible , Glycine max , África , Cambio Climático/estadística & datos numéricos , Producción de Cultivos/estadística & datos numéricos , Producción de Cultivos/tendencias , Grano Comestible/crecimiento & desarrollo , Fabaceae/crecimiento & desarrollo , Predicción , Modelos Estadísticos , Glycine max/crecimiento & desarrolloRESUMEN
Dormant or quiescent buds of woody perennials are often dense and in the case of grapevine (Vitis vinifera L.) have a low tissue oxygen status. The precise timing of the decision to resume growth is difficult to predict, but once committed, the increase in tissue oxygen status is rapid and developmentally regulated. Here, we show that more than a third of the grapevine homologues of widely conserved hypoxia-responsive genes and nearly a fifth of all grapevine genes possessing a plant hypoxia-responsive promoter element were differentially regulated during bud burst, in apparent harmony with resumption of meristem identity and cell-cycle gene regulation. We then investigated the molecular and biochemical properties of the grapevine ERF-VII homologues, which in other species are oxygen labile and function in transcriptional regulation of hypoxia-responsive genes. Each of the 3 VvERF-VIIs were substrates for oxygen-dependent proteolysis in vitro, as a function of the N-terminal cysteine. Collectively, these data support an important developmental function of oxygen-dependent signalling in determining the timing and effective coordination bud burst in grapevine. In addition, novel regulators, including GASA-, TCP-, MYB3R-, PLT-, and WUS-like transcription factors, were identified as hallmarks of the orderly and functional resumption of growth following quiescence in buds.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica , Oxígeno/metabolismo , Vitis/fisiología , Meristema/crecimiento & desarrollo , Meristema/fisiología , Latencia en las Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regiones Promotoras Genéticas/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Vitis/crecimiento & desarrolloRESUMEN
Background: Quiescence is a fundamental feature of plant life, which enables plasticity, renewal and fidelity of the somatic cell line. Cellular quiescence is defined by arrest in a particular phase of the cell cycle, typically G1 or G2; however, the regulation of quiescence and proliferation can also be considered across wider scales in space and time. As such, quiescence is a defining feature of plant development and phenology, from meristematic stem cell progenitors to terminally differentiated cells, as well as dormant or suppressed seeds and buds. While the physiology of each of these states differs considerably, each is referred to as 'cell cycle arrest' or 'G1 arrest'. Scope: Here the physiology and molecular regulation of (1) meristematic quiescence, (2) dormancy and (3) terminal differentiation (cell cycle exit) are considered in order to determine whether and how the molecular decisions guiding these nuclear states are distinct. A brief overview of the canonical cell cycle regulators is provided, and the genetic and genomic, as well as physiological, evidence is considered regarding two primary questions: (1) Are the canonical cell cycle regulators superior or subordinate in the regulation of quiescence? (2) Are these three modes of quiescence governed by distinct molecular controls? Conclusion: Meristematic quiescence, dormancy and terminal differentiation are each predominantly characterized by G1 arrest but regulated distinctly, at a level largely superior to the canonical cell cycle. Meristematic quiescence is intrinsically linked to non-cell-autonomous regulation of meristem cell identity, and particularly through the influence of ubiquitin-dependent proteolysis, in partnership with reactive oxygen species, abscisic acid and auxin. The regulation of terminal differentiation shares analogous features with meristematic quiescence, albeit with specific activators and a greater role for cytokinin signalling. Dormancy meanwhile appears to be regulated at the level of chromatin accessibility, by Polycomb group-type histone modifications of particular dormancy genes.
Asunto(s)
Puntos de Control del Ciclo Celular/fisiología , Desarrollo de la Planta/fisiología , Latencia en las Plantas/fisiología , Fase G1/fisiología , Meristema/crecimiento & desarrollo , Meristema/fisiología , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/fisiología , Brotes de la Planta/crecimiento & desarrollo , Brotes de la Planta/fisiología , Plantas , Fase de Descanso del Ciclo Celular/fisiología , Fase S/fisiologíaRESUMEN
The language of dormancy is rich and poetic, as researchers spanning disciplines and decades have attempted to understand the spell that entranced 'Sleeping Beauty', and how she was gently awoken. The misleading use of 'dormancy', applied to annual axillary buds, for example, has confounded progress. Language is increasingly important as genetic and genomic approaches become more accessible to species of agricultural and ecological importance. Here we examine how terminology has been applied to different eco-physiological states in plants, and with pertinent reference to quiescent states described in other domains of life, in order to place plant quiescence and dormancy in a more complete context than previously described. The physiological consensus defines latency or quiescence as opportunistic avoidance states, where growth resumes in favourable conditions. In contrast, the dormant state in higher plants is entrained in the life history of the organism. Competence to resume growth requires quantitative and specific conditioning. This definition applies only to the embryo of seeds and specialized meristems in higher plants; however, mechanistic control of dormancy extends to mobile signals from peripheral tissues and organs, such as the endosperm of seed or subtending leaf of buds. The distinction between dormancy, quiescence, and stress-hardiness remains poorly delineated, most particularly in buds of winter perennials, which comprise multiple meristems of differing organogenic states. Studies in seeds have shown that dormancy is not a monogenic trait, and limited study has thus far failed to canalize dormancy as seen in seeds and buds. We argue that a common language, based on physiology, is central to enable further dissection of the quiescent and dormant states in plants. We direct the topic largely to woody species showing a single cycle of growth and reproduction per year, as these bear the majority of global timber, fruit, and nut production, as well being of great ecological value. However, for context and hypotheses, we draw on knowledge from annuals and other specialized plant conditions, from a perspective of the major physical, metabolic, and molecular cues that regulate cellular activity.
Asunto(s)
Latencia en las Plantas , Árboles/fisiología , Botánica , Terminología como AsuntoRESUMEN
BACKGROUND AND AIMS: Reactive oxygen species (ROS) and reactive nitrogen species (RNS), such as nitric oxide (NO), play crucial roles in the signal transduction pathways that regulate plant growth, development and defence responses, providing a nexus of reduction/oxidation (redox) control that impacts on nearly every aspect of plant biology. Here we summarize current knowledge and concepts that lay the foundations of a new vision for ROS/RNS functions particularly through signalling hubs for the next decade. SCOPE: Plants have mastered the art of redox control using ROS and RNS as secondary messengers to regulate a diverse range of protein functions through redox-based, post-translational modifications that act as regulators of molecular master-switches. Much current focus concerns the impact of this regulation on local and systemic signalling pathways, as well as understanding how such reactive molecules can be effectively used in the control of plant growth and stress responses. CONCLUSIONS: The spectre of oxidative stress still overshadows much of our current philosophy and understanding of ROS and RNS functions. While many questions remain to be addressed for example regarding inter-organellar regulation and communication, the control of hypoxia and how ROS/RNS signalling is used in plant cells, not only to trigger acclimation responses but also to create molecular memories of stress it is clear that ROS and RNS function as vital signals of living cells.
Asunto(s)
Estrés Oxidativo , Plantas/metabolismo , Procesamiento Proteico-Postraduccional , Especies de Nitrógeno Reactivo/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Oxidación-Reducción , Transducción de SeñalRESUMEN
BACKGROUND AND AIMS: Plants regulate cellular oxygen partial pressures (pO2), together with reduction/oxidation (redox) state in order to manage rapid developmental transitions such as bud burst after a period of quiescence. However, our understanding of pO2 regulation in complex meristematic organs such as buds is incomplete and, in particular, lacks spatial resolution. METHODS: The gradients in pO2 from the outer scales to the primary meristem complex were measured in grapevine (Vitis vinifera) buds, together with respiratory CO2 production rates and the accumulation of superoxide and hydrogen peroxide, from ecodormancy through the first 72 h preceding bud burst, triggered by the transition from low to ambient temperatures. KEY RESULTS: Steep internal pO2 gradients were measured in dormant buds with values as low as 2·5 kPa found in the core of the bud prior to bud burst. Respiratory CO2 production rates increased soon after the transition from low to ambient temperatures and the bud tissues gradually became oxygenated in a patterned process. Within 3 h of the transition to ambient temperatures, superoxide accumulation was observed in the cambial meristem, co-localizing with lignified cellulose associated with pro-vascular tissues. Thereafter, superoxide accumulated in other areas subtending the apical meristem complex, in the absence of significant hydrogen peroxide accumulation, except in the cambial meristem. By 72 h, the internal pO2 gradient showed a biphasic profile, where the minimum pO2 was external to the core of the bud complex. CONCLUSIONS: Spatial and temporal control of the tissue oxygen environment occurs within quiescent buds, and the transition from quiescence to bud burst is accompanied by a regulated relaxation of the hypoxic state and accumulation of reactive oxygen species within the developing cambium and vascular tissues of the heterotrophic grapevine buds.
Asunto(s)
Oxígeno/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Vitis/crecimiento & desarrollo , Anaerobiosis , Meristema/metabolismo , Latencia en las Plantas , Vitis/metabolismoAsunto(s)
Oxígeno/metabolismo , Hojas de la Planta/fisiología , Brotes de la Planta/fisiología , Vitis/fisiología , Cloroplastos/fisiología , Metabolismo Energético , Regulación de la Expresión Génica de las Plantas , Luz , Meristema/fisiología , Brotes de la Planta/crecimiento & desarrollo , Plastidios/genética , Plastidios/metabolismo , Transducción de SeñalRESUMEN
Shoot-root communication is involved in plant stress responses, but its mechanism is largely unknown. To determine the role of roots in stress tolerance, cucumber (Cucumis sativus) shoots from plants with roots of their own or with figleaf gourd (Cucurbita ficifolia, a chilling-tolerant species) or luffa (Luffa cylindrica (L.) M. Roem., a heat-tolerant species) rootstocks were exposed to low (18/13°C), optimal (27/22°C) and high (36/31°C) temperatures, respectively. Grafting onto figleaf gourd and luffa rootstocks significantly alleviated chilling and heat-induced reductions, respectively, in biomass production and CO(2) assimilation capacity in the shoots, while levels of lipid peroxidation and protein oxidation were decreased. Figleaf gourd and luffa rootstocks upregulated a subset of stress-responsive genes involved in signal transduction (MAPK1 and RBOH), transcriptional regulation (MYB and MYC), protein protection (HSP45.9 and HSP70), the antioxidant response (Cu/Zn-SOD, cAPX and GR), and photosynthesis (RBCL, RBCS, RCA and FBPase) at low and high growth temperatures, respectively, and this was accompanied by increased activity of the encoded enzymes and reduced glutathione redox homeostasis in the leaves. Moreover, Heat Shock Protein 70 (HSP70) expression in cucumber leaves was strongly induced by the luffa rootstock at the high growth temperature but slightly induced by the figleaf gourd rootstock at low or high growth temperatures. These results indicate that rootstocks could induce significant changes in the transcripts of stress-responsive and defense-related genes, and the ROS scavenging activity via unknown signals, especially at stressful growth temperatures, and this is one of mechanisms involved in the grafting-induced stress tolerance.
Asunto(s)
Cucumis sativus/fisiología , Cucurbita/fisiología , Luffa/fisiología , Fotosíntesis , Antioxidantes/metabolismo , Biomasa , Dióxido de Carbono , Cucumis sativus/genética , Cucumis sativus/crecimiento & desarrollo , Cucurbita/genética , Cucurbita/crecimiento & desarrollo , Glutatión/metabolismo , Homeostasis , Peroxidación de Lípido , Luffa/genética , Luffa/crecimiento & desarrollo , Oxidación-Reducción , Estrés Oxidativo , Hojas de la Planta/genética , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/fisiología , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/fisiología , Transpiración de Plantas , Especies Reactivas de Oxígeno/metabolismo , Plantones/genética , Plantones/crecimiento & desarrollo , Plantones/fisiología , TemperaturaRESUMEN
Grapevine leafroll disease is a viral disease that affects grapevines (Vitis vinifera L.) and has a severe economic impact on viticulture. In this study, the effect of grapevine leafroll-associated viruses (GLRaV) on berry quality was investigated in clones of cultivar cv. Crimson Seedless table grapes infected with GLRaV. RT-PCR confirmed the identity of the clones: clone 3236, infected only with GLRaV-3 (termed single); clone 3215, infected with GLRaV-3, GLRaV-4 strain 9 and grapevine virus A (termed mixed); and a viral free clone of the same genetic background of the infected clones (termed control). The berry quality indices of size, sugar, acidity and anthocyanin content were measured at harvest maturity. RT-qPCR was used to determine the viral load. The study was repeated over 2 year. A two-way, multivariate analysis of variance was applied with clone and year as independent variables and the measured berry quality parameters as a dependent variable. All dependent variables were significantly affected by viral infection (Wilks, λ, (2,33) = 0.033895, P-value <0.001), while only titratable acidity was affected by year. The average berry dry mass decreased (P-value <0.001). The water content of both infected clones was greater than that of the control (P-value <0.001). Both infected clones displayed reduced sugar content as a fraction of the berry dry mass (P-value <0.001). The anthocyanin and the phenol content of the infected clones were significantly reduced compared with the control clone (P < 0.001, P < 0.05, clone 3236 and clone 3215, respectively). Finally, the viral load was highly variable, and no quantitative relationship between viral load and berry composition was found.
Asunto(s)
Closteroviridae , Frutas , Enfermedades de las Plantas , Carga Viral , Vitis , Vitis/virología , Vitis/crecimiento & desarrollo , Vitis/genética , Frutas/virología , Frutas/crecimiento & desarrollo , Closteroviridae/fisiología , Closteroviridae/genética , Enfermedades de las Plantas/virología , Antocianinas/metabolismo , Antocianinas/análisisRESUMEN
The regulation of DNA accessibility by histone modification has emerged as a paradigm of developmental and environmental programming. Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a versatile tool to investigate in vivo protein-DNA interaction and has enabled advances in mechanistic understanding of physiologies. The technique has been successfully demonstrated in several plant species and tissues; however, it has remained challenging in woody tissues, in particular complex structures such as perennating buds. Here we developed a ChIP method specifically for mature dormant buds of grapevine (Vitis vinifera cv. Cabernet Sauvignon). Each step of the protocol was systematically optimized, including crosslinking, chromatin extraction, sonication and antibody validation. Analysis of histone H3-enriched DNA was performed to evaluate the success of the protocol and identify occupancy of histone H3 along grapevine bud chromatin. To our best knowledge, this is the first ChIP experiment protocol optimized for the grapevine bud system.
Asunto(s)
Secuenciación de Inmunoprecipitación de Cromatina , Vitis , Histonas/genética , Madera , Cromatina , Vitis/genéticaRESUMEN
Hydrogen cyanamide (HC) has been widely used in horticulture to trigger bud burst following dormancy. Its use has been banned in some countries due to human health concerns, however the search for effective safe alternatives is delayed by lack of knowledge of the mechanism of HC action. Earlier studies demonstrate that HC stimulates the production of reactive oxygen species (ROS) and alters the rate of cell division. However, the relationships between HC effects on ROS, redox (reduction/oxidation) homeostasis and cell division are unknown. This study used Arabidopsis thaliana ((L.) Heynh.) seedlings expressing the redox reporter roGFP2 to measure the oxidation states of the nuclei and cytosol in response to HC treatment. The Cytrap dual cell cycle phase marker system and flow cytometry were used to study associated changes in cell proliferation. HC (1.5 mM) reversibly inhibited root growth during a 24 h treatment. Higher concentrations were not reversible. HC did not synchronise the cell cycle, in contrast to hydroxyurea. Rather, HC caused a gradual accumulation of cells in the G2/M phase and decline of G1/S phase cells, 16 to 24 h post-treatment. This was accompanied by increased oxidation of both the nuclei and cytosol. Taken together, these findings show that HC impairs proliferation of embryonic root meristem cells in a reversible manner through restriction of G2/M transition accompanied by increased cellular oxidation.
RESUMEN
Three-dimensional (3D) food printing is a rapidly emerging technology offering unprecedented potential for customised food design and personalised nutrition. Here, we evaluate the technological advances in extrusion-based 3D food printing and its possibilities to promote healthy and sustainable eating. We consider the challenges in implementing the technology in real-world applications. We propose viable applications for 3D food printing in health care, health promotion and food waste upcycling. Finally, we outline future work on 3D food printing in food safety, acceptability and economics, ethics and regulations.