RESUMEN
Stress-induced retrograde signal transmission from the plastids to the nucleus has long puzzled plant biologists. To address this, we performed a suppressor screen of the ceh1 mutant, which contains elevated 2-C-methyl-d-erythritol-2,4-cyclopyrophosphate (MEcPP) levels, and identified the gain-of-function mutant impα-9, which shows reversed dwarfism and suppressed expression of stress-response genes in the ceh1 background despite heightened MEcPP. Subsequent genetic and biochemical analyses established that the accumulation of MEcPP initiates an upsurge in Arabidopsis SKP1-like 1 (ASK1) abundance, a pivotal component in the proteasome degradation pathway. This increase in ASK1 prompts the degradation of IMPα-9. Moreover, we uncovered a protein-protein interaction between IMPα-9 and TPR2, a transcriptional co-suppressor and found that a reduction in IMPα-9 levels coincides with a decrease in TPR2 abundance. Significantly, the interaction between IMPα-9 and TPR2 was disrupted in impα-9 mutants, highlighting the critical role of a single amino acid alteration in maintaining their association. Disruption of their interaction results in the reversal of MEcPP-associated phenotypes. Chromatin immunoprecipitation coupled with sequencing analyses revealed that TPR2 binds globally to stress-response genes and suggested that IMPα-9 associates with the chromatin. They function together to suppress the expression of stress-response genes under normal conditions, but this suppression is alleviated in response to stress through the degradation of the suppressing machinery. The biological relevance of our discoveries was validated under high light stress, marked by MEcPP accumulation, elevated ASK1 levels, IMPα-9 degredation, reduced TPR2 abundance, and subsequent activation of a network of stress-response genes. In summary, our study collectively unveils fresh insights into plant adaptive mechanisms, highlighting intricate interactions among retrograde signaling, the proteasome, and nuclear transport machinery.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Regulación de la Expresión Génica de las Plantas , Transducción de Señal , Estrés Fisiológico , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Estrés Fisiológico/genética , Núcleo Celular/metabolismo , Carioferinas/metabolismo , Carioferinas/genética , Unión ProteicaRESUMEN
Low temperature conditions have been linked to a heightened susceptibility to lower respiratory infections (LRIs). Yet, our comprehension of the LRIs' disease burden due to such conditions remains limited, especially when considering the diverse socio-demographic indexes (SDIs) and climate types across various nations and regions. We examined the variations over time and space in the impact of LRIs due to low temperatures across a diverse set of 204 nations and regions, each with unique SDIs and climate types, spanning the years 1990 to 2019. Data from the Global Burden of Disease Study 2019 was used for this retrospective analysis. The burden of LRIs attributable to low temperatures was estimated by stratifying by sex, age, country, climate type, and SDI, including age-standardized mortality rate (ASMR) and age-standardized disability-adjusted life year rate (ASDR). We employed Joinpoint models to compute the annual average percent changes (AAPCs) in order to evaluate the trends in LRIs burden due to low temperatures from 1990 to 2019. Furthermore, we utilized Poisson age-period-cohort models to forecast the global and income-specific trends in LRIs burden due to low temperatures for the period 2020-2044. Generalized additive mixed models were used to fit changes in the disease burden of different climate regions. The relationship between SDI and both ASMR and ASDR was determined using models grounded in Gaussian process regression. In general, since the year 1990, there has been a significant reduction in the worldwide impact of LRIs due to low temperatures. This decrease is particularly noticeable among infants and the elderly, as well as in regions with a boreal climate and those with an average SDI. In 2019, LRIs induced by low temperatures showed an ASMR of 2.2 (95% CI: 1.34, 3.07) and an ASDR of 53.73 (95% CI: 17.5, 93.22) for every 100,000 individuals. A global reduction was observed in the ASMR and ASDR for LRIs over the period from 1990 to 2019, showing a decrease of 60.27% and 77.5%, in that order. For ASMR and ASDR, the AAPC values were found to be - 3.3 (95% CI: - 3.4, - 3.1) and - 5 (95% CI: - 5.2, - 4.9), in that order. However, a contrasting pattern was observed in southern Latin America, where an increase was noted in the ASMR for LRIs induced by low temperatures [AAPC: 0.5; 95% CI: (0.3, 0.8)]. Low temperature has decreased as an environmental risk factor for LRIs globally over 30 years, especially in middle SDI regions and boreal climates, but remains important for infants and the elderly population.
Asunto(s)
Clima , Infecciones del Sistema Respiratorio , Lactante , Humanos , Anciano , Estudios Retrospectivos , Temperatura , Frío , Productos Finales de Glicación Avanzada , Salud Global , Años de Vida Ajustados por Calidad de VidaRESUMEN
Root-knot nematodes (Meloidogyne spp., RKN) are responsible for extensive crop losses worldwide. During infection, they penetrate plant roots, migrate between plant cells, and establish feeding sites, known as giant cells, near the root vasculature. Previously, we found that nematode perception and early responses in plants were similar to those of microbial pathogens and required the BRI1-ASSOCIATED KINASE1/SOMATIC EMBRYOGENESIS RECEPTOR KINASE3 (BAK1/SERK3) coreceptor in Arabidopsis (Arabidopsis thaliana) and tomato (Solanum lycopersicum). Here, we implemented a reverse genetic screen for resistance or sensitivity to RKN using Arabidopsis T-DNA alleles of genes encoding transmembrane receptor-like kinases to identify additional receptors involved in this process. This screen identified a pair of allelic mutations with enhanced resistance to RKN in a gene we named ENHANCED RESISTANCE TO NEMATODES1 (ERN1). ERN1 encodes a G-type lectin receptor kinase (G-LecRK) with a single-pass transmembrane domain. Further characterization showed that ern1 mutants displayed stronger activation of MAP kinases, elevated levels of the defense marker MYB51, and enhanced H2O2 accumulation in roots upon RKN elicitor treatments. Elevated MYB51 expression and ROS bursts were also observed in leaves of ern1 mutants upon flg22 treatment. Complementation of ern1.1 with 35S- or native promoter-driven ERN1 rescued the RKN infection and enhanced defense phenotypes. Our results indicate that ERN1 is an important negative regulator of immunity.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Solanum lycopersicum , Tylenchoidea , Animales , Arabidopsis/fisiología , Proteínas Quinasas Dependientes de GMP Cíclico/metabolismo , Lectinas/metabolismo , Peróxido de Hidrógeno/metabolismo , Tylenchoidea/fisiología , Solanum lycopersicum/genética , Receptores Mitogénicos/metabolismo , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Enfermedades de las Plantas/genética , Factores de Transcripción/metabolismo , Proteínas de Arabidopsis/metabolismoRESUMEN
Epidemiological studies have confirmed salt sensitivity as a crucial risk factor for the development of hypertension. However, few studies have investigated the association between salt sensitivity of blood pressure (SSBP) and hypertension in Chinese Tibetan population. Therefore, we conducted a cross-sectional study based on a Tibetan population to evaluate the association between SSBP and the risk of hypertension. Seven hundred and eighty-four participants with hypertension and 645 participants without hypertension were included from five villages in Tibetan Autonomous Region of Gannan during 2013-2014. The assessment of salt sensitivity (SS) and non-salt sensitivity (NSS) was performed according to mean arterial pressure (MAP) changes by the modified Sullivan's acute oral saline load and diuresis shrinkage test (MSAOSL-DST). Logistic regression models and restricted cubic models were used to examine the association between SSBP and hypertension. There were 554 (70.5%) salt-sensitive participants with hypertension and 412 (63.9%) salt-sensitive participants without hypertension in this study. Compared with individuals with NSS, individuals with SS had a significantly increased risk of hypertension, and the multiple-adjusted odds ratios were 2.582 with 95% confidence interval of 1.357-4.912. Furthermore, a significant linear trend was found between MAP changes and hypertension. Subgroup analyses showed significant and stronger associations between SSBP and the risk of hypertension in the older (age ≥ 55 years old), males and participants who took exercise less than 1 time per week. Our results suggest that SS is associated with an increased risk of hypertension in Tibetan population, indicating a need for clinicians dealing with SSBP to decrease the risk of hypertension.
Asunto(s)
Hipertensión , Masculino , Humanos , Persona de Mediana Edad , Hipertensión/etiología , Hipertensión/inducido químicamente , Presión Sanguínea , Tibet/epidemiología , Estudios Transversales , Pueblos del Este de Asia , Cloruro de Sodio , Cloruro de Sodio Dietético/efectos adversosRESUMEN
Bacterial infection often induces chronic repair of wound healing owing to aggravated inflammation. Hydrogel dressing exhibiting intrinsic antibacterial activity may substantially reduce the use of antibiotics for infected wound management. Hence, a versatile hydrogel dressing (rGB/QCS/PDA-PAM) exhibiting skin adaptiveness on dynamic wounds and mild photothermal antibacterial activity is developed for safe and efficient infected wound treatment. Phenylboronic acid-functionalized graphene (rGB) and oxadiazole-decorated quaternary carboxymethyl chitosan (QCS) are incorporated into a polydopamine-polyacrylamide (PDA-PAM) network with multiple covalent and noncovalent bonds, which conferred the hydrogel with flexible mechanical properties, strong tissue adhesion and excellent self-healing ability on the dynamic wounds. Moreover, the glycocalyx-mimicking phenylboronic acid on the surface of rGB enables the hydrogel to specifically capture bacteria. The enhanced membrane permeability of QCS enhanced bacterial vulnerability to photothermal therapyï¼PTTï¼, which is demonstrated by efficient mild PTT antibacteria against methicillin-resistant Staphylococcus aureus in vitro and in vivo at temperatures of <49.6 °C. Consequently, the hydrogel demonstrate accelerated tissue regeneration on MRSA-infected wound in vivo, with an intact epidermis, abundant collagen deposition and prominent angiogenesis. Therefore, rGB/QCS/PDA-PAM is a versatile hydrogel dressing exhibiting inherent antibacterial activity and has considerable potential in treating wounds infected with drug-resistant bacteria.
Asunto(s)
Staphylococcus aureus Resistente a Meticilina , Hidrogeles , Antibacterianos/farmacología , Vendajes , Cicatrización de HeridasRESUMEN
The application of photodynamic therapy (PDT) has attracted remarkable interest in cancer treatment because of the advantages of noninvasiveness and spatiotemporal selectivity. However, the PDT efficiency is considerably limited by photosensitizer (PS) quenching and severe hypoxia in solid tumors. Herein, a kind of near infrared (NIR)-activatable methylene blue (MB) peptide nanocarrier was developed for codelivery of nitric oxide (NO) prodrug JSK, expecting a cascade of reactive oxygen species (ROS) amplification-mediated antitumor PDT. In detail, MB was conjugated to water-soluble polyethylene glycol-polylysine (PEG-PLL) through NIR-photocleavable 10-N-carbamoyl bonds, and the subsequent amphiphilic conjugates (mPEG-PLL-MB) self-assembled into nanoparticles (NPs), which allowed JSK codelivery via π-π stacking interactions. MB in quenched state in mPEG-PLL-MB/JSK NPs could be photoactivated by NIR light locoregionally in a controlled manner due to the photocleavage of carbamoyl bonds. Apart from ROS production, assembly disturbance and even disintegration of mPEG-PLL-MB/JSK occurred along with MB activation that subsequently freed JSK, which was further triggered by intracellularly overexpressed glutathione (GSH) and glutathione S-transferase (GST) to sustain the release of NO. NO then served as a hypoxia relief agent through inhibition of cellular respiration to economize O2, cooperating with MB activation and GSH depletion, which synergistically enabled a cascade of ROS amplification to augment PDT for mitochondrial apoptosis-mediated tumor inhibition in vitro and in vivo. Therefore, this pioneering strategy of cascade amplification of ROS addressed the key issues of PS inactivation, hypoxia resistance, and ROS neutralization in a three-pronged approach, which hold great promise in efficient antitumor PDT.
Asunto(s)
Nanopartículas , Neoplasias , Fotoquimioterapia , Profármacos , Humanos , Especies Reactivas de Oxígeno , Azul de Metileno/farmacología , Azul de Metileno/química , Óxido Nítrico , Profármacos/farmacología , Fármacos Fotosensibilizantes/química , Nanopartículas/química , Neoplasias/tratamiento farmacológico , Péptidos/farmacología , Hipoxia/tratamiento farmacológico , Línea Celular TumoralRESUMEN
Diosgenin saponins isolated from Dioscorea species such as D. zingiberensis exhibit a broad spectrum of pharmacological activities. Diosgenin, the aglycone of diosgenin saponins, is an important starting material for the production of steroidal drugs. However, how plants produce diosgenin saponins and the origin and evolution of the diosgenin saponin biosynthetic pathway remain a mystery. Here we report a high-quality, 629-Mb genome of D. zingiberensis anchored on 10 chromosomes with 30 322 protein-coding genes. We reveal that diosgenin is synthesized in leaves ('source'), then converted into diosgenin saponins, and finally transported to rhizomes ('sink') for storage in plants. By evaluating the distribution and evolutionary patterns of diosgenin saponins in Dioscorea species, we find that diosgenin saponin-containing may be an ancestral trait in Dioscorea and is selectively retained. The results of comparative genomic analysis indicate that tandem duplication coupled with a whole-genome duplication event provided key evolutionary resources for the diosgenin saponin biosynthetic pathway in the D. zingiberensis genome. Furthermore, comparative transcriptome and metabolite analysis among 13 Dioscorea species suggests that specific gene expression patterns of pathway genes promote the differential evolution of the diosgenin saponin biosynthetic pathway in Dioscorea species. Our study provides important insights and valuable resources for further understanding the biosynthesis, evolution, and utilization of plant specialized metabolites such as diosgenin saponins.
RESUMEN
Polar cell growth is a process that couples the establishment of cell polarity with growth and is extremely important in the growth, development, and reproduction of eukaryotic organisms, such as pollen tube growth during plant fertilization and neuronal axon growth in animals. Pollen tube growth requires dynamic but polarized distribution and activation of a signaling protein named ROP1 to the plasma membrane via three processes: positive feedback and negative feedback regulation of ROP1 activation and its lateral diffusion along the plasma membrane. In this paper, we introduce a mechanistic integro-differential equation (IDE) along with constrained semiparametric regression to quantitatively describe the interplay among these three processes that lead to the polar distribution of active ROP1 at a steady state. Moreover, we introduce a population variability by a constrained nonlinear mixed model. Our analysis of ROP1 activity distributions from multiple pollen tubes revealed that the equilibrium between the positive and negative feedbacks for pollen tubes with similar shapes are remarkably stable, permitting us to infer an inherent quantitative relationship between the positive and negative feedback loops that defines the tip growth of pollen tubes and the polarity of tip growth.
RESUMEN
As a universal second messenger, calcium (Ca2+) transmits specific cellular signals via a spatiotemporal signature generated from its extracellular source and internal stores. Our knowledge of the mechanisms underlying the generation of a Ca2+ signature is hampered by limited tools for simultaneously monitoring dynamic Ca2+ levels in multiple subcellular compartments. To overcome the limitation and to further improve spatiotemporal resolutions, we have assembled a molecular toolset (CamelliA lines) in Arabidopsis (Arabidopsis thaliana) that enables simultaneous and high-resolution monitoring of Ca2+ dynamics in multiple subcellular compartments through imaging different single-colored genetically encoded calcium indicators. We uncovered several Ca2+ signatures in three types of Arabidopsis cells in response to internal and external cues, including rapid oscillations of cytosolic Ca2+ and apical plasma membrane Ca2+ influx in fast-growing Arabidopsis pollen tubes, the spatiotemporal relationship of Ca2+ dynamics in four subcellular compartments of root epidermal cells challenged with salt, and a shockwave-like Ca2+ wave propagating in laser-wounded leaf epidermis. These observations serve as a testimony to the wide applicability of the CamelliA lines for elucidating the subcellular sources contributing to the Ca2+ signatures in plants.
Asunto(s)
Arabidopsis , Camellia , Arabidopsis/genética , Arabidopsis/metabolismo , Calcio/metabolismo , Camellia/genética , Camellia/metabolismo , Citosol/metabolismo , Tubo Polínico/metabolismoRESUMEN
During growth and morphogenesis, plant cells respond to mechanical stresses resulting from spatiotemporal changes in the cell wall that bear high internal turgor pressure. Microtubule (MT) arrays are reorganized to align in the direction of maximal tensile stress, presumably reinforcing the local cell wall by guiding the synthesis of cellulose. However, how mechanical forces regulate MT reorganization remains largely unknown. Here, we demonstrate that mechanical signaling that is based on the Catharanthus roseus RLK1-like kinase (CrRLK1L) subfamily receptor kinase FERONIA (FER) regulates the reorganization of cortical MT in cotyledon epidermal pavement cells (PCs) in Arabidopsis. Recessive mutations in FER compromised MT responses to mechanical perturbations, such as single-cell ablation, compression, and isoxaben treatment, in these PCs. These perturbations promoted the activation of ROP6 guanosine triphosphatase (GTPase) that acts directly downstream of FER. Furthermore, defects in the ROP6 signaling pathway negated the reorganization of cortical MTs induced by these stresses. Finally, reduction in highly demethylesterified pectin, which binds the extracellular malectin domains of FER and is required for FER-mediated ROP6 activation, also impacted mechanical induction of cortical MT reorganization. Taken together, our results suggest that the FER-pectin complex senses and/or transduces mechanical forces to regulate MT organization through activating the ROP6 signaling pathway in Arabidopsis.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , GTP Fosfohidrolasas/metabolismo , Morfogénesis , Pectinas/metabolismo , Fosfotransferasas/genética , Transducción de Señal/fisiologíaRESUMEN
Invasive or penetrative growth is critical for developmental and reproductive processes (e.g., pollen tube penetration of pistils) and disease progression (e.g., cancer metastasis and fungal hyphae invasion). The invading or penetrating cells experience drastic changes in mechanical pressure from the surroundings and must balance growth with cell integrity. Here, we show that Arabidopsis pollen tubes sense and/or respond to mechanical changes via a cell-surface receptor kinase Buddha's Paper Seal 1 (BUPS1) while emerging from compressing female tissues. BUPS1-defective pollen tubes fail to maintain cell integrity after emergence from these tissues. The mechano-transduction function of BUPS1 is established by using a microfluidic channel device mimicking the mechanical features of the in vivo growth path. BUPS1-based mechano-transduction activates Rho-like GTPase from Plant 1 (ROP1) GTPase to promote exocytosis that facilitates secretion of BUPS1's ligands for mechanical signal amplification and cell wall rigidification in pollen tubes. These findings uncover a membrane receptor-based mechano-transduction system for cells to cope with the physical challenges during invasive or penetrative growth.
Asunto(s)
Proteínas de Arabidopsis/fisiología , Arabidopsis/crecimiento & desarrollo , Mecanotransducción Celular , Tubo Polínico/crecimiento & desarrollo , Proteínas Serina-Treonina Quinasas/fisiología , Arabidopsis/anatomía & histología , Arabidopsis/enzimología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas Portadoras/metabolismo , Pared Celular , Flores/crecimiento & desarrollo , GTP Fosfohidrolasas/metabolismo , Proteínas de Unión al GTP/metabolismo , Tubo Polínico/anatomía & histología , Receptores de Superficie Celular/fisiología , Estrés FisiológicoRESUMEN
Cell polarity is fundamental to the development of both eukaryotes and prokaryotes, yet the mechanisms behind its formation are not well understood. Here we found that, phytohormone auxin-induced, sterol-dependent nanoclustering of cell surface transmembrane receptor kinase 1 (TMK1) is critical for the formation of polarized domains at the plasma membrane (PM) during the morphogenesis of cotyledon pavement cells (PC) in Arabidopsis. Auxin-induced TMK1 nanoclustering stabilizes flotillin1-associated ordered nanodomains, which in turn promote the nanoclustering of ROP6 GTPase that acts downstream of TMK1 to regulate cortical microtubule organization. In turn, cortical microtubules further stabilize TMK1- and flotillin1-containing nanoclusters at the PM. Hence, we propose a new paradigm for polarity formation: A diffusive signal triggers cell polarization by promoting cell surface receptor-mediated nanoclustering of signaling components and cytoskeleton-mediated positive feedback that reinforces these nanodomains into polarized domains.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/citología , Arabidopsis/metabolismo , Polaridad Celular/fisiología , Ácidos Indolacéticos/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/química , Membrana Celular/metabolismo , Polaridad Celular/genética , Metabolismo de los Lípidos , Proteínas de la Membrana/química , Proteínas de la Membrana/metabolismo , Modelos Biológicos , Proteínas de Unión al GTP Monoméricas/química , Proteínas de Unión al GTP Monoméricas/metabolismo , Mutación , Reguladores del Crecimiento de las Plantas/metabolismo , Plantas Modificadas Genéticamente , Agregado de Proteínas , Proteínas Serina-Treonina Quinasas/química , Estabilidad Proteica , Transducción de SeñalRESUMEN
Exocytosis is a fundamental process essential for many cellular functions by targeting signal peptides, proteins, and cell wall components to the plasma membrane (PM) or extracellular matrix. Conventional methods, such as FRAP, often underestimate the exocytosis rate of a specific molecule, because retrieval of the molecules from the PM by endocytosis can impact the measurement. To overcome this issue, we have previously established a novel method, corrected fluorescence recovery after photoconversion (cFRAPc), which allows us to accurately measure the exocytosis rate by monitoring both exocytosis-dependent and exocytosis-independent events. In this chapter, we provide detailed procedures for the cFRAPc method to measure the exocytosis rate of Arabidopsis receptor-like kinase PRK1 in pollen tubes. This method should be widely applicable to various cell types.
Asunto(s)
Exocitosis , Recuperación de Fluorescencia tras Fotoblanqueo/métodos , Tubo Polínico/metabolismo , Arabidopsis , Proteínas de Arabidopsis/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismoRESUMEN
Pollen tubes rapidly elongate, penetrate, and navigate through multiple female tissues to reach ovules for sperm delivery by utilizing a specialized form of polar growth known as tip growth. This process requires a battery of cellular activities differentially occurring at the apical growing region of the plasma membrane (PM), such as the differential cellular signaling involving calcium (Ca2+), phospholipids, and ROP-type Rho GTPases, fluctuation of ions and pH, exocytosis and endocytosis, and cell wall construction and remodeling. There is an emerging understanding of how at least some of these activities are coordinated and/or interconnected. The apical active ROP modulates exocytosis to the cell apex for PM and cell wall expansion differentially occurring at the tip. The differentiation of the cell wall involves at least the preferential distribution of deformable pectin polymers to the apex and non-deformable pectin polymers to the shank of pollen tubes, facilitating the apical cell expansion driven by high internal turgor pressure. Recent studies have generated inroads into how the ROP GTPase-based intracellular signaling is coordinated spatiotemporally with the external wall mechanics to maintain the tubular cell shape and how the apical cell wall mechanics are regulated to allow rapid tip growth while maintaining the cell wall integrity under the turgor pressure. Evidence suggests that exocytosis and endocytosis play crucial but distinct roles in this spatiotemporal coordination. In this review, we summarize recent advances in the regulation and coordination of the differential pectin distribution and the apical domain of active ROP by exocytosis and endocytosis in pollen tubes.
Asunto(s)
Exocitosis , Tubo Polínico , Endocitosis , Pectinas , Plantas , Tubo Polínico/metabolismo , Transducción de Señal , Proteínas de Unión al GTP rho/metabolismoRESUMEN
In plants, GABA plays a critical role in sexual plant reproduction; however, GABA receptors and the associated detailed signaling mechanisms remain to be elucidated. Our experiments show that the proposed technique is reliable and convenient for probing GABA-binding proteins and could be applicable in similar projects by covalently immobilizing the free carboxylic group of GABA on magnetic beads (SiMAG-Carboxyl). New probes produced by covalently immobilizing the free carboxylic group of GABA on magnetic beads (SiMAG-Carboxyl) can obtain useful information on GABA receptors in plants.
RESUMEN
A reaction-diffusion model is proposed to describe the mechanisms underlying the spatial distributions of ROP1 and calcium on the pollen tube tip. The model assumes that the plasma membrane ROP1 activates itself through positive feedback loop, while the cytosolic calcium ions inhibit ROP1 via a negative feedback loop. Furthermore it is proposed that lateral movement of molecules on the plasma membrane are depicted by diffusion. It is shown that bistable or oscillatory dynamics could exist even in the non-spatial model, and stationary and oscillatory spatiotemporal patterns are found in the full spatial model which resemble the experimental data of pollen tube tip growth.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Calcio/metabolismo , Modelos Teóricos , Tubo Polínico/metabolismo , Análisis Espacio-Temporal , Transducción de SeñalRESUMEN
As a universal energy generation pathway utilizing carbon metabolism, glycolysis plays an important housekeeping role in all organisms. Pollen tubes expand rapidly via a mechanism of polarized growth, known as tip growth, to deliver sperm for fertilization. Here, we report a novel and surprising role of glycolysis in the regulation of growth polarity in Arabidopsis pollen tubes via impingement of Rho GTPase-dependent signaling. We identified a cytosolic phosphoglycerate kinase (pgkc-1) mutant with accelerated pollen germination and compromised pollen tube growth polarity. pgkc-1 mutation greatly diminished apical exocytic vesicular distribution of REN1 RopGAP (Rop GTPase activating protein), leading to ROP1 hyper-activation at the apical plasma membrane. Consequently, pgkc-1 pollen tubes contained higher amounts of exocytic vesicles and actin microfilaments in the apical region, and showed reduced sensitivity to Brefeldin A and Latrunculin B, respectively. While inhibition of mitochondrial respiration could not explain the pgkc-1 phenotype, the glycolytic activity is indeed required for PGKc function in pollen tubes. Moreover, the pgkc-1 pollen tube phenotype was mimicked by the inhibition of another glycolytic enzyme. These findings highlight an unconventional regulatory function for a housekeeping metabolic pathway in the spatial control of a fundamental cellular process.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Glucólisis , Tubo Polínico/crecimiento & desarrollo , Tubo Polínico/metabolismo , Proteínas de Unión al GTP rho/metabolismo , Citoesqueleto de Actina/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/antagonistas & inhibidores , Proteínas de Arabidopsis/genética , Tipificación del Cuerpo/genética , Tipificación del Cuerpo/fisiología , Polaridad Celular/genética , Polaridad Celular/fisiología , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Técnicas de Inactivación de Genes , Genes de Plantas , Germinación/genética , Germinación/fisiología , Glucólisis/genética , Modelos Biológicos , Mutación , Fosfoglicerato Quinasa/antagonistas & inhibidores , Fosfoglicerato Quinasa/genética , Fosfoglicerato Quinasa/metabolismo , Plantas Modificadas Genéticamente , Polen/genética , Polen/crecimiento & desarrollo , Polen/metabolismo , Transducción de Señal/genética , Proteínas de Unión al GTP rho/genéticaRESUMEN
Many tip-growing cells are capable of responding to guidance cues, during which cells precisely steer their growth toward the source of guidance signals. Though several players in signal perception have been identified, little is known about the downstream signaling that controls growth direction during guidance. Here, using combined modeling and experimental studies, we demonstrate that the growth guidance of Arabidopsis pollen tubes is regulated by the signaling network that controls tip growth. Tip-localized exocytosis plays a key role in this network by integrating guidance signals with the ROP1 Rho GTPase signaling and coordinating intracellular signaling with cell wall mechanics. This model reproduces the high robustness and responsiveness of pollen tube guidance and explains the connection between guidance efficiency and the parameters of the tip growth system. Hence, our findings establish an exocytosis-coordinated mechanism underlying the cellular pathfinding guided by signal gradients and the mechanistic linkage between tip growth and guidance.
Asunto(s)
Arabidopsis/crecimiento & desarrollo , Tubo Polínico/crecimiento & desarrollo , Arabidopsis/genética , Arabidopsis/fisiología , Proteínas de Arabidopsis/fisiología , Fenómenos Biomecánicos , Forma de la Célula/fisiología , Pared Celular/fisiología , Exocitosis/fisiología , Proteínas de Unión al GTP/fisiología , Modelos Biológicos , Plantas Modificadas Genéticamente , Tubo Polínico/citología , Transducción de Señal/fisiologíaRESUMEN
In plants, active transport of auxin plays an essential role in root development. Localization of the PIN1 auxin transporters to the basal membrane of cells directs auxin flow and depends on the trafficking mediator GNOM. GNOM-dependent auxin transport is vital for root development and thus offers a useful tool for the investigation of a possible tissue-specific response to dynamic auxin transport. To avoid pleiotropic effects, DEX-inducible expression of GNOM antisense RNA was used to disrupt GNOM expression transiently or persistently during embryonic root development. It was found that the elongation zone and the pericycle layer are the most sensitive to GNOM-dependent auxin transport variations, which is shown by the phenotypes in cell elongation and the initiation of lateral root primordia, respectively. This suggests that auxin dynamics is critical to cell differentiation and cell fate transition, but not to cell division. The results also reveal that GNOM-dependent auxin transport could affect local auxin biosynthesis. This suggests that local auxin biosynthesis may also contribute to the establishment of GNOM-dependent auxin gradients in specific tissues, and that auxin transport and local auxin biosynthesis may function together in the regulatory network for initiation and development of lateral root primordia. Thus, the data reveal a tissue-specific response to auxin transport and modulation of local auxin biosynthesis by auxin transport.
Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Factores de Intercambio de Guanina Nucleótido/genética , Ácidos Indolacéticos/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Arabidopsis/citología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Transporte Biológico , Dexametasona , Técnicas de Silenciamiento del Gen , Genes Reporteros , Factores de Intercambio de Guanina Nucleótido/metabolismo , Especificidad de Órganos , Fenotipo , Raíces de Plantas/citología , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Plantas Modificadas Genéticamente , ARN sin Sentido/genética , ARN de Planta/genética , Plantones/citología , Plantones/genética , Plantones/metabolismoRESUMEN
Pollen tubes elongate rapidly at their tips through highly polarized cell growth known as tip growth. Tip growth requires intensive exocytosis at the tip, which is supported by a dynamic cytoskeleton and vesicle trafficking. Several signaling pathways have been demonstrated to coordinate pollen tube growth by regulating cellular activities such as actin dynamics, exocytosis, and endocytosis. These signaling pathways crosstalk to form a signaling network that coordinates the cellular processes required for tip growth. The homeostasis of key signaling molecules is critical for the proper elongation of the pollen tube tip, and is commonly fine-tuned by positive and negative regulations. In addition to the major signaling pathways, emerging evidence implies the roles of other signals in the regulation of pollen tube growth. Here we review and discuss how these signaling networks modulate the rapid growth of pollen tubes.