RESUMEN
KEY MESSAGE: The role of the root cap in the plant response to phosphate deprivation has been scarcely investigated. Here we describe early structural, physiological and molecular changes prior to the determinate growth program of the primary roots under low Pi and unveil a critical function of the transcription factor SOMBRERO in low Pi sensing. Mineral nutrient distribution in the soil is uneven and roots efficiently adapt to improve uptake and assimilation of sparingly available resources. Phosphate (Pi) accumulates in the upper layers and thus short and branched root systems proliferate to better exploit organic and inorganic Pi patches. Here we report an early adaptive response of the Arabidopsis primary root that precedes the entrance of the meristem into the determinate developmental program that is a hallmark of the low Pi sensing mechanism. In wild-type seedlings transferred to low Pi medium, the quiescent center domain in primary root tips increases as an early response, as revealed by WOX5:GFP expression and this correlates with a thicker root tip with extra root cap cell layers. The halted primary root growth in WT seedlings could be reversed upon transfer to medium supplemented with 250 µM Pi. Mutant and gene expression analysis indicates that auxin signaling negatively affects the cellular re-specification at the root tip and enabled identification of the transcription factor SOMBRERO as a critical element that orchestrates both the formation of extra root cap layers and primary root growth under Pi scarcity. Moreover, we provide evidence that low Pi-induced root thickening or the loss-of-function of SOMBRERO is associated with expression of phosphate transporters at the root tip. Our data uncover a developmental window where the root tip senses deprivation of a critical macronutrient to improve adaptation and surveillance.
Asunto(s)
Proteínas de Arabidopsis/fisiología , Ácidos Indolacéticos/metabolismo , Fosfatos/deficiencia , Reguladores del Crecimiento de las Plantas/fisiología , Cápsula de Raíz de Planta/crecimiento & desarrollo , Factores de Transcripción/fisiología , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Arabidopsis/fisiología , Regulación de la Expresión Génica de las Plantas , Meristema/crecimiento & desarrollo , Meristema/metabolismo , Meristema/fisiología , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/metabolismo , Transducción de SeñalRESUMEN
Plant roots secrete various substances with diverse functions against both plants and microbes in the rhizosphere. A major secretory substance is root-cap mucilage, whose functions have been well characterized, albeit mainly in crops. However, little is currently known about the developmental mechanisms of root-cap mucilage. Here, we show the accumulation and extrusion of root-cap mucilage in Arabidopsis. We found propidium iodide (PI) stainable structures between the plasma membrane and cell wall in the sixth layer of columella cells (c6) from the quiescent center. Ruthenium red staining and PI staining with calcium ions suggested that the structure comprises in part pectin polysaccharides. Electron microscopy revealed that the structure had a meshwork of electron-dense filaments that resembled periplasmic mucilage in other plants. In the c6 cells, we also observed many large vesicles with denser meshwork filaments to periplasmic mucilage, which likely mediate the transport of mucilage components. Extruded mucilage was observed outside a partially degraded cell wall in the c7 cells. Moreover, we found that the Class IIB NAC transcription factors BEARSKIN1 (BRN1) and BRN2, which are known to regulate the terminal differentiation of columella cells, were required for the efficient accumulation of root-cap mucilage in Arabidopsis. Taken together, our findings reveal the accumulation of and dynamic changes in periplasmic mucilage during columella cell development in Arabidopsis.
Asunto(s)
Arabidopsis/crecimiento & desarrollo , Periplasma/metabolismo , Mucílago de Planta/metabolismo , Cápsula de Raíz de Planta/metabolismo , Arabidopsis/citología , Arabidopsis/metabolismo , Arabidopsis/ultraestructura , Colorantes , Microscopía Electrónica de Transmisión , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/ultraestructura , PropidioRESUMEN
The root cap supports root growth by protecting the root meristem, sensing gravity and interacting with the rhizosphere through metabolite secretion and cell dispersal. Sustained root cap functions therefore rely on balanced proliferation of proximal stem cells and regulated detachment of distal mature cells. Although the gene regulatory network that governs stem cell activity in the root cap has been extensively studied in Arabidopsis, the mechanisms by which root cap cells mature and detach from the root tip are poorly understood. We performed a detailed expression analysis of three regulators of root cap differentiation, SOMBRERO, BEARSKIN1 and BEARSKIN2, and identified their downstream genes. Our results indicate that expression of BEARSKIN1 and BEARSKIN2 is associated with cell positioning on the root surface. We identified a glycosyl hydrolase 28 (GH28) family polygalacturonase (PG) gene as a direct target of BEARSKIN1. Overexpression and loss-of-function analyses demonstrated that the protein encoded by this PG gene facilitates cell detachment. We thus revealed a molecular link between the key regulators of root cap differentiation and the cellular events underlying root cap-specific functions.
Asunto(s)
Arabidopsis , Diferenciación Celular/genética , Movimiento Celular/genética , Cápsula de Raíz de Planta/crecimiento & desarrollo , Factores de Transcripción/fisiología , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/fisiología , Regulación de la Expresión Génica de las Plantas , Meristema/citología , Meristema/crecimiento & desarrollo , Meristema/metabolismo , Cápsula de Raíz de Planta/citología , Raíces de Plantas/citología , Raíces de Plantas/crecimiento & desarrollo , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Root tips of many plant species release a number of border, or border-like, cells that are thought to play a major role in the protection of root meristem. However, little is currently known on the structure and function of the cell wall components of such root cells. Here, we investigate the sugar composition of the cell wall of the root cap in two species: pea (Pisum sativum), which makes border cells, and Brassica napus, which makes border-like cells. We find that the cell walls are highly enriched in arabinose and galactose, two major residues of arabinogalactan proteins. We confirm the presence of arabinogalactan protein epitopes on root cap cell walls using immunofluorescence microscopy. We then focused on these proteoglycans by analyzing their carbohydrate moieties, linkages, and electrophoretic characteristics. The data reveal (1) significant structural differences between B. napus and pea root cap arabinogalactan proteins and (2) a cross-link between these proteoglycans and pectic polysaccharides. Finally, we assessed the impact of root cap arabinogalactan proteins on the behavior of zoospores of Aphanomyces euteiches, an oomycetous pathogen of pea roots. We find that although the arabinogalactan proteins of both species induce encystment and prevent germination, the effects of both species are similar. However, the arabinogalactan protein fraction from pea attracts zoospores far more effectively than that from B. napus. This suggests that root arabinogalactan proteins are involved in the control of early infection of roots and highlights a novel role for these proteoglycans in root-microbe interactions.
Asunto(s)
Aphanomyces/citología , Aphanomyces/crecimiento & desarrollo , Brassica napus/metabolismo , Quimiotaxis/efectos de los fármacos , Mucoproteínas/farmacología , Pisum sativum/metabolismo , Cápsula de Raíz de Planta/metabolismo , Aphanomyces/efectos de los fármacos , Brassica napus/citología , Brassica napus/efectos de los fármacos , Brassica napus/microbiología , Pared Celular/efectos de los fármacos , Pared Celular/metabolismo , Precipitación Química , Electroforesis en Gel de Poliacrilamida , Glucósidos/metabolismo , Microscopía Fluorescente , Monosacáridos/química , Monosacáridos/metabolismo , Mucoproteínas/química , Pisum sativum/citología , Pisum sativum/efectos de los fármacos , Pisum sativum/microbiología , Floroglucinol/análogos & derivados , Floroglucinol/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/farmacología , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/efectos de los fármacosRESUMEN
BACKGROUND AND AIMS: Root caps release border cells, which play central roles in microbe interaction and root protection against soil stresses. However, the number and connectivity of border cells differ widely among plant species. Better understanding of key border-cell phenotype across species will help define the total function of border cells and associated genes. METHODS: The spatio-temporal detachment of border cells in the leguminous tree Acacia mangium was investigated by using light and fluorescent microscopy with fluorescein diacetate, and their number and structural connectivity compared with that in soybean (Glycine max). KEY RESULTS: Border-like cells with a sheet structure peeled bilaterally from the lateral root cap of A. mangium. Hydroponic root elongation partially facilitated acropetal peeling of border-like cells, which accumulate as a sheath that covers the 0- to 4-mm tip within 1 week. Although root elongation under friction caused basipetal peeling, lateral root caps were minimally trimmed as compared with hydroponic roots. In the meantime, A. mangium columella caps simultaneously released single border cells with a number similar to those in soybean. CONCLUSIONS: These results suggest that cell type-specific inhibitory factors induce a distinct defective phenotype in single border-cell formation in A. mangium lateral root caps.
Asunto(s)
Acacia/citología , Acacia/crecimiento & desarrollo , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/crecimiento & desarrollo , Acacia/genética , Procesos de Crecimiento Celular/genética , Procesos de Crecimiento Celular/fisiología , Fenómenos Fisiológicos Celulares , Cápsula de Raíz de Planta/genética , Glycine max/citologíaRESUMEN
Results of the electron-microscopic investigation of root apices of Arabidopsis thaliana 3-, 5- and 7-days-old seedlings grown in the stationary conditions and under clinorotation are presented. It was shown the similarity in the root apex cell ultrastructure in control and under clinorotation. At the same time there were some differences in the ultrastructure of statocytes and the distal elongation zone under clinorotation. For the first time the sensitivity of ER-bodies, which are derivatives of GER and contain beta-glucosidase, to the influence of simulated microgravity was demonstrated by increased quantity and area of ER-bodies at the cell section as well as by higher variability of their form under clinorotation. A degree of these changes correlated with the duration of clinorotation. On the basis of experimental data a protective role of ER-bodies in adaptation of plants to microgravity is supposed.
Asunto(s)
Arabidopsis/ultraestructura , Cápsula de Raíz de Planta/ultraestructura , Arabidopsis/citología , Arabidopsis/crecimiento & desarrollo , Retículo Endoplásmico/ultraestructura , Gravitropismo , Microscopía Electrónica , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/crecimiento & desarrollo , Rotación , Plantones/citología , Plantones/crecimiento & desarrollo , Factores de Tiempo , Simulación de IngravidezRESUMEN
SUMMARY: The C(18) ketone (5E,7E)-6-methyl-8-(2,6,6-trimethylcyclohex-1-enyl)octa-5,7-dien-2-one (D'orenone) has been postulated to be an early cleavage product of beta-carotene en route to trisporic acids; these act as morphogenetic factors during the sexual reproduction of zygomycetes. Here we report that D'orenone blocks the highly polarized tip growth of root hairs, causing tip growth to stop completely within a few minutes. Importantly, external auxin reverses the effects of D'orenone on root hairs. Further analysis revealed that D'orenone lowers the auxin concentration in trichoblasts via PIN2-mediated auxin efflux to below the critical levels essential for root hair growth. D'orenone specifically increases PIN2 protein abundance without affecting PIN2 transcripts, and the PIN2 expression domain enlarges and shifts basipetally, resulting in more active auxin transport. The observation that D'orenone does not interfere with the root hair growth in roots of null mutant lines provides additional evidence that PIN2 is its specific target.
Asunto(s)
Ácidos Indolacéticos/metabolismo , Cetonas/farmacología , Cápsula de Raíz de Planta/fisiología , Raíces de Plantas/crecimiento & desarrollo , Arabidopsis/efectos de los fármacos , Arabidopsis/fisiología , Dimetilsulfóxido/farmacología , Ácidos Indolacéticos/farmacología , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/efectos de los fármacos , Raíces de Plantas/citología , Raíces de Plantas/efectos de los fármacos , Proteína 1 de Unión a Repeticiones Teloméricas/efectos de los fármacos , Proteína 1 de Unión a Repeticiones Teloméricas/fisiologíaRESUMEN
A two-dimensional micromorphometric analysis of a root cap in Allium cepa included the measurement of the areas of about six thousand cell profiles on both longitudinal and transversal sections. Basing on the results of this analysis, quantitative descriptions of two cell populations from the columella and periferic part of the root cap have been created. The scheme of arrangement of root cap cells, located at a distance of 250 microm or less from a root tip, has been developed.
Asunto(s)
Cebollas/citología , Cápsula de Raíz de Planta/citología , Cebollas/fisiología , Cápsula de Raíz de Planta/fisiologíaRESUMEN
Several studies on cytogenetic characterisation of passion flowers are helpful to elucidate doubts about taxa relationships, delimitation and classification into more coherent groups based on karyomorphological data. Molecular and conventional cytogenetic techniques were applied to three Passiflora species with red flowers, P. coccinea, P. vitifolia and P. tholozanii, for species karyotype relationships. Additionally, for descriptive morphology, were used flowers, leaves and seeds. Results describe for the first time the karyomorphological and chromosome number (2n = 18) for P. tholozanii. anova was performed (P < 0.05) and statistical significance for average chromosome size (CV: 16.53%) between species. Genomic in situ hybridisation (GISH) proved relationships between P. coccinea and P. tholozanii, which suggests a common origin, however, we could not identify hybridisation between genomic probes from P. vitifolia in P. tholozanii chromosomes. Among the species analysed, P. tholozanii has great similarity in karyotypic and morphology to P. coccinea but not to P. vitifolia. We suggest the inclusion of P. tholozanii in the same subgenus and section as P. coccinea based on the similarity in karyomorphological and morphological traits between the species. Additionally, GISH might indicate a common or hybrid origin of P. tholozanii.
Asunto(s)
Passiflora/citología , Colorantes Azulados , Cromosomas de las Plantas/genética , Análisis Citogenético , Cariotipificación , Passiflora/anatomía & histología , Passiflora/genética , Cápsula de Raíz de Planta/anatomía & histología , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/genéticaRESUMEN
Pea (Pisum sativum) root cap releases a large number of living border cells that secrete abundant mucilage into the extracellular medium. Mucilage contains a complex mixture of polysaccharides, proteins and secondary metabolites important for its structure and function in defense. Unlike xyloglucan and cellulose, pectin and arabinogalactan proteins have been investigated in pea root and shown to be major components of border cell walls and mucilage. In this study, we investigated the occurrence of xyloglucan and cellulose in pea border cells and mucilage using cytochemical staining, immunocytochemistry and laser scanning confocal microscopy. Our data show that i) unlike cellulose, xyloglucan is highly present in the released mucilage as a dense fibrillary network enclosing border cells and ii) that xyloglucan and cellulose form molecular cross-bridges that tether cells and maintain them attached together. These findings suggest that secreted xyloglucan is essential for mucilage strengthening and border cell attachment and functioning.
Asunto(s)
Celulosa/metabolismo , Glucanos/metabolismo , Pisum sativum/metabolismo , Raíces de Plantas/citología , Xilanos/metabolismo , Microscopía Confocal , Pisum sativum/ultraestructura , Mucílago de Planta/metabolismo , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/metabolismo , Raíces de Plantas/metabolismo , Raíces de Plantas/ultraestructuraRESUMEN
The root cap releases cells that produce massive amounts of mucilage containing polysaccharides, proteoglycans, extracellular DNA (exDNA) and a variety of antimicrobial compounds. The released cells - known as border cells or border-like cells - and mucilage secretions form networks that are defined as root extracellular traps (RETs). RETs are important players in root immunity. In animals, phagocytes are some of the most abundant white blood cells in circulation and are very important for immunity. These cells combat pathogens through multiple defence mechanisms, including the release of exDNA-containing extracellular traps (ETs). Traps of neutrophil origin are abbreviated herein as NETs. Similar to phagocytes, plant root cap-originating cells actively contribute to frontline defence against pathogens. RETs and NETs are thus components of the plant and animal immune systems, respectively, that exhibit similar compositional and functional properties. Herein, we describe and discuss the formation, molecular composition and functional similarities of these similar but different extracellular traps.
Asunto(s)
Trampas Extracelulares/fisiología , Neutrófilos/fisiología , Inmunidad de la Planta/fisiología , Raíces de Plantas/fisiología , Animales , ADN de Plantas/fisiología , Trampas Extracelulares/inmunología , Humanos , Neutrófilos/citología , Neutrófilos/inmunología , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/inmunología , Cápsula de Raíz de Planta/fisiología , Raíces de Plantas/inmunología , Especies Reactivas de Oxígeno/metabolismo , RizosferaRESUMEN
The results of light- and electron-microscopic investigations of root apices of Beta vulgaris 3-day-old seedlings grown in the stationary conditions and under clinorotation are presented. It was shown that ultrastructure and topography of organelles in root cap statocytes (graviperceptive cells) and in the cells of distal elongation zone clearly reflected the different direction in their growth and differentiation in space and time in dependence on specialization and functions. Cell growth and genetically determined differentiation occur similarly to control, although certain differences in ultrastructure are evident on metabolism changes.
Asunto(s)
Beta vulgaris , Diferenciación Celular , Raíces de Plantas , Beta vulgaris/citología , Beta vulgaris/crecimiento & desarrollo , Beta vulgaris/ultraestructura , Gravitropismo , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/crecimiento & desarrollo , Cápsula de Raíz de Planta/ultraestructura , Raíces de Plantas/citología , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/ultraestructura , Rotación , Simulación de IngravidezRESUMEN
Cell death mechanism triggered by aluminium (Al) ion was investigated at root apex of tobacco (cultivar Bright Yellow) and in cultured tobacco cell line BY-2 derived from Bright Yellow, focusing on VPE genes (NtVPE1a, NtVPE1b, NtVPE2, NtVPE3). Cell death was detected as a loss of integrity of the plasma membrane by vital staining with fluorescein diacetate (in root apex) and Evans blue (in BY-2), respectively. At root apex, the upregulation of gene expression of VPE1a and VPE1b was observed significantly after 9h of Al exposure in parallel with an enhancement of cell death, while the upregulation of VPE2 and VPE3 were observed later. Similarly, in BY-2 cells, the upregulation of VPE1a and VPE1b and the enhancement of cell death were synchronously observed after 3-h exposure to Al, while the upregulation of VPE2 and VPE3 occurred later. RNA interference (RNAi) lines of each of the VPEs were constructed in BY-2 cells. Comparative studies between wild-type and the RNAi lines indicated that both Al-enhanced VPE activity and Al-induced cell death were significantly suppressed in the RNAi lines of VPE1 (dual suppressor of VPE1a and VPE1b), but not in the RNAi lines of VPE2 and that of VPE3. Taken together, we conclude that the upregulation of VPE1 gene expression and following enhancement of VPE activity under Al stress cause cell death in actively growing or elongating cells of tobacco.
Asunto(s)
Aluminio/toxicidad , Cisteína Endopeptidasas/metabolismo , Nicotiana/efectos de los fármacos , Proteínas de Plantas/metabolismo , Cápsula de Raíz de Planta/efectos de los fármacos , Contaminantes del Suelo/toxicidad , Adsorción , Aluminio/química , Aluminio/metabolismo , Biomarcadores/metabolismo , Línea Celular , Membrana Celular/química , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Forma de la Célula/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cisteína Endopeptidasas/química , Cisteína Endopeptidasas/genética , Inducción Enzimática/efectos de los fármacos , Isoenzimas/antagonistas & inhibidores , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/metabolismo , Cinética , Meristema/citología , Meristema/efectos de los fármacos , Meristema/crecimiento & desarrollo , Meristema/metabolismo , Proteínas de Plantas/agonistas , Proteínas de Plantas/antagonistas & inhibidores , Proteínas de Plantas/genética , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/crecimiento & desarrollo , Cápsula de Raíz de Planta/metabolismo , Raíces de Plantas/citología , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Interferencia de ARN , Contaminantes del Suelo/química , Contaminantes del Suelo/metabolismo , Propiedades de Superficie , Nicotiana/citología , Nicotiana/crecimiento & desarrollo , Nicotiana/metabolismoRESUMEN
The survival of a plant depends upon the capacity of root tips to sense and move towards water and other nutrients in the soil. Perhaps because of the root tip's vital role in plant health, it is ensheathed by large populations of detached somatic cells - root 'border' cells - which have the ability to engineer the chemical and physical properties of the external environment. Of particular significance, is the production by border cells of specific chemicals that can dramatically alter the behavior of populations of soilborne microflora. Molecular approaches are being used to identify and manipulate the expression of plant genes that control the production and the specialized properties of border cells in transgenic plants. Such plants can be used to test the hypothesis that these unusual cells act as a phalanx of biological 'goalies', which neutralize dangers to newly generated root tissue as the root tip makes its way through soil.
Asunto(s)
Cápsula de Raíz de Planta/fisiología , Suelo , Cápsula de Raíz de Planta/anatomía & histología , Cápsula de Raíz de Planta/citología , Transducción de SeñalRESUMEN
The plant root cap, surrounding the very tip of the growing root, perceives and transmits environmental signals to the inner root tissues. In Arabidopsis thaliana, auxin released by the root cap contributes to the regular spacing of lateral organs along the primary root axis. Here, we show that the periodicity of lateral organ induction is driven by recurrent programmed cell death at the most distal edge of the root cap. We suggest that synchronous bursts of cell death in lateral root cap cells release pulses of auxin to surrounding root tissues, establishing the pattern for lateral root formation. The dynamics of root cap turnover may therefore coordinate primary root growth with root branching in order to optimize the uptake of water and nutrients from the soil.
Asunto(s)
Apoptosis , Arabidopsis/crecimiento & desarrollo , Ácidos Indolacéticos/metabolismo , Cápsula de Raíz de Planta/crecimiento & desarrollo , Arabidopsis/citología , Arabidopsis/metabolismo , Epidermis de la Planta/citología , Epidermis de la Planta/crecimiento & desarrollo , Epidermis de la Planta/metabolismo , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/metabolismo , Receptores del Ligando Inductor de Apoptosis Relacionado con TNF/genética , Receptores del Ligando Inductor de Apoptosis Relacionado con TNF/metabolismo , Transducción de Señal , Suelo , Agua/metabolismoRESUMEN
NASA: This review of plant gravity sensing examines sensing in organ gravitropism, sensing in single-cell gravitropism, and nongravitropic sensing. Topics related to sensing in organ gravitropism are (1) identification of the gravitropic susceptors, including intracellular asymmetry in equilibrium position and after reorientation, susceptor signal-to-noise ratio, signal integration over threshold stimulation periods, intracellular asymmetry and gravitropic competence, and starch deficiency and gravitropic competence; (2) possible root statocytes and receptors, including identification of presumptive statocytes, cytology, and possible receptors and models of sensing; and (3) negatively gravitropic organs, including identification and distribution of presumptive statocytes and cytology and possible receptors. Topics related to nongravitropic sensing include gravitaxis, reaction wood, gravimorphogenesis, other gravity-influenced organ movements, and cytoplasmic streaming.^ieng
Asunto(s)
Gravitropismo/fisiología , Sensación de Gravedad/fisiología , Desarrollo de la Planta , Cápsula de Raíz de Planta/ultraestructura , Plastidios/fisiología , Bryopsida/citología , Bryopsida/crecimiento & desarrollo , Bryopsida/fisiología , Bryopsida/ultraestructura , Chlorophyta/citología , Chlorophyta/crecimiento & desarrollo , Chlorophyta/fisiología , Chlorophyta/ultraestructura , Retículo Endoplásmico/fisiología , Células Vegetales , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/crecimiento & desarrollo , Cápsula de Raíz de Planta/fisiología , Raíces de Plantas/citología , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/fisiología , Raíces de Plantas/ultraestructura , Plantas/ultraestructuraRESUMEN
Root growth and cell differentiation were analysed in lentil seedlings grown (1) in microgravity (F microg), (2) on the 1 x g centrifuge (F1 x g), (3) in microgravity and placed on the 1 x g centrifuge for 4 h [F(microg + 1 x g)], (4) on the 1 x g centrifuge and placed in microgravity for 4 h [F(1 x g + microg)]. In microgravity, there were strong oscillations of the root tip, even when the seedlings were grown first on the 1 x g centrifuge [F(1 x g + microg)]. In the [F(microg + 1 x g)] sample, the roots grown in microgravity were oblique with respect to the 1 x g acceleration when the seedlings were placed on the centrifuge. They were therefore gravistimulated. However, root length was similar in the 4 samples after 29 h of growth and growth rate of the root was the same between 25 h and 29 h although it appeared to be slightly greater in the [F(microg + 1 x g)] sample. Cell elongation was analysed as a function of the distance from the root cap junction. Cell length was similar in the seedlings grown in microgravity or on the 1 x g centrifuge. The transfer from the 1 x g centrifuge to microgravity [F(1 x g + microg)] did not modify cell elongation in the roots. Cell length in the roots which were grown in microgravity and gravistimulated [F(microg + 1 x g)] was different from that observed in microgravity but this was only due to gravistimulation. Thus, gravity does not have an effect on cell elongation when the roots are strictly oriented in the vertical position but it does as soon as the root tip deviates from this orientation.
Asunto(s)
Gravitropismo/fisiología , Raíces de Plantas/crecimiento & desarrollo , Vuelo Espacial , Ingravidez , Centrifugación , Fabaceae/citología , Fabaceae/crecimiento & desarrollo , Cápsula de Raíz de Planta/citología , Raíces de Plantas/citología , Plantas MedicinalesRESUMEN
Microsomal and soluble protein fractions from Vicia faba root tips were used for SDS-PAGE and Western-immunoblot analysis with anti-ubiquitin antibodies after 9 h clinostat treatment of the plants. In contrast to soluble proteins omnilateral gravistimulation (9 h) resulted in an enhanced proteolytic capacity for microsomal proteins. The increase of vanadate-sensitive ATPase activity was 83% after 9 h clinostat treatment, when the enzyme activity was measured directly after membrane preparation. Enhanced ATPase activity was correlated with the appearance of a polypeptide of about 100 kDa and its fragments (93 and 80 kDa). ATPases are not the only membrane bound proteins, which are changed during clinostat treatment, as several ubiquitinated polypeptides were also affected. A 1 h storage of microsomal fractions led to a shift of band intensities on ubiquitin-specific Western-blots. The demonstrated effect could not be observed, when fractions were isolated in the presence of protease inhibitors. In accordance with the polypeptide analysis omnilateral gravistimulation resulted in an enhanced capacity to degrade specific microsomal ubiquitin-conjugates, whereas the soluble ubiquitin-pool was not visibly affected.
Asunto(s)
Adenosina Trifosfatasas/metabolismo , Fabaceae/metabolismo , Gravitación , Microsomas/metabolismo , Cápsula de Raíz de Planta/metabolismo , Plantas Medicinales , Rotación , Ubiquitinas/metabolismo , Fabaceae/citología , Fabaceae/enzimología , Membranas Intracelulares/enzimología , Membranas Intracelulares/metabolismo , Proteínas de la Membrana/metabolismo , Microsomas/enzimología , Proteínas de Plantas/metabolismo , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/enzimología , ATPasas de Translocación de Protón/metabolismoRESUMEN
The intra-S-phase checkpoint response to hydroxyurea (HU)-mediated arrest of DNA replication was analysed in root meristems of two legumes, Pisum sativum and Vicia faba. The obtained results suggest that a molecular signal which invokes mechanisms allowing the cells to override the S-M dependency control system may be generated by caffeine (CF) and a number of alternative, yet related chemical agents, benzyl-6-aminopurine (BAP), 2-aminopurine (2-AP), and 6-dimethylaminopurine (DMAP). A variety of aberrant mitotic divisions included chromosomal breaks and gaps, lost and lagging chromatids and chromosomes, acentric fragments, chromosome bridges and micronuclei. Furthermore, similar effects induced by sodium vanadate, an inhibitor of protein phosphatases, extend the number of inhibitors capable of inducing premature chromosome condensation (PCC) in root meristem cells, as well as the range of possible regulatory pathways leading to the transition from S-phase arrest towards abnormal mitosis. Until preprophase, FITC-conjugated monoclonal antibodies (alpha-Y(a)b-FITC) that specifically recognize phosphorylated form of threonine indicate no evident cell cycle-dependent changes in an overall phosphorylation status of root meristem cells in the control plants. Irrespective of the stage of interphase, alpha-Y(p)ab-FITC was localized basically in the cytoplasm, whereas nuclear staining was considerably weaker, with a significant fluorescence confined merely to nucleolar regions. The intensity of alpha-Y(p)ab-FITC staining in HU/CF-treated seedlings was found higher than that in the control plants (with the exception of G2 cells), suggesting a general increase in the level of protein phosphorylation, a physiological response mediated probably by an enhanced activity of the cdc-like protein kinase(s).
Asunto(s)
Cromosomas/metabolismo , ADN de Plantas/metabolismo , Mitosis/fisiología , Pisum sativum/crecimiento & desarrollo , Proteínas de Plantas/metabolismo , Cápsula de Raíz de Planta/crecimiento & desarrollo , Regulación hacia Arriba/genética , Anticuerpos Monoclonales/farmacología , Cafeína/farmacología , Núcleo Celular/efectos de los fármacos , Núcleo Celular/genética , Núcleo Celular/metabolismo , Aberraciones Cromosómicas/efectos de los fármacos , Cromosomas/efectos de los fármacos , ADN de Plantas/efectos de los fármacos , Colorantes Fluorescentes , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/fisiología , Genes cdc/efectos de los fármacos , Genes cdc/fisiología , Hidroxiurea/farmacología , Mitosis/efectos de los fármacos , Pisum sativum/citología , Pisum sativum/genética , Fosforilación/efectos de los fármacos , Proteínas de Plantas/efectos de los fármacos , Cápsula de Raíz de Planta/citología , Cápsula de Raíz de Planta/efectos de los fármacos , Purinas/farmacología , Regulación hacia Arriba/efectos de los fármacos , Vanadatos/farmacologíaRESUMEN
Axial force was applied to the root tip of corn (Zea mays L. cv. Merit) seedlings using a computerized, feedback-controlled mechanical device. The system's feedback capability allowed continuous control of a constant tip load, and the attached displacement transducer provided the time course of root elongation. Loads up to 7.5 g decreased the root elongation rate by 0.13 mm h-1 g-1, but loads 7.5 to 17.5 g decreased the growth rate by only 0.04 mm h-1 g-1. Loads higher than 18 g stopped root elongation completely. Measurement of the cross-sectional areas of the root tips indicated that the 18 g load had applied about 0.98 MPa of axial pressure to the root, thereby exceeding the root's ability to respond with increased turgor pressure. Recorded time-lapse images of loaded roots showed that radial thickening (swelling) occurred behind the root cap, whose cross-sectional area increased with tip load.