RESUMEN
In some species, a crucial role has been demonstrated for the seed endosperm during germination. The endosperm has been shown to integrate environmental cues with hormonal networks that underpin dormancy and seed germination, a process that involves the action of cell wall remodeling enzymes (CWREs). Here, we examine the cell wall architectures of the endosperms of two related Brassicaceae, Arabidopsis (Arabidopsis thaliana) and the close relative Lepidium (Lepidium sativum), and that of the Solanaceous species, tobacco (Nicotiana tabacum). The Brassicaceae species have a similar cell wall architecture that is rich in pectic homogalacturonan, arabinan, and xyloglucan. Distinctive features of the tobacco endosperm that are absent in the Brassicaceae representatives are major tissue asymmetries in cell wall structural components that reflect the future site of radicle emergence and abundant heteromannan. Cell wall architecture of the micropylar endosperm of tobacco seeds has structural components similar to those seen in Arabidopsis and Lepidium endosperms. In situ and biomechanical analyses were used to study changes in endosperms during seed germination and suggest a role for mannan degradation in tobacco. In the case of the Brassicaceae representatives, the structurally homogeneous cell walls of the endosperm can be acted on by spatially regulated CWRE expression. Genetic manipulations of cell wall components present in the Arabidopsis seed endosperm demonstrate the impact of cell wall architectural changes on germination kinetics.
Asunto(s)
Brassicaceae/anatomía & histología , Brassicaceae/citología , Pared Celular/química , Endospermo/anatomía & histología , Endospermo/citología , Solanaceae/anatomía & histología , Solanaceae/citología , Arabidopsis/anatomía & histología , Arabidopsis/citología , Celulosa/metabolismo , Endospermo/crecimiento & desarrollo , Germinación , Lepidium sativum/anatomía & histología , Lepidium sativum/citología , Mananos/metabolismo , Monosacáridos/química , Mutación/genética , Pectinas/metabolismo , Nicotiana/anatomía & histología , Nicotiana/citologíaRESUMEN
Fibrillarin and the plant nucleolin homolog NopA64 are two important nucleolar proteins involved in pre-rRNA processing. In order to determine the effects of the altered gravity environment on the nucleolus, we have investigated the location of fibrillarin and NopA64 in nucleolar subcomponents of cress (Lepidium sativum L.) root meristematic cells grown under clinorotation, which reproduces an important feature of microgravity, namely, the absence of the orienting action of a gravity vector, and compared it to the location in control cells grown in normal 1 g conditions. Prior to these experiments, we report here the characterization of cress fibrillarin as a 41 kDa protein which can be isolated from meristematic cells in three nuclear fractions, namely, the soluble ribonucleoprotein fraction, the chromatin fraction, and the nuclear-matrix fraction. Furthermore, as reported for other species, the location of both fibrillarin and NopA64 in the cress cell nucleolus was in zones known to contain complex ribonucleoprotein particles involved in early pre-rRNA processing, i.e., processomes. Under altered gravity, a decrease in the quantity of both fibrillarin and NopA64 compared to controls was observed in the transition zone between fibrillar centers and the dense fibrillar component, as well as in the bulk of the dense fibrillar component. These data suggest that altered (reduced) gravity results in a lowered level of functional activity in the nucleolus.
Asunto(s)
Proteínas Cromosómicas no Histona/análisis , Gravedad Alterada , Lepidium sativum/metabolismo , Meristema/metabolismo , Fosfoproteínas/análisis , Raíces de Plantas/metabolismo , Proteínas de Unión al ARN/análisis , Western Blotting , Nucléolo Celular/metabolismo , Nucléolo Celular/ultraestructura , Lepidium sativum/citología , Lepidium sativum/ultraestructura , Meristema/citología , Meristema/ultraestructura , Microscopía Inmunoelectrónica/métodos , Proteínas Nucleares/análisis , Raíces de Plantas/citología , Raíces de Plantas/ultraestructura , NucleolinaRESUMEN
The endosperm is a barrier for radicle protrusion of many angiosperm seeds. Rupture of the testa (seed coat) and rupture of the endosperm are two sequential events during the germination of Lepidium sativum L. and Arabidopsis thaliana (L.) Heyhn. Abscisic acid (ABA) specifically inhibits the endosperm rupture of these two closely related Brassicaceae species. Lepidium seeds are large enough to allow the direct measurement of endosperm weakening by the puncture force method. We found that the endosperm weakens prior to endosperm rupture and that ABA delays the onset and decreases the rate of this weakening process in a dose-dependent manner. An early embryo signal is required and sufficient to induce endosperm weakening, which afterwards appears to be an organ-autonomous process. Gibberellins can replace this embryo signal; de novo gibberellin biosynthesis occurs in the endosperm and weakening is regulated by the gibberellin/ABA ratio. Our results suggest that the control of radicle protrusion during the germination of Brassicaceae seeds is mediated, at least in part, by endosperm weakening. We propose that Lepidium is an emerging Brassicaceae model system for endosperm weakening and that the complementary advantages of Lepidium and Arabidopsis can be used in parallel experiments to investigate the molecular mechanisms of endosperm weakening.
Asunto(s)
Ácido Abscísico/farmacología , Arabidopsis/embriología , Brassicaceae/embriología , Germinación/efectos de los fármacos , Lepidium sativum/embriología , Reguladores del Crecimiento de las Plantas/farmacología , Arabidopsis/citología , Arabidopsis/crecimiento & desarrollo , Brassicaceae/citología , Brassicaceae/crecimiento & desarrollo , Relación Dosis-Respuesta a Droga , Germinación/fisiología , Giberelinas/farmacología , Lepidium sativum/citología , Lepidium sativum/crecimiento & desarrollo , Semillas/efectos de los fármacos , Semillas/crecimiento & desarrollo , Semillas/fisiologíaRESUMEN
For the first time, in altered gravity, with antifibrillarin antibodies and immunogold microscopy fibrillarin that is one of the most important proteins of rRNA processing was localized. The quantitative study of the density of gold particles in the nuclelous, under clinorotation both the transition zone FC-DFC and the dense fibrillar component were less labeled as compared to the control. Obtained data allow us to suppose the lowering of the rRNA processing level in the nucleoli under the influence of altered gravity.
Asunto(s)
Nucléolo Celular/fisiología , Proteínas Cromosómicas no Histona/metabolismo , Gravedad Alterada/efectos adversos , Lepidium sativum/fisiología , Ribonucleoproteínas/metabolismo , Anticuerpos/metabolismo , Nucléolo Celular/ultraestructura , Proteínas Cromosómicas no Histona/inmunología , Proteínas Cromosómicas no Histona/ultraestructura , Inmunohistoquímica , Lepidium sativum/citología , Procesamiento Postranscripcional del ARN , ARN Ribosómico/metabolismo , Ribonucleoproteínas/inmunología , Ribonucleoproteínas/ultraestructura , RotaciónRESUMEN
While there is ample evidence for a role of auxin in root gravitropism, the seeming rapidity of gravi-induced changes in electrical parameters has so far been an argument against auxin being a primary signal in gravitropic signal transmission. To address this problem, we re-investigated the effect of gravistimulation on membrane voltages of Lepidium sativum L. and Vigna mungo L. root cells. In our hands, gravistimulation did not induce changes in membrane voltage in cells of the root cap statenchyma, root meristem or apical elongation zone that can be correlated with the orientation of the cells relative to the gravity vector. While these results challenge a model of rapid electrically based signal transmission, there is evidence for a slower signal propagation along gravistimulated L. sativum roots. Using multiple proton-selective microelectrodes to simultaneously measure surface pH on opposite root flanks at different distances from the root tip, we observed gravi-induced asymmetric pH changes at the surface of all investigated root zones. Upon gravistimulation, the surface pH decreased on the physically upper root flank and increased on the lower flank. The pH asymmetry appeared first [2.1+/-0.4 min (mean +/- SD) after tilting] at the root cap and then - with incrementing lag times - at the meristem (after 2.5+/-0.3 min at 300 micro m from root tip; after 3.7+/-0.4 min at 700 micro m) and apical elongation zone (4.8+/-0.5 min at 1,000 micro m), suggesting a basipetal progression of differential surface acidification at a rate of 250-350 micro m min(-1), consistent with reported auxin transport rates.