RESUMEN
We have developed a methodology for the systematic generation of a large image dataset of macerated wood references, which we used to generate image data for nine hardwood genera. This is the basis for a substantial approach to automate, for the first time, the identification of hardwood species in microscopic images of fibrous materials by deep learning. Our methodology includes a flexible pipeline for easy annotation of vessel elements. We compare the performance of different neural network architectures and hyperparameters. Our proposed method performs similarly well to human experts. In the future, this will improve controls on global wood fiber product flows to protect forests.
Asunto(s)
Aprendizaje Profundo , Procesamiento de Imagen Asistido por Computador , Madera , Madera/anatomía & histología , Procesamiento de Imagen Asistido por Computador/métodos , Microscopía/métodos , Redes Neurales de la ComputaciónRESUMEN
Extrafloral nectaries secrete a sweet sugar cocktail that lures predator insects for protection from foraging herbivores. Apart from sugars and amino acids, the nectar contains the anions chloride and nitrate. Recent studies with Populus have identified a type of nectary covered by apical bipolar epidermal cells, reminiscent of the secretory brush border epithelium in animals. Border epithelia operate transepithelial anion transport, which is required for membrane potential and/or osmotic adjustment of the secretory cells. In search of anion transporters expressed in extrafloral nectaries, we identified PttSLAH3 (Populus tremula × Populus tremuloides SLAC1 Homologue3), an anion channel of the SLAC/SLAH family. When expressed in Xenopus oocytes, PttSLAH3 displayed the features of a voltage-dependent anion channel, permeable to both nitrate and chloride. In contrast to the Arabidopsis SLAC/SLAH family members, the poplar isoform PttSLAH3 is independent of phosphorylation activation by protein kinases. To understand the basis for the autonomous activity of the poplar SLAH3, we generated and expressed chimera between kinase-independent PttSLAH3 and kinase-dependent Arabidopsis AtSLAH3. We identified the N-terminal tail and, to a lesser extent, the C-terminal tail as responsible for PttSLAH3 kinase-(in)dependent action. This feature of PttSLAH3 may provide the secretory cell with a channel probably controlling long-term nectar secretion.
Asunto(s)
Aniones/metabolismo , Epitelio/metabolismo , Canales Iónicos/metabolismo , Proteínas de Plantas/metabolismo , Populus/metabolismo , Proteínas Quinasas/metabolismo , Arabidopsis/efectos de los fármacos , Arabidopsis/metabolismo , Epitelio/efectos de los fármacos , Flores/efectos de los fármacos , Flores/metabolismo , Activación del Canal Iónico/efectos de los fármacos , Nitratos/farmacología , Néctar de las Plantas , Proteínas de Plantas/química , Populus/efectos de los fármacos , Proteínas Recombinantes de Fusión/metabolismo , Relación Estructura-ActividadRESUMEN
Very long chain lipids are important components of the plant cuticle that establishes the boundary surface of aerial organs. In addition, these lipids were detected in the extracellular pollen coat (tryphine), where they play a crucial role in appropriate pollen-stigma communication. As such they are involved in the early interaction of pollen with the stigma. A substantial reduction in tryphine lipids was shown to compromise pollen germination and, consequently, resulted in male sterility. We investigated the role of two long-chain acyl-CoA synthetases (LACSs) in Arabidopsis with respect to their contribution to the production of tryphine lipids. LACS was shown to provide CoA-activated very long chain fatty acids (VLCFA-CoAs) to the pathways of wax biosynthesis. The allocation of sufficient quantities of VLCFA-CoA precursors should therefore be relevant to the generation of tryphine lipids. Here, we report on the identification of lacs1 lacs4 double knock-out mutant lines that were conditionally sterile and showed significant reductions in pollen coat lipids. Whereas the contributions of both LACS proteins to surface wax levels were roughly additive, their co-operation in tryphine lipid biosynthesis was clearly more complex. The inactivation of LACS4 resulted in increased levels of tryphine lipids accompanied by morphological anomalies of the pollen grains. The additional inactivation of LACS1 neutralized the morphological defects, decreased the tryphine lipids far below wild-type levels and resulted in conditionally sterile pollen.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Coenzima A Ligasas/metabolismo , Metabolismo de los Lípidos , Lípidos/biosíntesis , Polen/fisiología , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Coenzima A Ligasas/genética , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Técnicas de Inactivación de Genes , Infertilidad Vegetal , Polen/genética , Polen/ultraestructura , Ceras/metabolismoRESUMEN
To investigate whether leaves of plane trees (Platanus orientalis) are damaged by traffic pollution, trees from a megacity (Mashhad, Iran) and a rural area were investigated. Soil and air from the urban centre showed enrichment of several toxic elements, but only lead was enriched in leaves. Leaf size and stomata density were lower at the urban site. At the urban site leaf surfaces were heavily loaded by dust particles but the stomata were not occluded; the cuticle was thinner; other anatomical properties were unaffected suggesting that plane trees can cope with traffic exhaust in megacities.
Asunto(s)
Contaminantes Atmosféricos/toxicidad , Helechos/efectos de los fármacos , Hojas de la Planta/efectos de los fármacos , Árboles/efectos de los fármacos , Monitoreo del Ambiente , Helechos/citología , Helechos/metabolismo , Material Particulado/metabolismo , Material Particulado/toxicidad , Hojas de la Planta/citología , Hojas de la Planta/metabolismo , Estomas de Plantas/efectos de los fármacos , Estomas de Plantas/fisiología , Estomas de Plantas/ultraestructura , Árboles/citología , Árboles/metabolismoRESUMEN
Maintenance of ion concentration gradients is essential for the function of many organs, including the kidney, the cornea, and the inner ear. Ion concentrations and fluid content in the cornea are regulated by endothelial cells that separate the collagenous avascular corneal stroma from the anterior eye chamber. Failure to maintain correct ion concentrations leads to swelling and destruction of the cornea. In the inner ear, the stria vascularis is responsible for generating proper ion concentrations in the endolymph, which is essential for hearing. Mutations of SLC4A11 in humans lead to syndromes associated with corneal dystrophy and perceptive deafness. The molecular mechanisms underlying these symptoms are poorly understood, impeding therapeutic interventions. The ion transporter SLC4A11 mediates sodium-dependent transport of borate as well as flux of sodium and hydroxyl ions in vitro. Here, we show that SLC4A11 is expressed in the endothelial cells of the cornea where it prevents severe morphological changes of the cornea caused by increased sodium chloride concentrations in the stroma. In the inner ear, SLC4A11 is located in fibrocytes underlying the stria vascularis. Loss of SLC4A11 leads to morphological changes in the fibrocytes and deafness. We demonstrate that SLC4A11 is essential for the generation of the endocochlear potential but not for regulation of potassium concentrations in the endolymph. In the kidney, SLC4A11 is expressed in the thin descending limb of Henle loop. SLC4A11 is essential for urinary concentration, suggesting that SLC4A11 participates in the countercurrent multiplication that concentrates urine in the kidney medulla.
Asunto(s)
Proteínas de Transporte de Anión/fisiología , Sordera/prevención & control , Distrofia Endotelial de Fuchs/prevención & control , Ósmosis , Poliuria/prevención & control , Cloruro de Sodio/metabolismo , Simportadores/fisiología , Animales , Sordera/metabolismo , Oído Interno/metabolismo , Oído Interno/patología , Endolinfa/metabolismo , Femenino , Distrofia Endotelial de Fuchs/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Poliuria/metabolismo , Potasio/metabolismo , Fracciones SubcelularesRESUMEN
Using quantitative X-ray microanalysis in combination with CeCl3-based cytochemical staining of hydrogen peroxide (H2O2) we have developed a new solution for quantification of H2O2 at the subcellular level. Quantitative X-ray microanalysis of plastic-embedded leaves of Populus euphratica Oliv. showed that the obtained cerium precipitates by CeCl3 staining were the mixture of cerium perhydroxides and cerium phosphate, in which the fractions of CePO4 were: (1) 52-74% in cell walls of fresh leaf segments, and (2) 34-70% in the cytoplasm in 10 mM H2O2-treated leaf segments that were previously freeze-dried. Taking the concentration of cerium phosphate as staining background, we reached the cellular concentration of cerium perhydroxides and the corresponding concentration of H2O2. Results showed that H2O2 was present in the cytoplasm of rehydrated leaf segments (29-58 mM), but in fresh leaves, H2O2 was observed in the walls of all measured cell types (17-74 mM).
Asunto(s)
Microanálisis por Sonda Electrónica , Peróxido de Hidrógeno/análisis , Populus/química , Cerio , Hidróxidos , Espacio Intracelular/química , Fosfatos , Hojas de la Planta/química , Coloración y Etiquetado/métodosRESUMEN
GH3 genes related to the auxin-inducible Glycine max (L.) Merr. GmGH3 gene encode enzymes that conjugate amino acids to auxin. To investigate the role of GH3 enzymes in stress responses and normal wood development, Populus x canescens (Ait.) was transformed with the promoter-reporter construct GH3::GUS containing a GH3 promoter and the 5' UTR from soybean. beta-Glucuronidase (GUS) activity was present in the vascular tissues of leaves and in developing lateral roots and was inducible in silent tissues by external auxin application. A decrease in GUS activity from the stem apex to the bottom corresponded to decreases in auxin concentrations in these tissues. High auxin concentration and high GH3::GUS activity were present in the pith tissue, which may provide storage for auxin compounds. GH3 reporter was active in ray cells, paratracheal parenchyma cells, maturing vessels and in cells surrounding maturing phloem fibers but not in the cambium and immature phloem, despite high auxin concentrations in the latter tissues. However, the GH3 promoter in these tissues became active when the plants were exposed to abiotic stresses, like bending or salinity, causing changes in wood anatomy. We suggest that adjustment of the internal auxin balance in wood in response to environmental cues involves GH3 auxin conjugate synthases.
Asunto(s)
Ácidos Indolacéticos/metabolismo , Tallos de la Planta/metabolismo , Populus/fisiología , Proteínas de Soja/genética , Madera/crecimiento & desarrollo , Glucuronidasa/genética , Glucuronidasa/metabolismo , Familia de Multigenes , Plantas Modificadas Genéticamente/anatomía & histología , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Plantas Modificadas Genéticamente/metabolismo , Populus/anatomía & histología , Regiones Promotoras Genéticas , Salinidad , Estrés Mecánico , Madera/anatomía & histologíaRESUMEN
Plant cells are considered to possess functionally different types of vacuoles in the same cell. One of the papers cited in support of this concept reported that protein storage and lytic vacuoles in root tips of barley (Hordeum vulgare) and pea (Pisum sativum) seedlings were initially separate compartments that later fused to form a central vacuole during cell elongation. We have reinvestigated the situation in these two roots using immunogold electron microscopy as well as immunofluorescence microscopy of histological sections. Using antisera generated against the whole protein of alpha-tonoplast intrinsic protein (TIP) as well as specific C-terminal TIP peptide antisera against alpha-, gamma-, and delta-TIP, together with antisera against the storage proteins barley lectin and pea legumin and vicilin, we were unable to obtain evidence for separate vacuole populations. Instead, our observations point to the formation of a single type of vacuole in cells differentiating both proximally and distally from the root meristem. This is a hybrid-type vacuole containing storage proteins and having both alpha- and gamma-TIPs, but not delta-TIP, in its tonoplast. As cells differentiate toward the zone of elongation, their vacuoles are characterized by increasing amounts of gamma-TIP and decreasing amounts of alpha-TIP.