Ingeborg Lang

Endocytosis and vesicle trafficking during tip growth of root hairs

Summary.The directional elongation of root hairs, “tip growth”, depends on the coordinated and highly regulated trafficking of vesicles which fill the tip cytoplasm and are active in secretion of cell wall material. So far, little is known about the dynamics of endocytosis in living root hairs. We analyzed the motile behaviour of vesicles in the apical region of living root hairs of Arabidopsis thaliana and of Triticum aestivum by live cell microscopy. For direct observation of endocytosis and of the fate of endocytic vesicles, we used the fluorescent endocytosis marker dyes FM 1-43 and FM 4-64. Rapid endocytosis was detected mainly in the tip, where it caused a bright fluorescence of the apical cytoplasm. The internalized membranes proceeded through highly dynamic putative early endosomes in the clear zone to larger endosomal compartments in the subapical region that are excluded from the clear zone. The internalized cargo ended up in the dynamic vacuole by fusion of large endosomal compartments with the tonoplast. Before export to these lytic compartments, putative early endosomes remained in the apical zone, where they most probably recycled to the plasma membrane and back into the cytoplasm for more than 30 min. Endoplasmic reticulum was not involved in trafficking pathways of endosomes. Actin cytoskeleton was needed for the endocytosis itself, as well as for further membrane trafficking. The actin-depolymerizing drug latrunculin B modified the dynamic properties of vesicles and endosomes; they became immobilized and aggregated in the tip. Treatment with brefeldin A inhibited membrane trafficking and caused the disappearance of FM-containing vesicles and putative early endosomes from the clear zone; labelled structures accumulated in motile brefeldin A-induced compartments. These large endocytic compartments redispersed upon removal of the drug. Our results hence prove that endocytosis occurs in growing root hairs. We show the localization of endocytosis in the tip and indicate specific endomembrane compartments and their recycling.

Deadly Glue — Adhesive Traps of Carnivorous Plants

Carnivorous plants trap and utilize animals in order to improve their supply with mineral nutrients. One strategy for prey capture is the use of adhesive traps, i.e., leaves that produce sticky substances. Sticky shoots are widespread in the plant kingdom and serve to protect the plant, especially flowers and seeds. In some taxa, mechanisms have been developed to absorb nutrients from the decaying carcasses of animals killed by the glue. In carnivorous plants sensu stricto, additional digestive enzymes are secreted into the glue to accelerate degradation of prey organisms.

Plasmolysis: Loss of Turgor and Beyond

Plasmolysis is a typical response of plant cells exposed to hyperosmotic stress. The loss of turgor causes the violent detachment of the living protoplast from the cell wall. The plasmolytic process is mainly driven by the vacuole. Plasmolysis is reversible (deplasmolysis) and characteristic to living plant cells. Obviously, dramatic structural changes are required to fulfill a plasmolytic cycle. In the present paper, the fate of cortical microtubules and actin microfilaments is documented throughout a plasmolytic cycle in living cells of green fluorescent protein (GFP) tagged Arabidopsis lines. While the microtubules became wavy and highly bundled during plasmolysis, cortical filamentous actin remained in close vicinity to the plasma membrane lining the sites of concave plasmolysis and adjusting readily to the diminished size of the protoplast. During deplasmolysis, cortical microtubule re-organization progressed slowly and required up to 24 h to complete the restoration of the original pre-plasmolytic pattern. Actin microfilaments, again, recovered faster and organelle movement remained intact throughout the whole process. In summary, the hydrostatic skeleton resulting from the osmotic state of the plant vacuole “overrules” the stabilization by cortical cytoskeletal elements.

Glands of carnivorous plants as a model system in cell biological research

Abstract We use glands of carnivorous plants to investigate the cyto-architecture and the physiology of secreting and absorbing plant cells. Accordingly we apply life cell microscopy, e.g. video enhanced light microscopy and ultraviolet microscopy, and combine it with electron microscopy of cryo-fixed material. In Drosera capensis, Byblis liniflora and Nepenthes *coccinea, we analyse (1) the Golgi apparatus and its vesicles during formation and secretion of trapping mucilage, (2) the endoplasmic reticulum producing digestive enzymes, (3) digestive uptake and (4) the movement of organelles along the cytoskeleton. These observations improve our understanding of the structure and function of carnivorous plants; in addition, they advance our general conception of the cell biology of glandular cells in plants.

High-resolution densitometry and elemental analysis of tropical wood.

Key messageUnderstanding the mobility and distribution of chemical elements in wood is necessary to apply dendrochemistry. Crystals are likely stable and could be used to analyze changes in nutrient supply.AbstractDendrochemistry uses the variation in wood chemical composition to infer about past environmental conditions and possible effects on tree growth. Elemental or isotopic variation might also help to identify annual growth where tree rings are anatomically not distinct. However, most elements are—to a certain degree—mobile within wood and may be related to anatomical structures. Therefore, understanding what affects elemental distribution is important to make use of and critically assess the potential of dendrochemistry. We studied the variation of wood density and elements at high spatial resolution in wood of six species with anatomically distinct to rather indistinct tree rings from a Thai monsoon forest. Many elements had a higher concentration in parenchyma than in fiber cells, and the co-variation of elements differed strongly between elements and also between species. Strong wood density changes along the ring boundary were found only in Melia azedarach. In all species, the X-ray images showed crystals. EDX spectra showed that these consist of calcium or silicon (in Chukrasia tabularis) as major elemental components. A high concentration of heavy metals (Fe, Cu and Zn) was found in Vitex peduncularis. We conclude that at least for the species studied the radial variation of elemental concentration is unlikely to reveal annual rings that anatomy could not. However, if elements in crystals are more stable than in cell walls or living protoplasts, analyzing the distribution of elements present in crystals may show environmental conditions that, in turn, influence crystal formation and are little known.

The copper spoil heap Knappenberg, Austria, as a model for metal habitats - Vegetation, substrate and contamination.

Historic mining in the Eastern Alps has left us with a legacy of numerous spoil heaps hosting specific, metal tolerant vegetation. Such habitats are characterized by elevated concentrations of toxic elements but also by high irradiation, a poorly developed substrate or extreme pH of the soil. This study investigates the distribution of vascular plants, mosses and lichens on a copper spoil heap on the ore bearing Knappenberg formed by Prebichl Layers and Werfener Schist in Lower Austria. It serves as a model for discriminating between various ecological traits and their effects on vegetation. Five distinct clusters were distinguished: (1) The bare, metal rich Central Spoil Heap was only colonised by highly resistant specialists. (2) The Northern and (3) Southern Peripheries contained less copper; the contrasting vegetation was best explained by the different microclimate. (4) A forest over acidic bedrock hosted a vegetation overlapping with the periphery of the spoil heap. (5) A forest over calcareous bedrock was similar to the spoil heap with regard to pH and humus content but hosted a vegetation differing strongly to all other habitats. Among the multiple toxic elements at the spoil heap, only Cu seems to exert a crucial influence on the vegetation pattern. Besides metal concentrations, irradiation, humidity, humus, pH and grain size distribution are important for the establishment of a metal tolerant vegetation. The difference between the species poor Northern and the diverse Southern Periphery can be explained by the microclimate rather than by the substrate. All plant species penetrating from the forest into the periphery of the spoil heap originate from the acidic but not from the calcareous bedrock.

INDUCING SALT TOLERANCE ON GROWTH AND YIELD OF SUNFLOWER BY APPLYING DIFFERENT LEVELS OF ASCORBIC ACID

This experiment was conducted in the Department of Biological Sciences, University of Sargodha, Sargodha, Pakistan during the year 2008–2009 to evaluate the effect of ascorbic acid to minimize the toxicity of salinity on sunflower. For this purpose different concentrations of ascorbic acid (0, 50, 100 mg L−1) were applied through foliar spray at vegetative stage of sunflower plants. The experiment was laid out in completely randomized block design. In this study, growth, yield and physiological parameters of the sunflower plants were collected to investigate the effect of ascorbic acid under salt stress. Concentration of 100 mg L−1 of ascorbic acid applied as foliar spray significantly enhanced the fresh and dry weight of shoot of sunflower, photosynthetic pigments, ions uptake such as potassium (K+) and calcium (Ca2+) and 100 achene weight of sunflower. These findings led us to conclude that foliar applied ascorbic acid could mitigate the adverse effects of salinity on sunflower.

Plasmolysis-deplasmolysis causes changes in endoplasmic reticulum form, movement, flow, and cytoskeletal association

Upon plasmolysis, protoplast ER becomes more persistently cisternal without changing its actin association or internal flow, while wall-associated ER is tubular, associated with altered microtubules, and has reduced flow.

Usnic acid, as a biotic factor, changes the ploidy level in mosses

Abstract Lichens and mosses often share the same environmental conditions where they compete for substrate and other essential factors. Lichens use secondary metabolites as allelochemicals to repel surrounding plants and potential rivals. In mosses, endoreduplication leads to the occurrence of various ploidy levels in the same individual and has been suggested as an adaptation to abiotic stresses. Here, we show that also biotic factors such as usnic acid, an allelochemical produced by lichens, directly influenced the level of ploidy in mosses. Application of usnic acid changed the nuclei proportion and significantly enhanced the endoreduplication index in two moss species, Physcomitrella patens and Pohlia drummondii. These investigations add a new aspect on secondary metabolites of lichens which count as biotic factors and affect ploidy levels in mosses.

Effect of ellagic acid on growth and physiology of canola (Brassica napus L.) under saline conditions

ABSTRACT Salinity stress is limiting growth and productivity of plants in many areas of the world. Plants adopted different strategies to minimize the effect of salt stress. A pot experiment was conducted to investigate the morphological and physiological changes produced in Canola (Brassica napus) by exogenous application of ellagic acid (EA) under saline conditions. EA is an antioxidant, expected to reduce the effect of salinity stress. The seeds of two canola cultivars, Rainbow and Oscar, were soaked for 6 h with different concentrations of EA (0, 55 and 110 µg/ml). The soaked seeds were sown in small pots. Salt stress was imposed on the plants by applying NaCl solutions of different concentrations (0, 60 and 120 mM) and the duration of stress was for four weeks. Salinity stress reduced seed germination and disturbed the morphological and physiological attributes of B. napus. Application of EA as seed soaking reduced the effect of salinity and enhanced the growth of plants. Overall, we could confirm a significant role of EA by inducing salinity tolerance in B. napus.