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Although cell-anatomical variables are promising proxies reflecting seasonal as well as annual climate changes, their interdependencies are not yet fully understood. In the present study we assessed the changes in tree-ring width and various wood anatomical traits, including wall thickness, lumen diameter and tracheid diameter in the radial direction in saplings of Pinus sylvestris under six climatic conditions: 5°C warmer alone (ET) or combined with drought in June (ETJ) and in August (ETA) and CO2 enrichment alone (EC, 770 ppm) or combined with drought in June (ECJ) and in August (ECA). The experiments related to temperature conditions using 2-year saplings and CO2 conditions using 3-year saplings were completed in 2009 and 2010 in a greenhouse, respectively. Results showed that tree-ring width and tracheid diameter were not affected by any of the conditions applied, but the lumen diameter was larger and the wall thickness was thinner than those under control conditions. These reactions were verified under ETJ in the warming treatment and under all conditions under CO2 enrichment conditions. Our results indicated that drought counteracted the effects of elevated CO2 concentrations on wood anatomical properties, signifying complex interactions between the two major effects of climate change. Our comparison of wood parameters through experiments highlight the potential effect of climate change — increased drought stress due to higher temperatures and water shortage as well as elevated ambient CO2, on tracheid lumen diameter and wall thickness. Whereas the ring-width and tracheid diameter practically remained unaffected under the above-mentioned conditions.

The identification of wood species in paper products is highly important for the enforcement of the newly established timber regulations regarding the control of illegal logging. In the context of European Timber Regulation (EUTR), in the present paper a database has been established containing reference samples and anatomical descriptions for the identification of 23 tropical timbers from Southeast Asia in pulp and paper products. The vessel elements and fibers of these mixed tropical hardwoods (MTH) were characterized by light microscopy. The woods in focus were macerated, embedded and compared visually and statistically. The collected microscopic data and images are helpful for the identification of the wood species used for paper production.

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.

*SUMMARY 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.

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. peerReviewed

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.

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 α-tonoplast intrinsic protein (TIP) as well as specific C-terminal TIP peptide antisera against α-, γ-, and δ-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 α- and γ-TIPs, but not δ-TIP, in its tonoplast. As cells differentiate toward the zone of elongation, their vacuoles are characterized by increasing amounts of γ-TIP and decreasing amounts of α-TIP.

The growth of trees is a spectacular and exposed process based on a highly interlinked complex of hidden and cryptic metabolic and signaling pathways not yet fully understood. In this chapter, we focus on a sequence of studies on Scots pine as an example tree species during the past 10 years in the north of Finland. We particularly compare annual height growth and annual growth in girth in the long term. Moreover, we give attention to the chronological coherence between the growth in height and girth during a growing season. Finally, we go down on the cellular level and screen various variables of the water conducting cells for their suitability as climatic proxies.

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.

A new method for measuring annual growth rings in cork has been developed. It is based on the fact that excitation of Quercus suber L. cork cross-sectional planes with UV light and also with blue light results in enhanced autofluorescence at the growth ring borders. This distinct autofluorescence band is located in the cork produced at the end of the vegetation period, with its smaller cells and thicker cell walls. The enhanced autofluorescence of polyphenolics mainly results from a very thick suberin layer in the walls of the small late cork cells. The gradient in autofluorescence from late cork to first early cork is steep. The best visibility of cork annual rings was found under the epifluorescence microscope in 60 μm thick microtome cork cross sections. For fast screening of high sample numbers scanning the blue-excited (473 nm) fluorescence of water wetted polished cross-sectional surfaces of cork pieces with a laser-equipped fluorescence image analyzer was most suitable. Evaluation of visibility of cork rings showed a clear improvement in comparison with reflected light image analysis; thus data obtained with this new autofluorescence scanning method are excellent for modeling the yearly growth increment of cork. peerReviewed

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).